confocal laser scanning microscope  (Carl Zeiss)

 
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    Name:
    ZEN Module Confocal Topography Hardware License Key
    Description:
    ZEN Module Confocal Topography Hardware License Key Module for analysis of surface data and visualisation of measument results
    Catalog Number:
    410136-1038-230
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    Structured Review

    Carl Zeiss confocal laser scanning microscope
    <t>Confocal</t> <t>laser</t> <t>scanning</t> microscopic images of the cellular uptake of different coumarin-6-co-loaded etoposide (ETP) vehicles. (A) Cellular uptake of coumarin-6-co-loaded ETP/LMC solution, coumarin-6-co-loaded ETP/LMC in 0.3% NaCMC, coumarin-6-loaded ENE and ELNE#1, and coumarin-6-loaded ELNE incorporating an ionic complex of N α -deoxycholyl- l -lysyl-methylester and 1,2-didecanoyl-sn-glycero-3-phosphate (sodium salt) (ELNE#7) by Caco-2 cells. Cellular uptake of coumarin-6-co-loaded ETP/LMC solution, coumarin-6-co-loaded ETP/LMC in 0.3% NaCMC, coumarin-6-loaded ENE and ELNE#1, and coumarin-6-loaded ELNE incorporating an ionic complex of N α -deoxycholyl- l -lysyl-methylester and 1,2-didecanoyl-sn-glycero-3-phosphate (sodium salt) (ELNE#6 and #7) by (B) MDCK cells or (C) ASBT-transfected MDCK cells. Scale bar, 20 µm.
    ZEN Module Confocal Topography Hardware License Key Module for analysis of surface data and visualisation of measument results
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    Images

    1) Product Images from "Enhanced oral bioavailability of an etoposide multiple nanoemulsion incorporating a deoxycholic acid derivative–lipid complex"

    Article Title: Enhanced oral bioavailability of an etoposide multiple nanoemulsion incorporating a deoxycholic acid derivative–lipid complex

    Journal: Drug Delivery

    doi: 10.1080/10717544.2020.1837293

    Confocal laser scanning microscopic images of the cellular uptake of different coumarin-6-co-loaded etoposide (ETP) vehicles. (A) Cellular uptake of coumarin-6-co-loaded ETP/LMC solution, coumarin-6-co-loaded ETP/LMC in 0.3% NaCMC, coumarin-6-loaded ENE and ELNE#1, and coumarin-6-loaded ELNE incorporating an ionic complex of N α -deoxycholyl- l -lysyl-methylester and 1,2-didecanoyl-sn-glycero-3-phosphate (sodium salt) (ELNE#7) by Caco-2 cells. Cellular uptake of coumarin-6-co-loaded ETP/LMC solution, coumarin-6-co-loaded ETP/LMC in 0.3% NaCMC, coumarin-6-loaded ENE and ELNE#1, and coumarin-6-loaded ELNE incorporating an ionic complex of N α -deoxycholyl- l -lysyl-methylester and 1,2-didecanoyl-sn-glycero-3-phosphate (sodium salt) (ELNE#6 and #7) by (B) MDCK cells or (C) ASBT-transfected MDCK cells. Scale bar, 20 µm.
    Figure Legend Snippet: Confocal laser scanning microscopic images of the cellular uptake of different coumarin-6-co-loaded etoposide (ETP) vehicles. (A) Cellular uptake of coumarin-6-co-loaded ETP/LMC solution, coumarin-6-co-loaded ETP/LMC in 0.3% NaCMC, coumarin-6-loaded ENE and ELNE#1, and coumarin-6-loaded ELNE incorporating an ionic complex of N α -deoxycholyl- l -lysyl-methylester and 1,2-didecanoyl-sn-glycero-3-phosphate (sodium salt) (ELNE#7) by Caco-2 cells. Cellular uptake of coumarin-6-co-loaded ETP/LMC solution, coumarin-6-co-loaded ETP/LMC in 0.3% NaCMC, coumarin-6-loaded ENE and ELNE#1, and coumarin-6-loaded ELNE incorporating an ionic complex of N α -deoxycholyl- l -lysyl-methylester and 1,2-didecanoyl-sn-glycero-3-phosphate (sodium salt) (ELNE#6 and #7) by (B) MDCK cells or (C) ASBT-transfected MDCK cells. Scale bar, 20 µm.

    Techniques Used: Transfection

    2) Product Images from "The extracellular signal-regulated kinase 1/2 modulates the intracellular localization of DNA methyltransferase 3A to regulate erythrocytic differentiation"

    Article Title: The extracellular signal-regulated kinase 1/2 modulates the intracellular localization of DNA methyltransferase 3A to regulate erythrocytic differentiation

    Journal: American Journal of Translational Research

    doi:

    Either interaction with or phosphorylation of DNMT3A by ERK1/2 modulates intracellular localization of DNMT3A. The HA-DNMT3A-expressed cells were treated with NaB, then harvested at the indicated times for cytospin preparation. After fixation and blocking, the HA-DNMT3A proteins were detected using anti-HA antibodies, and then examined using confocal laser scanning microscopy. DAPI was used to label nuclear DNA. One represented figure for each treatment is shown.
    Figure Legend Snippet: Either interaction with or phosphorylation of DNMT3A by ERK1/2 modulates intracellular localization of DNMT3A. The HA-DNMT3A-expressed cells were treated with NaB, then harvested at the indicated times for cytospin preparation. After fixation and blocking, the HA-DNMT3A proteins were detected using anti-HA antibodies, and then examined using confocal laser scanning microscopy. DAPI was used to label nuclear DNA. One represented figure for each treatment is shown.

    Techniques Used: Blocking Assay, Confocal Laser Scanning Microscopy

    Induction of erythrocytic differentiation promotes the translocation of DNMT3A into the nucleus in K562 cells. A. The HA-DNMT3A-expressed cells were treated with NaB, then harvested at the indicated times for cytospin preparation. After fixation and blocking, the HA-DNMT3A proteins were detected using anti-HA antibodies, and then examined using confocal laser scanning microscopy. DAPI was used to label nuclear DNA. B. The quantified data were presented as means ± s.d. The two-way ANOVA followed by the Tukey’s multiple-comparison posttest was used for statistical analysis. * P
    Figure Legend Snippet: Induction of erythrocytic differentiation promotes the translocation of DNMT3A into the nucleus in K562 cells. A. The HA-DNMT3A-expressed cells were treated with NaB, then harvested at the indicated times for cytospin preparation. After fixation and blocking, the HA-DNMT3A proteins were detected using anti-HA antibodies, and then examined using confocal laser scanning microscopy. DAPI was used to label nuclear DNA. B. The quantified data were presented as means ± s.d. The two-way ANOVA followed by the Tukey’s multiple-comparison posttest was used for statistical analysis. * P

    Techniques Used: Translocation Assay, Blocking Assay, Confocal Laser Scanning Microscopy

    3) Product Images from "The Topographical Optimization of 3D Microgroove Pattern Intervals for Ligamentous Cell Orientations: In Vitro"

    Article Title: The Topographical Optimization of 3D Microgroove Pattern Intervals for Ligamentous Cell Orientations: In Vitro

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21249358

    Qualitative and quantitative assessments to identify different of microgroove pattern intervals. ( A ) Scanning electron microscope (SEM) showed surface morphologies with the statistically calculated microgroove intervals in four different groups (μG-25, μG-19, μG-12, and μG-6) by different slice thickness. ( B ) Based on the SEM images, microgroove patterns were quantitatively analyzed and profiled by crossing the pattern surfaces on PDL-guiding architectures. ( C ) Using the confocal microscope, surface topographies were characterized, as was surface roughness.
    Figure Legend Snippet: Qualitative and quantitative assessments to identify different of microgroove pattern intervals. ( A ) Scanning electron microscope (SEM) showed surface morphologies with the statistically calculated microgroove intervals in four different groups (μG-25, μG-19, μG-12, and μG-6) by different slice thickness. ( B ) Based on the SEM images, microgroove patterns were quantitatively analyzed and profiled by crossing the pattern surfaces on PDL-guiding architectures. ( C ) Using the confocal microscope, surface topographies were characterized, as was surface roughness.

    Techniques Used: Microscopy

    4) Product Images from "Procathepsin V Is Secreted in a TSH Regulated Manner from Human Thyroid Epithelial Cells and Is Accessible to an Activity-Based Probe"

    Article Title: Procathepsin V Is Secreted in a TSH Regulated Manner from Human Thyroid Epithelial Cells and Is Accessible to an Activity-Based Probe

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21239140

    Trafficking of eGFP-tagged full-length cathepsin V in thyroid epithelial cells. Confocal laser scanning micrographs of Nthyori-CV cells expressing hCV-eGFP chimeras (( A – C ), green signals) after immunolabeling with antibodies against PDI ( A ), GM130 ( B ) and Lamp1 ( C ) proteins residing in the ER, at the cytosolic face of Golgi cisternae and vesicles, and in endo-lysosomes, respectively (red signals). Yellow signals are indicative of co-localization. Nuclei were counter-stained with Draq5™ (blue signals). Single-channel fluorescence micrographs are depicted in the bottom panels as indicated. Scale bars represent 50 μm.
    Figure Legend Snippet: Trafficking of eGFP-tagged full-length cathepsin V in thyroid epithelial cells. Confocal laser scanning micrographs of Nthyori-CV cells expressing hCV-eGFP chimeras (( A – C ), green signals) after immunolabeling with antibodies against PDI ( A ), GM130 ( B ) and Lamp1 ( C ) proteins residing in the ER, at the cytosolic face of Golgi cisternae and vesicles, and in endo-lysosomes, respectively (red signals). Yellow signals are indicative of co-localization. Nuclei were counter-stained with Draq5™ (blue signals). Single-channel fluorescence micrographs are depicted in the bottom panels as indicated. Scale bars represent 50 μm.

    Techniques Used: Expressing, Immunolabeling, Staining, Fluorescence

    5) Product Images from "HistoMosaic Detecting KRAS G12V Mutation Across Colorectal Cancer Tissue Slices through in Situ PCR"

    Article Title: HistoMosaic Detecting KRAS G12V Mutation Across Colorectal Cancer Tissue Slices through in Situ PCR

    Journal: Analytical chemistry

    doi: 10.1021/acs.analchem.5b04460

    HistoMosaic and H E serial section comparison for CRC tissue tested G12V+ by conventional sequencing. (a) A digital slide scan of a thin section of CRC. (b) A serial, unstained section processed for HistoMosaic and captured as an automated tile scan by confocal laser scanning microscopy. PCR G12V probe (FAM, green) and KRAS probe (HEX, red) fluorescence are limited mostly to within wells. The approximate tumor border is delineated (—). The dual probe signal overlay is combined with autofluorescence background to demonstrate similar morphology of the two serial sections. Dashed boxes show the locations of (c) and (d). In (d), the symbols above the wells’ outlines indicate cancer (C), no cancer (N), no-call (?), KRAS+ or −, and G12V+ or − (well assignments from H E morphometry and PCR signal statistics). Well numbers 1–9 are referred to in the text.
    Figure Legend Snippet: HistoMosaic and H E serial section comparison for CRC tissue tested G12V+ by conventional sequencing. (a) A digital slide scan of a thin section of CRC. (b) A serial, unstained section processed for HistoMosaic and captured as an automated tile scan by confocal laser scanning microscopy. PCR G12V probe (FAM, green) and KRAS probe (HEX, red) fluorescence are limited mostly to within wells. The approximate tumor border is delineated (—). The dual probe signal overlay is combined with autofluorescence background to demonstrate similar morphology of the two serial sections. Dashed boxes show the locations of (c) and (d). In (d), the symbols above the wells’ outlines indicate cancer (C), no cancer (N), no-call (?), KRAS+ or −, and G12V+ or − (well assignments from H E morphometry and PCR signal statistics). Well numbers 1–9 are referred to in the text.

    Techniques Used: Sequencing, Confocal Laser Scanning Microscopy, Polymerase Chain Reaction, Fluorescence

    HistoMosaic and H E serial section comparison for CRC tissue tested G12V− by conventional sequencing. (a) A digital slide scan of a thin section of CRC. (b) A serial, unstained section processed for HistoMosaic and captured as an automated tile scan by confocal laser scanning microscopy. PCR G12V probe (FAM, green) and KRAS probe (HEX, red) fluorescence are limited mostly to within wells. The approximate tumor border is delineated (—). The dual probe signal overlay is combined with autofluorescence background to demonstrate similar morphology of the two serial sections.
    Figure Legend Snippet: HistoMosaic and H E serial section comparison for CRC tissue tested G12V− by conventional sequencing. (a) A digital slide scan of a thin section of CRC. (b) A serial, unstained section processed for HistoMosaic and captured as an automated tile scan by confocal laser scanning microscopy. PCR G12V probe (FAM, green) and KRAS probe (HEX, red) fluorescence are limited mostly to within wells. The approximate tumor border is delineated (—). The dual probe signal overlay is combined with autofluorescence background to demonstrate similar morphology of the two serial sections.

    Techniques Used: Sequencing, Confocal Laser Scanning Microscopy, Polymerase Chain Reaction, Fluorescence

    6) Product Images from "DEC-205 receptor-mediated long-circling nanoliposome as an antigen and Eucommia ulmoides polysaccharide delivery system enhances the immune response via facilitating dendritic cells maturation"

    Article Title: DEC-205 receptor-mediated long-circling nanoliposome as an antigen and Eucommia ulmoides polysaccharide delivery system enhances the immune response via facilitating dendritic cells maturation

    Journal: Drug Delivery

    doi: 10.1080/10717544.2020.1844343

    Laser confocal scanning microscopy of anti-DEC-205-EUPS-OVA-LPSM uptake by DCs. DCs were inoculated in a 6-well cell-culture plate with a round coverslip. After 24 h of culture, OVA, and anti-DEC-205-EUPS-OVA-LPSM were added separately. After incubation for 12 h, the slides were taken out and fixed and stained using DAPI and Phalloidin-iFluor 555. Blue fluorescence is the nucleus labeled by DAPI, while red fluorescence indicates the actin stained with Phalloidin-iFluor 555. Cells were mounted with 90% glycerol and photographed using a confocal laser scanning microscope. (A) Control, (B) OVA, (C) EUPS-OVA-LPSM, and (D) anti-DEC-205-EUPS-OVA-LPSM.
    Figure Legend Snippet: Laser confocal scanning microscopy of anti-DEC-205-EUPS-OVA-LPSM uptake by DCs. DCs were inoculated in a 6-well cell-culture plate with a round coverslip. After 24 h of culture, OVA, and anti-DEC-205-EUPS-OVA-LPSM were added separately. After incubation for 12 h, the slides were taken out and fixed and stained using DAPI and Phalloidin-iFluor 555. Blue fluorescence is the nucleus labeled by DAPI, while red fluorescence indicates the actin stained with Phalloidin-iFluor 555. Cells were mounted with 90% glycerol and photographed using a confocal laser scanning microscope. (A) Control, (B) OVA, (C) EUPS-OVA-LPSM, and (D) anti-DEC-205-EUPS-OVA-LPSM.

    Techniques Used: Confocal Laser Scanning Microscopy, Cell Culture, Incubation, Staining, Fluorescence, Labeling, Laser-Scanning Microscopy

    7) Product Images from "HAK/KUP/KT family potassium transporter genes are involved in potassium deficiency and stress responses in tea plants (Camellia sinensis L.): expression and functional analysis"

    Article Title: HAK/KUP/KT family potassium transporter genes are involved in potassium deficiency and stress responses in tea plants (Camellia sinensis L.): expression and functional analysis

    Journal: BMC Genomics

    doi: 10.1186/s12864-020-06948-6

    Tissue-specific expression pattern, subcellular localization and yeast complementation analysis of CsHAK7. a Expression pattern of CsHAK7 in indicated tissues of tea plant. MV, major vein; VB, vascular bundle, peeled from the stem. CsGAPDH was an internal control. Data are mean ± SE ( n = 3). b Confocal laser scanning microscopy images showed tobacco leaf epidermal cells transiently expressing either GFP or CsHAK7::GFP together with AtPIP2A:mCherry (plasma membrane maker, Nelson et al., 2007). (a), (e), Confocal images via the GFP channel only. (b), (f), Confocal images of the red mCherry fluorescence marking the PM position. (c), (j), Bright field. (d), (h), Merged images of GFP (green) and mCherry RFP (red) together with bright filed. Scale bar, 45 μm. c Yeast complementation assay of K + acquisition by the K + uptake-defective yeast mutant (R5421) complemented with CsHAK7. Growth status of R5421 cells expressing CsHAK7, empty vector (pDR196), on AP solid medium containing 10 or 0.1 mM K + . The 1:5 serial dilutions of yeast cells were spotted on the AP solid medium and then incubated at 30 °C for 3–5 d
    Figure Legend Snippet: Tissue-specific expression pattern, subcellular localization and yeast complementation analysis of CsHAK7. a Expression pattern of CsHAK7 in indicated tissues of tea plant. MV, major vein; VB, vascular bundle, peeled from the stem. CsGAPDH was an internal control. Data are mean ± SE ( n = 3). b Confocal laser scanning microscopy images showed tobacco leaf epidermal cells transiently expressing either GFP or CsHAK7::GFP together with AtPIP2A:mCherry (plasma membrane maker, Nelson et al., 2007). (a), (e), Confocal images via the GFP channel only. (b), (f), Confocal images of the red mCherry fluorescence marking the PM position. (c), (j), Bright field. (d), (h), Merged images of GFP (green) and mCherry RFP (red) together with bright filed. Scale bar, 45 μm. c Yeast complementation assay of K + acquisition by the K + uptake-defective yeast mutant (R5421) complemented with CsHAK7. Growth status of R5421 cells expressing CsHAK7, empty vector (pDR196), on AP solid medium containing 10 or 0.1 mM K + . The 1:5 serial dilutions of yeast cells were spotted on the AP solid medium and then incubated at 30 °C for 3–5 d

    Techniques Used: Expressing, Confocal Laser Scanning Microscopy, Fluorescence, Mutagenesis, Plasmid Preparation, Incubation

    8) Product Images from "Claudin-21 Has a Paracellular Channel Role at Tight Junctions"

    Article Title: Claudin-21 Has a Paracellular Channel Role at Tight Junctions

    Journal: Molecular and Cellular Biology

    doi: 10.1128/MCB.00758-15

    Mouse claudin-21 localization in MDCK I transfectant clones. (A) MDCK I cells or Venus-claudin-21-expressing transfected MDCK I cells (MDCK I-Venus-claudin-21 cells) were cultured to confluence on glass coverslips and examined by confocal laser scanning
    Figure Legend Snippet: Mouse claudin-21 localization in MDCK I transfectant clones. (A) MDCK I cells or Venus-claudin-21-expressing transfected MDCK I cells (MDCK I-Venus-claudin-21 cells) were cultured to confluence on glass coverslips and examined by confocal laser scanning

    Techniques Used: Transfection, Expressing, Cell Culture

    9) Product Images from "Effects of Thrombomodulin in Reducing Lethality and Suppressing Neutrophil Extracellular Trap Formation in the Lungs and Liver in a Lipopolysaccharide-Induced Murine Septic Shock Model"

    Article Title: Effects of Thrombomodulin in Reducing Lethality and Suppressing Neutrophil Extracellular Trap Formation in the Lungs and Liver in a Lipopolysaccharide-Induced Murine Septic Shock Model

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms22094933

    Confirmation of NETs formation in the liver using confocal microscopy and SEM images. Saline is administered to LPS-induced septic shock model mice 1 h after the LPS administration, and the livers are observed after 8 h using confocal microscopy and Scanning Electron Microscope (SEM). Immunofluorescence staining for MPO (green) and histone H2A.X (red) are performed, and nuclei are stained with DAPI (blue); the three colored images are merged. ( a ) Magnification: ×100; scale bar = 20 μm; ( b ) magnification: ×400; scale bar = 80 μm. Sections are sputter-coated with gold palladium and observed using SEM after confocal microscopy. ( c ) Magnification: ×7000; scale bar = 2 μm. ( d ) The SEM and confocal images are merged). Magnification: ×7000; scale bar = 250 nm. The yellow box indicates enlargement.
    Figure Legend Snippet: Confirmation of NETs formation in the liver using confocal microscopy and SEM images. Saline is administered to LPS-induced septic shock model mice 1 h after the LPS administration, and the livers are observed after 8 h using confocal microscopy and Scanning Electron Microscope (SEM). Immunofluorescence staining for MPO (green) and histone H2A.X (red) are performed, and nuclei are stained with DAPI (blue); the three colored images are merged. ( a ) Magnification: ×100; scale bar = 20 μm; ( b ) magnification: ×400; scale bar = 80 μm. Sections are sputter-coated with gold palladium and observed using SEM after confocal microscopy. ( c ) Magnification: ×7000; scale bar = 2 μm. ( d ) The SEM and confocal images are merged). Magnification: ×7000; scale bar = 250 nm. The yellow box indicates enlargement.

    Techniques Used: Confocal Microscopy, Mouse Assay, Microscopy, Immunofluorescence, Staining

    10) Product Images from "Localization of the signal of dystonia-associated protein torsinA near the Golgi apparatus in cultured central neurons"

    Article Title: Localization of the signal of dystonia-associated protein torsinA near the Golgi apparatus in cultured central neurons

    Journal: bioRxiv

    doi: 10.1101/2019.12.11.872804

    Localization of overexpressed torsinA proteins in hippocampal neurons of rats and ΔE-torsinA knock-in mice. Cultured neurons were transfected with a construct encoding either WT-torsinA-GFP ( A ) or ΔE-torsinA-GFP ( B ). A modification of the calcium-phosphate method in Fig. 1 was used to enhance the efficiency of transfection ( Jiang and Chen, 2006 ). A confocal laser-scanning microscope was used to observe the neurons. Neurons are shown both as single optical sections at the somatic level (Single) and as maximum intensity projection (MIP) images that represent the overall structure at all heights. Neurons were cultured from WT rats (top row), WT mice (2 nd row), heterozygous (HET, Tor1a +/ΔE , 3 rd row) or homozygous (HOM, Tor1a ΔE/ΔE , bottom row) ΔE-torsinA knock-in mice. Arrows point to representative examples of the diffuse distribution of WT-torsinA-GFP in the cytoplasm ( A ), and ΔE-torsinA-GFP in cytoplasmic inclusion bodies ( B ).
    Figure Legend Snippet: Localization of overexpressed torsinA proteins in hippocampal neurons of rats and ΔE-torsinA knock-in mice. Cultured neurons were transfected with a construct encoding either WT-torsinA-GFP ( A ) or ΔE-torsinA-GFP ( B ). A modification of the calcium-phosphate method in Fig. 1 was used to enhance the efficiency of transfection ( Jiang and Chen, 2006 ). A confocal laser-scanning microscope was used to observe the neurons. Neurons are shown both as single optical sections at the somatic level (Single) and as maximum intensity projection (MIP) images that represent the overall structure at all heights. Neurons were cultured from WT rats (top row), WT mice (2 nd row), heterozygous (HET, Tor1a +/ΔE , 3 rd row) or homozygous (HOM, Tor1a ΔE/ΔE , bottom row) ΔE-torsinA knock-in mice. Arrows point to representative examples of the diffuse distribution of WT-torsinA-GFP in the cytoplasm ( A ), and ΔE-torsinA-GFP in cytoplasmic inclusion bodies ( B ).

    Techniques Used: Knock-In, Mouse Assay, Cell Culture, Transfection, Construct, Modification, Laser-Scanning Microscopy

    11) Product Images from "Guanidine modifications enhance the anti‐herpes simplex virus activity of (E,E)‐4,6‐bis(styryl)‐pyrimidine derivatives in vitro and in vivo, et al. Guanidine modifications enhance the anti‐herpes simplex virus activity of (E,E)‐4,6‐bis(styryl)‐pyrimidine derivatives in vitro and in vivo"

    Article Title: Guanidine modifications enhance the anti‐herpes simplex virus activity of (E,E)‐4,6‐bis(styryl)‐pyrimidine derivatives in vitro and in vivo, et al. Guanidine modifications enhance the anti‐herpes simplex virus activity of (E,E)‐4,6‐bis(styryl)‐pyrimidine derivatives in vitro and in vivo

    Journal: British Journal of Pharmacology

    doi: 10.1111/bph.14918

    Subcellular localization of BS‐pyrimidine derivatives in Vero and HeLa cells. (a) Vero cells were incubated with compounds 3a, 5a, or 5d (30 μM) for 1 hr at 37°C. The fluorescence was then detected by confocal laser scanning microscopy. The scale bar represents 50 μm. An enlarged view of part of one field (highlighted by the red rectangle) is shown to indicate the localization of compound 5d within the Vero cells. The scale bar represents 10 μm in this case. (b‐d) HeLa cells were transfected with expression plasmids for red fluorescent protein (RFP)‐coupled (b) Rab5, (c) Rab7, or (d) LAMP‐1. At 24 hr post‐transfection, cells were added with compound 5d (20 μM) and incubated for 1 hr at 37°C. After that, the fluorescence was detected via confocal microscopy. The scale bar represents 10 μm. (e, f) Herpes simplex virus (HSV)‐2 (multiplicity of infection = 1.0) was pretreated with (e) compound 3a or (f) 5d at 37°C for 1 hr before infection. Then, after removal of the virus inoculum, media containing compound 3a or 5d (20 μM) were added to cells. At 1 hr p.i., the localization of virus gB protein was evaluated via immunofluorescence assay. The fluorescence was detected via confocal microscopy. The scale bar represents 20 μm
    Figure Legend Snippet: Subcellular localization of BS‐pyrimidine derivatives in Vero and HeLa cells. (a) Vero cells were incubated with compounds 3a, 5a, or 5d (30 μM) for 1 hr at 37°C. The fluorescence was then detected by confocal laser scanning microscopy. The scale bar represents 50 μm. An enlarged view of part of one field (highlighted by the red rectangle) is shown to indicate the localization of compound 5d within the Vero cells. The scale bar represents 10 μm in this case. (b‐d) HeLa cells were transfected with expression plasmids for red fluorescent protein (RFP)‐coupled (b) Rab5, (c) Rab7, or (d) LAMP‐1. At 24 hr post‐transfection, cells were added with compound 5d (20 μM) and incubated for 1 hr at 37°C. After that, the fluorescence was detected via confocal microscopy. The scale bar represents 10 μm. (e, f) Herpes simplex virus (HSV)‐2 (multiplicity of infection = 1.0) was pretreated with (e) compound 3a or (f) 5d at 37°C for 1 hr before infection. Then, after removal of the virus inoculum, media containing compound 3a or 5d (20 μM) were added to cells. At 1 hr p.i., the localization of virus gB protein was evaluated via immunofluorescence assay. The fluorescence was detected via confocal microscopy. The scale bar represents 20 μm

    Techniques Used: Incubation, Fluorescence, Confocal Laser Scanning Microscopy, Transfection, Expressing, Confocal Microscopy, Infection, Immunofluorescence

    12) Product Images from "Potential involvement of Streptococcus mutans possessing collagen binding protein Cnm in infective endocarditis"

    Article Title: Potential involvement of Streptococcus mutans possessing collagen binding protein Cnm in infective endocarditis

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-75933-6

    Effect of silencing genes involved in HUVEC invasion by S. mutans . ( A ) RT-PCR analysis of HUVECs transfected with siRNAs targeting ARHGAP9 , ARHGEF38 , or GPR179 . GAPDH mRNA expression was analyzed as an internal control. ( B ) Representative confocal laser scanning microscopy images of HUVECs invaded by S. mutans in the presence or absence of serum. Nuclei are stained blue (DAPI) and each transcript ( ARHGAP9 , ARHGEF38 , and GPR179 ) is stained red (Alexa Fluor 555). Arrowheads indicate the expression of each transcript. ( C ) HUVECs were transfected with ARHGEF38 , ARHGAP9 , GPR179, or control siRNAs. Invasion ratios of these HUVECs after their incubation with S. mutans strain TW295 for 2 h at a multiplicity of infection of 100. Data are presented as the means ± SD of four technical replicates. * p
    Figure Legend Snippet: Effect of silencing genes involved in HUVEC invasion by S. mutans . ( A ) RT-PCR analysis of HUVECs transfected with siRNAs targeting ARHGAP9 , ARHGEF38 , or GPR179 . GAPDH mRNA expression was analyzed as an internal control. ( B ) Representative confocal laser scanning microscopy images of HUVECs invaded by S. mutans in the presence or absence of serum. Nuclei are stained blue (DAPI) and each transcript ( ARHGAP9 , ARHGEF38 , and GPR179 ) is stained red (Alexa Fluor 555). Arrowheads indicate the expression of each transcript. ( C ) HUVECs were transfected with ARHGEF38 , ARHGAP9 , GPR179, or control siRNAs. Invasion ratios of these HUVECs after their incubation with S. mutans strain TW295 for 2 h at a multiplicity of infection of 100. Data are presented as the means ± SD of four technical replicates. * p

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Transfection, Expressing, Confocal Laser Scanning Microscopy, Staining, Incubation, Infection

    13) Product Images from "PEGylated lipid nanocarrier for enhancing photodynamic therapy of skin carcinoma using curcumin: in-vitro/in-vivo studies and histopathological examination"

    Article Title: PEGylated lipid nanocarrier for enhancing photodynamic therapy of skin carcinoma using curcumin: in-vitro/in-vivo studies and histopathological examination

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-67349-z

    Confocal laser scanning microscopy photographs of dorsal mice skin of control, group treated with drug suspension (Cur) and group treated with the selected Cur-loaded PEGylated lipidic nanoparticles (PLN3).
    Figure Legend Snippet: Confocal laser scanning microscopy photographs of dorsal mice skin of control, group treated with drug suspension (Cur) and group treated with the selected Cur-loaded PEGylated lipidic nanoparticles (PLN3).

    Techniques Used: Confocal Laser Scanning Microscopy, Mouse Assay

    14) Product Images from "Functional Characterization of PsnNAC036 under Salinity and High Temperature Stresses"

    Article Title: Functional Characterization of PsnNAC036 under Salinity and High Temperature Stresses

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms22052656

    Subcellular localization of the PsnNAC036 protein. ( A ) Schematic map of the T-DNA inserted in the 35S::GFP binary vector. ( B ) The 35S::PsnNAC036-GFP fusion construct and the positive control 35S::GFP plasmid were introduced into onion epidermal cells by particle bombardment. GFP fluorescence was observed by confocal laser scanning microscopy. ( a , e ) are fluorescence images observed in a dark field (green); ( b , f ) are 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) staining, which is specific for the nucleus (blue); ( c , g ) are light images observed in bright field; ( d , h ) are merged images of dark field and bright field. Scale bar = 20 μm.
    Figure Legend Snippet: Subcellular localization of the PsnNAC036 protein. ( A ) Schematic map of the T-DNA inserted in the 35S::GFP binary vector. ( B ) The 35S::PsnNAC036-GFP fusion construct and the positive control 35S::GFP plasmid were introduced into onion epidermal cells by particle bombardment. GFP fluorescence was observed by confocal laser scanning microscopy. ( a , e ) are fluorescence images observed in a dark field (green); ( b , f ) are 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) staining, which is specific for the nucleus (blue); ( c , g ) are light images observed in bright field; ( d , h ) are merged images of dark field and bright field. Scale bar = 20 μm.

    Techniques Used: Plasmid Preparation, Construct, Positive Control, Fluorescence, Confocal Laser Scanning Microscopy, Staining

    15) Product Images from "Hexarelin Modulation of MAPK and PI3K/Akt Pathways in Neuro-2A Cells Inhibits Hydrogen Peroxide—Induced Apoptotic Toxicity"

    Article Title: Hexarelin Modulation of MAPK and PI3K/Akt Pathways in Neuro-2A Cells Inhibits Hydrogen Peroxide—Induced Apoptotic Toxicity

    Journal: Pharmaceuticals

    doi: 10.3390/ph14050444

    Effects of hexarelin on morphology of Neuro-2A cells stimulated with H 2 O 2 . Representative photomicrographs of Neuro-2A cells incubated for 24 h with or without hexarelin and 100 µM H 2 O 2 , and examples of cell binarized and outlined. Cells were seeded on poly-D-lysine pre-treated coverslips and, at the end of the treatments, were fixed and stained with phalloidin and DAPI. Images were captured with a confocal laser scan microscope. Scale bar: 20 µm.
    Figure Legend Snippet: Effects of hexarelin on morphology of Neuro-2A cells stimulated with H 2 O 2 . Representative photomicrographs of Neuro-2A cells incubated for 24 h with or without hexarelin and 100 µM H 2 O 2 , and examples of cell binarized and outlined. Cells were seeded on poly-D-lysine pre-treated coverslips and, at the end of the treatments, were fixed and stained with phalloidin and DAPI. Images were captured with a confocal laser scan microscope. Scale bar: 20 µm.

    Techniques Used: Incubation, Staining, Microscopy

    Hexarelin reduces Neuro-2A cells de-ramification induced by H 2 O 2 . ( A ) Neuro-2A cells were seeded on poly-D-lysine pre-treated coverslips and incubated for 24 h with or without hexarelin and 100 µM H 2 O 2 . At the end of the treatment, cells were fixed and stained for phalloidin and DAPI. Images were captured with confocal laser scan microscope. Scale bar: 20 µm. ( B ) Graphical representation of the number of cells in the same areas per each treatment, ( C ) of the process endpoints/cells and ( D ) process length/cells. Data are expressed as mean ± SEM of 3 replicates (total number of cells analyzed = 160). Statistical significance: ** p
    Figure Legend Snippet: Hexarelin reduces Neuro-2A cells de-ramification induced by H 2 O 2 . ( A ) Neuro-2A cells were seeded on poly-D-lysine pre-treated coverslips and incubated for 24 h with or without hexarelin and 100 µM H 2 O 2 . At the end of the treatment, cells were fixed and stained for phalloidin and DAPI. Images were captured with confocal laser scan microscope. Scale bar: 20 µm. ( B ) Graphical representation of the number of cells in the same areas per each treatment, ( C ) of the process endpoints/cells and ( D ) process length/cells. Data are expressed as mean ± SEM of 3 replicates (total number of cells analyzed = 160). Statistical significance: ** p

    Techniques Used: Incubation, Staining, Microscopy

    16) Product Images from "The Recombinant Fragment of Human κ-Casein Induces Cell Death by Targeting the Proteins of Mitochondrial Import in Breast Cancer Cells"

    Article Title: The Recombinant Fragment of Human κ-Casein Induces Cell Death by Targeting the Proteins of Mitochondrial Import in Breast Cancer Cells

    Journal: Cancers

    doi: 10.3390/cancers12061427

    RL2 induces loss of mitochondrial membrane potential. ( A – D ) MDA-MB-231 and MCF-7 cells were stimulated with 300 µg/mL RL2 for 6 and 24 h. Cells were subsequently stained with 20 µM TMRM. The reduced TMRM signal indicated the loss of mitochondrial membrane potential. CCCP treatment served as positive control for TMRM reduction. ( A ) Cells were analyzed with FlowSight ® for TMRM-signal. Representative cells from three independent experiments are shown. ( B ) Mean and standard deviations for the amounts of TMRM negative cells (in %) from three independent experiments are shown. The statistical analysis was performed by paired student t-tests. ( C , D ) RL2-treated MDA-MB-231 cells were stained with 5 mg/mL cell membrane stain (CellMask™ Deep Red Plasma membrane Stain, C10046, Thermo Fisher, Walham, MA, USA) and introduced to confocal laser scanning microscopy. Membrane- and TMRM-stained cells are shown in merge for single cells ( C , scale: 10 µM) and cell populations ( D , scale: 100 µM). The red color corresponds to the plasma membrane staining. The green ( C ) and yellow ( A , B ) colors correspond to TMRM staining. * (significant; p
    Figure Legend Snippet: RL2 induces loss of mitochondrial membrane potential. ( A – D ) MDA-MB-231 and MCF-7 cells were stimulated with 300 µg/mL RL2 for 6 and 24 h. Cells were subsequently stained with 20 µM TMRM. The reduced TMRM signal indicated the loss of mitochondrial membrane potential. CCCP treatment served as positive control for TMRM reduction. ( A ) Cells were analyzed with FlowSight ® for TMRM-signal. Representative cells from three independent experiments are shown. ( B ) Mean and standard deviations for the amounts of TMRM negative cells (in %) from three independent experiments are shown. The statistical analysis was performed by paired student t-tests. ( C , D ) RL2-treated MDA-MB-231 cells were stained with 5 mg/mL cell membrane stain (CellMask™ Deep Red Plasma membrane Stain, C10046, Thermo Fisher, Walham, MA, USA) and introduced to confocal laser scanning microscopy. Membrane- and TMRM-stained cells are shown in merge for single cells ( C , scale: 10 µM) and cell populations ( D , scale: 100 µM). The red color corresponds to the plasma membrane staining. The green ( C ) and yellow ( A , B ) colors correspond to TMRM staining. * (significant; p

    Techniques Used: Multiple Displacement Amplification, Staining, Positive Control, Confocal Laser Scanning Microscopy

    17) Product Images from "A complement component C1q-mediated mechanism of antibody-dependent enhancement of Ebola virus infection"

    Article Title: A complement component C1q-mediated mechanism of antibody-dependent enhancement of Ebola virus infection

    Journal: PLoS Neglected Tropical Diseases

    doi: 10.1371/journal.pntd.0008602

    Enhanced VLP attachment to the cell surface in the presence of ZGP12/1.1 and C1q. (a, b, c) Fluorescent images of attachment and trafficking to late endosomes of VLPs. (d, e, f, g) Quantified fluorescent signals of VLPs, Rab7, and Dx10. HEK293 cells expressing eGFP-Rab7 (a, b) or HEK293 cells (c) were inoculated DiI-labeled VLPs mixed with PBS (Untreated), or treated with C1q (20 μg/ml) alone, ZGP12/1.1 (10 μg/ml) alone, CTR-IgG (10 μg/ml) and C1q, or ZGP12/1.1 and C1q. After adsorption, cells were incubated with Alexa647-labeled Dx10 (0.5 mg/ml) for 2 h at 37˚C (c). VLPs (red) on the cell surface at 0 h (a, d), VLPs (red) and eGFP-Rab7 (green) in the cytoplasm at 2 h (b, e, f), and VLPs (red) and Alexa647-labeled Dx10 (green) in the cytoplasm at 2 h (c, g) after adsorption were monitored by confocal laser scanning microscopy. Top panels show merged image and bottom panels show VLPs (b, c). Scale bars represent 10 μm (a, b, c). Nuclei of cells are visualized with DAPI (blue). The number of VLPs on the cell surface (d) and incorporated into the cells (e), the colocalization of VLPs (DiI) and eGFP-Rab7 signals (f), and the colocalization of VLP (DiI) and Dx10 (Alexa647) signals (g) were quantified and percentages of DiI-labeled VLPs that colocalized with eGFP-Rab7 (f) and Alexa647 labeled Dx10 (g) were determined as described in Methods. The mean and standard deviation of three independent experiments are shown. Statistical analysis was performed using Student’s t -test (* p
    Figure Legend Snippet: Enhanced VLP attachment to the cell surface in the presence of ZGP12/1.1 and C1q. (a, b, c) Fluorescent images of attachment and trafficking to late endosomes of VLPs. (d, e, f, g) Quantified fluorescent signals of VLPs, Rab7, and Dx10. HEK293 cells expressing eGFP-Rab7 (a, b) or HEK293 cells (c) were inoculated DiI-labeled VLPs mixed with PBS (Untreated), or treated with C1q (20 μg/ml) alone, ZGP12/1.1 (10 μg/ml) alone, CTR-IgG (10 μg/ml) and C1q, or ZGP12/1.1 and C1q. After adsorption, cells were incubated with Alexa647-labeled Dx10 (0.5 mg/ml) for 2 h at 37˚C (c). VLPs (red) on the cell surface at 0 h (a, d), VLPs (red) and eGFP-Rab7 (green) in the cytoplasm at 2 h (b, e, f), and VLPs (red) and Alexa647-labeled Dx10 (green) in the cytoplasm at 2 h (c, g) after adsorption were monitored by confocal laser scanning microscopy. Top panels show merged image and bottom panels show VLPs (b, c). Scale bars represent 10 μm (a, b, c). Nuclei of cells are visualized with DAPI (blue). The number of VLPs on the cell surface (d) and incorporated into the cells (e), the colocalization of VLPs (DiI) and eGFP-Rab7 signals (f), and the colocalization of VLP (DiI) and Dx10 (Alexa647) signals (g) were quantified and percentages of DiI-labeled VLPs that colocalized with eGFP-Rab7 (f) and Alexa647 labeled Dx10 (g) were determined as described in Methods. The mean and standard deviation of three independent experiments are shown. Statistical analysis was performed using Student’s t -test (* p

    Techniques Used: Expressing, Labeling, Adsorption, Incubation, Confocal Laser Scanning Microscopy, Standard Deviation

    18) Product Images from "Amyloid-Beta Induces Different Expression Pattern of Tissue Transglutaminase and Its Isoforms on Olfactory Ensheathing Cells: Modulatory Effect of Indicaxanthin"

    Article Title: Amyloid-Beta Induces Different Expression Pattern of Tissue Transglutaminase and Its Isoforms on Olfactory Ensheathing Cells: Modulatory Effect of Indicaxanthin

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms22073388

    Confocal laser scanning microscopy of labeling immunocytochemistry for anti-TG2 in OECs. Images show different conditions: control, DMSO, 100 µM indicaxanthin (INDI), 10 μM Aβ(1-42) or Aβ(25-35) or Aβ(35-25) both in the absence and presence of 100 µM INDI for 24 h. Scale bar 20 µm.
    Figure Legend Snippet: Confocal laser scanning microscopy of labeling immunocytochemistry for anti-TG2 in OECs. Images show different conditions: control, DMSO, 100 µM indicaxanthin (INDI), 10 μM Aβ(1-42) or Aβ(25-35) or Aβ(35-25) both in the absence and presence of 100 µM INDI for 24 h. Scale bar 20 µm.

    Techniques Used: Confocal Laser Scanning Microscopy, Labeling, Immunocytochemistry

    19) Product Images from "F-Box Gene D5RF Is Regulated by Agrobacterium Virulence Protein VirD5 and Essential for Agrobacterium-Mediated Plant Transformation"

    Article Title: F-Box Gene D5RF Is Regulated by Agrobacterium Virulence Protein VirD5 and Essential for Agrobacterium-Mediated Plant Transformation

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21186731

    Subcellular localization assay for D5RF.1 and D5RF.2 proteins. The D5RF.1-YFP or D5RF.2-YFP vectors were separately co-transferred with the 35S::CFP-Ghd7 vector into Arabidopsis protoplast cells ( a , b ). The 35S::CFP-Ghd7 was used as the nucleic marker. The confocal image was observed using a confocal laser scanning microscope.
    Figure Legend Snippet: Subcellular localization assay for D5RF.1 and D5RF.2 proteins. The D5RF.1-YFP or D5RF.2-YFP vectors were separately co-transferred with the 35S::CFP-Ghd7 vector into Arabidopsis protoplast cells ( a , b ). The 35S::CFP-Ghd7 was used as the nucleic marker. The confocal image was observed using a confocal laser scanning microscope.

    Techniques Used: Plasmid Preparation, Marker, Laser-Scanning Microscopy

    20) Product Images from "Foreign Cry1Ab/c Delays Flowering in Insect-Resistant Transgenic Rice via Interaction With Hd3a Florigen"

    Article Title: Foreign Cry1Ab/c Delays Flowering in Insect-Resistant Transgenic Rice via Interaction With Hd3a Florigen

    Journal: Frontiers in Plant Science

    doi: 10.3389/fpls.2021.608721

    Co-expression of Hd3a-GFP4, Cry1Ab/c-mCherry, and E3s-mCherry inhibits Hd3a protein expression in tobacco mesophyll cells. (A) Co-IP analysis of interactions between Hd3a-GFP4, Cry1Ab/c-mCherry, and E3s-mCherry. Protein extracts (input) were immunoprecipitated with GFP-trap beads (IP) and resolved by SDS-PAGE. Immunoblots were developed with the GFP antibody (AbGFP) to detect Hd3a-GFP (45 kDa) and with mCherry antibody (AbmCherry) to detect Cry1Ab/c-mCherry (94 kDa) and E3s-mCherry (87 kDa) fusion proteins. * indicates the target band. The co-IP assay was repeated three times. (B) The fluorescence intensity of Hd3a-GFP was detected in tobacco mesophyll cells co-expressing Hd3a-GFP4, Cry1Ab/c-mCherry, and E3s-mCherry using a laser confocal microscope. The co-expression of Hd3a-GFP and Cry1Ab/c-mCherry, Hd3a-GFP and mCherry were used as the controls. (C) The expression of Hd3a proteins and co-expression of Hd3a-GFP4, Cry1Ab/c-mCherry, and E3s-mCherry was detected quantitatively (20 μg) by western blotting; co-expressed Hd3a-GFP and Cry1Ab/c-mCherry was used as the control; Rubisco (bottom) was used as loading sample control. (D) Western blot band density of Hd3a protein expression (relative to actin) was analyzed using ImageJ software; * indicates significant differences between treatment means ( P ≤ 0.05; Student’s t -test). The western blot assays were repeated three times.
    Figure Legend Snippet: Co-expression of Hd3a-GFP4, Cry1Ab/c-mCherry, and E3s-mCherry inhibits Hd3a protein expression in tobacco mesophyll cells. (A) Co-IP analysis of interactions between Hd3a-GFP4, Cry1Ab/c-mCherry, and E3s-mCherry. Protein extracts (input) were immunoprecipitated with GFP-trap beads (IP) and resolved by SDS-PAGE. Immunoblots were developed with the GFP antibody (AbGFP) to detect Hd3a-GFP (45 kDa) and with mCherry antibody (AbmCherry) to detect Cry1Ab/c-mCherry (94 kDa) and E3s-mCherry (87 kDa) fusion proteins. * indicates the target band. The co-IP assay was repeated three times. (B) The fluorescence intensity of Hd3a-GFP was detected in tobacco mesophyll cells co-expressing Hd3a-GFP4, Cry1Ab/c-mCherry, and E3s-mCherry using a laser confocal microscope. The co-expression of Hd3a-GFP and Cry1Ab/c-mCherry, Hd3a-GFP and mCherry were used as the controls. (C) The expression of Hd3a proteins and co-expression of Hd3a-GFP4, Cry1Ab/c-mCherry, and E3s-mCherry was detected quantitatively (20 μg) by western blotting; co-expressed Hd3a-GFP and Cry1Ab/c-mCherry was used as the control; Rubisco (bottom) was used as loading sample control. (D) Western blot band density of Hd3a protein expression (relative to actin) was analyzed using ImageJ software; * indicates significant differences between treatment means ( P ≤ 0.05; Student’s t -test). The western blot assays were repeated three times.

    Techniques Used: Expressing, Co-Immunoprecipitation Assay, Immunoprecipitation, SDS Page, Western Blot, Fluorescence, Microscopy, Software

    21) Product Images from "Engineered exosomes delivering specific tumor-suppressive RNAi attenuate oral cancer progression"

    Article Title: Engineered exosomes delivering specific tumor-suppressive RNAi attenuate oral cancer progression

    Journal: Scientific Reports

    doi: 10.1038/s41598-021-85242-1

    Uptake of visualized exosomes under a confocal laser scanning microscope. ( A ) Exosomes from NB1RGB cells were stained with the SYTO RNA Select Reagent (SYTO). HaCaT cells and NB1RGB cells were treated with exosomes derived from NB1RGB cells (NB1RGB exo). Note that immuno-reaction for SYTO (in green) was not evident in these cells. ( B ) NB1RGB exo or EBI3 exo were stained with SYTO. Note that accumulated SYTO staining was detected in EBI3 exo.
    Figure Legend Snippet: Uptake of visualized exosomes under a confocal laser scanning microscope. ( A ) Exosomes from NB1RGB cells were stained with the SYTO RNA Select Reagent (SYTO). HaCaT cells and NB1RGB cells were treated with exosomes derived from NB1RGB cells (NB1RGB exo). Note that immuno-reaction for SYTO (in green) was not evident in these cells. ( B ) NB1RGB exo or EBI3 exo were stained with SYTO. Note that accumulated SYTO staining was detected in EBI3 exo.

    Techniques Used: Laser-Scanning Microscopy, Staining, Derivative Assay

    22) Product Images from "Efficient Transfection of Large Plasmids Encoding HIV-1 into Human Cells—A High Potential Transfection System Based on a Peptide Mimicking Cationic Lipid"

    Article Title: Efficient Transfection of Large Plasmids Encoding HIV-1 into Human Cells—A High Potential Transfection System Based on a Peptide Mimicking Cationic Lipid

    Journal: Pharmaceutics

    doi: 10.3390/pharmaceutics12090805

    Confocal fluorescence microscopy images of HEK293T cells at 8 h ( A ) and 24 h ( B , C ) post-transfection using a lipid mixture containing OH4/DOPE 1/1 (n/n) in a complex with pDNA of the molecular clone HIV-1 JR-FL Gag-iGFP at an N/P ratio of 4. To show typical three-color merged images, Rhod-DOPE (red), pDNA labelled with PoPo ® -1 iodide (blue), and iGFP (green) are selected. Colocalization of Rhod-DOPE and pDNA-PoPo ® -1 iodide in the cell membrane (white arrow) is displayed; Rhod-DOPE cell inclusions were marked with yellow arrows. Observed omega-like structures are marked with a dot line. After 24 h ( B , C ) post-transfection, the expression of the molecular clone HIV-1 JR-FL Gag-iGFP is detectable (green), and viral budding formations can be observed on the surface (cut-out from C). The images were obtained with an inverted confocal laser microscope (Zeiss cLSM 780). For image ( C ), a z-stack of optical sections through the samples was visualized using Zeiss ZEN2011 software to generate a 3D projection. The scale bars represent a length of 10 µm, except in the image cut-outs, where they represent a length of 1 µm.
    Figure Legend Snippet: Confocal fluorescence microscopy images of HEK293T cells at 8 h ( A ) and 24 h ( B , C ) post-transfection using a lipid mixture containing OH4/DOPE 1/1 (n/n) in a complex with pDNA of the molecular clone HIV-1 JR-FL Gag-iGFP at an N/P ratio of 4. To show typical three-color merged images, Rhod-DOPE (red), pDNA labelled with PoPo ® -1 iodide (blue), and iGFP (green) are selected. Colocalization of Rhod-DOPE and pDNA-PoPo ® -1 iodide in the cell membrane (white arrow) is displayed; Rhod-DOPE cell inclusions were marked with yellow arrows. Observed omega-like structures are marked with a dot line. After 24 h ( B , C ) post-transfection, the expression of the molecular clone HIV-1 JR-FL Gag-iGFP is detectable (green), and viral budding formations can be observed on the surface (cut-out from C). The images were obtained with an inverted confocal laser microscope (Zeiss cLSM 780). For image ( C ), a z-stack of optical sections through the samples was visualized using Zeiss ZEN2011 software to generate a 3D projection. The scale bars represent a length of 10 µm, except in the image cut-outs, where they represent a length of 1 µm.

    Techniques Used: Fluorescence, Microscopy, Transfection, Expressing, Confocal Laser Scanning Microscopy, Software

    23) Product Images from "Analysis of Pigment-Dispersing Factor Neuropeptides and Their Receptor in a Velvet Worm"

    Article Title: Analysis of Pigment-Dispersing Factor Neuropeptides and Their Receptor in a Velvet Worm

    Journal: Frontiers in Endocrinology

    doi: 10.3389/fendo.2020.00273

    Combined immunolocalization of Er-PDF-I and Er-PDF-I in the brain of E. rowelli . Confocal laser scanning micrographs of vibratome sections. Dorsal is up in all images and anterior is left in (B,G) . Er-PDF-I-ir (magenta), Er-PDF-II-ir (green) and DNA (gray). (A,B) Er-PDF-I and Er-PDF-II are co-localized in neuropil and somata, albeit at different intensities. (C) Somata (arrowheads) and processes (arrows) of dorsal cell group exhibit equal intensity levels of Er-PDF-I-ir and Er-PDF-II-ir. (D) In contrast, somata and processes of ventral cell group occur in three variants: (i) Er-PDF-I-ir and Er-PDF-II-ir at equal levels (open arrowheads), (ii) Er-PDF-I-ir at higher level (arrows), and (iii) Er-PDF-II-ir at higher level (filled arrowheads). (E–H) Differences in expression levels of Er-PDF-I-ir and Er-PDF-I-ir are also seen in optic neuropil (E) , inner lobe (black asterisk) as opposed to remaining lobes of the mushroom bodies (white asterisks) (F) , somata of connecting cords (G) , and somata and neuropil of nerve cords (H) . an, anterior neuropil; ad, anterior division of central body; at, antennal tract; cb, central body; cc, connecting cord; cn, central neuropil; dl, dorsal perikaryal layer; ey, eye; mb, mushroom body; ol, olfactory lobe; on, optic neuropil; ot, optic tract; pd, posterior division of the central body; vl, ventral perikaryal layer; vn, neuropil of ventral nerve cord; vp, perikaryal layer of ventral nerve cord. Scale bars: 50 μm (A,B) and 20 μm (C–H) .
    Figure Legend Snippet: Combined immunolocalization of Er-PDF-I and Er-PDF-I in the brain of E. rowelli . Confocal laser scanning micrographs of vibratome sections. Dorsal is up in all images and anterior is left in (B,G) . Er-PDF-I-ir (magenta), Er-PDF-II-ir (green) and DNA (gray). (A,B) Er-PDF-I and Er-PDF-II are co-localized in neuropil and somata, albeit at different intensities. (C) Somata (arrowheads) and processes (arrows) of dorsal cell group exhibit equal intensity levels of Er-PDF-I-ir and Er-PDF-II-ir. (D) In contrast, somata and processes of ventral cell group occur in three variants: (i) Er-PDF-I-ir and Er-PDF-II-ir at equal levels (open arrowheads), (ii) Er-PDF-I-ir at higher level (arrows), and (iii) Er-PDF-II-ir at higher level (filled arrowheads). (E–H) Differences in expression levels of Er-PDF-I-ir and Er-PDF-I-ir are also seen in optic neuropil (E) , inner lobe (black asterisk) as opposed to remaining lobes of the mushroom bodies (white asterisks) (F) , somata of connecting cords (G) , and somata and neuropil of nerve cords (H) . an, anterior neuropil; ad, anterior division of central body; at, antennal tract; cb, central body; cc, connecting cord; cn, central neuropil; dl, dorsal perikaryal layer; ey, eye; mb, mushroom body; ol, olfactory lobe; on, optic neuropil; ot, optic tract; pd, posterior division of the central body; vl, ventral perikaryal layer; vn, neuropil of ventral nerve cord; vp, perikaryal layer of ventral nerve cord. Scale bars: 50 μm (A,B) and 20 μm (C–H) .

    Techniques Used: Expressing

    Combined immunolocalization of Er-PDF-II and Er-PDFR in E. rowelli . Confocal laser scanning micrographs of vibratome sections. Dorsal is up in all images. Er-PDF-II (yellow), Er-PDFR (cyan), and DNA (gray). Note that cuticle is autofluorescent. (A) Overview of protocerebrum. (B) Detailed view of dorsal perikaryal layer. (C) Detailed view of ventral perikaryal layer. Note that Er-PDF-II and Er-PDFR are co-localized in some cells of the ventral group (arrowheads). at, antennal tract; cn, central neuropil; dl, dorsal perikaryal layer; ey, eye; mb, mushroom body; ol, olfactory lobe; on, optic neuropil; vl, ventral perikaryal layer; Scale bars: 50 μm (A) and 10 μm (B,C) .
    Figure Legend Snippet: Combined immunolocalization of Er-PDF-II and Er-PDFR in E. rowelli . Confocal laser scanning micrographs of vibratome sections. Dorsal is up in all images. Er-PDF-II (yellow), Er-PDFR (cyan), and DNA (gray). Note that cuticle is autofluorescent. (A) Overview of protocerebrum. (B) Detailed view of dorsal perikaryal layer. (C) Detailed view of ventral perikaryal layer. Note that Er-PDF-II and Er-PDFR are co-localized in some cells of the ventral group (arrowheads). at, antennal tract; cn, central neuropil; dl, dorsal perikaryal layer; ey, eye; mb, mushroom body; ol, olfactory lobe; on, optic neuropil; vl, ventral perikaryal layer; Scale bars: 50 μm (A) and 10 μm (B,C) .

    Techniques Used:

    Combined immunolocalization of Er-PDF-I and Er-PDF-II in the brain of E. rowelli . Maximum projection of a substack of confocal laser scanning micrographs. Dotted lines indicate outline of brain. Anterior is up in all images. Er-PDF-I-ir (magenta), Er-PDF-II-ir (green), and co-localization (white). (A,B) Dorsal Er-PDF-I-ir /Er-PDF-II-ir cell group is subdivided in anterior (filled arrowheads) and median cell groups (open arrowheads). Ventrally located somata with strong Er-PDF-I immunoreactivity occur anteriorly and laterally (A ′ ,C ′ ) , whereas those with strong Er-PDF-II immunoreactivity are located further posteriorly (B ′ ,C ′ ) . (C) Note that all Er-PDF-I-ir and Er-PDF-II-ir somata of dorsal group appear in white, indicating co-localization of both peptides at similar levels. Scale bar: 50 μm.
    Figure Legend Snippet: Combined immunolocalization of Er-PDF-I and Er-PDF-II in the brain of E. rowelli . Maximum projection of a substack of confocal laser scanning micrographs. Dotted lines indicate outline of brain. Anterior is up in all images. Er-PDF-I-ir (magenta), Er-PDF-II-ir (green), and co-localization (white). (A,B) Dorsal Er-PDF-I-ir /Er-PDF-II-ir cell group is subdivided in anterior (filled arrowheads) and median cell groups (open arrowheads). Ventrally located somata with strong Er-PDF-I immunoreactivity occur anteriorly and laterally (A ′ ,C ′ ) , whereas those with strong Er-PDF-II immunoreactivity are located further posteriorly (B ′ ,C ′ ) . (C) Note that all Er-PDF-I-ir and Er-PDF-II-ir somata of dorsal group appear in white, indicating co-localization of both peptides at similar levels. Scale bar: 50 μm.

    Techniques Used:

    Combined immunolocalization of either Er-PDF-I or Er-PDF-I with Er-PDFR in the vascular system of E. rowelli . Confocal laser scanning micrographs of vibratome sections. Anterior is left in all images. Er-PDF-I (magenta), Er-PDF-II (green), Er-PDFR (cyan), and DNA (gray). (A) Er-PDF-I and Er-PDF-II are co-localized in heart nerve with varying intensities, especially within fine branches (arrowheads). (B,C) PDFR immunoreactivity in cell membranes of hemocytes found in heart lumen (B) and body cavity (C) . he, hemocyte; hn, heart nerve. Scale bars: 25 μm (A) , 10 μm (B) , and 5 μm (C) .
    Figure Legend Snippet: Combined immunolocalization of either Er-PDF-I or Er-PDF-I with Er-PDFR in the vascular system of E. rowelli . Confocal laser scanning micrographs of vibratome sections. Anterior is left in all images. Er-PDF-I (magenta), Er-PDF-II (green), Er-PDFR (cyan), and DNA (gray). (A) Er-PDF-I and Er-PDF-II are co-localized in heart nerve with varying intensities, especially within fine branches (arrowheads). (B,C) PDFR immunoreactivity in cell membranes of hemocytes found in heart lumen (B) and body cavity (C) . he, hemocyte; hn, heart nerve. Scale bars: 25 μm (A) , 10 μm (B) , and 5 μm (C) .

    Techniques Used:

    Combined immunolocalization of either Er-PDF-I or Er-PDF-I with Er-PDFR in the visual system E. rowelli . Confocal laser scanning micrographs of vibratome sections. Dorsal is up in all images. Er-PDF-I (magenta), Er-PDF-II (yellow), Er-PDFR (cyan) and DNA (gray). (A–D) PDFR immunoreactivity occurs in optic neuropil, few somata of optic ganglion (arrowheads), rhabdomeric layer ( A–C and insets for detail), and pigment granules (C,D) . Square in (C) indicates region of (D) . Note the lack of co-localization od Er-PDFR with Er-PDF-I (A) or ER-PDF-II (B) . on, optic neuropil; pe, perikaryal layer of eye; pg, pigment granules rh, rhabdomeric layer; layer. Scale bars: 20 μm (A–C) and 2 μm (insets of A , B,D ).
    Figure Legend Snippet: Combined immunolocalization of either Er-PDF-I or Er-PDF-I with Er-PDFR in the visual system E. rowelli . Confocal laser scanning micrographs of vibratome sections. Dorsal is up in all images. Er-PDF-I (magenta), Er-PDF-II (yellow), Er-PDFR (cyan) and DNA (gray). (A–D) PDFR immunoreactivity occurs in optic neuropil, few somata of optic ganglion (arrowheads), rhabdomeric layer ( A–C and insets for detail), and pigment granules (C,D) . Square in (C) indicates region of (D) . Note the lack of co-localization od Er-PDFR with Er-PDF-I (A) or ER-PDF-II (B) . on, optic neuropil; pe, perikaryal layer of eye; pg, pigment granules rh, rhabdomeric layer; layer. Scale bars: 20 μm (A–C) and 2 μm (insets of A , B,D ).

    Techniques Used:

    Immunolocalization of Er-PDF-I and Er-PDF-II in E. rowelli . Confocal laser scanning micrographs of vibratome sections. Dorsal is up in all images. Hatched line indicates median regions. Er-PDF-I-ir (magenta), Er-PDF-II-ir (green) and DNA-labeling (gray) from anterior to posterior through head (A–C) and trunk (D) . Note similar distribution of either peptide in brain and ventral nerve cords. (A) Arrows indicate dorsal groups of somata in protocerebrum. Insets show large varicosities in PDF-immunoreactive fibers. (B) Arrowheads point to large ventral groups of somata in protocerebrum. Asterisks indicate four lobes of mushroom bodies. (C) Filled arrowheads point to somata in deutocerebrum. Empty arrowheads demarcate somata in connecting cords. (D) Cross sections of ventral nerve cords. at, antennal tract; cb, central body; cc, connecting cord; cn, central neuropil; dc, deutocerebrum; dl, dorsal perikaryal layer; ol, olfactory lobe; on, optic neuropil; ot, optic tract; vl, ventral perikaryal layer; vn, neuropil of ventral nerve cord. Scale bars: 50 μm (A–D) and 500 nm (insets).
    Figure Legend Snippet: Immunolocalization of Er-PDF-I and Er-PDF-II in E. rowelli . Confocal laser scanning micrographs of vibratome sections. Dorsal is up in all images. Hatched line indicates median regions. Er-PDF-I-ir (magenta), Er-PDF-II-ir (green) and DNA-labeling (gray) from anterior to posterior through head (A–C) and trunk (D) . Note similar distribution of either peptide in brain and ventral nerve cords. (A) Arrows indicate dorsal groups of somata in protocerebrum. Insets show large varicosities in PDF-immunoreactive fibers. (B) Arrowheads point to large ventral groups of somata in protocerebrum. Asterisks indicate four lobes of mushroom bodies. (C) Filled arrowheads point to somata in deutocerebrum. Empty arrowheads demarcate somata in connecting cords. (D) Cross sections of ventral nerve cords. at, antennal tract; cb, central body; cc, connecting cord; cn, central neuropil; dc, deutocerebrum; dl, dorsal perikaryal layer; ol, olfactory lobe; on, optic neuropil; ot, optic tract; vl, ventral perikaryal layer; vn, neuropil of ventral nerve cord. Scale bars: 50 μm (A–D) and 500 nm (insets).

    Techniques Used: DNA Labeling

    Combined immunolocalization of Er-PDF-I and Er-PDFR in E. rowelli . Confocal laser scanning micrographs of vibratome sections. Dorsal is up in all images. Er-PDF-I (magenta), Er-PDFR (cyan), and DNA (gray). Note that cuticle is autofluorescent. (A) Overview of protocerebrum. (B) Detailed view of dorsal perikaryal layer. (C) Detailed view of ventral perikaryal layer. Note that Er-PDF-I and Er-PDFR are co-localized only in some cells of ventral group (arrowheads). at, antennal tract; cn, central neuropil; dl, dorsal perikaryal layer; ey, eye; mb, mushroom body; ol, olfactory lobe; on, optic neuropil; vl, ventral perikaryal layer; Scale bars: 50 μm (A) and 10 μm (B,C) .
    Figure Legend Snippet: Combined immunolocalization of Er-PDF-I and Er-PDFR in E. rowelli . Confocal laser scanning micrographs of vibratome sections. Dorsal is up in all images. Er-PDF-I (magenta), Er-PDFR (cyan), and DNA (gray). Note that cuticle is autofluorescent. (A) Overview of protocerebrum. (B) Detailed view of dorsal perikaryal layer. (C) Detailed view of ventral perikaryal layer. Note that Er-PDF-I and Er-PDFR are co-localized only in some cells of ventral group (arrowheads). at, antennal tract; cn, central neuropil; dl, dorsal perikaryal layer; ey, eye; mb, mushroom body; ol, olfactory lobe; on, optic neuropil; vl, ventral perikaryal layer; Scale bars: 50 μm (A) and 10 μm (B,C) .

    Techniques Used:

    24) Product Images from "Anemoside B4 ameliorates TNBS-induced colitis through S100A9/MAPK/NF-κB signaling pathway"

    Article Title: Anemoside B4 ameliorates TNBS-induced colitis through S100A9/MAPK/NF-κB signaling pathway

    Journal: Chinese Medicine

    doi: 10.1186/s13020-020-00410-1

    Anemoside B4 attenuates the recruitment of inflammatory cells to colon. a Sections of colon tissue were immunostained with FITC-S100A9 (red) and FITC-CD11b (green) and the immunofluorescence was detected by confocal laser-scanning microscope. Shown were data from three rats in one experiment (n = 3 per group). (B) MPO in colon tissues (U/g) was determined by ELISA kits. Data were shown as mean ± SD (n = 3)
    Figure Legend Snippet: Anemoside B4 attenuates the recruitment of inflammatory cells to colon. a Sections of colon tissue were immunostained with FITC-S100A9 (red) and FITC-CD11b (green) and the immunofluorescence was detected by confocal laser-scanning microscope. Shown were data from three rats in one experiment (n = 3 per group). (B) MPO in colon tissues (U/g) was determined by ELISA kits. Data were shown as mean ± SD (n = 3)

    Techniques Used: Immunofluorescence, Laser-Scanning Microscopy, Enzyme-linked Immunosorbent Assay

    Inhibitory effect of anemoside B4 on the proliferation, apoptosis or inflammation induced by TNBS or LPS. a Sections of colon tissue (n = 3) were immunostained with Edu and observed by confocal laser-scanning microscope. b Fluorescence intensity of Edu stained points. c Sections of colon tissue were immunostained with TUNEL and observed by confocal laser-scanning microscope. d Fluorescence intensity of TUNEL stained points. e p53, Bcl-2, Bax and cleaved caspase 3 was measured by Western blotting. f Statistical analysis of p53, Bcl-2, Bax and cleaved caspase 3 protein level. g LPS-induced NF-κB activation. Caco-2 cells were treated with anemoside B4 as indicated concentration and LPS (1 µg/ml) for 60 min. Cells were harvested and total cell extracts were prepared. p-ERK, p-JNK, p-p38 and p65 were detected by Western blotting analysis. Total ERK, JNK, p38, p65 and GAPDH were used as internal standards. Each experiment was repeated for three times. h Western blotting analysis of proteins in the MAPK/NF-κB pathway. # : P
    Figure Legend Snippet: Inhibitory effect of anemoside B4 on the proliferation, apoptosis or inflammation induced by TNBS or LPS. a Sections of colon tissue (n = 3) were immunostained with Edu and observed by confocal laser-scanning microscope. b Fluorescence intensity of Edu stained points. c Sections of colon tissue were immunostained with TUNEL and observed by confocal laser-scanning microscope. d Fluorescence intensity of TUNEL stained points. e p53, Bcl-2, Bax and cleaved caspase 3 was measured by Western blotting. f Statistical analysis of p53, Bcl-2, Bax and cleaved caspase 3 protein level. g LPS-induced NF-κB activation. Caco-2 cells were treated with anemoside B4 as indicated concentration and LPS (1 µg/ml) for 60 min. Cells were harvested and total cell extracts were prepared. p-ERK, p-JNK, p-p38 and p65 were detected by Western blotting analysis. Total ERK, JNK, p38, p65 and GAPDH were used as internal standards. Each experiment was repeated for three times. h Western blotting analysis of proteins in the MAPK/NF-κB pathway. # : P

    Techniques Used: Laser-Scanning Microscopy, Fluorescence, Staining, TUNEL Assay, Western Blot, Activation Assay, Concentration Assay

    25) Product Images from "Reversal of Multidrug Resistance by Apolipoprotein A1-Modified Doxorubicin Liposome for Breast Cancer Treatment"

    Article Title: Reversal of Multidrug Resistance by Apolipoprotein A1-Modified Doxorubicin Liposome for Breast Cancer Treatment

    Journal: Molecules

    doi: 10.3390/molecules26051280

    Cellular uptake profile of Dox-loaded liposomes. ( A , B ): Intracellular fluorescence of Dox measured by flow cytometry and a confocal laser scanning microscope (CLSM). The cells were co-incubated with different Dox liposomal formulations for 2 h. ( C ) Confocal observation after treating cells with free Dox, Lip/Dox, ApoA1-lip/Dox after pretreatment of 0.5 mg/mL anti-SR-B1 (scavenger receptor, class B type 1) antibody and ApoA1 for 2 h. ( D ) Effect of endocytosis inhibitors on cellular uptake. The percentage of cellular uptake was calculated by the Median Fluorescence Intensity (MFI) value of the inhibitor group normalized with that of the control group (100%). Cells without inhibitors were regarded as a control ( n = 3; * p
    Figure Legend Snippet: Cellular uptake profile of Dox-loaded liposomes. ( A , B ): Intracellular fluorescence of Dox measured by flow cytometry and a confocal laser scanning microscope (CLSM). The cells were co-incubated with different Dox liposomal formulations for 2 h. ( C ) Confocal observation after treating cells with free Dox, Lip/Dox, ApoA1-lip/Dox after pretreatment of 0.5 mg/mL anti-SR-B1 (scavenger receptor, class B type 1) antibody and ApoA1 for 2 h. ( D ) Effect of endocytosis inhibitors on cellular uptake. The percentage of cellular uptake was calculated by the Median Fluorescence Intensity (MFI) value of the inhibitor group normalized with that of the control group (100%). Cells without inhibitors were regarded as a control ( n = 3; * p

    Techniques Used: Fluorescence, Flow Cytometry, Laser-Scanning Microscopy, Confocal Laser Scanning Microscopy, Incubation

    26) Product Images from "Guanidine modifications enhance the anti‐herpes simplex virus activity of (E,E)‐4,6‐bis(styryl)‐pyrimidine derivatives in vitro and in vivo, et al. Guanidine modifications enhance the anti‐herpes simplex virus activity of (E,E)‐4,6‐bis(styryl)‐pyrimidine derivatives in vitro and in vivo"

    Article Title: Guanidine modifications enhance the anti‐herpes simplex virus activity of (E,E)‐4,6‐bis(styryl)‐pyrimidine derivatives in vitro and in vivo, et al. Guanidine modifications enhance the anti‐herpes simplex virus activity of (E,E)‐4,6‐bis(styryl)‐pyrimidine derivatives in vitro and in vivo

    Journal: British Journal of Pharmacology

    doi: 10.1111/bph.14918

    Subcellular localization of BS‐pyrimidine derivatives in Vero and HeLa cells. (a) Vero cells were incubated with compounds 3a, 5a, or 5d (30 μM) for 1 hr at 37°C. The fluorescence was then detected by confocal laser scanning microscopy. The scale bar represents 50 μm. An enlarged view of part of one field (highlighted by the red rectangle) is shown to indicate the localization of compound 5d within the Vero cells. The scale bar represents 10 μm in this case. (b‐d) HeLa cells were transfected with expression plasmids for red fluorescent protein (RFP)‐coupled (b) Rab5, (c) Rab7, or (d) LAMP‐1. At 24 hr post‐transfection, cells were added with compound 5d (20 μM) and incubated for 1 hr at 37°C. After that, the fluorescence was detected via confocal microscopy. The scale bar represents 10 μm. (e, f) Herpes simplex virus (HSV)‐2 (multiplicity of infection = 1.0) was pretreated with (e) compound 3a or (f) 5d at 37°C for 1 hr before infection. Then, after removal of the virus inoculum, media containing compound 3a or 5d (20 μM) were added to cells. At 1 hr p.i., the localization of virus gB protein was evaluated via immunofluorescence assay. The fluorescence was detected via confocal microscopy. The scale bar represents 20 μm
    Figure Legend Snippet: Subcellular localization of BS‐pyrimidine derivatives in Vero and HeLa cells. (a) Vero cells were incubated with compounds 3a, 5a, or 5d (30 μM) for 1 hr at 37°C. The fluorescence was then detected by confocal laser scanning microscopy. The scale bar represents 50 μm. An enlarged view of part of one field (highlighted by the red rectangle) is shown to indicate the localization of compound 5d within the Vero cells. The scale bar represents 10 μm in this case. (b‐d) HeLa cells were transfected with expression plasmids for red fluorescent protein (RFP)‐coupled (b) Rab5, (c) Rab7, or (d) LAMP‐1. At 24 hr post‐transfection, cells were added with compound 5d (20 μM) and incubated for 1 hr at 37°C. After that, the fluorescence was detected via confocal microscopy. The scale bar represents 10 μm. (e, f) Herpes simplex virus (HSV)‐2 (multiplicity of infection = 1.0) was pretreated with (e) compound 3a or (f) 5d at 37°C for 1 hr before infection. Then, after removal of the virus inoculum, media containing compound 3a or 5d (20 μM) were added to cells. At 1 hr p.i., the localization of virus gB protein was evaluated via immunofluorescence assay. The fluorescence was detected via confocal microscopy. The scale bar represents 20 μm

    Techniques Used: Incubation, Fluorescence, Confocal Laser Scanning Microscopy, Transfection, Expressing, Confocal Microscopy, Infection, Immunofluorescence

    27) Product Images from "Targeted Nanobubbles Carrying Indocyanine Green for Ultrasound, Photoacoustic and Fluorescence Imaging of Prostate Cancer"

    Article Title: Targeted Nanobubbles Carrying Indocyanine Green for Ultrasound, Photoacoustic and Fluorescence Imaging of Prostate Cancer

    Journal: International Journal of Nanomedicine

    doi: 10.2147/IJN.S243548

    Verification of the coupling of PSMA-binding peptides to the surfaces of the PSMAP/ICG NBs When observed under a confocal laser scanning microscope, the NBs with encapsulated ICG in the lipid shell showed red fluorescence ( A ), and the FITC-labeled PSMA-binding peptides showed green fluorescence ( B ). The complete overlap of the two types of fluorescence resulted in yellow fluorescence ( C ).
    Figure Legend Snippet: Verification of the coupling of PSMA-binding peptides to the surfaces of the PSMAP/ICG NBs When observed under a confocal laser scanning microscope, the NBs with encapsulated ICG in the lipid shell showed red fluorescence ( A ), and the FITC-labeled PSMA-binding peptides showed green fluorescence ( B ). The complete overlap of the two types of fluorescence resulted in yellow fluorescence ( C ).

    Techniques Used: Binding Assay, Laser-Scanning Microscopy, Fluorescence, Labeling

    28) Product Images from "Metastasis-associated gene, mag-1 improves tumour microenvironmental adaptation and potentiates tumour metastasis"

    Article Title: Metastasis-associated gene, mag-1 improves tumour microenvironmental adaptation and potentiates tumour metastasis

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/j.1582-4934.2012.01633.x

    Identification and expression profile of mag-1 gene. (A) Verification of differential expression of mag-1 in PLA801C and D cells. Upper two lines indicate semi-quantitative RT-PCR analysis of mag-1 mRNAs. Middle two lines are Western blot analysis of Mag-1 protein levels using β-actin as internal control between PLA801C and D. Lower two lines indicate northern blot analysis of mag-1 mRNAs using GAPDH as internal control between PLA801C and PLA801D. (B) Molecular weight of Mag-1 protein analysed using Western blot analysis. (C) Subcellular localization of Mag-1 within cells. A immunofluorescent staining of COS-7 and H1299 cells overexpressing His-Tagged Mag-1. Mag-1 was stained with anti-His antibody (Green) and visualized co-localization with ER specific protein, Calnexin (Red). Cells were double stained with Hoechst 33258 (blue) to identify the nuclei in the corresponding fields. All cell samples were visualized using Confocal laser-scanning microscope (Zeiss 510 META, Oberkochen, Germany). (D) Western blot analysis of cell membrane and cytoplasma fraction from H1299 and COS-7 cells, stably transfected mag-1 or vector control. (E) Northern blot analysis of mag-1 transcription in eight human tissues. A human multiple tissue northern blot membrane was hybridized with 32 P-labelled mag-1 specific or β-actin cDNA probes (bottom) respectively. Hybridization with β-actin served as a loading control. Size marker is indicated on the left. (F) Western blot analysis of mag-1 protein in multiple cancer cell lines. β-Actin was used as an internal control.
    Figure Legend Snippet: Identification and expression profile of mag-1 gene. (A) Verification of differential expression of mag-1 in PLA801C and D cells. Upper two lines indicate semi-quantitative RT-PCR analysis of mag-1 mRNAs. Middle two lines are Western blot analysis of Mag-1 protein levels using β-actin as internal control between PLA801C and D. Lower two lines indicate northern blot analysis of mag-1 mRNAs using GAPDH as internal control between PLA801C and PLA801D. (B) Molecular weight of Mag-1 protein analysed using Western blot analysis. (C) Subcellular localization of Mag-1 within cells. A immunofluorescent staining of COS-7 and H1299 cells overexpressing His-Tagged Mag-1. Mag-1 was stained with anti-His antibody (Green) and visualized co-localization with ER specific protein, Calnexin (Red). Cells were double stained with Hoechst 33258 (blue) to identify the nuclei in the corresponding fields. All cell samples were visualized using Confocal laser-scanning microscope (Zeiss 510 META, Oberkochen, Germany). (D) Western blot analysis of cell membrane and cytoplasma fraction from H1299 and COS-7 cells, stably transfected mag-1 or vector control. (E) Northern blot analysis of mag-1 transcription in eight human tissues. A human multiple tissue northern blot membrane was hybridized with 32 P-labelled mag-1 specific or β-actin cDNA probes (bottom) respectively. Hybridization with β-actin served as a loading control. Size marker is indicated on the left. (F) Western blot analysis of mag-1 protein in multiple cancer cell lines. β-Actin was used as an internal control.

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Northern Blot, Molecular Weight, Staining, Laser-Scanning Microscopy, Stable Transfection, Transfection, Plasmid Preparation, Hybridization, Marker

    29) Product Images from "Guanidine modifications enhance the anti‐herpes simplex virus activity of (E,E)‐4,6‐bis(styryl)‐pyrimidine derivatives in vitro and in vivo, et al. Guanidine modifications enhance the anti‐herpes simplex virus activity of (E,E)‐4,6‐bis(styryl)‐pyrimidine derivatives in vitro and in vivo"

    Article Title: Guanidine modifications enhance the anti‐herpes simplex virus activity of (E,E)‐4,6‐bis(styryl)‐pyrimidine derivatives in vitro and in vivo, et al. Guanidine modifications enhance the anti‐herpes simplex virus activity of (E,E)‐4,6‐bis(styryl)‐pyrimidine derivatives in vitro and in vivo

    Journal: British Journal of Pharmacology

    doi: 10.1111/bph.14918

    Subcellular localization of BS‐pyrimidine derivatives in Vero and HeLa cells. (a) Vero cells were incubated with compounds 3a, 5a, or 5d (30 μM) for 1 hr at 37°C. The fluorescence was then detected by confocal laser scanning microscopy. The scale bar represents 50 μm. An enlarged view of part of one field (highlighted by the red rectangle) is shown to indicate the localization of compound 5d within the Vero cells. The scale bar represents 10 μm in this case. (b‐d) HeLa cells were transfected with expression plasmids for red fluorescent protein (RFP)‐coupled (b) Rab5, (c) Rab7, or (d) LAMP‐1. At 24 hr post‐transfection, cells were added with compound 5d (20 μM) and incubated for 1 hr at 37°C. After that, the fluorescence was detected via confocal microscopy. The scale bar represents 10 μm. (e, f) Herpes simplex virus (HSV)‐2 (multiplicity of infection = 1.0) was pretreated with (e) compound 3a or (f) 5d at 37°C for 1 hr before infection. Then, after removal of the virus inoculum, media containing compound 3a or 5d (20 μM) were added to cells. At 1 hr p.i., the localization of virus gB protein was evaluated via immunofluorescence assay. The fluorescence was detected via confocal microscopy. The scale bar represents 20 μm
    Figure Legend Snippet: Subcellular localization of BS‐pyrimidine derivatives in Vero and HeLa cells. (a) Vero cells were incubated with compounds 3a, 5a, or 5d (30 μM) for 1 hr at 37°C. The fluorescence was then detected by confocal laser scanning microscopy. The scale bar represents 50 μm. An enlarged view of part of one field (highlighted by the red rectangle) is shown to indicate the localization of compound 5d within the Vero cells. The scale bar represents 10 μm in this case. (b‐d) HeLa cells were transfected with expression plasmids for red fluorescent protein (RFP)‐coupled (b) Rab5, (c) Rab7, or (d) LAMP‐1. At 24 hr post‐transfection, cells were added with compound 5d (20 μM) and incubated for 1 hr at 37°C. After that, the fluorescence was detected via confocal microscopy. The scale bar represents 10 μm. (e, f) Herpes simplex virus (HSV)‐2 (multiplicity of infection = 1.0) was pretreated with (e) compound 3a or (f) 5d at 37°C for 1 hr before infection. Then, after removal of the virus inoculum, media containing compound 3a or 5d (20 μM) were added to cells. At 1 hr p.i., the localization of virus gB protein was evaluated via immunofluorescence assay. The fluorescence was detected via confocal microscopy. The scale bar represents 20 μm

    Techniques Used: Incubation, Fluorescence, Confocal Laser Scanning Microscopy, Transfection, Expressing, Confocal Microscopy, Infection, Immunofluorescence

    30) Product Images from "Tubule-specific protein nanocages potentiate targeted renal fibrosis therapy"

    Article Title: Tubule-specific protein nanocages potentiate targeted renal fibrosis therapy

    Journal: Journal of Nanobiotechnology

    doi: 10.1186/s12951-021-00900-w

    Confocal laser scanning microscope images of HK-2 cells incubated with ( A ) HBc-183/CLT and ( B ) K3-HBc/CLT at different time points (0.5 and 1 h). Representative fluorescence images of nuclei counterstained with DAPI (blue) and HBc-183/CLT and K3-HBc/CLT labeled with Cy5.5 (red), Scale bars, 20 μm. C – E Flow cytometric analyses of HK-2 cells treated with FITC-labeled HBc-183/CLT or K3-HBc/CLT NCs for 0.5, 1, 2 and 4 h at 37 °C (n = 3) (*** p
    Figure Legend Snippet: Confocal laser scanning microscope images of HK-2 cells incubated with ( A ) HBc-183/CLT and ( B ) K3-HBc/CLT at different time points (0.5 and 1 h). Representative fluorescence images of nuclei counterstained with DAPI (blue) and HBc-183/CLT and K3-HBc/CLT labeled with Cy5.5 (red), Scale bars, 20 μm. C – E Flow cytometric analyses of HK-2 cells treated with FITC-labeled HBc-183/CLT or K3-HBc/CLT NCs for 0.5, 1, 2 and 4 h at 37 °C (n = 3) (*** p

    Techniques Used: Laser-Scanning Microscopy, Incubation, Fluorescence, Labeling

    31) Product Images from "In vivo vaccination with cell line-derived whole tumor lysates: neoantigen quality, not quantity matters"

    Article Title: In vivo vaccination with cell line-derived whole tumor lysates: neoantigen quality, not quantity matters

    Journal: Journal of Translational Medicine

    doi: 10.1186/s12967-020-02570-y

    Tumor microenvironment. Immunofluorescence was done on 4 µM slides stained with mAbs as stated in the material and methods section. Cell nuclei were stained with DAPI. Representative images are given showing either GIT in which treatment failed (left panel) or, as in the case of A7450 T1 M1, treatment succeeded. Differences in leukocytic infiltration are evident. Analyses were done on a laser scanning microscope (Zeiss) using 20× objectives
    Figure Legend Snippet: Tumor microenvironment. Immunofluorescence was done on 4 µM slides stained with mAbs as stated in the material and methods section. Cell nuclei were stained with DAPI. Representative images are given showing either GIT in which treatment failed (left panel) or, as in the case of A7450 T1 M1, treatment succeeded. Differences in leukocytic infiltration are evident. Analyses were done on a laser scanning microscope (Zeiss) using 20× objectives

    Techniques Used: Immunofluorescence, Staining, Laser-Scanning Microscopy

    32) Product Images from "PD-L1 expression in bone marrow plasma cells as a biomarker to predict multiple myeloma prognosis: developing a nomogram-based prognostic model"

    Article Title: PD-L1 expression in bone marrow plasma cells as a biomarker to predict multiple myeloma prognosis: developing a nomogram-based prognostic model

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-69616-5

    Immunofluorescence analysis of PD-L1 expression in bone marrow-aspirated plasma cells from patients with multiple myeloma. ( A ) Formalin-fixed, paraffin-embedded bone marrow aspirate specimens (clot section) from myeloma patients were sectioned at 4–5 µm. The sections were then incubated with antibodies to CD138 (1:100) and PD-L1 (1:100) overnight at 4 °C, followed by incubation with the appropriate secondary antibodies (Alexa Fluor 488, 1:200 and Alexa Fluor 647, 1:200) at room temperature for one hour. Nuclei were counterstained using DAPI, and all images were captured using a confocal laser scanning microscope (CLSM 800, Carl Zeiss Microscopy GmbH). Original magnification × 200. ( B ) Representative immunofluorescence images for the PD-L1 expression scores and groups based on the predetermined cut-off value of 7.65. ( C ) The distribution of the PD-L1 expression scores in patients with multiple myeloma. PD-L1 programmed death-ligand 1.
    Figure Legend Snippet: Immunofluorescence analysis of PD-L1 expression in bone marrow-aspirated plasma cells from patients with multiple myeloma. ( A ) Formalin-fixed, paraffin-embedded bone marrow aspirate specimens (clot section) from myeloma patients were sectioned at 4–5 µm. The sections were then incubated with antibodies to CD138 (1:100) and PD-L1 (1:100) overnight at 4 °C, followed by incubation with the appropriate secondary antibodies (Alexa Fluor 488, 1:200 and Alexa Fluor 647, 1:200) at room temperature for one hour. Nuclei were counterstained using DAPI, and all images were captured using a confocal laser scanning microscope (CLSM 800, Carl Zeiss Microscopy GmbH). Original magnification × 200. ( B ) Representative immunofluorescence images for the PD-L1 expression scores and groups based on the predetermined cut-off value of 7.65. ( C ) The distribution of the PD-L1 expression scores in patients with multiple myeloma. PD-L1 programmed death-ligand 1.

    Techniques Used: Immunofluorescence, Expressing, Formalin-fixed Paraffin-Embedded, Incubation, Laser-Scanning Microscopy, Confocal Laser Scanning Microscopy, Microscopy

    33) Product Images from "Poly I:C Activated Microglia Disrupt Perineuronal Nets and Modulate Synaptic Balance in Primary Hippocampal Neurons in vitro"

    Article Title: Poly I:C Activated Microglia Disrupt Perineuronal Nets and Modulate Synaptic Balance in Primary Hippocampal Neurons in vitro

    Journal: Frontiers in Synaptic Neuroscience

    doi: 10.3389/fnsyn.2021.637549

    (A–F) Confocal laser-scanning microscopy of immunocytochemically visualized presynaptic vGAT (red) and postsynaptic gephyrin puncta (green) on cultured hippocampal neurons incubated for 24 h with microglia conditioned medium or hippocampus medium. Neurons were previously kept for 12 days in vitro ; (E–G) A colocalization of vGAT and gephyrin puncta led to yellow signals in maximum intensity projection (indicated by arrows) which were defined as structural GABAergic synapses; (H) Analysis of presynaptic vGAT puncta revealed a significant reduction in neuronal cultures treated for 24 h with microglia conditioned medium in comparison to neuronal cultures treated with hippocampus medium; (I) No significant changes were observed with regard to the number of postsynaptic gephyrin puncta between both conditions; (J) The number of colocalized inhibitory puncta was significantly lower when neurons were incubated with conditioned medium of Poly I:C activated microglia (scale bar: 50 μm). Statistics: Three independent experimental repetitions ( N = 3) were performed for the analysis of synaptic puncta. For the quantification 20 neurons ( n = 20) were recorded per repetition. Data are shown as mean ± SD (Mann-Whitney U -test, * p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001). Dots in diagram (H–J) stand for puncta numbers of single neurons, as previously described in figure legend 5.
    Figure Legend Snippet: (A–F) Confocal laser-scanning microscopy of immunocytochemically visualized presynaptic vGAT (red) and postsynaptic gephyrin puncta (green) on cultured hippocampal neurons incubated for 24 h with microglia conditioned medium or hippocampus medium. Neurons were previously kept for 12 days in vitro ; (E–G) A colocalization of vGAT and gephyrin puncta led to yellow signals in maximum intensity projection (indicated by arrows) which were defined as structural GABAergic synapses; (H) Analysis of presynaptic vGAT puncta revealed a significant reduction in neuronal cultures treated for 24 h with microglia conditioned medium in comparison to neuronal cultures treated with hippocampus medium; (I) No significant changes were observed with regard to the number of postsynaptic gephyrin puncta between both conditions; (J) The number of colocalized inhibitory puncta was significantly lower when neurons were incubated with conditioned medium of Poly I:C activated microglia (scale bar: 50 μm). Statistics: Three independent experimental repetitions ( N = 3) were performed for the analysis of synaptic puncta. For the quantification 20 neurons ( n = 20) were recorded per repetition. Data are shown as mean ± SD (Mann-Whitney U -test, * p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001). Dots in diagram (H–J) stand for puncta numbers of single neurons, as previously described in figure legend 5.

    Techniques Used: Confocal Laser Scanning Microscopy, Cell Culture, Incubation, In Vitro, MANN-WHITNEY

    (A–F) Confocal laser-scanning microscopy of immunocytochemically visualized presynaptic vGlut (red) and postsynaptic PSD-95 puncta (green) on hippocampal neurons treated for 24 h with conditioned medium of Poly I:C activated microglia or hippocampus medium as control. Before treatment, neuronal networks were cultured for 12 days; (E–G) A colocalization of presynaptic vGlut and postsynaptic PSD-95 puncta yielded in yellow signals (indicated by arrows) which were defined as structural glutamatergic synapses; (H) Quantification of vGlut puncta revealed a significant reduction when neurons were treated with microglia conditioned medium; (I) With regard to PSD-95 puncta no significant changes were observed after treatment; (J) The number of colocalized synaptic puncta was significantly lower in neuronal cultures treated with microglia conditioned medium (scale bar: 50 μm). Statistics: Three independent experimental repetitions ( N = 3) were performed for the analysis of synaptic puncta. For the quantification 20 neurons ( n = 20) were recorded per repetition. Data are shown as mean ± SD (t-test, * p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001). Single dots in diagram (H–J) are representing puncta numbers of individually analyzed neurons.
    Figure Legend Snippet: (A–F) Confocal laser-scanning microscopy of immunocytochemically visualized presynaptic vGlut (red) and postsynaptic PSD-95 puncta (green) on hippocampal neurons treated for 24 h with conditioned medium of Poly I:C activated microglia or hippocampus medium as control. Before treatment, neuronal networks were cultured for 12 days; (E–G) A colocalization of presynaptic vGlut and postsynaptic PSD-95 puncta yielded in yellow signals (indicated by arrows) which were defined as structural glutamatergic synapses; (H) Quantification of vGlut puncta revealed a significant reduction when neurons were treated with microglia conditioned medium; (I) With regard to PSD-95 puncta no significant changes were observed after treatment; (J) The number of colocalized synaptic puncta was significantly lower in neuronal cultures treated with microglia conditioned medium (scale bar: 50 μm). Statistics: Three independent experimental repetitions ( N = 3) were performed for the analysis of synaptic puncta. For the quantification 20 neurons ( n = 20) were recorded per repetition. Data are shown as mean ± SD (t-test, * p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001). Single dots in diagram (H–J) are representing puncta numbers of individually analyzed neurons.

    Techniques Used: Confocal Laser Scanning Microscopy, Cell Culture, T-Test

    (A,B) Representative confocal laser scanning microscopy images of structural glutamatergic synapses on perineuronal net wearing neurons after a treatment with fresh hippocampus medium (controls). Neurons were immunocytochemically stained against vGlut (red), PSD-95 (green), and Aggrecan (blue), a major PNN component; (C,D) Grayscale images of Aggrecan positive PNNs. PNNs were evenly covering the soma and proximal dendrites of cultured hippocampal neurons (scale bar: 100 μm); (E–H) Representative immunostainings of excitatory synapses on PNN positive neurons after a 24 h treatment with conditioned medium of activated microglia. Grayscale depictions show a clear disruption of Aggrecan containing PNNs as indicated by arrows; (I–K) Quantification of PNN parameters showed a significant reduction in PNN area and staining intensity; (L–N) Analysis of synaptic puncta showed no changes with regard to presynaptic vGlut signals. However, a significantly raised number of postsynaptic PSD-95 and colocalized puncta could be verified. Statistics: Three independent experimental repetitions ( N = 3) were performed for the analysis of synaptic puncta PNN parameters. 20 neurons ( n = 20) were recorded and quantified per repetition. Data are shown as mean ± SD ( t -test, * p ≤ 0.05, ** p ≤ 0.01, and *** p ≤ 0.001). The data dots in (I–N) represent individual values of single neurons.
    Figure Legend Snippet: (A,B) Representative confocal laser scanning microscopy images of structural glutamatergic synapses on perineuronal net wearing neurons after a treatment with fresh hippocampus medium (controls). Neurons were immunocytochemically stained against vGlut (red), PSD-95 (green), and Aggrecan (blue), a major PNN component; (C,D) Grayscale images of Aggrecan positive PNNs. PNNs were evenly covering the soma and proximal dendrites of cultured hippocampal neurons (scale bar: 100 μm); (E–H) Representative immunostainings of excitatory synapses on PNN positive neurons after a 24 h treatment with conditioned medium of activated microglia. Grayscale depictions show a clear disruption of Aggrecan containing PNNs as indicated by arrows; (I–K) Quantification of PNN parameters showed a significant reduction in PNN area and staining intensity; (L–N) Analysis of synaptic puncta showed no changes with regard to presynaptic vGlut signals. However, a significantly raised number of postsynaptic PSD-95 and colocalized puncta could be verified. Statistics: Three independent experimental repetitions ( N = 3) were performed for the analysis of synaptic puncta PNN parameters. 20 neurons ( n = 20) were recorded and quantified per repetition. Data are shown as mean ± SD ( t -test, * p ≤ 0.05, ** p ≤ 0.01, and *** p ≤ 0.001). The data dots in (I–N) represent individual values of single neurons.

    Techniques Used: Confocal Laser Scanning Microscopy, Staining, Cell Culture

    (A,B) Representative confocal laser scanning microscopy images of structural GABAergic synapses on perineuronal net wearing neurons after a 24 h incubation with fresh hippocampus medium (controls). Neurons were immunostained with antibodies against vGAT (red), gephyrin (green) and Aggrecan (blue), which can be found in all mature PNNs; (C,D) Grayscale images of Aggrecan positive PNNs that were evenly covering the soma and proximal dendrites of cultured neurons (scale bar: 100 μm); (E–H) Representative recordings of inhibitory synapses on PNN positive neurons after a 24 h treatment with conditioned medium of Poly I:C activated microglia. Grayscale depictions show a clear disruption of Aggrecan containing PNNs as indicated by arrows; (I) Quantification of presynaptic vGAT puncta revealed a significant reduction when neuronal networks were treated with microglia conditioned medium; (J) In contrast, postsynaptic gephyrin puncta were significantly increased on neurons with disrupted perineuronal nets; (K) Number of structural inhibitory synapses was significantly decreased after the treatment with conditioned medium of activated microglia; (L) For the analysis of excitatory and inhibitory balance, a ratio was formed between the colocalized excitatory and colocalized inhibitory fluorescence of PNN negative neurons. Here, no significant alteration could be observed in the excitatory: inhibitory ratio; (M) The excitatory: inhibitory ratio was also determined for PNN wearing neurons and showed a significant increase when neurons were treated for 24 h with microglia conditioned medium of activated microglia. This indicates a shift toward a higher excitatory input compared to control neurons treated with the freshly prepared medium. Statistics: Three independent experimental repetitions ( N = 3) were performed for the analysis of synaptic puncta parameters and ratio formation. 20 neurons ( n = 20) were recorded and quantified per repetition. Data of puncta analysis are shown as mean ± SD (Mann-Whitney U -test, * p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001) and data of ratio analysis are shown as mean ± SEM (Mann-Whitney U-test, p ≤ 0.05). Single data dots in (I–K) are representing puncta numbers of single neurons.
    Figure Legend Snippet: (A,B) Representative confocal laser scanning microscopy images of structural GABAergic synapses on perineuronal net wearing neurons after a 24 h incubation with fresh hippocampus medium (controls). Neurons were immunostained with antibodies against vGAT (red), gephyrin (green) and Aggrecan (blue), which can be found in all mature PNNs; (C,D) Grayscale images of Aggrecan positive PNNs that were evenly covering the soma and proximal dendrites of cultured neurons (scale bar: 100 μm); (E–H) Representative recordings of inhibitory synapses on PNN positive neurons after a 24 h treatment with conditioned medium of Poly I:C activated microglia. Grayscale depictions show a clear disruption of Aggrecan containing PNNs as indicated by arrows; (I) Quantification of presynaptic vGAT puncta revealed a significant reduction when neuronal networks were treated with microglia conditioned medium; (J) In contrast, postsynaptic gephyrin puncta were significantly increased on neurons with disrupted perineuronal nets; (K) Number of structural inhibitory synapses was significantly decreased after the treatment with conditioned medium of activated microglia; (L) For the analysis of excitatory and inhibitory balance, a ratio was formed between the colocalized excitatory and colocalized inhibitory fluorescence of PNN negative neurons. Here, no significant alteration could be observed in the excitatory: inhibitory ratio; (M) The excitatory: inhibitory ratio was also determined for PNN wearing neurons and showed a significant increase when neurons were treated for 24 h with microglia conditioned medium of activated microglia. This indicates a shift toward a higher excitatory input compared to control neurons treated with the freshly prepared medium. Statistics: Three independent experimental repetitions ( N = 3) were performed for the analysis of synaptic puncta parameters and ratio formation. 20 neurons ( n = 20) were recorded and quantified per repetition. Data of puncta analysis are shown as mean ± SD (Mann-Whitney U -test, * p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001) and data of ratio analysis are shown as mean ± SEM (Mann-Whitney U-test, p ≤ 0.05). Single data dots in (I–K) are representing puncta numbers of single neurons.

    Techniques Used: Confocal Laser Scanning Microscopy, Incubation, Cell Culture, Fluorescence, MANN-WHITNEY

    34) Product Images from "Antimicrobial Photodynamic Therapy Combined With Antibiotic in the Treatment of Rats With Third-Degree Burns"

    Article Title: Antimicrobial Photodynamic Therapy Combined With Antibiotic in the Treatment of Rats With Third-Degree Burns

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2021.622410

    Confocal laser scanning microscopy images of (a) Pseudomonas aeruginosa , (b) Escherichia coli , and (c) methicillin-resistant Staphylococcus aureus (MRSA) and optical images of (d) P. aeruginosa , (e) E. coli , and (f) MRSA. The strains were imaged after incubation for 30 min with PPIX-MED (15.6 μM) in phosphate-buffered saline. Excitation was at 405 nm. Scale bars = 2 μm.
    Figure Legend Snippet: Confocal laser scanning microscopy images of (a) Pseudomonas aeruginosa , (b) Escherichia coli , and (c) methicillin-resistant Staphylococcus aureus (MRSA) and optical images of (d) P. aeruginosa , (e) E. coli , and (f) MRSA. The strains were imaged after incubation for 30 min with PPIX-MED (15.6 μM) in phosphate-buffered saline. Excitation was at 405 nm. Scale bars = 2 μm.

    Techniques Used: Confocal Laser Scanning Microscopy, Incubation

    35) Product Images from "The role of PQL genes in response to salinity tolerance in Arabidopsis and barley, et al. The role of PQL genes in response to salinity tolerance in Arabidopsis and barley"

    Article Title: The role of PQL genes in response to salinity tolerance in Arabidopsis and barley, et al. The role of PQL genes in response to salinity tolerance in Arabidopsis and barley

    Journal: Plant Direct

    doi: 10.1002/pld3.301

    AtPQL1a, AtPQL1b, AtPQL1c, and HvPQL1 co‐localize with the tonoplast marker during transient co‐expression in N. benthamiana leaf epidermal cells. Transient expression of UBQ10p::AtPQL1a‐eGFP, UBQ10p::AtPQL1b‐eGFP, UBQ10p::eGFP‐AtPQL1c, and UBQ10p::HvPQL1‐eGFP was observed using confocal laser scanning microscopy. For all transformants, the PQL proteins (left panel) were co‐infiltrated with tonoplast marker (CD3‐975:: mCherry , middle panel), and the overlay of the two was used to study co‐localization. The red arrows indicate the circular structures (bulbs) that formed in the lumen of the vacuole. The yellow stars indicate the separation between the two neighboring cells. The yellow arrowheads represent the transvacuolar strands. The images are representative of 15 replicates, visualized during 3 independent microscopy sessions
    Figure Legend Snippet: AtPQL1a, AtPQL1b, AtPQL1c, and HvPQL1 co‐localize with the tonoplast marker during transient co‐expression in N. benthamiana leaf epidermal cells. Transient expression of UBQ10p::AtPQL1a‐eGFP, UBQ10p::AtPQL1b‐eGFP, UBQ10p::eGFP‐AtPQL1c, and UBQ10p::HvPQL1‐eGFP was observed using confocal laser scanning microscopy. For all transformants, the PQL proteins (left panel) were co‐infiltrated with tonoplast marker (CD3‐975:: mCherry , middle panel), and the overlay of the two was used to study co‐localization. The red arrows indicate the circular structures (bulbs) that formed in the lumen of the vacuole. The yellow stars indicate the separation between the two neighboring cells. The yellow arrowheads represent the transvacuolar strands. The images are representative of 15 replicates, visualized during 3 independent microscopy sessions

    Techniques Used: Marker, Expressing, Confocal Laser Scanning Microscopy, Microscopy

    AtPQL1b localizes in the internal membrane compartments in stably transformed Arabidopsis root tips. The root tips of stably transformed Arabidopsis plants were imaged using confocal laser scanning microscopy. The left panel shows the root epidermal cells stably transformed with UBQ10p::AtPQL1b‐GFP . The middle panel shows the propidium iodide staining of the same cells, while the right panel is the merge between the two channels. Similar results were observed in 12 replicates, visualized during 3 independent microscopy sessions
    Figure Legend Snippet: AtPQL1b localizes in the internal membrane compartments in stably transformed Arabidopsis root tips. The root tips of stably transformed Arabidopsis plants were imaged using confocal laser scanning microscopy. The left panel shows the root epidermal cells stably transformed with UBQ10p::AtPQL1b‐GFP . The middle panel shows the propidium iodide staining of the same cells, while the right panel is the merge between the two channels. Similar results were observed in 12 replicates, visualized during 3 independent microscopy sessions

    Techniques Used: Stable Transfection, Transformation Assay, Confocal Laser Scanning Microscopy, Staining, Microscopy

    36) Product Images from "Heterotrophic Nitrogen Fixation at the Hyper-Eutrophic Qishon River and Estuary System"

    Article Title: Heterotrophic Nitrogen Fixation at the Hyper-Eutrophic Qishon River and Estuary System

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2020.01370

    Visualization of the natural microbial population in the stream (A–E) and estuary (F–M) captured by a confocal laser scanning microscope during September 2017 and January 2018 ( Supplementary Table S1 ) at T 48 . (A,F) active diazotrophs tagged by immunolabeling (green); (B,G) cyanobacteria phycoerythrin autofluorescence (orange); (C,H) , total bacteria stained with DAPI (dark blue); and (D,I) polysaccharides stained with ConA (light blue). (E,M) The 3D images show the superimposed signals of the different stains. (J–L) 3D images show the zoom in of the aggregates in different locations. The axes of the superimposed images are reported in micrometers. For additional magnified confocal images see Supplementary Figure S5 .
    Figure Legend Snippet: Visualization of the natural microbial population in the stream (A–E) and estuary (F–M) captured by a confocal laser scanning microscope during September 2017 and January 2018 ( Supplementary Table S1 ) at T 48 . (A,F) active diazotrophs tagged by immunolabeling (green); (B,G) cyanobacteria phycoerythrin autofluorescence (orange); (C,H) , total bacteria stained with DAPI (dark blue); and (D,I) polysaccharides stained with ConA (light blue). (E,M) The 3D images show the superimposed signals of the different stains. (J–L) 3D images show the zoom in of the aggregates in different locations. The axes of the superimposed images are reported in micrometers. For additional magnified confocal images see Supplementary Figure S5 .

    Techniques Used: Laser-Scanning Microscopy, Immunolabeling, Staining

    37) Product Images from "Arabidopsis exoribonuclease USB1 interacts with the PPR-domain protein SOAR1 to negatively regulate abscisic acid signaling"

    Article Title: Arabidopsis exoribonuclease USB1 interacts with the PPR-domain protein SOAR1 to negatively regulate abscisic acid signaling

    Journal: Journal of Experimental Botany

    doi: 10.1093/jxb/eraa315

    USB1 co-localizes and physically interacts with SOAR1 in both the nucleus and cytoplasm. (A) In vitro pull-down assay to test the direct interaction between SOAR1 and USB1 protein. The SOAR1-His proteins were incubated with immobilized GST or USB1–GST proteins, and the antibody against GST tag (IP:anti-GST) was used to pull down the purified USB1–GST or GST protein. Both the immunoprecipitated fractions and input were tested by immunoblotting with anti-His and anti-GST antibodies, respectively. (B) Co-immunoprecipitation (Co-IP) assay to test the interaction of USB1 with SOAR1 in Arabidopsis. Total proteins, extracted from the homozygous transgenic plants expressing GFP or USB1–GFP fusion protein (USB1–OE3), were immunoprecipitated with anti-GFP antibody (IP:anti-GFP). The immunoprecipitates and the input were tested by immunoblotting with anti-SOAR1 and anti-GFP antibodies. (C) Luciferase complementation imaging (LCI) assay to test the interaction between USB1 and SOAR1 using the N. benthamiana system. The coding sequences of USB1 or SOAR1 were cloned into the N-terminal fragment of Luc (NLuc) and the C-terminal fragment of Luc (CLuc) to form USB1–NLuc(or SOAR1–NLuc) and CLuc–USB1 (or CLuc–SOAR1), respectively. The constructs pairs of USB1–NLuc/CLuc–SOAR1 and CLuc–USB1/SOAR1–NLuc were co-injected into N. benthamiana leaves, and the Luc signals were observed 72 h after infiltration. The combinations of USB1–NLuc/CLuc and SOAR1–NLuc/CLuc vectors were used as negative controls. (D) Bimolecular fluorescent complementation imaging (BiFC) assay using yellow fluorescent protein (YFP) to test the interaction between USB1 and SOAR1 in Arabidopsis mesophyll protoplasts. The coding region of USB1 or SOAR1 was fused to the N-terminus of YFP (USB1–NYFP or SOAR1–NYFP), or to the C-terminus of YFP (USB1–CYFP or SOAR1–CYFP). The constructs pairs (as indicated) were co-transformed into Arabidopsis wild-type (Col-0) mesophyll protoplasts. The combinations of NYFP /SOAR1–CYFP or USB1–CYFP were used as negative controls. The YFP fluorescence was imaged under a confocal laser scanning microscope. YFP, YFP signal; Auto, chloroplast autofluorescent signal; Merged, merged image of the YFP signal with the chloroplast autofluorescent signal. (E) Transgenic expression of the USB1–GFP fusion protein in whole Arabidopsis plants, showing that the USB1–GFP fusion protein (left) is localized to both the nucleus and cytoplasm in the root of a transgenic complementation line (the line Com11-1, see Supplementary Fig. S5 ) expressing 35S:: USB1-GFP in the usb1-1 background. Note that the expression level of USB1 in the Com11-1 transgenic line is slightly higher than that in the wild-type plants ( Supplementary Fig. S5 ). The nuclei are indicated by DAPI staining. Bright, bright field; Merged, merged image of the DAPI signal with the USB1–GFP signal in the bright field. (F) Transient expression of the USB1–GFP fusion protein in the Arabidopsis protoplasts, showing the nuclear–cytoplasmic dual localization of USB1. Left panels: the signal of the USB1–GFP fusion protein overlaps the signal of a nuclear-localized bHLH (basic helix–loop-helix) transcription factor FBI1/HFR1 (FBI1–RFP) tagged with mCherry (a red fluorescent protein) in the nuclear portion (Merged, merged image of USB1–GFP with FBI1–RFP in the bright field). Right panels: the signal of the USB1–GFP fusion protein completely overlaps the signal of the cytosolic–nuclear dual-localized SOAR1–RFP (SOAR1 tagged with mCherry) fusion protein (Merged, merged image of USB1–GFP with SOAR1–RFP in the bright field). Bright, bright-field. (G) BiFC assays in the N. benthamiana leaves, indicating that USB1 interacts with SOAR1 in both the nucleus and cytoplasm. The coding regions of USB1 and SOAR1 were fused to the N-terminus of YFP and the C-terminus of YFP to form USB1–NYFP and SOAR1–CYFP, respectively. The construct pairs (as indicated) were co-infiltrated into N. benthamiana leaves. YFP fluorescence was detected in N. benthamiana leaves co-infiltrated with combinations of USB1–NYFP and SOAR1–CYFP. The combination of NYFP/SOAR1–CYFP was used as a negative control. The YFP fluorescence was imaged under a confocal laser scanning microscope. YFP, YFP signal; DAPI, staining for the nucleus; Merged, merged image of the YFP signal with the DAPI signal. All the experiments were repeated five times with similar results.
    Figure Legend Snippet: USB1 co-localizes and physically interacts with SOAR1 in both the nucleus and cytoplasm. (A) In vitro pull-down assay to test the direct interaction between SOAR1 and USB1 protein. The SOAR1-His proteins were incubated with immobilized GST or USB1–GST proteins, and the antibody against GST tag (IP:anti-GST) was used to pull down the purified USB1–GST or GST protein. Both the immunoprecipitated fractions and input were tested by immunoblotting with anti-His and anti-GST antibodies, respectively. (B) Co-immunoprecipitation (Co-IP) assay to test the interaction of USB1 with SOAR1 in Arabidopsis. Total proteins, extracted from the homozygous transgenic plants expressing GFP or USB1–GFP fusion protein (USB1–OE3), were immunoprecipitated with anti-GFP antibody (IP:anti-GFP). The immunoprecipitates and the input were tested by immunoblotting with anti-SOAR1 and anti-GFP antibodies. (C) Luciferase complementation imaging (LCI) assay to test the interaction between USB1 and SOAR1 using the N. benthamiana system. The coding sequences of USB1 or SOAR1 were cloned into the N-terminal fragment of Luc (NLuc) and the C-terminal fragment of Luc (CLuc) to form USB1–NLuc(or SOAR1–NLuc) and CLuc–USB1 (or CLuc–SOAR1), respectively. The constructs pairs of USB1–NLuc/CLuc–SOAR1 and CLuc–USB1/SOAR1–NLuc were co-injected into N. benthamiana leaves, and the Luc signals were observed 72 h after infiltration. The combinations of USB1–NLuc/CLuc and SOAR1–NLuc/CLuc vectors were used as negative controls. (D) Bimolecular fluorescent complementation imaging (BiFC) assay using yellow fluorescent protein (YFP) to test the interaction between USB1 and SOAR1 in Arabidopsis mesophyll protoplasts. The coding region of USB1 or SOAR1 was fused to the N-terminus of YFP (USB1–NYFP or SOAR1–NYFP), or to the C-terminus of YFP (USB1–CYFP or SOAR1–CYFP). The constructs pairs (as indicated) were co-transformed into Arabidopsis wild-type (Col-0) mesophyll protoplasts. The combinations of NYFP /SOAR1–CYFP or USB1–CYFP were used as negative controls. The YFP fluorescence was imaged under a confocal laser scanning microscope. YFP, YFP signal; Auto, chloroplast autofluorescent signal; Merged, merged image of the YFP signal with the chloroplast autofluorescent signal. (E) Transgenic expression of the USB1–GFP fusion protein in whole Arabidopsis plants, showing that the USB1–GFP fusion protein (left) is localized to both the nucleus and cytoplasm in the root of a transgenic complementation line (the line Com11-1, see Supplementary Fig. S5 ) expressing 35S:: USB1-GFP in the usb1-1 background. Note that the expression level of USB1 in the Com11-1 transgenic line is slightly higher than that in the wild-type plants ( Supplementary Fig. S5 ). The nuclei are indicated by DAPI staining. Bright, bright field; Merged, merged image of the DAPI signal with the USB1–GFP signal in the bright field. (F) Transient expression of the USB1–GFP fusion protein in the Arabidopsis protoplasts, showing the nuclear–cytoplasmic dual localization of USB1. Left panels: the signal of the USB1–GFP fusion protein overlaps the signal of a nuclear-localized bHLH (basic helix–loop-helix) transcription factor FBI1/HFR1 (FBI1–RFP) tagged with mCherry (a red fluorescent protein) in the nuclear portion (Merged, merged image of USB1–GFP with FBI1–RFP in the bright field). Right panels: the signal of the USB1–GFP fusion protein completely overlaps the signal of the cytosolic–nuclear dual-localized SOAR1–RFP (SOAR1 tagged with mCherry) fusion protein (Merged, merged image of USB1–GFP with SOAR1–RFP in the bright field). Bright, bright-field. (G) BiFC assays in the N. benthamiana leaves, indicating that USB1 interacts with SOAR1 in both the nucleus and cytoplasm. The coding regions of USB1 and SOAR1 were fused to the N-terminus of YFP and the C-terminus of YFP to form USB1–NYFP and SOAR1–CYFP, respectively. The construct pairs (as indicated) were co-infiltrated into N. benthamiana leaves. YFP fluorescence was detected in N. benthamiana leaves co-infiltrated with combinations of USB1–NYFP and SOAR1–CYFP. The combination of NYFP/SOAR1–CYFP was used as a negative control. The YFP fluorescence was imaged under a confocal laser scanning microscope. YFP, YFP signal; DAPI, staining for the nucleus; Merged, merged image of the YFP signal with the DAPI signal. All the experiments were repeated five times with similar results.

    Techniques Used: In Vitro, Pull Down Assay, Incubation, Purification, Immunoprecipitation, Co-Immunoprecipitation Assay, Transgenic Assay, Expressing, Luciferase, Imaging, Clone Assay, Construct, Injection, Bimolecular Fluorescence Complementation Assay, Transformation Assay, Fluorescence, Laser-Scanning Microscopy, Staining, Negative Control

    38) Product Images from "Molecular dissection of the photoreceptor ribbon synapse"

    Article Title: Molecular dissection of the photoreceptor ribbon synapse

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.200408157

    Screening the photoreceptor ribbon synaptic complex for ribbon-associated proteins in wild-type (+/+) and Bassoon mutant (−/−) retinae. (Left) Confocal laser-scanning micrographs of photoreceptor ribbons in the +/+ retina double labeled for RIBEYE combined with the following CAZ proteins: Piccolo, KIF3A, RIM1, RIM2, Munc13-1, Ca 2+ channel α1 subunit, and ERC2/CAST1. All CAZ proteins colocalize with RIBEYE as seen in the merge of the stainings. (Right) In the −/− retina, Piccolo, KIF3A, and RIM1 colocalize with RIBEYE, but not RIM2, Munc13-1, Ca 2+ channel α1 subunit, and ERC2/CAST1. Bars, 5 μm.
    Figure Legend Snippet: Screening the photoreceptor ribbon synaptic complex for ribbon-associated proteins in wild-type (+/+) and Bassoon mutant (−/−) retinae. (Left) Confocal laser-scanning micrographs of photoreceptor ribbons in the +/+ retina double labeled for RIBEYE combined with the following CAZ proteins: Piccolo, KIF3A, RIM1, RIM2, Munc13-1, Ca 2+ channel α1 subunit, and ERC2/CAST1. All CAZ proteins colocalize with RIBEYE as seen in the merge of the stainings. (Right) In the −/− retina, Piccolo, KIF3A, and RIM1 colocalize with RIBEYE, but not RIM2, Munc13-1, Ca 2+ channel α1 subunit, and ERC2/CAST1. Bars, 5 μm.

    Techniques Used: Mutagenesis, Labeling

    CtBP1, a RIBEYE homologue, is expressed at ribbon and conventional synapses. (A–C) Confocal laser-scanning micrographs of a vertical section through mouse retina double labeled for CtBP1 (A) and Piccolo (B), which marks ribbon and conventional synapses in the retina. As seen in the merge of the two stainings (C), CtBP1 and Piccolo immunoreactivities completely colocalize at the synapses in the outer plexiform layer (OPL) and inner plexiform layer (IPL) of the retina. (D and E) Electron micrographs of a photoreceptor (D) and an amacrine cell synapse (E) immunogold labeled for CtBP1. At the ribbon synapse the gold particles for CtBP1 decorate the ribbon (arrowheads), at the amacrine cell synapse they are located some distance from the active zone (arrowheads) at the edge of the electron-dense CAZ material. (F) Western blots of retina homogenate probed with antibodies against CtBP1 and the RIBEYE A- and CtBP2/RIBEYE B-domain. The antibody against CtBP1 recognizes the 50-kD CtBP1 protein and does not cross-react with RIBEYE. (G–I) Micrographs of cultured hippocampal neurons double labeled for CtBP1 (G) and Piccolo (H). In hippocampal neurons, like in retinal neurons, a fraction of CtBP1 immunoreactivity is localized at synapses where it colocalizes with Piccolo as seen in the merge of the two stainings (I). (J and K) CtBP2 (J) is not present at hippocampal synapses labeled with Piccolo (K). (L) Immunoblots showing that CtBP1 can be coimmunoprecipitated from brain synaptosomes with a monoclonal anti-Bassoon antibody. The boxes in G–K mark the regions that are shown at higher magnification. INL, inner nuclear layer; GCL, ganglion cell layer; Ext., brain synaptosomal extract; IP, immunoprecipitate; S, supernatant. Bars: 20 μm (A–C), 0.2 μm (D and E), and 10 μm (G–K).
    Figure Legend Snippet: CtBP1, a RIBEYE homologue, is expressed at ribbon and conventional synapses. (A–C) Confocal laser-scanning micrographs of a vertical section through mouse retina double labeled for CtBP1 (A) and Piccolo (B), which marks ribbon and conventional synapses in the retina. As seen in the merge of the two stainings (C), CtBP1 and Piccolo immunoreactivities completely colocalize at the synapses in the outer plexiform layer (OPL) and inner plexiform layer (IPL) of the retina. (D and E) Electron micrographs of a photoreceptor (D) and an amacrine cell synapse (E) immunogold labeled for CtBP1. At the ribbon synapse the gold particles for CtBP1 decorate the ribbon (arrowheads), at the amacrine cell synapse they are located some distance from the active zone (arrowheads) at the edge of the electron-dense CAZ material. (F) Western blots of retina homogenate probed with antibodies against CtBP1 and the RIBEYE A- and CtBP2/RIBEYE B-domain. The antibody against CtBP1 recognizes the 50-kD CtBP1 protein and does not cross-react with RIBEYE. (G–I) Micrographs of cultured hippocampal neurons double labeled for CtBP1 (G) and Piccolo (H). In hippocampal neurons, like in retinal neurons, a fraction of CtBP1 immunoreactivity is localized at synapses where it colocalizes with Piccolo as seen in the merge of the two stainings (I). (J and K) CtBP2 (J) is not present at hippocampal synapses labeled with Piccolo (K). (L) Immunoblots showing that CtBP1 can be coimmunoprecipitated from brain synaptosomes with a monoclonal anti-Bassoon antibody. The boxes in G–K mark the regions that are shown at higher magnification. INL, inner nuclear layer; GCL, ganglion cell layer; Ext., brain synaptosomal extract; IP, immunoprecipitate; S, supernatant. Bars: 20 μm (A–C), 0.2 μm (D and E), and 10 μm (G–K).

    Techniques Used: Labeling, Western Blot, Cell Culture

    Characterization of the photoreceptor ribbon protein RIBEYE in wild-type and Bassoon mutant mice. (A) Immunoblot of mouse retinal homogenate probed with antibodies against RIBEYE A- and B-domain. Two protein bands (double arrow) of ∼110 kD and 120 kD, representing RIBEYE, are recognized by both antibodies. In addition, the anti-RIBEYE B-domain antibody labels the 50-kD CtBP2 protein band (arrow). Protein bands above and below the RIBEYE doublet are unspecific and appeared only occasionally. (B) Preincubation of the anti-GST-RIBEYE A-domain antiserum with an excess of the MBP-RIB179-448 fusion protein (+) completely prevents the immunodetection of RIBEYE. −, no RIBEYE antigen included. Incubation of the same blot with the anti-RIBEYE B-domain antibody shows the presence of RIBEYE in both lanes. (C) In a vertical cryostat section through mouse retina, the anti-RIBEYE A-domain antiserum stains the photoreceptor ribbons in the outer plexiform layer (OPL) and the bipolar cell ribbons in the inner plexiform layer (IPL). (D) Preadsorption of the anti-RIBEYE antiserum with the antigen results in a complete loss of RIBEYE staining. (E and F) EM and postembedding immunogold labeling shows that photoreceptor ribbons are decorated with gold particles for RIBEYE A- (E) and B-domain (F). Note the absence of gold particles at the base of the ribbons (arrowheads), the region of the arciform density. (G) Confocal laser-scanning micrograph of a region of the OPL double labeled for RIBEYE A- (green) and B-domain (red) demonstrate the colocalization of the two immunoreactivities at the photoreceptor ribbons. (H) An aggregate of free-floating ribbons in a cone photoreceptor terminal of Bassoon mutant retina decorated with RIBEYE immunoreactivity (preembedding labeling). ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer. Bars: 10 μm (C and D); 0.1 μm (E and F); 5 μm (G); and 0.4 μm (H).
    Figure Legend Snippet: Characterization of the photoreceptor ribbon protein RIBEYE in wild-type and Bassoon mutant mice. (A) Immunoblot of mouse retinal homogenate probed with antibodies against RIBEYE A- and B-domain. Two protein bands (double arrow) of ∼110 kD and 120 kD, representing RIBEYE, are recognized by both antibodies. In addition, the anti-RIBEYE B-domain antibody labels the 50-kD CtBP2 protein band (arrow). Protein bands above and below the RIBEYE doublet are unspecific and appeared only occasionally. (B) Preincubation of the anti-GST-RIBEYE A-domain antiserum with an excess of the MBP-RIB179-448 fusion protein (+) completely prevents the immunodetection of RIBEYE. −, no RIBEYE antigen included. Incubation of the same blot with the anti-RIBEYE B-domain antibody shows the presence of RIBEYE in both lanes. (C) In a vertical cryostat section through mouse retina, the anti-RIBEYE A-domain antiserum stains the photoreceptor ribbons in the outer plexiform layer (OPL) and the bipolar cell ribbons in the inner plexiform layer (IPL). (D) Preadsorption of the anti-RIBEYE antiserum with the antigen results in a complete loss of RIBEYE staining. (E and F) EM and postembedding immunogold labeling shows that photoreceptor ribbons are decorated with gold particles for RIBEYE A- (E) and B-domain (F). Note the absence of gold particles at the base of the ribbons (arrowheads), the region of the arciform density. (G) Confocal laser-scanning micrograph of a region of the OPL double labeled for RIBEYE A- (green) and B-domain (red) demonstrate the colocalization of the two immunoreactivities at the photoreceptor ribbons. (H) An aggregate of free-floating ribbons in a cone photoreceptor terminal of Bassoon mutant retina decorated with RIBEYE immunoreactivity (preembedding labeling). ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer. Bars: 10 μm (C and D); 0.1 μm (E and F); 5 μm (G); and 0.4 μm (H).

    Techniques Used: Mutagenesis, Mouse Assay, Immunodetection, Incubation, Staining, Labeling

    Bassoon–RIBEYE interaction at the base of the photoreceptor ribbon. (A–C) Confocal laser-scanning micrographs of photoreceptor ribbon synapses double labeled for RIBEYE (A) and Bassoon (B). In the merge of the two stainings (C), the horseshoe-like appearance of the photoreceptor ribbon colabeled for RIBEYE and Bassoon is clearly visible. Bassoon labeling is surrounded by the RIBEYE labeling. (D) Electron micrographs of photoreceptor ribbons, en face and cross-section view (inset), postembedding immunogold double labeled for Bassoon (large gold particles) and RIBEYE (small gold particles). The gold particles for Bassoon are located closest to the active zone (arrowheads). (E) Immunoblots of 25 μg total protein per lane from wild-type (+/+) and Bassoon mutant (−/−) retina homogenates show a 50% reduction of RIBEYE immunoreactivity in the −/− retina. Data are normalized to the mean value (100%) obtained from wild-type samples. Statistical analysis by the unpaired t test shows a significant difference in the RIBEYE immunoreactivity between the two genotypes (asterisk, P
    Figure Legend Snippet: Bassoon–RIBEYE interaction at the base of the photoreceptor ribbon. (A–C) Confocal laser-scanning micrographs of photoreceptor ribbon synapses double labeled for RIBEYE (A) and Bassoon (B). In the merge of the two stainings (C), the horseshoe-like appearance of the photoreceptor ribbon colabeled for RIBEYE and Bassoon is clearly visible. Bassoon labeling is surrounded by the RIBEYE labeling. (D) Electron micrographs of photoreceptor ribbons, en face and cross-section view (inset), postembedding immunogold double labeled for Bassoon (large gold particles) and RIBEYE (small gold particles). The gold particles for Bassoon are located closest to the active zone (arrowheads). (E) Immunoblots of 25 μg total protein per lane from wild-type (+/+) and Bassoon mutant (−/−) retina homogenates show a 50% reduction of RIBEYE immunoreactivity in the −/− retina. Data are normalized to the mean value (100%) obtained from wild-type samples. Statistical analysis by the unpaired t test shows a significant difference in the RIBEYE immunoreactivity between the two genotypes (asterisk, P

    Techniques Used: Labeling, Western Blot, Mutagenesis

    39) Product Images from "Transient Receptor Potential 1 Regulates Capacitative Ca2+ Entry and Ca2+ Release from Endoplasmic Reticulum in B Lymphocytes 〉"

    Article Title: Transient Receptor Potential 1 Regulates Capacitative Ca2+ Entry and Ca2+ Release from Endoplasmic Reticulum in B Lymphocytes 〉

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20011758

    Targeted disruption of the TRP1 gene in DT40 B lymphocytes. Partial restriction map of chicken TRP1 gene (A), targeting construct (B), and expected structure of the disrupted allele (C). (D) Southern blot analysis of genomic DNAs from DT40 cells. Genomic DNAs were prepared from WT (+/+), neo -targeted (+/−), and neo / his -targeted (−/−) clones, digested with Xba I, and hybridized with a 3′-flanking probe. The restriction endonuclease cleavage site of Xba I is abbreviated as X. (E) Northern blot analysis of WT and TRP1-deficient DT40 cells (clone TRP1 − -14). (F) Immunolocalization of TRP1 in WT cells, and loss of its expression in TRP1 − -14 cells. The fluorescence images were acquired with a confocal laser microscope. (G) BCR expression on TRP1-deficient DT40 cells. DT40 cells were stained with FITC-conjugated anti–chicken IgM Ab.
    Figure Legend Snippet: Targeted disruption of the TRP1 gene in DT40 B lymphocytes. Partial restriction map of chicken TRP1 gene (A), targeting construct (B), and expected structure of the disrupted allele (C). (D) Southern blot analysis of genomic DNAs from DT40 cells. Genomic DNAs were prepared from WT (+/+), neo -targeted (+/−), and neo / his -targeted (−/−) clones, digested with Xba I, and hybridized with a 3′-flanking probe. The restriction endonuclease cleavage site of Xba I is abbreviated as X. (E) Northern blot analysis of WT and TRP1-deficient DT40 cells (clone TRP1 − -14). (F) Immunolocalization of TRP1 in WT cells, and loss of its expression in TRP1 − -14 cells. The fluorescence images were acquired with a confocal laser microscope. (G) BCR expression on TRP1-deficient DT40 cells. DT40 cells were stained with FITC-conjugated anti–chicken IgM Ab.

    Techniques Used: Construct, Southern Blot, Clone Assay, Northern Blot, Expressing, Fluorescence, Microscopy, Staining

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    Carl Zeiss lsm 510 meta confocal laser scanning microscope
    iPLA 2 localizes to mitochondria . A , colocalization of iPLA 2 -GFP with mitochondria. pEGFP-iPLA 2 -transfected INS-1 cells were incubated for 15 min in Mito Tracker Red CMXRos, fixed with 3.8% paraformaldehyde, stained with 4′,6-diamidino-2-phenylindole, and analyzed on a Zeiss <t>LSM</t> 510 <t>META</t> confocal laser scanning microscope. Images of red (Mito Tracker) and green (iPLA 2 -GFP) fluorescence were collected by confocal microscopy. Colocalization of iPLA 2 -GFP with mitochondria appears as yellow to orange spots, depending on the ratio of the merged red and green fluorescence. B , resistance of cells expressing GFP-iPLA 2 to STS-induced apoptosis. pEGFP-iPLA 2 -transfected INS-1 cells were treated with STS and incubated with Mito Tracker Red CMXrox as in A . The cells with green and yellow to orange dots are iPLA 2 -GFP-transfected INS-1 cells. Those with red color only are non-transfected cells. C , localization of iPLA 2 -GFP in mitochondria during STS-induced apoptosis in individual cells. Merged images of red (mitochondria) and green (iPLA 2 -GFP) fluorescence were collected by confocal microscopy. The images represent at least four independent experiments with similar results.
    Lsm 510 Meta Confocal Laser Scanning Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Carl Zeiss laser scanning microscopy lsm
    Appearance of phosphoethanolamine-enriched PFCE nanoparticles. (a) Cryogenic transmission electron microscopy (TEM) images of DSPE-PEG2000, Pluronic-basic, Pluronic-DPPE and Pluronic-DPPE-Rhodamine nanoparticles encapsulating PFCE fluorine compound (size-bar for TEM images: 50 nm). (b) Upper two panels and lower left panel show ultrathin sections of <t>DCs</t> labeled with DSPE-PEG2000, basic and DPPE-enriched PFCE nanoparticles (size-bar for EM images: 2 μm). Lower right panel shows a laser scanning microscopy image of DCs labeled with DPPE-Rhodamine-enriched 19 F nanoparticles (size-bar for <t>LSM</t> image: 10 μm). (c) DCs were labeled with different nanoparticle preparations (DSPE-PEG2000 NP, basic NP, DPPE-NP and DPPE-Rhodamine-NP) using a PFCE concentration of 10 μmol per 10 7 , fixed in 2% PFA and transferred (10 6 ) to NMR tubes. 19 F signal was acquired using a 90° block excitation pulse and the PFCE amount per 10 6 calculated using a 500 mM PFCE standard.
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    Carl Zeiss confocal laser scanning microscopy clsm
    Arabidopsis UAP56 localises to the nucleus in protoplasts and root cells. Tobacco BY2 cell protoplasts were transformed with constructs driving the expression of the indicated <t>GFP</t> fusion proteins and GFP fluorescence was visualised by <t>CLSM</t> ( A,B ; size bar: 10 µm). GFP fluorescence of a nuclear HMGB protein and overlay with the corresponding bright field image ( A ). GFP fluorescence of a UAP56-GFP fusion ( B ). Analysis of Arabidopsis Col-0 root tip cells by immunofluorescence microscopy using different antibodies and DAPI staining ( E – H , size bar: 5 µm). Immunostaining of fibrillarin and UAP56, as well as visualisation of the DNA by DAPI staining of the same nucleus ( C – E ). A merge of the three images is shown in ( F ) and examples of brightly DAPI-stained heterochromatic chromocenters are indicated by arrows. Immunostaining of RNAPII and UAP56 of the same nucleus ( G,H ).
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    iPLA 2 localizes to mitochondria . A , colocalization of iPLA 2 -GFP with mitochondria. pEGFP-iPLA 2 -transfected INS-1 cells were incubated for 15 min in Mito Tracker Red CMXRos, fixed with 3.8% paraformaldehyde, stained with 4′,6-diamidino-2-phenylindole, and analyzed on a Zeiss LSM 510 META confocal laser scanning microscope. Images of red (Mito Tracker) and green (iPLA 2 -GFP) fluorescence were collected by confocal microscopy. Colocalization of iPLA 2 -GFP with mitochondria appears as yellow to orange spots, depending on the ratio of the merged red and green fluorescence. B , resistance of cells expressing GFP-iPLA 2 to STS-induced apoptosis. pEGFP-iPLA 2 -transfected INS-1 cells were treated with STS and incubated with Mito Tracker Red CMXrox as in A . The cells with green and yellow to orange dots are iPLA 2 -GFP-transfected INS-1 cells. Those with red color only are non-transfected cells. C , localization of iPLA 2 -GFP in mitochondria during STS-induced apoptosis in individual cells. Merged images of red (mitochondria) and green (iPLA 2 -GFP) fluorescence were collected by confocal microscopy. The images represent at least four independent experiments with similar results.

    Journal: The Journal of biological chemistry

    Article Title: Calcium-independent Phospholipase A2 Localizes in and Protects Mitochondria during Apoptotic Induction by Staurosporine *

    doi: 10.1074/jbc.M604330200

    Figure Lengend Snippet: iPLA 2 localizes to mitochondria . A , colocalization of iPLA 2 -GFP with mitochondria. pEGFP-iPLA 2 -transfected INS-1 cells were incubated for 15 min in Mito Tracker Red CMXRos, fixed with 3.8% paraformaldehyde, stained with 4′,6-diamidino-2-phenylindole, and analyzed on a Zeiss LSM 510 META confocal laser scanning microscope. Images of red (Mito Tracker) and green (iPLA 2 -GFP) fluorescence were collected by confocal microscopy. Colocalization of iPLA 2 -GFP with mitochondria appears as yellow to orange spots, depending on the ratio of the merged red and green fluorescence. B , resistance of cells expressing GFP-iPLA 2 to STS-induced apoptosis. pEGFP-iPLA 2 -transfected INS-1 cells were treated with STS and incubated with Mito Tracker Red CMXrox as in A . The cells with green and yellow to orange dots are iPLA 2 -GFP-transfected INS-1 cells. Those with red color only are non-transfected cells. C , localization of iPLA 2 -GFP in mitochondria during STS-induced apoptosis in individual cells. Merged images of red (mitochondria) and green (iPLA 2 -GFP) fluorescence were collected by confocal microscopy. The images represent at least four independent experiments with similar results.

    Article Snippet: Confocal fluorescence microscopy was performed using a Zeiss LSM 510 META confocal laser scanning microscope (Carl Zeiss MicroImaging, Inc. Thornwood, NY).

    Techniques: Transfection, Incubation, Staining, Laser-Scanning Microscopy, Fluorescence, Confocal Microscopy, Expressing

    Co-localization of GH43A-YFP and GH43B-YFP with mCherry Golgi markers. (A) Confocal laser scanning microscope images of root epidermis cells in Arabidopsis seedlings stably expressing pGH43A::GH43A-cYFP or pGH43B::GH43B-cYFP and the cis-Golgi marker SYP32-mCherry or the Trans Golgi Network (TGN) marker SYP43-mCherry. Images were captured with a Zeiss LSM880 confocal microscope. Scale bars = 5µm. (B) Degree of co-localization of pGH43A::GH43A-cYFP with SYP32-mCherry or SYP43-mCherry in the root elongation zone epidermis. Images were captured from three independent GH43A-cYFP lines (L1-L3) and five seedlings per line. The values represent the degree of Pearson’s correlation between the YFP and RFP channel. (C) Degree of co-localization of pGH43B::GH43B-cYFP with SYP32-mCherry or SYP43-mCherry in the root elongation zone epidermis. Images were captured from three independent GH43A-cYFP lines (L1-L3) and five seedlings per line. The values represent the degree of Pearson’s correlation between the YFP and RFP channel.

    Journal: bioRxiv

    Article Title: Golgi-localized exo-β1,3-galactosidases involved in AGP modification and root cell expansion in Arabidopsis

    doi: 10.1101/2020.02.13.947820

    Figure Lengend Snippet: Co-localization of GH43A-YFP and GH43B-YFP with mCherry Golgi markers. (A) Confocal laser scanning microscope images of root epidermis cells in Arabidopsis seedlings stably expressing pGH43A::GH43A-cYFP or pGH43B::GH43B-cYFP and the cis-Golgi marker SYP32-mCherry or the Trans Golgi Network (TGN) marker SYP43-mCherry. Images were captured with a Zeiss LSM880 confocal microscope. Scale bars = 5µm. (B) Degree of co-localization of pGH43A::GH43A-cYFP with SYP32-mCherry or SYP43-mCherry in the root elongation zone epidermis. Images were captured from three independent GH43A-cYFP lines (L1-L3) and five seedlings per line. The values represent the degree of Pearson’s correlation between the YFP and RFP channel. (C) Degree of co-localization of pGH43B::GH43B-cYFP with SYP32-mCherry or SYP43-mCherry in the root elongation zone epidermis. Images were captured from three independent GH43A-cYFP lines (L1-L3) and five seedlings per line. The values represent the degree of Pearson’s correlation between the YFP and RFP channel.

    Article Snippet: A Zeiss LSM880 confocal laser scanning microscope with an Airyscan detector and a LD LCI Plan-Apochromat 40x/1.2 Imm AutoCorr DIC M27 water immersion objective was used.

    Techniques: Laser-Scanning Microscopy, Stable Transfection, Expressing, Marker, Microscopy

    Appearance of phosphoethanolamine-enriched PFCE nanoparticles. (a) Cryogenic transmission electron microscopy (TEM) images of DSPE-PEG2000, Pluronic-basic, Pluronic-DPPE and Pluronic-DPPE-Rhodamine nanoparticles encapsulating PFCE fluorine compound (size-bar for TEM images: 50 nm). (b) Upper two panels and lower left panel show ultrathin sections of DCs labeled with DSPE-PEG2000, basic and DPPE-enriched PFCE nanoparticles (size-bar for EM images: 2 μm). Lower right panel shows a laser scanning microscopy image of DCs labeled with DPPE-Rhodamine-enriched 19 F nanoparticles (size-bar for LSM image: 10 μm). (c) DCs were labeled with different nanoparticle preparations (DSPE-PEG2000 NP, basic NP, DPPE-NP and DPPE-Rhodamine-NP) using a PFCE concentration of 10 μmol per 10 7 , fixed in 2% PFA and transferred (10 6 ) to NMR tubes. 19 F signal was acquired using a 90° block excitation pulse and the PFCE amount per 10 6 calculated using a 500 mM PFCE standard.

    Journal: Scientific Reports

    Article Title: Anchoring Dipalmitoyl Phosphoethanolamine to Nanoparticles Boosts Cellular Uptake and Fluorine-19 Magnetic Resonance Signal

    doi: 10.1038/srep08427

    Figure Lengend Snippet: Appearance of phosphoethanolamine-enriched PFCE nanoparticles. (a) Cryogenic transmission electron microscopy (TEM) images of DSPE-PEG2000, Pluronic-basic, Pluronic-DPPE and Pluronic-DPPE-Rhodamine nanoparticles encapsulating PFCE fluorine compound (size-bar for TEM images: 50 nm). (b) Upper two panels and lower left panel show ultrathin sections of DCs labeled with DSPE-PEG2000, basic and DPPE-enriched PFCE nanoparticles (size-bar for EM images: 2 μm). Lower right panel shows a laser scanning microscopy image of DCs labeled with DPPE-Rhodamine-enriched 19 F nanoparticles (size-bar for LSM image: 10 μm). (c) DCs were labeled with different nanoparticle preparations (DSPE-PEG2000 NP, basic NP, DPPE-NP and DPPE-Rhodamine-NP) using a PFCE concentration of 10 μmol per 10 7 , fixed in 2% PFA and transferred (10 6 ) to NMR tubes. 19 F signal was acquired using a 90° block excitation pulse and the PFCE amount per 10 6 calculated using a 500 mM PFCE standard.

    Article Snippet: Laser Scanning Microscopy (LSM) of DCs Intracellular fluorescence in DCs labeled with fluorescently-labeled nanoparticles was investigated using an LSM780 laser scanning microscope (Carl Zeiss MicroImaging GmbH, Jena, Germany).

    Techniques: Transmission Assay, Electron Microscopy, Transmission Electron Microscopy, Labeling, Laser-Scanning Microscopy, Concentration Assay, Nuclear Magnetic Resonance, Blocking Assay

    Arabidopsis UAP56 localises to the nucleus in protoplasts and root cells. Tobacco BY2 cell protoplasts were transformed with constructs driving the expression of the indicated GFP fusion proteins and GFP fluorescence was visualised by CLSM ( A,B ; size bar: 10 µm). GFP fluorescence of a nuclear HMGB protein and overlay with the corresponding bright field image ( A ). GFP fluorescence of a UAP56-GFP fusion ( B ). Analysis of Arabidopsis Col-0 root tip cells by immunofluorescence microscopy using different antibodies and DAPI staining ( E – H , size bar: 5 µm). Immunostaining of fibrillarin and UAP56, as well as visualisation of the DNA by DAPI staining of the same nucleus ( C – E ). A merge of the three images is shown in ( F ) and examples of brightly DAPI-stained heterochromatic chromocenters are indicated by arrows. Immunostaining of RNAPII and UAP56 of the same nucleus ( G,H ).

    Journal: PLoS ONE

    Article Title: Arabidopsis DEAD-Box RNA Helicase UAP56 Interacts with Both RNA and DNA as well as with mRNA Export Factors

    doi: 10.1371/journal.pone.0060644

    Figure Lengend Snippet: Arabidopsis UAP56 localises to the nucleus in protoplasts and root cells. Tobacco BY2 cell protoplasts were transformed with constructs driving the expression of the indicated GFP fusion proteins and GFP fluorescence was visualised by CLSM ( A,B ; size bar: 10 µm). GFP fluorescence of a nuclear HMGB protein and overlay with the corresponding bright field image ( A ). GFP fluorescence of a UAP56-GFP fusion ( B ). Analysis of Arabidopsis Col-0 root tip cells by immunofluorescence microscopy using different antibodies and DAPI staining ( E – H , size bar: 5 µm). Immunostaining of fibrillarin and UAP56, as well as visualisation of the DNA by DAPI staining of the same nucleus ( C – E ). A merge of the three images is shown in ( F ) and examples of brightly DAPI-stained heterochromatic chromocenters are indicated by arrows. Immunostaining of RNAPII and UAP56 of the same nucleus ( G,H ).

    Article Snippet: Transient protoplast transformation assays with GFP fusion constructs Protoplasts of dark-grown tobacco BY-2 cells were transiently transformed using PEG-mediated transformation and analysed for the localisation of GFP fusion proteins by confocal laser scanning microscopy (CLSM) as previously described , using a LSM 510 microscope (Zeiss).

    Techniques: Transformation Assay, Construct, Expressing, Fluorescence, Confocal Laser Scanning Microscopy, Immunofluorescence, Microscopy, Staining, Immunostaining