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Nikon inverted fluorescence microscope
Inverted Fluorescence Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 99/100, based on 1190 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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inverted fluorescence microscope - by Bioz Stars, 2019-10
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Centrifugation:

Article Title: A multifunctional-targeted nanoagent for dual-mode image-guided therapeutic effects on ovarian cancer cells
Article Snippet: The phase-transition ability after being heated by a temperature control heating-plate or irradiated by a NIR laser exciter was observed by inverted fluorescence microscope (Eclipse Ti, Nikon Corporation, Tokyo, Japan). .. The entrapment efficiency (EE) and the ICG-loading efficiency (LE) were determined in triplicate by an ultraviolet–visible (UV–Vis) spectrophotometer (260-Bio; Thermo Fisher Scientific).

Immunocytochemistry:

Article Title: Nephron organoids derived from human pluripotent stem cells model kidney development and injury
Article Snippet: Paragraph title: Immunocytochemistry ... Immunofluorescence was visualized using an inverted fluorescence microscope (Nikon Eclipse Ti).

Mass Spectrometry:

Article Title: Heart repair by reprogramming non-myocytes with cardiac transcription factors
Article Snippet: Tomato+ cardiomyocytes were identified under a Nikon inverted fluorescence microscope with a red filter. .. Tomato+ cardiomyocytes were identified under a Nikon inverted fluorescence microscope with a red filter.

Stable Transfection:

Article Title: Cardamonin inhibits colonic neoplasia through modulation of MicroRNA expression
Article Snippet: The cells homogeneously and stably expressing the probes were seeded in 96 well glass bottom plates (BD Biosciences, #353219) and were incubated at 37 °C for 24 h in a humidified 5% CO2 incubator. .. Imaging was carried out using a ×40 Plan Apo 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan).

Cytometry:

Article Title: Synergistic suppression effect on tumor growth of acute myeloid leukemia by combining cytarabine with an engineered oncolytic vaccinia virus
Article Snippet: After 1 week, the tumors of two mice from each group were collected and digested with the help of enzymes into cell suspensions, and then subjected to flow cytometry with Annexin V-FITC/PI double staining. .. Images were photographed by an inverted fluorescence microscope (Nikon E300; Tokyo, Japan).

Immunostaining:

Article Title: Nephron organoids derived from human pluripotent stem cells model kidney development and injury
Article Snippet: For immunostaining with biotinylated LTL (Vector Labs, #B-1325), Streptavidin/Biotin Blocking Kit (Vector Labs, #SP-2002) and Alexa Fluor 488- or 647-conjugated streptavidin (Lafe Technologies) were used according to manufacturer’s instructions. .. Immunofluorescence was visualized using an inverted fluorescence microscope (Nikon Eclipse Ti).

Modification:

Article Title: Self-Assembly DNA Polyplex Vaccine inside Dissolving Microneedles for High-Potency Intradermal Vaccination
Article Snippet: HEK293 cells were added to 24-well plates at a density of about 1 × 105 cells per well in 0.5 mL of Dulbecco's modified Eagle's medium (DMEM) containing 2.2 mg/mL sodium carbonate, 10% fetal bovine serum (FBS), 50 μg/mL gentamicin, and 50 μg/mL penicillin, then incubated overnight before transfection; all incubations were performed at 37°C in 5% CO2 . .. Fluorescence images of GFP expression in cells were captured using an inverted fluorescence microscope (Eclipse Ti-S; Nikon Corp., Tokyo, Japan).

Incubation:

Article Title: Subepithelial telocytes are an important source of Wnts that supports intestinal crypts
Article Snippet: For immunofluorescence, a goat anti-PDGFRα antibody was diluted in hybridization buffer (1:100) added to the FISH probes, and incubated overnight at 30°C. .. Secondary antibody Cy2-conjugated donkey anti-goat (1:200) was added to glucose oxidase (GLOX) buffer for 20 minutes in room temperature. smFISH imaging was performed on a Nikon-Ti-E inverted fluorescence microscope with a 100x oil-immersion objective and a Photometrics Pixis 1024 CCD camera using MetaMorph software as previously described , .

Article Title: Nephron organoids derived from human pluripotent stem cells model kidney development and injury
Article Snippet: Cells were then washed three times in PBS and incubated with Alexa Fluor 488-, 555-, or 647-conjugated secondary antibodies (1:500) (Life Technologies) in antibody dilution buffer for 1 hour at RT. .. Immunofluorescence was visualized using an inverted fluorescence microscope (Nikon Eclipse Ti).

Article Title: Assembly of Functional Forebrain Spheroids from Human Pluripotent Cells
Article Snippet: Dissociated hSS (day 62) or hCS (day 123) derived from control and TS lines were cultured on poly-L-ornithine and laminin (Sigma) coated coverslips for 4–5 days. .. The cultures were incubated with 1 μM Fura-2 acetoxymethyl ester (Fura-2AM; Invitrogen) for 25 min at 37°C in NM medium, washed for 5 min and placed in a perfusion chamber on the stage of an inverted fluorescence microscope (TE2000U; Nikon). .. Cells were then stimulated with high-KCl Tyrode’s solution (67 mM KCl, 67 mM NaCl2 mM CaCl2 , 1 mM MgCl2 , 30 mM glucose and 25 mM HEPES, pH 7.4).

Article Title: Characteristics of Pulmonary Vascular Remodeling in a Novel Model of Shunt-Associated Pulmonary Arterial Hypertension
Article Snippet: After slightly drying, 4′,6-diamidino-2-phenylindole dye was added to the slices, and the samples were incubated in the dark at room temperature for 10 min. Each slide received PBS (pH, 7.4) and was placed on the shaker for 3 cycles of 5 min each to wash out the stain. .. The samples were analyzed under a Nikon inverted fluorescence microscope to acquire images and perform image processing.

Article Title: Streptomyces sp metabolite(s) promotes Bax mediated intrinsic apoptosis and autophagy involving inhibition of mTOR pathway in cervical cancer cell lines
Article Snippet: SEA4 was added at a concentration of 20ug/mL, incubated for 36 hours and the cells were imaged. .. The plates were imaged using a 40× Plan Apo λ 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan).

Article Title: Cardamonin inhibits colonic neoplasia through modulation of MicroRNA expression
Article Snippet: The cells homogeneously and stably expressing the probes were seeded in 96 well glass bottom plates (BD Biosciences, #353219) and were incubated at 37 °C for 24 h in a humidified 5% CO2 incubator. .. Imaging was carried out using a ×40 Plan Apo 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan).

Standard Deviation:

Article Title: Subepithelial telocytes are an important source of Wnts that supports intestinal crypts
Article Snippet: Secondary antibody Cy2-conjugated donkey anti-goat (1:200) was added to glucose oxidase (GLOX) buffer for 20 minutes in room temperature. smFISH imaging was performed on a Nikon-Ti-E inverted fluorescence microscope with a 100x oil-immersion objective and a Photometrics Pixis 1024 CCD camera using MetaMorph software as previously described , . .. Single cell segmentation and automatic transcript quantification were performed in a custom Matlab program (MATLAB Release 2013b, the Mathworks Inc., USA) using TransQuant software .

Expressing:

Article Title: A CRISPR-Cas9 Gene Drive System Targeting Female Reproduction in the Malaria Mosquito vector Anopheles gambiae
Article Snippet: Females were blood fed on an anaesthetized mouse on the 6th day and a minimum of 40 mosquitoes were isolated individually into 300ml beakers and allowed to lay 3 days later into a 25ml cup filled with water and lined with filter paper . .. For each female, eggs and larvae were counted and 16 larvae were screened for GFP expression using a Nikon inverted fluorescence microscope (Eclipse TE200) to confirm parental hetero/homozygosity at the HDR locus. .. Heterozygotes were confirmed by the presence of GFP- progeny.

Article Title: Self-Assembly DNA Polyplex Vaccine inside Dissolving Microneedles for High-Potency Intradermal Vaccination
Article Snippet: Cells were washed three times with serum-free DMEM, then their nuclei were stained using Hoechst. .. Fluorescence images of GFP expression in cells were captured using an inverted fluorescence microscope (Eclipse Ti-S; Nikon Corp., Tokyo, Japan). .. All images were collected using the same imaging parameters to facilitate direct comparison between figures.

Article Title: Streptomyces sp metabolite(s) promotes Bax mediated intrinsic apoptosis and autophagy involving inhibition of mTOR pathway in cervical cancer cell lines
Article Snippet: SiHa cells that were transfected and expressing 53BP1 fused with mCherry (pLPC-Puro plasmid procured from Addgene (#19835)) was used to monitor DNA damage induced by the active fraction. .. The plates were imaged using a 40× Plan Apo λ 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan).

Article Title: Cardamonin inhibits colonic neoplasia through modulation of MicroRNA expression
Article Snippet: The cells homogeneously and stably expressing the probes were seeded in 96 well glass bottom plates (BD Biosciences, #353219) and were incubated at 37 °C for 24 h in a humidified 5% CO2 incubator. .. Imaging was carried out using a ×40 Plan Apo 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan).

Knock-In:

Article Title: A CRISPR-Cas9 Gene Drive System Targeting Female Reproduction in the Malaria Mosquito vector Anopheles gambiae
Article Snippet: Offspring were screened for homozygous or heterozygous knock-in mutations by “strong” or “intermediate” intensity of GFP fluorescence respectively. .. For each female, eggs and larvae were counted and 16 larvae were screened for GFP expression using a Nikon inverted fluorescence microscope (Eclipse TE200) to confirm parental hetero/homozygosity at the HDR locus.

Translocation Assay:

Article Title: Streptomyces sp metabolite(s) promotes Bax mediated intrinsic apoptosis and autophagy involving inhibition of mTOR pathway in cervical cancer cell lines
Article Snippet: The plates were imaged using a 40× Plan Apo λ 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan). .. The images were captured using EMCCD camera Andor iXON and analysed using NIS Elements AR 3.2 software.

Derivative Assay:

Article Title: Assembly of Functional Forebrain Spheroids from Human Pluripotent Cells
Article Snippet: Dissociated hSS (day 62) or hCS (day 123) derived from control and TS lines were cultured on poly-L-ornithine and laminin (Sigma) coated coverslips for 4–5 days. .. The cultures were incubated with 1 μM Fura-2 acetoxymethyl ester (Fura-2AM; Invitrogen) for 25 min at 37°C in NM medium, washed for 5 min and placed in a perfusion chamber on the stage of an inverted fluorescence microscope (TE2000U; Nikon).

Hybridization:

Article Title: Subepithelial telocytes are an important source of Wnts that supports intestinal crypts
Article Snippet: For immunofluorescence, a goat anti-PDGFRα antibody was diluted in hybridization buffer (1:100) added to the FISH probes, and incubated overnight at 30°C. .. Secondary antibody Cy2-conjugated donkey anti-goat (1:200) was added to glucose oxidase (GLOX) buffer for 20 minutes in room temperature. smFISH imaging was performed on a Nikon-Ti-E inverted fluorescence microscope with a 100x oil-immersion objective and a Photometrics Pixis 1024 CCD camera using MetaMorph software as previously described , .

TUNEL Assay:

Article Title: Synergistic suppression effect on tumor growth of acute myeloid leukemia by combining cytarabine with an engineered oncolytic vaccinia virus
Article Snippet: Images were photographed by an inverted fluorescence microscope (Nikon E300; Tokyo, Japan). .. Images were photographed by an inverted fluorescence microscope (Nikon E300; Tokyo, Japan).

Electron Microscopy:

Article Title: Streptomyces sp metabolite(s) promotes Bax mediated intrinsic apoptosis and autophagy involving inhibition of mTOR pathway in cervical cancer cell lines
Article Snippet: 53BP1-mcherry was imaged using the filter combination of ex: 545 ± 30, em: 620 ± 60 and dichroic 570LP. .. The plates were imaged using a 40× Plan Apo λ 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan).

Article Title: Cardamonin inhibits colonic neoplasia through modulation of MicroRNA expression
Article Snippet: Imaging was carried out using a ×40 Plan Apo 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan). .. For imaging Bax EGFP, excitation filter of 470 ± 40, emission filter of 525 ± 50 and dichroic 495LP were used.

Transfection:

Article Title: Self-Assembly DNA Polyplex Vaccine inside Dissolving Microneedles for High-Potency Intradermal Vaccination
Article Snippet: Paragraph title: In-vitro gene transfection and expression study ... Fluorescence images of GFP expression in cells were captured using an inverted fluorescence microscope (Eclipse Ti-S; Nikon Corp., Tokyo, Japan).

Article Title: Streptomyces sp metabolite(s) promotes Bax mediated intrinsic apoptosis and autophagy involving inhibition of mTOR pathway in cervical cancer cell lines
Article Snippet: SiHa cells that were transfected and expressing 53BP1 fused with mCherry (pLPC-Puro plasmid procured from Addgene (#19835)) was used to monitor DNA damage induced by the active fraction. .. The plates were imaged using a 40× Plan Apo λ 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan).

Immunohistochemistry:

Article Title: Synergistic suppression effect on tumor growth of acute myeloid leukemia by combining cytarabine with an engineered oncolytic vaccinia virus
Article Snippet: For the immunohistochemical (IHC) study, the sections were stained with anti-Caspase-3 or anti-ING4 (1:100 dilutions) primary antibodies overnight at 4°C, added with the avidin-biotin-peroxidase complex, and the slides were detected with diaminobenzidine Kit (Thermo Fisher Scientific). .. Images were photographed by an inverted fluorescence microscope (Nikon E300; Tokyo, Japan).

Sublimation:

Article Title: A multifunctional-targeted nanoagent for dual-mode image-guided therapeutic effects on ovarian cancer cells
Article Snippet: The size distribution, zeta potential, and polydispersity index (PDI) of NPs were determined by dynamic light scattering (DLS) using a Malvern Zetasizer Nano ZS (Malvern Instruments, Malvern, UK). .. The phase-transition ability after being heated by a temperature control heating-plate or irradiated by a NIR laser exciter was observed by inverted fluorescence microscope (Eclipse Ti, Nikon Corporation, Tokyo, Japan). .. And the size stability of the TOI_LNPs, OI_PNPs, and TOI_HNPs in water was observed after storing at 4°C varying from day 0 to day 30.

Cell Culture:

Article Title: Assembly of Functional Forebrain Spheroids from Human Pluripotent Cells
Article Snippet: Dissociated hSS (day 62) or hCS (day 123) derived from control and TS lines were cultured on poly-L-ornithine and laminin (Sigma) coated coverslips for 4–5 days. .. The cultures were incubated with 1 μM Fura-2 acetoxymethyl ester (Fura-2AM; Invitrogen) for 25 min at 37°C in NM medium, washed for 5 min and placed in a perfusion chamber on the stage of an inverted fluorescence microscope (TE2000U; Nikon).

Article Title: Synergistic suppression effect on tumor growth of acute myeloid leukemia by combining cytarabine with an engineered oncolytic vaccinia virus
Article Snippet: Images were photographed by an inverted fluorescence microscope (Nikon E300; Tokyo, Japan). .. Moreover, the mice were randomly divided into four groups (three per group), and intraperitoneally injected with PBS (100 µL), oVV-ING4 (1×107 pfu), a dose of cytarabine (50 mg/Kg), respectively, or combination of oVV-ING4 with cytarabine.

Confocal Laser Scanning Microscopy:

Article Title: A multifunctional-targeted nanoagent for dual-mode image-guided therapeutic effects on ovarian cancer cells
Article Snippet: The morphology of TOI_HNPs was observed by confocal laser scanning microscopy (CLSM, Leica TCS SP5, Heidelberg, Germany) and the structure was characterized by using transmission electron microscope (TEM, FEI Tecnai G2 F20, FEI Company, Hillsboro, OR, USA). .. The phase-transition ability after being heated by a temperature control heating-plate or irradiated by a NIR laser exciter was observed by inverted fluorescence microscope (Eclipse Ti, Nikon Corporation, Tokyo, Japan).

Imaging:

Article Title: Subepithelial telocytes are an important source of Wnts that supports intestinal crypts
Article Snippet: For immunofluorescence, a goat anti-PDGFRα antibody was diluted in hybridization buffer (1:100) added to the FISH probes, and incubated overnight at 30°C. .. Secondary antibody Cy2-conjugated donkey anti-goat (1:200) was added to glucose oxidase (GLOX) buffer for 20 minutes in room temperature. smFISH imaging was performed on a Nikon-Ti-E inverted fluorescence microscope with a 100x oil-immersion objective and a Photometrics Pixis 1024 CCD camera using MetaMorph software as previously described , . .. Single cell segmentation and automatic transcript quantification were performed in a custom Matlab program (MATLAB Release 2013b, the Mathworks Inc., USA) using TransQuant software .

Article Title: Multiple Dynamin family members collaborate to drive mitochondrial division
Article Snippet: Imaging was performed with an inverted fluorescence microscope (TE2000-U; Nikon) equipped with an electron-multiplying charge-coupled device camera (Cascade II; Photometrics) and a Yokogawa spinning disc confocal system (CSU-Xm2; Nikon). .. Super-resolution microscopy (SIM) was performed with an inverted fluorescence microscope (TI-E; Nikon) equipped with an electron-multiplying charge-coupled device camera (iXon3 897; Andor).

Article Title: Assembly of Functional Forebrain Spheroids from Human Pluripotent Cells
Article Snippet: Paragraph title: Fura-2 calcium imaging of hSS or hCS cultures ... The cultures were incubated with 1 μM Fura-2 acetoxymethyl ester (Fura-2AM; Invitrogen) for 25 min at 37°C in NM medium, washed for 5 min and placed in a perfusion chamber on the stage of an inverted fluorescence microscope (TE2000U; Nikon).

Article Title: Multiple Dynamin family members collaborate to drive mitochondrial division
Article Snippet: Immunofluorescence was accomplished using monoclonal antibodies against cytochrome C clone 6H2.B4 (12963S, Cell Signaling Tech) and Bax6A7 (B8429, Sigma Aldrich), and the polyclonal rabbit antibody against TOM20 (sc-11415, Santa Cruz Biotech). .. Imaging was performed with an inverted fluorescence microscope (TE2000-U; Nikon) equipped with an electron-multiplying charge-coupled device camera (Cascade II; Photometrics) and a Yokogawa spinning disc confocal system (CSU-Xm2; Nikon). .. Live-cell imaging was performed at 37°C.

Article Title: Cardamonin inhibits colonic neoplasia through modulation of MicroRNA expression
Article Snippet: For imaging of TMRM the respective cells were stained with 100 nM of TMRM (Molecular Probes, #T-668, Eugene, OR, USA) followed by imaging in drug and 20 nM of TMRM containing medium. .. Imaging was carried out using a ×40 Plan Apo 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan). .. The excitation and emission filter wheels were independently and automatically controlled through NIS element software.

Transmission Assay:

Article Title: A CRISPR-Cas9 Gene Drive System Targeting Female Reproduction in the Malaria Mosquito vector Anopheles gambiae
Article Snippet: For each female, eggs and larvae were counted and 16 larvae were screened for GFP expression using a Nikon inverted fluorescence microscope (Eclipse TE200) to confirm parental hetero/homozygosity at the HDR locus. .. For each female, eggs and larvae were counted and 16 larvae were screened for GFP expression using a Nikon inverted fluorescence microscope (Eclipse TE200) to confirm parental hetero/homozygosity at the HDR locus.

Article Title: A multifunctional-targeted nanoagent for dual-mode image-guided therapeutic effects on ovarian cancer cells
Article Snippet: The morphology of TOI_HNPs was observed by confocal laser scanning microscopy (CLSM, Leica TCS SP5, Heidelberg, Germany) and the structure was characterized by using transmission electron microscope (TEM, FEI Tecnai G2 F20, FEI Company, Hillsboro, OR, USA). .. The phase-transition ability after being heated by a temperature control heating-plate or irradiated by a NIR laser exciter was observed by inverted fluorescence microscope (Eclipse Ti, Nikon Corporation, Tokyo, Japan).

Polymerase Chain Reaction:

Article Title: A CRISPR-Cas9 Gene Drive System Targeting Female Reproduction in the Malaria Mosquito vector Anopheles gambiae
Article Snippet: For each female, eggs and larvae were counted and 16 larvae were screened for GFP expression using a Nikon inverted fluorescence microscope (Eclipse TE200) to confirm parental hetero/homozygosity at the HDR locus. .. For each female, eggs and larvae were counted and 16 larvae were screened for GFP expression using a Nikon inverted fluorescence microscope (Eclipse TE200) to confirm parental hetero/homozygosity at the HDR locus.

Injection:

Article Title: Synergistic suppression effect on tumor growth of acute myeloid leukemia by combining cytarabine with an engineered oncolytic vaccinia virus
Article Snippet: When the volume of tumors approximately grew up to 200 mm, the mice were injected with PBS, cytarabine, oVV, oVV-ING4 or cytarabine, and oVV-ING4. .. Images were photographed by an inverted fluorescence microscope (Nikon E300; Tokyo, Japan).

Software:

Article Title: Subepithelial telocytes are an important source of Wnts that supports intestinal crypts
Article Snippet: For immunofluorescence, a goat anti-PDGFRα antibody was diluted in hybridization buffer (1:100) added to the FISH probes, and incubated overnight at 30°C. .. Secondary antibody Cy2-conjugated donkey anti-goat (1:200) was added to glucose oxidase (GLOX) buffer for 20 minutes in room temperature. smFISH imaging was performed on a Nikon-Ti-E inverted fluorescence microscope with a 100x oil-immersion objective and a Photometrics Pixis 1024 CCD camera using MetaMorph software as previously described , . .. Single cell segmentation and automatic transcript quantification were performed in a custom Matlab program (MATLAB Release 2013b, the Mathworks Inc., USA) using TransQuant software .

Article Title: Self-Assembly DNA Polyplex Vaccine inside Dissolving Microneedles for High-Potency Intradermal Vaccination
Article Snippet: The ImageJ software was used to analyze the area of residual needle region then calculated how many percentage of needle region was dissolved in skin compared with the original one. .. The skin sections were finally viewed using an inverted fluorescence microscope (Eclipse Ti-S; Nikon Corp., Tokyo, Japan).

Article Title: Assembly of Functional Forebrain Spheroids from Human Pluripotent Cells
Article Snippet: The cultures were incubated with 1 μM Fura-2 acetoxymethyl ester (Fura-2AM; Invitrogen) for 25 min at 37°C in NM medium, washed for 5 min and placed in a perfusion chamber on the stage of an inverted fluorescence microscope (TE2000U; Nikon). .. Imaging was performed at room temperature (25°C) on an epifluorescence microscope equipped with an excitation filter wheel and an automated stage.

Article Title: Streptomyces sp metabolite(s) promotes Bax mediated intrinsic apoptosis and autophagy involving inhibition of mTOR pathway in cervical cancer cell lines
Article Snippet: The plates were imaged using a 40× Plan Apo λ 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan). .. After indicated treatment, cells were imaged with NIKON A1R confocal microscope using 488 and 562 nm laser excitation .

Immunofluorescence:

Article Title: Subepithelial telocytes are an important source of Wnts that supports intestinal crypts
Article Snippet: Paragraph title: Single Molecule FISH and Immunofluorescence ... Secondary antibody Cy2-conjugated donkey anti-goat (1:200) was added to glucose oxidase (GLOX) buffer for 20 minutes in room temperature. smFISH imaging was performed on a Nikon-Ti-E inverted fluorescence microscope with a 100x oil-immersion objective and a Photometrics Pixis 1024 CCD camera using MetaMorph software as previously described , .

Article Title: Nephron organoids derived from human pluripotent stem cells model kidney development and injury
Article Snippet: Nuclei were counterstained with DAPI (Sigma, #D8417). .. Immunofluorescence was visualized using an inverted fluorescence microscope (Nikon Eclipse Ti). .. Quantification was performed using ImageJ by counting three to five representative fields per experiment at 20X magnification.

Article Title: Characteristics of Pulmonary Vascular Remodeling in a Novel Model of Shunt-Associated Pulmonary Arterial Hypertension
Article Snippet: Paragraph title: Immunofluorescence assays on the lung tissue sections ... The samples were analyzed under a Nikon inverted fluorescence microscope to acquire images and perform image processing.

Transmission Electron Microscopy:

Article Title: A multifunctional-targeted nanoagent for dual-mode image-guided therapeutic effects on ovarian cancer cells
Article Snippet: The morphology of TOI_HNPs was observed by confocal laser scanning microscopy (CLSM, Leica TCS SP5, Heidelberg, Germany) and the structure was characterized by using transmission electron microscope (TEM, FEI Tecnai G2 F20, FEI Company, Hillsboro, OR, USA). .. The phase-transition ability after being heated by a temperature control heating-plate or irradiated by a NIR laser exciter was observed by inverted fluorescence microscope (Eclipse Ti, Nikon Corporation, Tokyo, Japan).

Fluorescence:

Article Title: Heart repair by reprogramming non-myocytes with cardiac transcription factors
Article Snippet: Action potentials of beating iCLMs with di-4-ANEPPS (Invitrogen) were assessed, as described previously . .. Tomato+ cardiomyocytes were identified under a Nikon inverted fluorescence microscope with a red filter. .. Whole-cell current clamp experiments (Axopatch 200B, Molecular Devices) were conducted to measure cardiomyocyte membrane action potentials (AP).

Article Title: A CRISPR-Cas9 Gene Drive System Targeting Female Reproduction in the Malaria Mosquito vector Anopheles gambiae
Article Snippet: Females were blood fed on an anaesthetized mouse on the 6th day and a minimum of 40 mosquitoes were isolated individually into 300ml beakers and allowed to lay 3 days later into a 25ml cup filled with water and lined with filter paper . .. For each female, eggs and larvae were counted and 16 larvae were screened for GFP expression using a Nikon inverted fluorescence microscope (Eclipse TE200) to confirm parental hetero/homozygosity at the HDR locus. .. Heterozygotes were confirmed by the presence of GFP- progeny.

Article Title: Subepithelial telocytes are an important source of Wnts that supports intestinal crypts
Article Snippet: For immunofluorescence, a goat anti-PDGFRα antibody was diluted in hybridization buffer (1:100) added to the FISH probes, and incubated overnight at 30°C. .. Secondary antibody Cy2-conjugated donkey anti-goat (1:200) was added to glucose oxidase (GLOX) buffer for 20 minutes in room temperature. smFISH imaging was performed on a Nikon-Ti-E inverted fluorescence microscope with a 100x oil-immersion objective and a Photometrics Pixis 1024 CCD camera using MetaMorph software as previously described , . .. Single cell segmentation and automatic transcript quantification were performed in a custom Matlab program (MATLAB Release 2013b, the Mathworks Inc., USA) using TransQuant software .

Article Title: Self-Assembly DNA Polyplex Vaccine inside Dissolving Microneedles for High-Potency Intradermal Vaccination
Article Snippet: The frozen OCT-skin samples were subsequently sliced into 50-μm thick sections using a cryotome and collected by silane coated glass slides. .. The skin sections were finally viewed using an inverted fluorescence microscope (Eclipse Ti-S; Nikon Corp., Tokyo, Japan). .. MN patches containing pcDNA4-PCV2 were stored at 45°C for 1-121 days, and then dissolved in DI water.

Article Title: Multiple Dynamin family members collaborate to drive mitochondrial division
Article Snippet: Images were captured with a 100x NA 1.4 oil objective and acquired using MetaMorph (version 7.0; MDS Analytical Technologies). .. Super-resolution microscopy (SIM) was performed with an inverted fluorescence microscope (TI-E; Nikon) equipped with an electron-multiplying charge-coupled device camera (iXon3 897; Andor). .. Z-stack images were captured every 200 nm and reconstructed using NIS elements with SIM (Nikon).

Article Title: Nephron organoids derived from human pluripotent stem cells model kidney development and injury
Article Snippet: Nuclei were counterstained with DAPI (Sigma, #D8417). .. Immunofluorescence was visualized using an inverted fluorescence microscope (Nikon Eclipse Ti). .. Quantification was performed using ImageJ by counting three to five representative fields per experiment at 20X magnification.

Article Title: Assembly of Functional Forebrain Spheroids from Human Pluripotent Cells
Article Snippet: Dissociated hSS (day 62) or hCS (day 123) derived from control and TS lines were cultured on poly-L-ornithine and laminin (Sigma) coated coverslips for 4–5 days. .. The cultures were incubated with 1 μM Fura-2 acetoxymethyl ester (Fura-2AM; Invitrogen) for 25 min at 37°C in NM medium, washed for 5 min and placed in a perfusion chamber on the stage of an inverted fluorescence microscope (TE2000U; Nikon). .. Cells were then stimulated with high-KCl Tyrode’s solution (67 mM KCl, 67 mM NaCl2 mM CaCl2 , 1 mM MgCl2 , 30 mM glucose and 25 mM HEPES, pH 7.4).

Article Title: Characteristics of Pulmonary Vascular Remodeling in a Novel Model of Shunt-Associated Pulmonary Arterial Hypertension
Article Snippet: After slightly drying, the antifade solution was added to the slices, and the samples were mounted and sealed. .. The samples were analyzed under a Nikon inverted fluorescence microscope to acquire images and perform image processing. .. SPSS version 16.0 statistical software (SPSS, Inc.) was used to perform the statistical analyses.

Article Title: Effect of bone marrow mesenchymal stem cells on the polarization of macrophages
Article Snippet: Whole lung samples were fixed in 10% formalin for 24 h at room temperature and were sent to Google Biological Technology Co., Ltd. (Wuhan, China) for further procedure, where they were subjected to conventional methods of dehydration, paraffin-embedding and hematoxylin and eosin (H & E) staining. .. All the lung tissues (thickness: 5 µm) were observed with an inverted fluorescence microscope (TI-S; Nikon Corporation, Tokyo, Japan). .. Flow cytometry was used to assess the cell surface or intracellular marker, CD68, of BMSCs and macrophages, as described in previous literature ( – ).

Article Title: Multiple Dynamin family members collaborate to drive mitochondrial division
Article Snippet: Immunofluorescence was accomplished using monoclonal antibodies against cytochrome C clone 6H2.B4 (12963S, Cell Signaling Tech) and Bax6A7 (B8429, Sigma Aldrich), and the polyclonal rabbit antibody against TOM20 (sc-11415, Santa Cruz Biotech). .. Imaging was performed with an inverted fluorescence microscope (TE2000-U; Nikon) equipped with an electron-multiplying charge-coupled device camera (Cascade II; Photometrics) and a Yokogawa spinning disc confocal system (CSU-Xm2; Nikon). .. Live-cell imaging was performed at 37°C.

Article Title: A multifunctional-targeted nanoagent for dual-mode image-guided therapeutic effects on ovarian cancer cells
Article Snippet: The size distribution, zeta potential, and polydispersity index (PDI) of NPs were determined by dynamic light scattering (DLS) using a Malvern Zetasizer Nano ZS (Malvern Instruments, Malvern, UK). .. The phase-transition ability after being heated by a temperature control heating-plate or irradiated by a NIR laser exciter was observed by inverted fluorescence microscope (Eclipse Ti, Nikon Corporation, Tokyo, Japan). .. And the size stability of the TOI_LNPs, OI_PNPs, and TOI_HNPs in water was observed after storing at 4°C varying from day 0 to day 30.

Article Title: pH-sensitive doxorubicin-loaded polymeric nanocomplex based on β-cyclodextrin for liver cancer-targeted therapy
Article Snippet: Then, cells were washed with PBS and stained with DAPI in the dark at room temperature for 10 minutes. .. Images were obtained using an inverted fluorescence microscope (DM505, Nikon Corporation, Tokyo, Japan). .. A549 and HepG2 cells were plated in 6-well plates in complete medium and cultivated overnight.

Article Title: The Extracts of Artemisia absinthium L. Suppress the Growth of Hepatocellular Carcinoma Cells through Induction of Apoptosis via Endoplasmic Reticulum Stress and Mitochondrial-Dependent Pathway
Article Snippet: The relative cell viability was calculated according to the formula Cell viability (%) = (ODtreated /ODuntreated ) × 100%. .. H22 and BEL-7404 cells were seeded in 96-well plates and were treated with different concentrations of AAEE, AAEE-Pe, and AAEE-Ea for 24 h. After treatment, the morphology of H22 and BEL-7404 cells was observed by inverted fluorescence microscope (Nikon Eclipse Ti-E, Tokyo, Japan). .. H22 and BEL-7404 cells were treated with different concentrations of AAEE, AAEE-Pe, and AAEE-Ea for 24 h, and then stained with an Annexin V-FITC/propidium iodide (PI) Apoptosis Detection Kit (YEASEN, Shanghai, China) according to the manufacturer’s instructions.

Article Title: Chemical composition, antioxidant and antitumor activities of sub-fractions of wild and cultivated Pleurotus ferulae ethanol extracts
Article Snippet: After treatment with 400 μg/ml PFEE-W/C and subfractions or 0.4% DMSO for 24 h, HeLa cells were washed with PBS and fixed with 4% ice–cold paraformaldehyde at 4 °C for 10 min. Then cells were stained with Hoechst 33342 (Beyotime, Shanghai, China) at 4 °C for 10 min in the dark after washing with PBS. .. Samples were observed by inverted fluorescence microscope (Nikon Eclipse Ti-E, Tokyo, Japan). .. Intracellular reactive oxygen species (ROS) production was measured using DCFH-DA probes.

Article Title: Streptomyces sp metabolite(s) promotes Bax mediated intrinsic apoptosis and autophagy involving inhibition of mTOR pathway in cervical cancer cell lines
Article Snippet: 53BP1-mcherry was imaged using the filter combination of ex: 545 ± 30, em: 620 ± 60 and dichroic 570LP. .. The plates were imaged using a 40× Plan Apo λ 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan). .. To analyze cytochrome c EGFP release from mitochondria, cells expressing cytochrome c EGFP and MitoDsRed, were grown on 8 well chambered cover glass.

Article Title: Cardamonin inhibits colonic neoplasia through modulation of MicroRNA expression
Article Snippet: For imaging of TMRM the respective cells were stained with 100 nM of TMRM (Molecular Probes, #T-668, Eugene, OR, USA) followed by imaging in drug and 20 nM of TMRM containing medium. .. Imaging was carried out using a ×40 Plan Apo 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan). .. The excitation and emission filter wheels were independently and automatically controlled through NIS element software.

Article Title: Synergistic suppression effect on tumor growth of acute myeloid leukemia by combining cytarabine with an engineered oncolytic vaccinia virus
Article Snippet: For the immunohistochemical (IHC) study, the sections were stained with anti-Caspase-3 or anti-ING4 (1:100 dilutions) primary antibodies overnight at 4°C, added with the avidin-biotin-peroxidase complex, and the slides were detected with diaminobenzidine Kit (Thermo Fisher Scientific). .. Images were photographed by an inverted fluorescence microscope (Nikon E300; Tokyo, Japan). .. Moreover, the mice were randomly divided into four groups (three per group), and intraperitoneally injected with PBS (100 µL), oVV-ING4 (1×107 pfu), a dose of cytarabine (50 mg/Kg), respectively, or combination of oVV-ING4 with cytarabine.

Isolation:

Article Title: Heart repair by reprogramming non-myocytes with cardiac transcription factors
Article Snippet: Paragraph title: Action potential recordings of iCLMs isolated from adult mouse hearts ... Tomato+ cardiomyocytes were identified under a Nikon inverted fluorescence microscope with a red filter.

Article Title: A CRISPR-Cas9 Gene Drive System Targeting Female Reproduction in the Malaria Mosquito vector Anopheles gambiae
Article Snippet: For each female, eggs and larvae were counted and 16 larvae were screened for GFP expression using a Nikon inverted fluorescence microscope (Eclipse TE200) to confirm parental hetero/homozygosity at the HDR locus. .. For each female, eggs and larvae were counted and 16 larvae were screened for GFP expression using a Nikon inverted fluorescence microscope (Eclipse TE200) to confirm parental hetero/homozygosity at the HDR locus.

Article Title: Synergistic suppression effect on tumor growth of acute myeloid leukemia by combining cytarabine with an engineered oncolytic vaccinia virus
Article Snippet: Images were photographed by an inverted fluorescence microscope (Nikon E300; Tokyo, Japan). .. Moreover, the mice were randomly divided into four groups (three per group), and intraperitoneally injected with PBS (100 µL), oVV-ING4 (1×107 pfu), a dose of cytarabine (50 mg/Kg), respectively, or combination of oVV-ING4 with cytarabine.

Flow Cytometry:

Article Title: Synergistic suppression effect on tumor growth of acute myeloid leukemia by combining cytarabine with an engineered oncolytic vaccinia virus
Article Snippet: After 1 week, the tumors of two mice from each group were collected and digested with the help of enzymes into cell suspensions, and then subjected to flow cytometry with Annexin V-FITC/PI double staining. .. Images were photographed by an inverted fluorescence microscope (Nikon E300; Tokyo, Japan).

Microscopy:

Article Title: Heart repair by reprogramming non-myocytes with cardiac transcription factors
Article Snippet: Action potentials of beating iCLMs with di-4-ANEPPS (Invitrogen) were assessed, as described previously . .. Tomato+ cardiomyocytes were identified under a Nikon inverted fluorescence microscope with a red filter. .. Whole-cell current clamp experiments (Axopatch 200B, Molecular Devices) were conducted to measure cardiomyocyte membrane action potentials (AP).

Article Title: A CRISPR-Cas9 Gene Drive System Targeting Female Reproduction in the Malaria Mosquito vector Anopheles gambiae
Article Snippet: Females were blood fed on an anaesthetized mouse on the 6th day and a minimum of 40 mosquitoes were isolated individually into 300ml beakers and allowed to lay 3 days later into a 25ml cup filled with water and lined with filter paper . .. For each female, eggs and larvae were counted and 16 larvae were screened for GFP expression using a Nikon inverted fluorescence microscope (Eclipse TE200) to confirm parental hetero/homozygosity at the HDR locus. .. Heterozygotes were confirmed by the presence of GFP- progeny.

Article Title: Subepithelial telocytes are an important source of Wnts that supports intestinal crypts
Article Snippet: For immunofluorescence, a goat anti-PDGFRα antibody was diluted in hybridization buffer (1:100) added to the FISH probes, and incubated overnight at 30°C. .. Secondary antibody Cy2-conjugated donkey anti-goat (1:200) was added to glucose oxidase (GLOX) buffer for 20 minutes in room temperature. smFISH imaging was performed on a Nikon-Ti-E inverted fluorescence microscope with a 100x oil-immersion objective and a Photometrics Pixis 1024 CCD camera using MetaMorph software as previously described , . .. Single cell segmentation and automatic transcript quantification were performed in a custom Matlab program (MATLAB Release 2013b, the Mathworks Inc., USA) using TransQuant software .

Article Title: Self-Assembly DNA Polyplex Vaccine inside Dissolving Microneedles for High-Potency Intradermal Vaccination
Article Snippet: The frozen OCT-skin samples were subsequently sliced into 50-μm thick sections using a cryotome and collected by silane coated glass slides. .. The skin sections were finally viewed using an inverted fluorescence microscope (Eclipse Ti-S; Nikon Corp., Tokyo, Japan). .. MN patches containing pcDNA4-PCV2 were stored at 45°C for 1-121 days, and then dissolved in DI water.

Article Title: Multiple Dynamin family members collaborate to drive mitochondrial division
Article Snippet: Images were captured with a 100x NA 1.4 oil objective and acquired using MetaMorph (version 7.0; MDS Analytical Technologies). .. Super-resolution microscopy (SIM) was performed with an inverted fluorescence microscope (TI-E; Nikon) equipped with an electron-multiplying charge-coupled device camera (iXon3 897; Andor). .. Z-stack images were captured every 200 nm and reconstructed using NIS elements with SIM (Nikon).

Article Title: Nephron organoids derived from human pluripotent stem cells model kidney development and injury
Article Snippet: Nuclei were counterstained with DAPI (Sigma, #D8417). .. Immunofluorescence was visualized using an inverted fluorescence microscope (Nikon Eclipse Ti). .. Quantification was performed using ImageJ by counting three to five representative fields per experiment at 20X magnification.

Article Title: Assembly of Functional Forebrain Spheroids from Human Pluripotent Cells
Article Snippet: Dissociated hSS (day 62) or hCS (day 123) derived from control and TS lines were cultured on poly-L-ornithine and laminin (Sigma) coated coverslips for 4–5 days. .. The cultures were incubated with 1 μM Fura-2 acetoxymethyl ester (Fura-2AM; Invitrogen) for 25 min at 37°C in NM medium, washed for 5 min and placed in a perfusion chamber on the stage of an inverted fluorescence microscope (TE2000U; Nikon). .. Cells were then stimulated with high-KCl Tyrode’s solution (67 mM KCl, 67 mM NaCl2 mM CaCl2 , 1 mM MgCl2 , 30 mM glucose and 25 mM HEPES, pH 7.4).

Article Title: Characteristics of Pulmonary Vascular Remodeling in a Novel Model of Shunt-Associated Pulmonary Arterial Hypertension
Article Snippet: After slightly drying, the antifade solution was added to the slices, and the samples were mounted and sealed. .. The samples were analyzed under a Nikon inverted fluorescence microscope to acquire images and perform image processing. .. SPSS version 16.0 statistical software (SPSS, Inc.) was used to perform the statistical analyses.

Article Title: Effect of bone marrow mesenchymal stem cells on the polarization of macrophages
Article Snippet: Whole lung samples were fixed in 10% formalin for 24 h at room temperature and were sent to Google Biological Technology Co., Ltd. (Wuhan, China) for further procedure, where they were subjected to conventional methods of dehydration, paraffin-embedding and hematoxylin and eosin (H & E) staining. .. All the lung tissues (thickness: 5 µm) were observed with an inverted fluorescence microscope (TI-S; Nikon Corporation, Tokyo, Japan). .. Flow cytometry was used to assess the cell surface or intracellular marker, CD68, of BMSCs and macrophages, as described in previous literature ( – ).

Article Title: Multiple Dynamin family members collaborate to drive mitochondrial division
Article Snippet: Immunofluorescence was accomplished using monoclonal antibodies against cytochrome C clone 6H2.B4 (12963S, Cell Signaling Tech) and Bax6A7 (B8429, Sigma Aldrich), and the polyclonal rabbit antibody against TOM20 (sc-11415, Santa Cruz Biotech). .. Imaging was performed with an inverted fluorescence microscope (TE2000-U; Nikon) equipped with an electron-multiplying charge-coupled device camera (Cascade II; Photometrics) and a Yokogawa spinning disc confocal system (CSU-Xm2; Nikon). .. Live-cell imaging was performed at 37°C.

Article Title: A multifunctional-targeted nanoagent for dual-mode image-guided therapeutic effects on ovarian cancer cells
Article Snippet: The size distribution, zeta potential, and polydispersity index (PDI) of NPs were determined by dynamic light scattering (DLS) using a Malvern Zetasizer Nano ZS (Malvern Instruments, Malvern, UK). .. The phase-transition ability after being heated by a temperature control heating-plate or irradiated by a NIR laser exciter was observed by inverted fluorescence microscope (Eclipse Ti, Nikon Corporation, Tokyo, Japan). .. And the size stability of the TOI_LNPs, OI_PNPs, and TOI_HNPs in water was observed after storing at 4°C varying from day 0 to day 30.

Article Title: pH-sensitive doxorubicin-loaded polymeric nanocomplex based on β-cyclodextrin for liver cancer-targeted therapy
Article Snippet: Then, cells were washed with PBS and stained with DAPI in the dark at room temperature for 10 minutes. .. Images were obtained using an inverted fluorescence microscope (DM505, Nikon Corporation, Tokyo, Japan). .. A549 and HepG2 cells were plated in 6-well plates in complete medium and cultivated overnight.

Article Title: The Extracts of Artemisia absinthium L. Suppress the Growth of Hepatocellular Carcinoma Cells through Induction of Apoptosis via Endoplasmic Reticulum Stress and Mitochondrial-Dependent Pathway
Article Snippet: The relative cell viability was calculated according to the formula Cell viability (%) = (ODtreated /ODuntreated ) × 100%. .. H22 and BEL-7404 cells were seeded in 96-well plates and were treated with different concentrations of AAEE, AAEE-Pe, and AAEE-Ea for 24 h. After treatment, the morphology of H22 and BEL-7404 cells was observed by inverted fluorescence microscope (Nikon Eclipse Ti-E, Tokyo, Japan). .. H22 and BEL-7404 cells were treated with different concentrations of AAEE, AAEE-Pe, and AAEE-Ea for 24 h, and then stained with an Annexin V-FITC/propidium iodide (PI) Apoptosis Detection Kit (YEASEN, Shanghai, China) according to the manufacturer’s instructions.

Article Title: Chemical composition, antioxidant and antitumor activities of sub-fractions of wild and cultivated Pleurotus ferulae ethanol extracts
Article Snippet: After treatment with 400 μg/ml PFEE-W/C and subfractions or 0.4% DMSO for 24 h, HeLa cells were washed with PBS and fixed with 4% ice–cold paraformaldehyde at 4 °C for 10 min. Then cells were stained with Hoechst 33342 (Beyotime, Shanghai, China) at 4 °C for 10 min in the dark after washing with PBS. .. Samples were observed by inverted fluorescence microscope (Nikon Eclipse Ti-E, Tokyo, Japan). .. Intracellular reactive oxygen species (ROS) production was measured using DCFH-DA probes.

Article Title: Streptomyces sp metabolite(s) promotes Bax mediated intrinsic apoptosis and autophagy involving inhibition of mTOR pathway in cervical cancer cell lines
Article Snippet: 53BP1-mcherry was imaged using the filter combination of ex: 545 ± 30, em: 620 ± 60 and dichroic 570LP. .. The plates were imaged using a 40× Plan Apo λ 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan). .. To analyze cytochrome c EGFP release from mitochondria, cells expressing cytochrome c EGFP and MitoDsRed, were grown on 8 well chambered cover glass.

Article Title: Cardamonin inhibits colonic neoplasia through modulation of MicroRNA expression
Article Snippet: For imaging of TMRM the respective cells were stained with 100 nM of TMRM (Molecular Probes, #T-668, Eugene, OR, USA) followed by imaging in drug and 20 nM of TMRM containing medium. .. Imaging was carried out using a ×40 Plan Apo 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan). .. The excitation and emission filter wheels were independently and automatically controlled through NIS element software.

Article Title: Synergistic suppression effect on tumor growth of acute myeloid leukemia by combining cytarabine with an engineered oncolytic vaccinia virus
Article Snippet: For the immunohistochemical (IHC) study, the sections were stained with anti-Caspase-3 or anti-ING4 (1:100 dilutions) primary antibodies overnight at 4°C, added with the avidin-biotin-peroxidase complex, and the slides were detected with diaminobenzidine Kit (Thermo Fisher Scientific). .. Images were photographed by an inverted fluorescence microscope (Nikon E300; Tokyo, Japan). .. Moreover, the mice were randomly divided into four groups (three per group), and intraperitoneally injected with PBS (100 µL), oVV-ING4 (1×107 pfu), a dose of cytarabine (50 mg/Kg), respectively, or combination of oVV-ING4 with cytarabine.

Avidin-Biotin Assay:

Article Title: Synergistic suppression effect on tumor growth of acute myeloid leukemia by combining cytarabine with an engineered oncolytic vaccinia virus
Article Snippet: For the immunohistochemical (IHC) study, the sections were stained with anti-Caspase-3 or anti-ING4 (1:100 dilutions) primary antibodies overnight at 4°C, added with the avidin-biotin-peroxidase complex, and the slides were detected with diaminobenzidine Kit (Thermo Fisher Scientific). .. Images were photographed by an inverted fluorescence microscope (Nikon E300; Tokyo, Japan).

Transgenic Assay:

Article Title: A CRISPR-Cas9 Gene Drive System Targeting Female Reproduction in the Malaria Mosquito vector Anopheles gambiae
Article Snippet: For each female, eggs and larvae were counted and 16 larvae were screened for GFP expression using a Nikon inverted fluorescence microscope (Eclipse TE200) to confirm parental hetero/homozygosity at the HDR locus. .. For each female, eggs and larvae were counted and 16 larvae were screened for GFP expression using a Nikon inverted fluorescence microscope (Eclipse TE200) to confirm parental hetero/homozygosity at the HDR locus.

Blocking Assay:

Article Title: Nephron organoids derived from human pluripotent stem cells model kidney development and injury
Article Snippet: For immunostaining with biotinylated LTL (Vector Labs, #B-1325), Streptavidin/Biotin Blocking Kit (Vector Labs, #SP-2002) and Alexa Fluor 488- or 647-conjugated streptavidin (Lafe Technologies) were used according to manufacturer’s instructions. .. Immunofluorescence was visualized using an inverted fluorescence microscope (Nikon Eclipse Ti).

Confocal Microscopy:

Article Title: Streptomyces sp metabolite(s) promotes Bax mediated intrinsic apoptosis and autophagy involving inhibition of mTOR pathway in cervical cancer cell lines
Article Snippet: Paragraph title: Confocal microscopy ... The plates were imaged using a 40× Plan Apo λ 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan).

Staining:

Article Title: Characteristics of Pulmonary Vascular Remodeling in a Novel Model of Shunt-Associated Pulmonary Arterial Hypertension
Article Snippet: After slightly drying, 4′,6-diamidino-2-phenylindole dye was added to the slices, and the samples were incubated in the dark at room temperature for 10 min. Each slide received PBS (pH, 7.4) and was placed on the shaker for 3 cycles of 5 min each to wash out the stain. .. The samples were analyzed under a Nikon inverted fluorescence microscope to acquire images and perform image processing.

Article Title: Effect of bone marrow mesenchymal stem cells on the polarization of macrophages
Article Snippet: Whole lung samples were fixed in 10% formalin for 24 h at room temperature and were sent to Google Biological Technology Co., Ltd. (Wuhan, China) for further procedure, where they were subjected to conventional methods of dehydration, paraffin-embedding and hematoxylin and eosin (H & E) staining. .. All the lung tissues (thickness: 5 µm) were observed with an inverted fluorescence microscope (TI-S; Nikon Corporation, Tokyo, Japan).

Article Title: Self-Assembly DNA Polyplex Vaccine inside Dissolving Microneedles for High-Potency Intradermal Vaccination
Article Snippet: Cells were washed three times with serum-free DMEM, then their nuclei were stained using Hoechst. .. Fluorescence images of GFP expression in cells were captured using an inverted fluorescence microscope (Eclipse Ti-S; Nikon Corp., Tokyo, Japan).

Article Title: pH-sensitive doxorubicin-loaded polymeric nanocomplex based on β-cyclodextrin for liver cancer-targeted therapy
Article Snippet: Then, cells were washed with PBS and stained with DAPI in the dark at room temperature for 10 minutes. .. Images were obtained using an inverted fluorescence microscope (DM505, Nikon Corporation, Tokyo, Japan).

Article Title: Chemical composition, antioxidant and antitumor activities of sub-fractions of wild and cultivated Pleurotus ferulae ethanol extracts
Article Snippet: Paragraph title: Hoechst 33342 staining ... Samples were observed by inverted fluorescence microscope (Nikon Eclipse Ti-E, Tokyo, Japan).

Article Title: Cardamonin inhibits colonic neoplasia through modulation of MicroRNA expression
Article Snippet: For imaging of TMRM the respective cells were stained with 100 nM of TMRM (Molecular Probes, #T-668, Eugene, OR, USA) followed by imaging in drug and 20 nM of TMRM containing medium. .. Imaging was carried out using a ×40 Plan Apo 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan).

Article Title: Synergistic suppression effect on tumor growth of acute myeloid leukemia by combining cytarabine with an engineered oncolytic vaccinia virus
Article Snippet: For the immunohistochemical (IHC) study, the sections were stained with anti-Caspase-3 or anti-ING4 (1:100 dilutions) primary antibodies overnight at 4°C, added with the avidin-biotin-peroxidase complex, and the slides were detected with diaminobenzidine Kit (Thermo Fisher Scientific). .. Images were photographed by an inverted fluorescence microscope (Nikon E300; Tokyo, Japan).

Mouse Assay:

Article Title: Synergistic suppression effect on tumor growth of acute myeloid leukemia by combining cytarabine with an engineered oncolytic vaccinia virus
Article Snippet: Images were photographed by an inverted fluorescence microscope (Nikon E300; Tokyo, Japan). .. Images were photographed by an inverted fluorescence microscope (Nikon E300; Tokyo, Japan).

Plasmid Preparation:

Article Title: Streptomyces sp metabolite(s) promotes Bax mediated intrinsic apoptosis and autophagy involving inhibition of mTOR pathway in cervical cancer cell lines
Article Snippet: SiHa cells that were transfected and expressing 53BP1 fused with mCherry (pLPC-Puro plasmid procured from Addgene (#19835)) was used to monitor DNA damage induced by the active fraction. .. The plates were imaged using a 40× Plan Apo λ 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan).

Double Staining:

Article Title: Synergistic suppression effect on tumor growth of acute myeloid leukemia by combining cytarabine with an engineered oncolytic vaccinia virus
Article Snippet: After 1 week, the tumors of two mice from each group were collected and digested with the help of enzymes into cell suspensions, and then subjected to flow cytometry with Annexin V-FITC/PI double staining. .. Images were photographed by an inverted fluorescence microscope (Nikon E300; Tokyo, Japan).

Irradiation:

Article Title: A multifunctional-targeted nanoagent for dual-mode image-guided therapeutic effects on ovarian cancer cells
Article Snippet: The size distribution, zeta potential, and polydispersity index (PDI) of NPs were determined by dynamic light scattering (DLS) using a Malvern Zetasizer Nano ZS (Malvern Instruments, Malvern, UK). .. The phase-transition ability after being heated by a temperature control heating-plate or irradiated by a NIR laser exciter was observed by inverted fluorescence microscope (Eclipse Ti, Nikon Corporation, Tokyo, Japan). .. And the size stability of the TOI_LNPs, OI_PNPs, and TOI_HNPs in water was observed after storing at 4°C varying from day 0 to day 30.

In Vitro:

Article Title: Self-Assembly DNA Polyplex Vaccine inside Dissolving Microneedles for High-Potency Intradermal Vaccination
Article Snippet: Paragraph title: In-vitro gene transfection and expression study ... Fluorescence images of GFP expression in cells were captured using an inverted fluorescence microscope (Eclipse Ti-S; Nikon Corp., Tokyo, Japan).

Live Cell Imaging:

Article Title: Multiple Dynamin family members collaborate to drive mitochondrial division
Article Snippet: Super-resolution microscopy (SIM) was performed with an inverted fluorescence microscope (TI-E; Nikon) equipped with an electron-multiplying charge-coupled device camera (iXon3 897; Andor). .. Super-resolution microscopy (SIM) was performed with an inverted fluorescence microscope (TI-E; Nikon) equipped with an electron-multiplying charge-coupled device camera (iXon3 897; Andor).

Spectrophotometry:

Article Title: A multifunctional-targeted nanoagent for dual-mode image-guided therapeutic effects on ovarian cancer cells
Article Snippet: The phase-transition ability after being heated by a temperature control heating-plate or irradiated by a NIR laser exciter was observed by inverted fluorescence microscope (Eclipse Ti, Nikon Corporation, Tokyo, Japan). .. The phase-transition ability after being heated by a temperature control heating-plate or irradiated by a NIR laser exciter was observed by inverted fluorescence microscope (Eclipse Ti, Nikon Corporation, Tokyo, Japan).

Concentration Assay:

Article Title: A multifunctional-targeted nanoagent for dual-mode image-guided therapeutic effects on ovarian cancer cells
Article Snippet: The phase-transition ability after being heated by a temperature control heating-plate or irradiated by a NIR laser exciter was observed by inverted fluorescence microscope (Eclipse Ti, Nikon Corporation, Tokyo, Japan). .. The entrapment efficiency (EE) and the ICG-loading efficiency (LE) were determined in triplicate by an ultraviolet–visible (UV–Vis) spectrophotometer (260-Bio; Thermo Fisher Scientific).

Article Title: Streptomyces sp metabolite(s) promotes Bax mediated intrinsic apoptosis and autophagy involving inhibition of mTOR pathway in cervical cancer cell lines
Article Snippet: SEA4 was added at a concentration of 20ug/mL, incubated for 36 hours and the cells were imaged. .. The plates were imaged using a 40× Plan Apo λ 0.95 NA objective under an inverted fluorescence microscope (Nikon Eclipse, TE2000-E, Tokyo, Japan).

Marker:

Article Title: Characteristics of Pulmonary Vascular Remodeling in a Novel Model of Shunt-Associated Pulmonary Arterial Hypertension
Article Snippet: After drying, the tissue surroundings were outlined with a marker, and 5% bovine serum albumin diluted rabbit anti-mouse CD31 antibody and rabbit anti-mouse α-SMA antibody were added to cover the tissues. .. The samples were analyzed under a Nikon inverted fluorescence microscope to acquire images and perform image processing.

FACS:

Article Title: pH-sensitive doxorubicin-loaded polymeric nanocomplex based on β-cyclodextrin for liver cancer-targeted therapy
Article Snippet: The DOX content in the cells was detected and analyzed by FACS (ACEA Novocyte, San Diego, CA, USA). .. Images were obtained using an inverted fluorescence microscope (DM505, Nikon Corporation, Tokyo, Japan).

Fluorescence In Situ Hybridization:

Article Title: Subepithelial telocytes are an important source of Wnts that supports intestinal crypts
Article Snippet: Paragraph title: Single Molecule FISH and Immunofluorescence ... Secondary antibody Cy2-conjugated donkey anti-goat (1:200) was added to glucose oxidase (GLOX) buffer for 20 minutes in room temperature. smFISH imaging was performed on a Nikon-Ti-E inverted fluorescence microscope with a 100x oil-immersion objective and a Photometrics Pixis 1024 CCD camera using MetaMorph software as previously described , .

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    Nikon te2000u inverted fluorescence microscope
    4-OH-E2-induced cell transformation is inhibited by AKT1 silencing. Inhibition of AKT expression by its silencing detected by Western Bloting (A) and confocal microscopy (B). (C) Detection of inhibition of 4-OH-E2-induced cell transformation by AKT1 silencing by anchorage-independent growth assay. The MCF-10A cells were transfected with pre-designed and verified human shRNA for AKT1 and control shRNA plasmid consisting of scrambled shRNA sequence that does not lead to the specific degradation of AKT1 (OriGene Technologies, Inc. Rockville, MD). These cells were exposed to a carcinogenic dose of 4-OH-E2 (10 ng/ml) as described in Fig. 2. The cellular extracts from treated and controls cells were separated on SDS-PAGE, transferred to the membrane, and followed by Western detection of AKT. Images of 40x of AKT immuno-reactivity of 4-OH-E2 treated wild type and Akt silenced MCF-10A cells were acquired by immunofluorescence confocal microscopy using Alexafluor 488. Anchorage independent growth, an indicator of neoplastic transformation of cells, was assessed in soft agar. Images were acquired by using an Olympus C-5060 digital camera attached to the Nikon <t>TE2000U</t> inverted microscope with a 4x objective. Colony efficiency was determined by a count of the number of colonies > 63 um in diameter and data expressed as mean of five wells +/− S.D.
    Te2000u Inverted Fluorescence Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/te2000u inverted fluorescence microscope/product/Nikon
    Average 90 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    te2000u inverted fluorescence microscope - by Bioz Stars, 2019-10
    90/100 stars
      Buy from Supplier

    78
    Nikon inverted fluorescent widefield microscope
    Unlike the attenuated BCG vaccine, virulent M. tuberculosis is able to induce phagosomal rupture in THP-1 macrophages. THP-1 cells were infected with BCG DsRed (A,C) or M. tuberculosis DsRed (B,D) at a MOI of 1 for the indicated time and then loaded with CCF-4 molecule for 2 h. After PFA fixation, cells were imaged using a fluorescence <t>widefield</t> microscope (Nikon Ti) with an 40X objective (A, B). Picture acquisition was achieved randomly and automatically for each condition on 49 fields in duplicates and further 450/535 nm intensity ratio measurements shown in panels C and D were obtained through analysis by a specialized algorithm using the Metamorph software. The plots are representative of 3 independent experiments.
    Inverted Fluorescent Widefield Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/inverted fluorescent widefield microscope/product/Nikon
    Average 78 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    inverted fluorescent widefield microscope - by Bioz Stars, 2019-10
    78/100 stars
      Buy from Supplier

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    4-OH-E2-induced cell transformation is inhibited by AKT1 silencing. Inhibition of AKT expression by its silencing detected by Western Bloting (A) and confocal microscopy (B). (C) Detection of inhibition of 4-OH-E2-induced cell transformation by AKT1 silencing by anchorage-independent growth assay. The MCF-10A cells were transfected with pre-designed and verified human shRNA for AKT1 and control shRNA plasmid consisting of scrambled shRNA sequence that does not lead to the specific degradation of AKT1 (OriGene Technologies, Inc. Rockville, MD). These cells were exposed to a carcinogenic dose of 4-OH-E2 (10 ng/ml) as described in Fig. 2. The cellular extracts from treated and controls cells were separated on SDS-PAGE, transferred to the membrane, and followed by Western detection of AKT. Images of 40x of AKT immuno-reactivity of 4-OH-E2 treated wild type and Akt silenced MCF-10A cells were acquired by immunofluorescence confocal microscopy using Alexafluor 488. Anchorage independent growth, an indicator of neoplastic transformation of cells, was assessed in soft agar. Images were acquired by using an Olympus C-5060 digital camera attached to the Nikon TE2000U inverted microscope with a 4x objective. Colony efficiency was determined by a count of the number of colonies > 63 um in diameter and data expressed as mean of five wells +/− S.D.

    Journal: PLoS ONE

    Article Title: Reactive Oxygen Species via Redox Signaling to PI3K/AKT Pathway Contribute to the Malignant Growth of 4-Hydroxy Estradiol-Transformed Mammary Epithelial Cells

    doi: 10.1371/journal.pone.0054206

    Figure Lengend Snippet: 4-OH-E2-induced cell transformation is inhibited by AKT1 silencing. Inhibition of AKT expression by its silencing detected by Western Bloting (A) and confocal microscopy (B). (C) Detection of inhibition of 4-OH-E2-induced cell transformation by AKT1 silencing by anchorage-independent growth assay. The MCF-10A cells were transfected with pre-designed and verified human shRNA for AKT1 and control shRNA plasmid consisting of scrambled shRNA sequence that does not lead to the specific degradation of AKT1 (OriGene Technologies, Inc. Rockville, MD). These cells were exposed to a carcinogenic dose of 4-OH-E2 (10 ng/ml) as described in Fig. 2. The cellular extracts from treated and controls cells were separated on SDS-PAGE, transferred to the membrane, and followed by Western detection of AKT. Images of 40x of AKT immuno-reactivity of 4-OH-E2 treated wild type and Akt silenced MCF-10A cells were acquired by immunofluorescence confocal microscopy using Alexafluor 488. Anchorage independent growth, an indicator of neoplastic transformation of cells, was assessed in soft agar. Images were acquired by using an Olympus C-5060 digital camera attached to the Nikon TE2000U inverted microscope with a 4x objective. Colony efficiency was determined by a count of the number of colonies > 63 um in diameter and data expressed as mean of five wells +/− S.D.

    Article Snippet: The confocal fluorescence images were scanned on a Nikon TE2000U inverted fluorescence microscope equipped with a Nikon D-Eclipse C1 laser scanning confocal microscope system as described above (Nikon Corp., USA).

    Techniques: Transformation Assay, Inhibition, Expressing, Western Blot, Confocal Microscopy, Growth Assay, Transfection, shRNA, Plasmid Preparation, Sequencing, SDS Page, Immunofluorescence, Inverted Microscopy

    ROS modifiers inhibited estrogen-induced colony formation. For investigating inhibition of 4-OH-E2-induced cell transformation by ROS modifiers, MCF-10A cells were transfected with 100 MOI adenovirus expressing catalase or MnSOD or treated with an antioxidant Ebselen (40 uM). Cells overexpressing catalase or MnSOD or treated with Ebselen were exposed to a carcinogenic regimen of estrogen as described in the legend of figure 2 . Anchorage independent growth was assessed in soft agar after 21 days. Images were acquired by using an Olympus C-5060 digital camera attached to the Nikon TE2000U inverted microscope with a 4x objective (bottom panel shows representative pictures of colonies in soft agar in both 2A and 2B). Colony efficiency was determined by a count of the number of colonies > 63 um in diameter and data expressed as mean of five wells +/− S.D. *P

    Journal: PLoS ONE

    Article Title: Reactive Oxygen Species via Redox Signaling to PI3K/AKT Pathway Contribute to the Malignant Growth of 4-Hydroxy Estradiol-Transformed Mammary Epithelial Cells

    doi: 10.1371/journal.pone.0054206

    Figure Lengend Snippet: ROS modifiers inhibited estrogen-induced colony formation. For investigating inhibition of 4-OH-E2-induced cell transformation by ROS modifiers, MCF-10A cells were transfected with 100 MOI adenovirus expressing catalase or MnSOD or treated with an antioxidant Ebselen (40 uM). Cells overexpressing catalase or MnSOD or treated with Ebselen were exposed to a carcinogenic regimen of estrogen as described in the legend of figure 2 . Anchorage independent growth was assessed in soft agar after 21 days. Images were acquired by using an Olympus C-5060 digital camera attached to the Nikon TE2000U inverted microscope with a 4x objective (bottom panel shows representative pictures of colonies in soft agar in both 2A and 2B). Colony efficiency was determined by a count of the number of colonies > 63 um in diameter and data expressed as mean of five wells +/− S.D. *P

    Article Snippet: The confocal fluorescence images were scanned on a Nikon TE2000U inverted fluorescence microscope equipped with a Nikon D-Eclipse C1 laser scanning confocal microscope system as described above (Nikon Corp., USA).

    Techniques: Inhibition, Transformation Assay, Transfection, Expressing, Inverted Microscopy

    Exposure of MCF-10A cells to E2 and its hydroxy metabolites induced dose-dependent AIG positive colony formation (2A) and ROS modifiers inhibited estrogen-induced colony formation (2B). The cell transformation was carried out by a modified protocol of Russo's group (18). Briefly, MCF-10A cells were seeded at 30% density in a 10 cm dish. After 24 hrs of seeding, cells were exposed to E2 or its hydroxy metabolites. A treatment cycle includes a 48 hr starvation period, 48 hr treatment period (100 ng/ml of E2, 2-OH-E2, or 4-OH-E2), and 48 hr recovery period in DMEM-F12 media containing 10% horse serum (HS) and no growth supplements. Benzo(a)pyrene (BaP) was used as a positive control to show AIG positive colony formation. For inhibition of 4-OH-E2-induced cell transformation by ROS modifiers, MCF-10A cells were transfected with 50 MOI adenovirus expressing catalase or MnSOD or treated with an antioxidant Ebselen (40 uM). Cells overexpressing catalase or MnSOD or treated with Ebselen were exposed to a carcinogenic regimen of estrogen as described above. Anchorage independent growth, an indicator of neoplastic transformation of cells, was assessed in soft agar as previously described by Zhang et al (31) after 21 days. Images were acquired by using an Olympus C-5060 digital camera attached to the Nikon TE2000U inverted microscope with a 4x objective (bottom panel shows representative pictures of colonies in soft agar in both 2A and 2B). Colony efficiency was determined by a count of the number of colonies > 63 um in diameter and data expressed as mean of five wells +/− S.D.

    Journal: PLoS ONE

    Article Title: Reactive Oxygen Species via Redox Signaling to PI3K/AKT Pathway Contribute to the Malignant Growth of 4-Hydroxy Estradiol-Transformed Mammary Epithelial Cells

    doi: 10.1371/journal.pone.0054206

    Figure Lengend Snippet: Exposure of MCF-10A cells to E2 and its hydroxy metabolites induced dose-dependent AIG positive colony formation (2A) and ROS modifiers inhibited estrogen-induced colony formation (2B). The cell transformation was carried out by a modified protocol of Russo's group (18). Briefly, MCF-10A cells were seeded at 30% density in a 10 cm dish. After 24 hrs of seeding, cells were exposed to E2 or its hydroxy metabolites. A treatment cycle includes a 48 hr starvation period, 48 hr treatment period (100 ng/ml of E2, 2-OH-E2, or 4-OH-E2), and 48 hr recovery period in DMEM-F12 media containing 10% horse serum (HS) and no growth supplements. Benzo(a)pyrene (BaP) was used as a positive control to show AIG positive colony formation. For inhibition of 4-OH-E2-induced cell transformation by ROS modifiers, MCF-10A cells were transfected with 50 MOI adenovirus expressing catalase or MnSOD or treated with an antioxidant Ebselen (40 uM). Cells overexpressing catalase or MnSOD or treated with Ebselen were exposed to a carcinogenic regimen of estrogen as described above. Anchorage independent growth, an indicator of neoplastic transformation of cells, was assessed in soft agar as previously described by Zhang et al (31) after 21 days. Images were acquired by using an Olympus C-5060 digital camera attached to the Nikon TE2000U inverted microscope with a 4x objective (bottom panel shows representative pictures of colonies in soft agar in both 2A and 2B). Colony efficiency was determined by a count of the number of colonies > 63 um in diameter and data expressed as mean of five wells +/− S.D.

    Article Snippet: The confocal fluorescence images were scanned on a Nikon TE2000U inverted fluorescence microscope equipped with a Nikon D-Eclipse C1 laser scanning confocal microscope system as described above (Nikon Corp., USA).

    Techniques: Transformation Assay, Modification, Positive Control, Inhibition, Transfection, Expressing, Inverted Microscopy

    Phenotypic assessment of transformed cells. Clones from p21 cells were assessed for ductulogenesis (A), invasiveness (B) and seeding phenotypes (C). A ) Ductulogenic assay was conducted on the collagen matrix. B) Invasive characteristic of clone was compared to MDA-MB 231 cells using invasion assay in BD BioCoat™ Matrigel™ Invasion Chambers. Histogram of invaded cells expressed as mean invaded cells per field. C ) Phenotype of transformed clone (MCF-10AT) and wild type MCF-10A cells were captured through acquiring images of these cells after 3hrs and 24 hrs of seeding in a monlayer culture using an Olympus C-5060 digital camera attached to the Nikon TE2000U inverted microscope with 20x and 40x objectives.

    Journal: PLoS ONE

    Article Title: Reactive Oxygen Species via Redox Signaling to PI3K/AKT Pathway Contribute to the Malignant Growth of 4-Hydroxy Estradiol-Transformed Mammary Epithelial Cells

    doi: 10.1371/journal.pone.0054206

    Figure Lengend Snippet: Phenotypic assessment of transformed cells. Clones from p21 cells were assessed for ductulogenesis (A), invasiveness (B) and seeding phenotypes (C). A ) Ductulogenic assay was conducted on the collagen matrix. B) Invasive characteristic of clone was compared to MDA-MB 231 cells using invasion assay in BD BioCoat™ Matrigel™ Invasion Chambers. Histogram of invaded cells expressed as mean invaded cells per field. C ) Phenotype of transformed clone (MCF-10AT) and wild type MCF-10A cells were captured through acquiring images of these cells after 3hrs and 24 hrs of seeding in a monlayer culture using an Olympus C-5060 digital camera attached to the Nikon TE2000U inverted microscope with 20x and 40x objectives.

    Article Snippet: The confocal fluorescence images were scanned on a Nikon TE2000U inverted fluorescence microscope equipped with a Nikon D-Eclipse C1 laser scanning confocal microscope system as described above (Nikon Corp., USA).

    Techniques: Transformation Assay, Clone Assay, Multiple Displacement Amplification, Invasion Assay, Inverted Microscopy

    Unlike the attenuated BCG vaccine, virulent M. tuberculosis is able to induce phagosomal rupture in THP-1 macrophages. THP-1 cells were infected with BCG DsRed (A,C) or M. tuberculosis DsRed (B,D) at a MOI of 1 for the indicated time and then loaded with CCF-4 molecule for 2 h. After PFA fixation, cells were imaged using a fluorescence widefield microscope (Nikon Ti) with an 40X objective (A, B). Picture acquisition was achieved randomly and automatically for each condition on 49 fields in duplicates and further 450/535 nm intensity ratio measurements shown in panels C and D were obtained through analysis by a specialized algorithm using the Metamorph software. The plots are representative of 3 independent experiments.

    Journal: PLoS Pathogens

    Article Title: Phagosomal Rupture by Mycobacterium tuberculosis Results in Toxicity and Host Cell Death

    doi: 10.1371/journal.ppat.1002507

    Figure Lengend Snippet: Unlike the attenuated BCG vaccine, virulent M. tuberculosis is able to induce phagosomal rupture in THP-1 macrophages. THP-1 cells were infected with BCG DsRed (A,C) or M. tuberculosis DsRed (B,D) at a MOI of 1 for the indicated time and then loaded with CCF-4 molecule for 2 h. After PFA fixation, cells were imaged using a fluorescence widefield microscope (Nikon Ti) with an 40X objective (A, B). Picture acquisition was achieved randomly and automatically for each condition on 49 fields in duplicates and further 450/535 nm intensity ratio measurements shown in panels C and D were obtained through analysis by a specialized algorithm using the Metamorph software. The plots are representative of 3 independent experiments.

    Article Snippet: Samples were imaged using an automated inverted fluorescent widefield microscope Nikon Ti with 20X or 40X objective driven by Metamorph.

    Techniques: Infection, Fluorescence, Microscopy, Software

    Analysis of ESX-1 deletion-, truncation- and complementation-mutants highlights the link between phagosomal rupture and functional ESX-1 secretion. THP-1 cells were infected with M. tuberculosis ΔRD1 (A,D), BCG::RD1 (B,E) or BCG::RD1-ESAT-6Δ84–95 (C,F) at a MOI of 1 for the indicated time and then loaded with CCF-4 molecule for 2 h. After PFA fixation, cells were imaged using a fluorescence widefield microscope (Nikon Ti) equipped with a 40X objective (A,B,C). Picture acquisition was achieved randomly and automatically for each condition on 49 fields in duplicates and further 450/535 nm intensity ratio measurement (D,E,F) was obtained through analysis by a specialized algorithm using the Metamorph software. The plots are representative of 3 independent experiments.

    Journal: PLoS Pathogens

    Article Title: Phagosomal Rupture by Mycobacterium tuberculosis Results in Toxicity and Host Cell Death

    doi: 10.1371/journal.ppat.1002507

    Figure Lengend Snippet: Analysis of ESX-1 deletion-, truncation- and complementation-mutants highlights the link between phagosomal rupture and functional ESX-1 secretion. THP-1 cells were infected with M. tuberculosis ΔRD1 (A,D), BCG::RD1 (B,E) or BCG::RD1-ESAT-6Δ84–95 (C,F) at a MOI of 1 for the indicated time and then loaded with CCF-4 molecule for 2 h. After PFA fixation, cells were imaged using a fluorescence widefield microscope (Nikon Ti) equipped with a 40X objective (A,B,C). Picture acquisition was achieved randomly and automatically for each condition on 49 fields in duplicates and further 450/535 nm intensity ratio measurement (D,E,F) was obtained through analysis by a specialized algorithm using the Metamorph software. The plots are representative of 3 independent experiments.

    Article Snippet: Samples were imaged using an automated inverted fluorescent widefield microscope Nikon Ti with 20X or 40X objective driven by Metamorph.

    Techniques: Functional Assay, Infection, Fluorescence, Microscopy, Software

    Evolution of cell numbers during the time course of infection with different M. tuberculosis complex members. THP-1 cells were infected with M. tuberculosis PFA killed, BCG, BCG::RD1, M. tuberculosis or M. tuberculosis ΔRD1 at a MOI of 1 for the indicated time. Arrows indicate the decrease of the number of cells with virulent strains. After PFA fixation, cells were stained by incubation with WGA (wheat germ agglutinin) and imaged using a Nikon Ti widefield microscope. Pictures represent a mosaic of 49 individual images from 96 well plates acquired with a 20X objective (A). Cytofluorometry analysis of THP-1 cells infected with various wild-type and recombinant M. tuberculosis and BCG strains at day 7 post-infection. THP-1 cells were stained with FLICA Poly-Caspases SR-VAD-FMK (apoptosis) and Live/Dead Pacific Blue (necrosis) (B).

    Journal: PLoS Pathogens

    Article Title: Phagosomal Rupture by Mycobacterium tuberculosis Results in Toxicity and Host Cell Death

    doi: 10.1371/journal.ppat.1002507

    Figure Lengend Snippet: Evolution of cell numbers during the time course of infection with different M. tuberculosis complex members. THP-1 cells were infected with M. tuberculosis PFA killed, BCG, BCG::RD1, M. tuberculosis or M. tuberculosis ΔRD1 at a MOI of 1 for the indicated time. Arrows indicate the decrease of the number of cells with virulent strains. After PFA fixation, cells were stained by incubation with WGA (wheat germ agglutinin) and imaged using a Nikon Ti widefield microscope. Pictures represent a mosaic of 49 individual images from 96 well plates acquired with a 20X objective (A). Cytofluorometry analysis of THP-1 cells infected with various wild-type and recombinant M. tuberculosis and BCG strains at day 7 post-infection. THP-1 cells were stained with FLICA Poly-Caspases SR-VAD-FMK (apoptosis) and Live/Dead Pacific Blue (necrosis) (B).

    Article Snippet: Samples were imaged using an automated inverted fluorescent widefield microscope Nikon Ti with 20X or 40X objective driven by Metamorph.

    Techniques: Infection, Staining, Incubation, Whole Genome Amplification, Microscopy, Recombinant

    Necrosis induction does not lead to the release of mycobacteria from the phagolysosome to the cytosol. THP-1 cells were infected with BCG at a MOI of 1 for 2 days before necrosis induction using 100 µg/ml concanavalin A for 24 h or 48 h. After CCF-4 loading for 2 h, cells were incubated 5 minutes in the presence of 2 µg/ml propidium iodide (PI) and then subjected to PFA fixation. Cells were imaged using fluorescence widefield Nikon Ti microscope with 40X objective (A). Picture acquisition was achieved randomly and automatically for each condition on 49 fields and further 450/535 nm intensity ratio measurements (B,C) were obtained through analysis by a specialized algorithm using the Metamorph software. The plots were representative of 2 independent experiments.

    Journal: PLoS Pathogens

    Article Title: Phagosomal Rupture by Mycobacterium tuberculosis Results in Toxicity and Host Cell Death

    doi: 10.1371/journal.ppat.1002507

    Figure Lengend Snippet: Necrosis induction does not lead to the release of mycobacteria from the phagolysosome to the cytosol. THP-1 cells were infected with BCG at a MOI of 1 for 2 days before necrosis induction using 100 µg/ml concanavalin A for 24 h or 48 h. After CCF-4 loading for 2 h, cells were incubated 5 minutes in the presence of 2 µg/ml propidium iodide (PI) and then subjected to PFA fixation. Cells were imaged using fluorescence widefield Nikon Ti microscope with 40X objective (A). Picture acquisition was achieved randomly and automatically for each condition on 49 fields and further 450/535 nm intensity ratio measurements (B,C) were obtained through analysis by a specialized algorithm using the Metamorph software. The plots were representative of 2 independent experiments.

    Article Snippet: Samples were imaged using an automated inverted fluorescent widefield microscope Nikon Ti with 20X or 40X objective driven by Metamorph.

    Techniques: Infection, Incubation, Fluorescence, Microscopy, Software

    M. marinum is able to rupture the phagosome of THP-1 cells unlike an ESAT-6/CFP-10 secretion-deficient strain. THP-1 cells were infected with M. marinum expressing DsRed, M. smegmatis or M. marinum M VU at a MOI of 1 for the indicated time and then loaded with CCF-4 molecule for 2 h. After PFA fixation, cells were imaged by fluorescence widefield microscope Nikon Ti with 20X objective (A). Picture acquisition was done randomly and automatically for each condition on 36 fields to follow the FRET signals and the fluorescent bacteria. Data represented in panels B and C were obtained via specialized algorithms using the Metamorph software. The plots are representative of 3 independent experiments.

    Journal: PLoS Pathogens

    Article Title: Phagosomal Rupture by Mycobacterium tuberculosis Results in Toxicity and Host Cell Death

    doi: 10.1371/journal.ppat.1002507

    Figure Lengend Snippet: M. marinum is able to rupture the phagosome of THP-1 cells unlike an ESAT-6/CFP-10 secretion-deficient strain. THP-1 cells were infected with M. marinum expressing DsRed, M. smegmatis or M. marinum M VU at a MOI of 1 for the indicated time and then loaded with CCF-4 molecule for 2 h. After PFA fixation, cells were imaged by fluorescence widefield microscope Nikon Ti with 20X objective (A). Picture acquisition was done randomly and automatically for each condition on 36 fields to follow the FRET signals and the fluorescent bacteria. Data represented in panels B and C were obtained via specialized algorithms using the Metamorph software. The plots are representative of 3 independent experiments.

    Article Snippet: Samples were imaged using an automated inverted fluorescent widefield microscope Nikon Ti with 20X or 40X objective driven by Metamorph.

    Techniques: Infection, Expressing, Fluorescence, Microscopy, Software