Structured Review

Hitachi Ltd ultrathin sections
HS1 is involved in cytosol-to-chloroplast transport of HPT. Subcellular localization of HPT-GFP in the WT (A) and hs1-1 (B) backgrounds in Arabidopsis. HPT-GFP constructs were transformed transiently into plant cells. Arrowheads indicate higher chloroplast GFP fluorescence intensity in a mesophyll protoplast of WT compared to hs1-1 . Bars = 10 μm. (C , D) Western blot of HPT in WT/ HPT (empty vector transgenic lines), HS1 RNAi/ HPT , and hs1-1 / HPT (empty vector transformed into hs1-1 ) transgenic plants. HPT accumulation was reduced significantly or undetectable in HS1 RNAi/ HPT and hs1-1/HPT chloroplasts compared with the control (C) , while total HPT varied slightly (D) . The large subunit of RUBISCO (RBCL) was probed as a protein loading control. Approximately 20 μg (C) and 10 μg (D) of protein were loaded in each lane. Similar results were obtained in two additional independent experiments. (E–H) Transmission electron microscopic images of immunogold localization of HPT in transgenic Arabidopsis . HPT in <t>ultrathin</t> leaf sections reacted with anti-HPT antibody and a gold-conjugated secondary antibody. Gold particles were detected by transmission electron microscopy. Immunogold labeling of HPT in leaves from WT/ HPT (E) , hs1-1 / HPT (F) , hs1-1 / HPT / HS1 (G) , negative control (H) transgenic Arabidopsis plants, and negative control without anti-HPT antibody in blocking buffer showed no gold particles. Arrowheads show typical gold particles (10 nm, black dots). HPT present inside of chloroplasts (chl, the dark area) in WT / HPT , hs1-1 / HPT / HS1 , and outside of chloroplasts in hs1-1 / HPT . Three different leaf samples and more than 30 immunogold-labeled positive cells were observed with similar results. Images represent typical observations in different leaf samples. chl, chloroplast; cyt, cytosol; va, vacuole; cw, cell wall. Bars = 1 μm.
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1) Product Images from "HS1 Is Involved in Hygromycin Resistance Through Facilitating Hygromycin Phosphotransferase Transportation From Cytosol to Chloroplast"

Article Title: HS1 Is Involved in Hygromycin Resistance Through Facilitating Hygromycin Phosphotransferase Transportation From Cytosol to Chloroplast

Journal: Frontiers in Plant Science

doi: 10.3389/fpls.2020.00613

HS1 is involved in cytosol-to-chloroplast transport of HPT. Subcellular localization of HPT-GFP in the WT (A) and hs1-1 (B) backgrounds in Arabidopsis. HPT-GFP constructs were transformed transiently into plant cells. Arrowheads indicate higher chloroplast GFP fluorescence intensity in a mesophyll protoplast of WT compared to hs1-1 . Bars = 10 μm. (C , D) Western blot of HPT in WT/ HPT (empty vector transgenic lines), HS1 RNAi/ HPT , and hs1-1 / HPT (empty vector transformed into hs1-1 ) transgenic plants. HPT accumulation was reduced significantly or undetectable in HS1 RNAi/ HPT and hs1-1/HPT chloroplasts compared with the control (C) , while total HPT varied slightly (D) . The large subunit of RUBISCO (RBCL) was probed as a protein loading control. Approximately 20 μg (C) and 10 μg (D) of protein were loaded in each lane. Similar results were obtained in two additional independent experiments. (E–H) Transmission electron microscopic images of immunogold localization of HPT in transgenic Arabidopsis . HPT in ultrathin leaf sections reacted with anti-HPT antibody and a gold-conjugated secondary antibody. Gold particles were detected by transmission electron microscopy. Immunogold labeling of HPT in leaves from WT/ HPT (E) , hs1-1 / HPT (F) , hs1-1 / HPT / HS1 (G) , negative control (H) transgenic Arabidopsis plants, and negative control without anti-HPT antibody in blocking buffer showed no gold particles. Arrowheads show typical gold particles (10 nm, black dots). HPT present inside of chloroplasts (chl, the dark area) in WT / HPT , hs1-1 / HPT / HS1 , and outside of chloroplasts in hs1-1 / HPT . Three different leaf samples and more than 30 immunogold-labeled positive cells were observed with similar results. Images represent typical observations in different leaf samples. chl, chloroplast; cyt, cytosol; va, vacuole; cw, cell wall. Bars = 1 μm.
Figure Legend Snippet: HS1 is involved in cytosol-to-chloroplast transport of HPT. Subcellular localization of HPT-GFP in the WT (A) and hs1-1 (B) backgrounds in Arabidopsis. HPT-GFP constructs were transformed transiently into plant cells. Arrowheads indicate higher chloroplast GFP fluorescence intensity in a mesophyll protoplast of WT compared to hs1-1 . Bars = 10 μm. (C , D) Western blot of HPT in WT/ HPT (empty vector transgenic lines), HS1 RNAi/ HPT , and hs1-1 / HPT (empty vector transformed into hs1-1 ) transgenic plants. HPT accumulation was reduced significantly or undetectable in HS1 RNAi/ HPT and hs1-1/HPT chloroplasts compared with the control (C) , while total HPT varied slightly (D) . The large subunit of RUBISCO (RBCL) was probed as a protein loading control. Approximately 20 μg (C) and 10 μg (D) of protein were loaded in each lane. Similar results were obtained in two additional independent experiments. (E–H) Transmission electron microscopic images of immunogold localization of HPT in transgenic Arabidopsis . HPT in ultrathin leaf sections reacted with anti-HPT antibody and a gold-conjugated secondary antibody. Gold particles were detected by transmission electron microscopy. Immunogold labeling of HPT in leaves from WT/ HPT (E) , hs1-1 / HPT (F) , hs1-1 / HPT / HS1 (G) , negative control (H) transgenic Arabidopsis plants, and negative control without anti-HPT antibody in blocking buffer showed no gold particles. Arrowheads show typical gold particles (10 nm, black dots). HPT present inside of chloroplasts (chl, the dark area) in WT / HPT , hs1-1 / HPT / HS1 , and outside of chloroplasts in hs1-1 / HPT . Three different leaf samples and more than 30 immunogold-labeled positive cells were observed with similar results. Images represent typical observations in different leaf samples. chl, chloroplast; cyt, cytosol; va, vacuole; cw, cell wall. Bars = 1 μm.

Techniques Used: Construct, Transformation Assay, Fluorescence, Western Blot, Plasmid Preparation, Transgenic Assay, Transmission Assay, Electron Microscopy, Labeling, Negative Control, Blocking Assay

2) Product Images from "Vaccinia Virus Mutations in the L4R Gene Encoding a Virion Structural Protein Produce Abnormal Mature Particles Lacking a Nucleocapsid"

Article Title: Vaccinia Virus Mutations in the L4R Gene Encoding a Virion Structural Protein Produce Abnormal Mature Particles Lacking a Nucleocapsid

Journal: Journal of Virology

doi: 10.1128/JVI.02126-14

Immunogold labeling of core proteins. Cells were infected with WR or Ets85 at an MOI of 10 and incubated at 31°C or 39.7°C. At 24 h postinfection, cells were processed for immunoelectron microscopy. Ultrathin sections were probed with
Figure Legend Snippet: Immunogold labeling of core proteins. Cells were infected with WR or Ets85 at an MOI of 10 and incubated at 31°C or 39.7°C. At 24 h postinfection, cells were processed for immunoelectron microscopy. Ultrathin sections were probed with

Techniques Used: Labeling, Infection, Incubation, Immuno-Electron Microscopy

3) Product Images from "Three-dimensional reconstruction of electron micrographs reveals intrabulbar circuit differences between accessory and main olfactory bulbs"

Article Title: Three-dimensional reconstruction of electron micrographs reveals intrabulbar circuit differences between accessory and main olfactory bulbs

Journal: Frontiers in Neuroanatomy

doi: 10.3389/fnana.2013.00005

A 3D image of an AOB Gr cell. (A) A drawing of a prepared rat AOB slice (upper panel) and the position of a biocytin-injected Gr cell (red cross, upper panel), and a joint image of differential interference and fluorescence of the slice reacted with avidin-FITC-gold (lower panel). Scale bar = 200 μm. (B) A confocal fluorescent image of the injected cell. The red dotted line indicates the border between the MTL, the olfactory tract layer (OTL), and the GRL. The red box indicates the area observed with electron microscopy corresponding to the 3D image of C . Scale bar = 50 μm. (C) 3D reconstruction of the apical dendrite in B by serial electron micrograph. The green is the injected dendrite, and the yellow structures indicate the neuron's dendritic spines. The white structures are filopodia-like structures. Arrows indicate synaptic transmission direction. iSSs (mainly excitatory synapses) are labeled in red, oSSs (all inhibitory synapses) are labeled in blue, and bidirectional arrows indicate RSs. Orange boxes indicate the areas shown in D,E . Scale bar = 10 μm. (D and E) An example of 3D morphology of the spine containing the iSS (iSS3 in C ) and the gemmule containing the RS (RS10 in C ). The 3D image (left panels) was reconstructed with the serial electron micrograph of a labeled spine by silver-enhances gold particles (right panels). Numbers on the right panels indicate ultrathin section sequence. The asterisk indicates the same section of the low-magnification panel (upper). Arrows indicate synaptic transmission direction. Red and blue areas indicate the locations and sizes of synaptic sites (red, PSD of the labeled cell; blue, PSD of a paired MT cell). Ax: axon-like structure. White scale bar = 500 nm (in D ) and 1 μm (in E ). Black scale bar = 500 nm (in D ), 1 μm (low-magnification photo in E ), and 500 nm (high-magnification photos in E ).
Figure Legend Snippet: A 3D image of an AOB Gr cell. (A) A drawing of a prepared rat AOB slice (upper panel) and the position of a biocytin-injected Gr cell (red cross, upper panel), and a joint image of differential interference and fluorescence of the slice reacted with avidin-FITC-gold (lower panel). Scale bar = 200 μm. (B) A confocal fluorescent image of the injected cell. The red dotted line indicates the border between the MTL, the olfactory tract layer (OTL), and the GRL. The red box indicates the area observed with electron microscopy corresponding to the 3D image of C . Scale bar = 50 μm. (C) 3D reconstruction of the apical dendrite in B by serial electron micrograph. The green is the injected dendrite, and the yellow structures indicate the neuron's dendritic spines. The white structures are filopodia-like structures. Arrows indicate synaptic transmission direction. iSSs (mainly excitatory synapses) are labeled in red, oSSs (all inhibitory synapses) are labeled in blue, and bidirectional arrows indicate RSs. Orange boxes indicate the areas shown in D,E . Scale bar = 10 μm. (D and E) An example of 3D morphology of the spine containing the iSS (iSS3 in C ) and the gemmule containing the RS (RS10 in C ). The 3D image (left panels) was reconstructed with the serial electron micrograph of a labeled spine by silver-enhances gold particles (right panels). Numbers on the right panels indicate ultrathin section sequence. The asterisk indicates the same section of the low-magnification panel (upper). Arrows indicate synaptic transmission direction. Red and blue areas indicate the locations and sizes of synaptic sites (red, PSD of the labeled cell; blue, PSD of a paired MT cell). Ax: axon-like structure. White scale bar = 500 nm (in D ) and 1 μm (in E ). Black scale bar = 500 nm (in D ), 1 μm (low-magnification photo in E ), and 500 nm (high-magnification photos in E ).

Techniques Used: Injection, Fluorescence, Avidin-Biotin Assay, Electron Microscopy, Transmission Assay, Labeling, Sequencing

4) Product Images from "Mode of Action and Functional Significance of Estrogen-Inducing Dendritic Growth, Spinogenesis, and Synaptogenesis in the Developing Purkinje Cell"

Article Title: Mode of Action and Functional Significance of Estrogen-Inducing Dendritic Growth, Spinogenesis, and Synaptogenesis in the Developing Purkinje Cell

Journal: The Journal of Neuroscience

doi: 10.1523/JNEUROSCI.0710-07.2007

A , B , Higher magnification of synaptic terminals in the molecular layers of vermal cerebella (lobe IX). Arrows indicate postsynaptic density (psd), synaptic vesicle (sv), synaptic cleft (sc), and presynaptic dense projections (pdp). PD, Purkinje cell dendrite. Scale bars, 500 nm. C , D , Quantitative electron microscopic analysis of Purkinje dendritic synapse density of newborn WT mice treated with vehicle, EB, tamoxifen, or EB plus tamoxifen. C , Axospinous synapse density. D , Shaft synapse density. Ultrathin sections (60 nm in thickness) containing calbindin-immunoreactive Purkinje dendrites in lobe IX at 6 d of age were analyzed. Each column and error bar represent the mean ± SEM ( n = 5 in each group). * p
Figure Legend Snippet: A , B , Higher magnification of synaptic terminals in the molecular layers of vermal cerebella (lobe IX). Arrows indicate postsynaptic density (psd), synaptic vesicle (sv), synaptic cleft (sc), and presynaptic dense projections (pdp). PD, Purkinje cell dendrite. Scale bars, 500 nm. C , D , Quantitative electron microscopic analysis of Purkinje dendritic synapse density of newborn WT mice treated with vehicle, EB, tamoxifen, or EB plus tamoxifen. C , Axospinous synapse density. D , Shaft synapse density. Ultrathin sections (60 nm in thickness) containing calbindin-immunoreactive Purkinje dendrites in lobe IX at 6 d of age were analyzed. Each column and error bar represent the mean ± SEM ( n = 5 in each group). * p

Techniques Used: Mouse Assay

Quantitative electron microscopic analysis of Purkinje dendritic synapse density of WT, ArKO, and EB-treated ArKO (ArKO + Estradiol benzoate) newborn mice. A , Axospinous synapse density. B , Shaft synapse density. Ultrathin sections (60 nm in thickness) containing calbindin-immunoreactive Purkinje dendrites in lobe IX at 6 d of age were analyzed. Each column and error bar represent the mean ± SEM ( n = 5 in each group). ** p
Figure Legend Snippet: Quantitative electron microscopic analysis of Purkinje dendritic synapse density of WT, ArKO, and EB-treated ArKO (ArKO + Estradiol benzoate) newborn mice. A , Axospinous synapse density. B , Shaft synapse density. Ultrathin sections (60 nm in thickness) containing calbindin-immunoreactive Purkinje dendrites in lobe IX at 6 d of age were analyzed. Each column and error bar represent the mean ± SEM ( n = 5 in each group). ** p

Techniques Used: Mouse Assay

5) Product Images from "Co-transmission of acetylcholine and GABA regulates hippocampal states"

Article Title: Co-transmission of acetylcholine and GABA regulates hippocampal states

Journal: Nature Communications

doi: 10.1038/s41467-018-05136-1

Cholinergic cells express the molecular machinery required for GABA release. a , b The cholinergic neurons of the MS are GABAergic. White box in a contains area enlarged in b . Images show neurons stained for ChAT in red, while the green labelling marks the vGAT-expressing neurons in vGAT-ZsGreen reporter mouse. c , d AAV-eYFP virus-traced septo-hippocampal fibres express GAD65 ( c ). AAV-eYFP virus-traced septo-hippocampal fibres express vGAT and vAChT ( d ). Insets show xz and yz projections of the terminal labelled with an arrow. Arrowhead points to another terminal. Green line marks the fibre outline. (Scale bar on d is 210, 14, 2 and 1 μm for a , b , c and d , respectively.) e , f Hippocampal cholinergic terminals contain GABA. Three consecutive EM sections of a vAChT-positive terminal ( e , red pseudocolor) are shown. vAChT was visualised by pre-embedding immunogold method (the first panel of e , silver-intensified gold particles, large arrows), whereas on the next ultrathin sections (the second and third panels of e ) postembedding GABA immunostaining was performed (smaller gold particles, thin arrows, some GABA molecules penetrate into mitochondria during fixation). vAChT signal is absent in postembedding images, because of the etching procedure. Scale bar is 200 nm for all EM images. f Cholinergic terminals contained significantly higher immunogold signal than glutamatergic ones ( p
Figure Legend Snippet: Cholinergic cells express the molecular machinery required for GABA release. a , b The cholinergic neurons of the MS are GABAergic. White box in a contains area enlarged in b . Images show neurons stained for ChAT in red, while the green labelling marks the vGAT-expressing neurons in vGAT-ZsGreen reporter mouse. c , d AAV-eYFP virus-traced septo-hippocampal fibres express GAD65 ( c ). AAV-eYFP virus-traced septo-hippocampal fibres express vGAT and vAChT ( d ). Insets show xz and yz projections of the terminal labelled with an arrow. Arrowhead points to another terminal. Green line marks the fibre outline. (Scale bar on d is 210, 14, 2 and 1 μm for a , b , c and d , respectively.) e , f Hippocampal cholinergic terminals contain GABA. Three consecutive EM sections of a vAChT-positive terminal ( e , red pseudocolor) are shown. vAChT was visualised by pre-embedding immunogold method (the first panel of e , silver-intensified gold particles, large arrows), whereas on the next ultrathin sections (the second and third panels of e ) postembedding GABA immunostaining was performed (smaller gold particles, thin arrows, some GABA molecules penetrate into mitochondria during fixation). vAChT signal is absent in postembedding images, because of the etching procedure. Scale bar is 200 nm for all EM images. f Cholinergic terminals contained significantly higher immunogold signal than glutamatergic ones ( p

Techniques Used: Mass Spectrometry, Staining, Expressing, Immunostaining

6) Product Images from "Removal of Abnormal Myofilament O-GlcNAcylation Restores Ca2+ Sensitivity in Diabetic Cardiac Muscle"

Article Title: Removal of Abnormal Myofilament O-GlcNAcylation Restores Ca2+ Sensitivity in Diabetic Cardiac Muscle

Journal: Diabetes

doi: 10.2337/db14-1107

Differential OGT and OGA subcellular localization and myofilament interactions in control and diabetic myocardium. Representative transmission electron microscopy images of control ( A ) and STZ diabetic rat myocardium ( B ). Ultrathin sections were examined with immunoelectron microscopy with primary antibodies (anti-OGT AL-28 and anti-OGA), and secondary antibodies gold-labeled (anti-rabbit, 12 nm; anti-chicken, 6 nm). Z-line, green arrowhead; OGT, purple circles; OGA, red circles; OGT and OGA in close vicinity, pink circles; OGT, purple arrowhead; and OGA, red arrowhead. Quantification of OGT ( C ) and OGA ( D ) number of particles/field in nine fields shows an increase for OGT (2 ± 0.6 vs. 8.6 ± 2.6, * P ≤ 0.028) and OGA (9.4 ± 2.9 vs 37.6 ± 12.6, * P ≤ 0.05) immunoelectron microscopy in STZ diabetic myocardium. Images were analyzed in ImageJ software (NIH). Representative coimmunoprecipitations (co-IP) of OGT ( E ) and OGA ( F ), followed by Western blots (WB) for α-sarcomeric actin, Tm, and MLC 1. A fraction of the inputs from controls (C1, C2), STZ (S1, S2), and agarose beads with no antibody (M) or isotype-specific normal antibody + agarose beads (1°) were included. G : Analysis of integrated signal density of myofilament immunoreactivity normalized to total IP OGT displayed a trend toward increased interactions with Tm and MLC 1. H : OGA co-IP shows that diabetic STZ rats display several fold increased associations with α-actin, α-Tm, and MLC 1. * P ≤ 0.05.
Figure Legend Snippet: Differential OGT and OGA subcellular localization and myofilament interactions in control and diabetic myocardium. Representative transmission electron microscopy images of control ( A ) and STZ diabetic rat myocardium ( B ). Ultrathin sections were examined with immunoelectron microscopy with primary antibodies (anti-OGT AL-28 and anti-OGA), and secondary antibodies gold-labeled (anti-rabbit, 12 nm; anti-chicken, 6 nm). Z-line, green arrowhead; OGT, purple circles; OGA, red circles; OGT and OGA in close vicinity, pink circles; OGT, purple arrowhead; and OGA, red arrowhead. Quantification of OGT ( C ) and OGA ( D ) number of particles/field in nine fields shows an increase for OGT (2 ± 0.6 vs. 8.6 ± 2.6, * P ≤ 0.028) and OGA (9.4 ± 2.9 vs 37.6 ± 12.6, * P ≤ 0.05) immunoelectron microscopy in STZ diabetic myocardium. Images were analyzed in ImageJ software (NIH). Representative coimmunoprecipitations (co-IP) of OGT ( E ) and OGA ( F ), followed by Western blots (WB) for α-sarcomeric actin, Tm, and MLC 1. A fraction of the inputs from controls (C1, C2), STZ (S1, S2), and agarose beads with no antibody (M) or isotype-specific normal antibody + agarose beads (1°) were included. G : Analysis of integrated signal density of myofilament immunoreactivity normalized to total IP OGT displayed a trend toward increased interactions with Tm and MLC 1. H : OGA co-IP shows that diabetic STZ rats display several fold increased associations with α-actin, α-Tm, and MLC 1. * P ≤ 0.05.

Techniques Used: Transmission Assay, Electron Microscopy, Immuno-Electron Microscopy, Labeling, Software, Co-Immunoprecipitation Assay, Western Blot

7) Product Images from "Derlin-1 Regulates Mutant VCP-Linked Pathogenesis and Endoplasmic Reticulum Stress-Induced Apoptosis"

Article Title: Derlin-1 Regulates Mutant VCP-Linked Pathogenesis and Endoplasmic Reticulum Stress-Induced Apoptosis

Journal: PLoS Genetics

doi: 10.1371/journal.pgen.1004675

Derlin-1 overexpression impairs ER homeostasis and produces mitochondrial abnormality. (A and B) Confocal images of larval eye discs expressing CD3δ-YFP (A) and Xbp1-eGFP (B) probes (green), stained with anti-Elav antibodies (red) to label neuronal nuclei. The genotypes of eye discs include GMR > LacZ (control), GMR > derlin-1 ( derlin-1 overexpression), GMR > derlin-1 > TER94 WT (overexpression of both derlin-1 and TER94 ), and GMR > derlin-1 > TER94 A229E (overexpression of derlin-1 and TER94 A229E ). (C–F) TEM micrographs of 18-day-old Rh1 > LacZ control (C) and Rh1 > derlin-1 eyes (D–F). Unlike Rh1 > LacZ (C), Derlin-1-overexpressing photoreceptors (D) contain an elevated level of ER-resembling tubular membranes. (E) Another TEM section shows that Rh1 > derlin-1 outer photoreceptors contain excessive ER membrane, as well as abnormal mitochondria with intracristal swelling (white arrowheads) and discontinuous membrane (red arrowhead). Inset shows higher magnification of mitochondrion pointed by red arrowhead. (F) A representative TEM micrograph shows that Derlin-1-overexpressing photoreceptors contain smaller mitochondria (white arrowheads). As Rh1 promoter is active only in the outer photoreceptors, the mitochondria (arrows) in the inner photoreceptor (outlined in yellow) serve as an internal control. Scale bars: 10 µm (confocal). 1 µm (TEM). (G and H) Scatter dot plots of individual mitochondria in photoreceptor cells from four ultrathin sections. Mitochondrial size in outer photoreceptor cells of Rh1 > LacZ and Rh1 > derlin-1 (G, n = 50 mitochondria per genotype), and in both inner and outer photoreceptor cells of Rh1 > derlin-1 (H, n = 21 and 37 mitochondria for inner and outer cells, respectively) were manually outlined to measure the size by ImageJ. Magenta bar and blue line represent mean ± SE in each group. *** p
Figure Legend Snippet: Derlin-1 overexpression impairs ER homeostasis and produces mitochondrial abnormality. (A and B) Confocal images of larval eye discs expressing CD3δ-YFP (A) and Xbp1-eGFP (B) probes (green), stained with anti-Elav antibodies (red) to label neuronal nuclei. The genotypes of eye discs include GMR > LacZ (control), GMR > derlin-1 ( derlin-1 overexpression), GMR > derlin-1 > TER94 WT (overexpression of both derlin-1 and TER94 ), and GMR > derlin-1 > TER94 A229E (overexpression of derlin-1 and TER94 A229E ). (C–F) TEM micrographs of 18-day-old Rh1 > LacZ control (C) and Rh1 > derlin-1 eyes (D–F). Unlike Rh1 > LacZ (C), Derlin-1-overexpressing photoreceptors (D) contain an elevated level of ER-resembling tubular membranes. (E) Another TEM section shows that Rh1 > derlin-1 outer photoreceptors contain excessive ER membrane, as well as abnormal mitochondria with intracristal swelling (white arrowheads) and discontinuous membrane (red arrowhead). Inset shows higher magnification of mitochondrion pointed by red arrowhead. (F) A representative TEM micrograph shows that Derlin-1-overexpressing photoreceptors contain smaller mitochondria (white arrowheads). As Rh1 promoter is active only in the outer photoreceptors, the mitochondria (arrows) in the inner photoreceptor (outlined in yellow) serve as an internal control. Scale bars: 10 µm (confocal). 1 µm (TEM). (G and H) Scatter dot plots of individual mitochondria in photoreceptor cells from four ultrathin sections. Mitochondrial size in outer photoreceptor cells of Rh1 > LacZ and Rh1 > derlin-1 (G, n = 50 mitochondria per genotype), and in both inner and outer photoreceptor cells of Rh1 > derlin-1 (H, n = 21 and 37 mitochondria for inner and outer cells, respectively) were manually outlined to measure the size by ImageJ. Magenta bar and blue line represent mean ± SE in each group. *** p

Techniques Used: Over Expression, Expressing, Staining, Transmission Electron Microscopy

8) Product Images from "Activated N"

Article Title: Activated N

Journal: Molecular Biology of the Cell

doi: 10.1091/mbc.E05-11-1073

Ultrathin sections examined by TEM show that FPR and FcεRI can occupy the same preendocytic structures within 2-min addition of both ligands (FITC-fMLF and DNP-gold). After stimulus, cells in B–G were fixed with paraformaldehyde and incubated with anti-FITC gold to label the FPR. Cells in A were prefixed and then incubated with ligand–gold reagents to establish that FPR (5-nm gold) and FcεRI (10-nm gold) are not colocalized under resting conditions. Images in B and C show the presence of both sizes of gold (arrows) in deeply cupped membrane invaginations. Bold arrow in C shows large gold marking FcεRI in an endosome. Small gold particles in D, F, and G (arrows) mark FPR–ligand–gold probe complexes within lightly coated invaginations on the plasma membrane. Bold arrows in C, E, F, and G point to internalized FcεRI. (H) Clathrin was immunoprecipitated from RBL FPR-GFP cells treated for specified times with 1 μg/ml DNP-BSA to cross-link IgE-primed FcεRI (top) or with 10 nM fMLF (bottom). Proteins in the immunoprecipitates were separated by SDS-PAGE and electrophoretically transferred to nitrocellulose. Blots were probed with anti-phosphotyrosine antibodies. Gels were routinely stripped and reprobed with anti-clathrin as a loading control (data not shown). Bar (left), 100 nm.
Figure Legend Snippet: Ultrathin sections examined by TEM show that FPR and FcεRI can occupy the same preendocytic structures within 2-min addition of both ligands (FITC-fMLF and DNP-gold). After stimulus, cells in B–G were fixed with paraformaldehyde and incubated with anti-FITC gold to label the FPR. Cells in A were prefixed and then incubated with ligand–gold reagents to establish that FPR (5-nm gold) and FcεRI (10-nm gold) are not colocalized under resting conditions. Images in B and C show the presence of both sizes of gold (arrows) in deeply cupped membrane invaginations. Bold arrow in C shows large gold marking FcεRI in an endosome. Small gold particles in D, F, and G (arrows) mark FPR–ligand–gold probe complexes within lightly coated invaginations on the plasma membrane. Bold arrows in C, E, F, and G point to internalized FcεRI. (H) Clathrin was immunoprecipitated from RBL FPR-GFP cells treated for specified times with 1 μg/ml DNP-BSA to cross-link IgE-primed FcεRI (top) or with 10 nM fMLF (bottom). Proteins in the immunoprecipitates were separated by SDS-PAGE and electrophoretically transferred to nitrocellulose. Blots were probed with anti-phosphotyrosine antibodies. Gels were routinely stripped and reprobed with anti-clathrin as a loading control (data not shown). Bar (left), 100 nm.

Techniques Used: Transmission Electron Microscopy, Incubation, Immunoprecipitation, SDS Page

9) Product Images from "Improved Serial Sectioning Techniques for Correlative Light-Electron Microscopy Mapping of Human Langerhans Islets"

Article Title: Improved Serial Sectioning Techniques for Correlative Light-Electron Microscopy Mapping of Human Langerhans Islets

Journal: Acta Histochemica et Cytochemica

doi: 10.1267/ahc.17020

Correlative light-electron microscopy mapping of a diffuse islet using serial Epon sections from a human pancreatic tissue specimen. Ultrastructural observation and double-immunofluorescence staining for insulin, glucagon, and REG1α were carried out. Granular immunoreactivities for insulin ( A , C , red) and glucagon ( B , C , green) are sparse and dense respectively, while the immunostaining patterns of insulin ( D , F , red) and REG1α ( E , F , green) largely overlap. G: A light microscopy image of the same thick section stained with toluidine blue (TB) after microscopy observation of fluorescence immunostaining ( A–C ). The diffuse islet is composed of a mass of endocrine cells in contact with adjacent exocrine acinar-like cell clusters without a clear capsule (red arrowheads). H: Superimposed image of ( C ) and ( G ). I: An electron microscopy image of the black-boxed area shown in ( G ) obtained from a serial ultrathin section demonstrating the ultrastructural features of insulin- and glucagon-positive endocrine cells in a diffuse pancreatic islet. The diffuse islet is composed of a mass of endocrine cells interspersed between adjacent exocrine acinar-like cell clusters without a clear capsule (red arrowheads). J: An electron microscopy montage showing a pancreatic islet corresponding to the rectangle of TB staining shown in ( G ). Endoplasmic reticulum and Golgi apparatus are indicated by blue and red arrowheads, respectively. K: A higher magnification image of the respective boxed area in ( J ) illustrating an α cell containing α-cell granules (red arrowheads) in contact with a β cell containing β-cell granules (blue arrowheads) and condensing small vacuoles (cyan arrowheads). L: A higher magnification image of the respective boxed area in ( J ) illustrating a β cell with β-cell granules (blue arrowheads) and spherical and smaller granules with small halo (green arrowheads) in contact with an α cell containing α-cell granules (red arrowheads). M: A higher magnification image of the respective boxed area in ( J ) illustrating a β cell with β-cell granules (blue arrowheads) and a zymogen-like granule (yellow arrow) in cell-to-cell contact with an exocrine acinar-like cell containing zymogen-like granules. N: Interdigitation of cell membranes (white arrows) containing β-cell granules (blue arrowheads) and 200–500-nm condensing vacuoles (cyan arrowheads) between two β cells. Bars = 50 μm ( A, D, G ); 5 μm ( I ); 2 μm ( J ); 500 nm ( K, N ); 1 μm ( L, M ).
Figure Legend Snippet: Correlative light-electron microscopy mapping of a diffuse islet using serial Epon sections from a human pancreatic tissue specimen. Ultrastructural observation and double-immunofluorescence staining for insulin, glucagon, and REG1α were carried out. Granular immunoreactivities for insulin ( A , C , red) and glucagon ( B , C , green) are sparse and dense respectively, while the immunostaining patterns of insulin ( D , F , red) and REG1α ( E , F , green) largely overlap. G: A light microscopy image of the same thick section stained with toluidine blue (TB) after microscopy observation of fluorescence immunostaining ( A–C ). The diffuse islet is composed of a mass of endocrine cells in contact with adjacent exocrine acinar-like cell clusters without a clear capsule (red arrowheads). H: Superimposed image of ( C ) and ( G ). I: An electron microscopy image of the black-boxed area shown in ( G ) obtained from a serial ultrathin section demonstrating the ultrastructural features of insulin- and glucagon-positive endocrine cells in a diffuse pancreatic islet. The diffuse islet is composed of a mass of endocrine cells interspersed between adjacent exocrine acinar-like cell clusters without a clear capsule (red arrowheads). J: An electron microscopy montage showing a pancreatic islet corresponding to the rectangle of TB staining shown in ( G ). Endoplasmic reticulum and Golgi apparatus are indicated by blue and red arrowheads, respectively. K: A higher magnification image of the respective boxed area in ( J ) illustrating an α cell containing α-cell granules (red arrowheads) in contact with a β cell containing β-cell granules (blue arrowheads) and condensing small vacuoles (cyan arrowheads). L: A higher magnification image of the respective boxed area in ( J ) illustrating a β cell with β-cell granules (blue arrowheads) and spherical and smaller granules with small halo (green arrowheads) in contact with an α cell containing α-cell granules (red arrowheads). M: A higher magnification image of the respective boxed area in ( J ) illustrating a β cell with β-cell granules (blue arrowheads) and a zymogen-like granule (yellow arrow) in cell-to-cell contact with an exocrine acinar-like cell containing zymogen-like granules. N: Interdigitation of cell membranes (white arrows) containing β-cell granules (blue arrowheads) and 200–500-nm condensing vacuoles (cyan arrowheads) between two β cells. Bars = 50 μm ( A, D, G ); 5 μm ( I ); 2 μm ( J ); 500 nm ( K, N ); 1 μm ( L, M ).

Techniques Used: Electron Microscopy, Double Immunofluorescence Staining, Immunostaining, Light Microscopy, Staining, Microscopy, Fluorescence

Schematic representation of serial sectioning techniques for correlative light-electron microscopy mapping of human Langerhans islets. A: Schematic images illustrating an ultrathin section of a Langerhans islet mounted on a Φ1 × 2 mm single slit copper grid with a formvar film covered by evaporated carbon. B: Flow chart of pre-treatments and immunohistochemical staining procedures applied to serial thick sections after ultrathin sectioning of Epon blocks.
Figure Legend Snippet: Schematic representation of serial sectioning techniques for correlative light-electron microscopy mapping of human Langerhans islets. A: Schematic images illustrating an ultrathin section of a Langerhans islet mounted on a Φ1 × 2 mm single slit copper grid with a formvar film covered by evaporated carbon. B: Flow chart of pre-treatments and immunohistochemical staining procedures applied to serial thick sections after ultrathin sectioning of Epon blocks.

Techniques Used: Electron Microscopy, Flow Cytometry, Immunohistochemistry, Staining

10) Product Images from "Microglia control the spread of neurotropic virus infection via P2Y12 signalling and recruit monocytes through P2Y12-independent mechanisms"

Article Title: Microglia control the spread of neurotropic virus infection via P2Y12 signalling and recruit monocytes through P2Y12-independent mechanisms

Journal: Acta Neuropathologica

doi: 10.1007/s00401-018-1885-0

Microglia rapidly isolate virus-infected neurons in the brain. a Microglia (Iba1, yellow) are recruited to neurons infected with PRV expressing GFP with immediate-early kinetics (BDG) in the hypothalamic paraventricular nucleus (PVN). Note that microglia recruitment starts after the expression of the immediate-early marker GFP (phase I), when low levels of viral structural proteins become detectable (phase II). High levels of viral structural proteins indicate a late stage of viral infection (phase III), which is associated with higher number of microglia recruited to infected cells. b Microglial numbers increase significantly around virus-infected neurons in parallel with the propagation of infection c In vivo two-photon imaging reveals the recruitment of microglia (green) to virus-infected neurons (red) in real-time. Retrograde transsynaptic infection was induced by the virus BDR in Cx3Cr1 +/GFP microglia reporter mice 7 days prior to imaging to visualize infected cells in the cerebral cortex based on the immediate-early DSRed expression. d Merged Z -planes of microglial cells around a PRV-DSRed-positive neuron (arrows indicate recruited microglia, arrowheads indicate microglial processes contacting the infected cell). e Microglial process velocity increases significantly in response to viral infection compared to that seen in control mice. f – i Correlated CLSM, electron microscopy and electron tomography confirms direct microglia–neuron contact with intercellular molecular links in early phase of viral infection. f , 3D-reconstruction from deconvolved confocal stack of a recruited microglia (green) engulfing a PRV-positive neuronal cell body (cyan) with CD68-positive phagolysosomes (magenta) within the microglia, arranged around infected neuron. Upper insert: single image plane of the same confocal stack. Middle insert: 3D-reconstruction of the same microglial cell, rotated 180° around the vertical axis. Asterisk labels the bay engulfing the soma of the infected neuron. Lower insert: 3D-reconstruction of the infected neuronal cell body with surrounding phagolysosomes located within the microglia. g Transmission electron micrograph shows the same cells on an ultrathin section matching the confocal image plane in the upper insert in f . Note that microglial processes surround the apical dendrites of the infected neuron. h Part of g (in white box) enlarged. Note that the nucleus is void of viral capsids and the membrane of the neuron is intact (white arrowheads), confirming the early phase of infection. i 3 nm thick electron tomographic section and corresponding 3D-reconstruction shows the very close cell–cell contact between the same microglia (green pseudocolor) and infected neuron (cyan pseudocolor, mitochondria are in yellow). White arrows point to putative contact sites, where the distance between the membranes is the smallest, and several filament-like structures (magenta) can be observed connecting the two cells. b *** p
Figure Legend Snippet: Microglia rapidly isolate virus-infected neurons in the brain. a Microglia (Iba1, yellow) are recruited to neurons infected with PRV expressing GFP with immediate-early kinetics (BDG) in the hypothalamic paraventricular nucleus (PVN). Note that microglia recruitment starts after the expression of the immediate-early marker GFP (phase I), when low levels of viral structural proteins become detectable (phase II). High levels of viral structural proteins indicate a late stage of viral infection (phase III), which is associated with higher number of microglia recruited to infected cells. b Microglial numbers increase significantly around virus-infected neurons in parallel with the propagation of infection c In vivo two-photon imaging reveals the recruitment of microglia (green) to virus-infected neurons (red) in real-time. Retrograde transsynaptic infection was induced by the virus BDR in Cx3Cr1 +/GFP microglia reporter mice 7 days prior to imaging to visualize infected cells in the cerebral cortex based on the immediate-early DSRed expression. d Merged Z -planes of microglial cells around a PRV-DSRed-positive neuron (arrows indicate recruited microglia, arrowheads indicate microglial processes contacting the infected cell). e Microglial process velocity increases significantly in response to viral infection compared to that seen in control mice. f – i Correlated CLSM, electron microscopy and electron tomography confirms direct microglia–neuron contact with intercellular molecular links in early phase of viral infection. f , 3D-reconstruction from deconvolved confocal stack of a recruited microglia (green) engulfing a PRV-positive neuronal cell body (cyan) with CD68-positive phagolysosomes (magenta) within the microglia, arranged around infected neuron. Upper insert: single image plane of the same confocal stack. Middle insert: 3D-reconstruction of the same microglial cell, rotated 180° around the vertical axis. Asterisk labels the bay engulfing the soma of the infected neuron. Lower insert: 3D-reconstruction of the infected neuronal cell body with surrounding phagolysosomes located within the microglia. g Transmission electron micrograph shows the same cells on an ultrathin section matching the confocal image plane in the upper insert in f . Note that microglial processes surround the apical dendrites of the infected neuron. h Part of g (in white box) enlarged. Note that the nucleus is void of viral capsids and the membrane of the neuron is intact (white arrowheads), confirming the early phase of infection. i 3 nm thick electron tomographic section and corresponding 3D-reconstruction shows the very close cell–cell contact between the same microglia (green pseudocolor) and infected neuron (cyan pseudocolor, mitochondria are in yellow). White arrows point to putative contact sites, where the distance between the membranes is the smallest, and several filament-like structures (magenta) can be observed connecting the two cells. b *** p

Techniques Used: Infection, Expressing, Marker, In Vivo, Imaging, Mouse Assay, Confocal Laser Scanning Microscopy, Electron Microscopy, Transmission Assay

11) Product Images from "ATP-Binding Cassette Transporter ABC2/ABCA2 in the Rat Brain: A Novel Mammalian Lysosome-Associated Membrane Protein and a Specific Marker for Oligodendrocytes But Not for Myelin Sheaths"

Article Title: ATP-Binding Cassette Transporter ABC2/ABCA2 in the Rat Brain: A Novel Mammalian Lysosome-Associated Membrane Protein and a Specific Marker for Oligodendrocytes But Not for Myelin Sheaths

Journal: The Journal of Neuroscience

doi: 10.1523/JNEUROSCI.21-03-00849.2001

ABC2 in oligodendrocytes by immunoelectron microscopy. The ultrathin section was cut from the medulla of the cerebellum. The peroxidase–DAB reaction product from ABC2 immunohistochemistry was intensified with the silver–gold particles. A , Most of the intense labeling was observed around lysosomes ( arrows ), but only particular labeling or none at all was observed in other organelles at a magnification of 7000×. Note that there is no specific labeling in the myelin sheath or the axon around the cell body. AX , Axon; N , nucleus; V , blood vessel. B , High-magnification (10,000×) graphs for organellar localization of the ABC2 in oligodendrocytes. The silver–gold particles representing ABC2 immunoreactivities localized densely in lysosome membranes and particularly in the trans face of the Golgi apparatus ( arrow ). ER , Endoplasmic reticulum; G , Golgi body; L , lysosome; M , mitochondria. Scale bar, 500 nm.
Figure Legend Snippet: ABC2 in oligodendrocytes by immunoelectron microscopy. The ultrathin section was cut from the medulla of the cerebellum. The peroxidase–DAB reaction product from ABC2 immunohistochemistry was intensified with the silver–gold particles. A , Most of the intense labeling was observed around lysosomes ( arrows ), but only particular labeling or none at all was observed in other organelles at a magnification of 7000×. Note that there is no specific labeling in the myelin sheath or the axon around the cell body. AX , Axon; N , nucleus; V , blood vessel. B , High-magnification (10,000×) graphs for organellar localization of the ABC2 in oligodendrocytes. The silver–gold particles representing ABC2 immunoreactivities localized densely in lysosome membranes and particularly in the trans face of the Golgi apparatus ( arrow ). ER , Endoplasmic reticulum; G , Golgi body; L , lysosome; M , mitochondria. Scale bar, 500 nm.

Techniques Used: Immuno-Electron Microscopy, Immunohistochemistry, Labeling

12) Product Images from "Testicular regulation of seasonal change in apocrine glands in the back skin of the brown bear (Ursus arctos)"

Article Title: Testicular regulation of seasonal change in apocrine glands in the back skin of the brown bear (Ursus arctos)

Journal: The Journal of Veterinary Medical Science

doi: 10.1292/jvms.17-0689

Localization of VVA reaction in apocrine cells of castrated males during the breeding season. Semi-thin sections with VVA staining (A). Square (A) corresponds to panel (B). Arrowheads, positive VVA staining. Transmission electron microscopy (TEM) image of ultrathin section (B) adjacent to panel (A). TEM image without VVA staining from same individual (C). Square (B) and (C) correspond to panel (D) and (E), respectively. Asterisk, secretory granule. G, Golgi apparatus. Scale bar: 5 µ m.
Figure Legend Snippet: Localization of VVA reaction in apocrine cells of castrated males during the breeding season. Semi-thin sections with VVA staining (A). Square (A) corresponds to panel (B). Arrowheads, positive VVA staining. Transmission electron microscopy (TEM) image of ultrathin section (B) adjacent to panel (A). TEM image without VVA staining from same individual (C). Square (B) and (C) correspond to panel (D) and (E), respectively. Asterisk, secretory granule. G, Golgi apparatus. Scale bar: 5 µ m.

Techniques Used: Staining, Transmission Assay, Electron Microscopy, Transmission Electron Microscopy

13) Product Images from "Insect’s intestinal organ for symbiont sorting"

Article Title: Insect’s intestinal organ for symbiont sorting

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.1511454112

Fine structure of the constricted region in second instar nymphs of R. pedestris . ( A ) Semiultrathin section stained with toluidine blue and observed by light microscopy. ( B–E ) Ultrathin sections observed by transmission electron microscopy. (
Figure Legend Snippet: Fine structure of the constricted region in second instar nymphs of R. pedestris . ( A ) Semiultrathin section stained with toluidine blue and observed by light microscopy. ( B–E ) Ultrathin sections observed by transmission electron microscopy. (

Techniques Used: Staining, Light Microscopy, Transmission Assay, Electron Microscopy

14) Product Images from "The complete genome sequence of an alphabaculovirus from Spodoptera exempta, an agricultural pest of major economic significance in Africa"

Article Title: The complete genome sequence of an alphabaculovirus from Spodoptera exempta, an agricultural pest of major economic significance in Africa

Journal: PLoS ONE

doi: 10.1371/journal.pone.0209937

Occlusion bodies (OBs) of baculovirus isolate SpexNPV-244.1. (A, B) Scanning electron micrographs of SpexNPV-244.1 OBs. (C, D) Transmission electron micrographs of ultrathin sections through SpexNPV-244.1 OBs. The red arrow in (C) indicates a longitudinal section through a virion containing a single nucleocapsid, while the red arrow in (D) indicates a cross-section through a virion containing four nucleocapsids. The lengths of scale bars are given beneath each scale bar.
Figure Legend Snippet: Occlusion bodies (OBs) of baculovirus isolate SpexNPV-244.1. (A, B) Scanning electron micrographs of SpexNPV-244.1 OBs. (C, D) Transmission electron micrographs of ultrathin sections through SpexNPV-244.1 OBs. The red arrow in (C) indicates a longitudinal section through a virion containing a single nucleocapsid, while the red arrow in (D) indicates a cross-section through a virion containing four nucleocapsids. The lengths of scale bars are given beneath each scale bar.

Techniques Used: Transmission Assay

15) Product Images from "Removal of Abnormal Myofilament O-GlcNAcylation Restores Ca2+ Sensitivity in Diabetic Cardiac Muscle"

Article Title: Removal of Abnormal Myofilament O-GlcNAcylation Restores Ca2+ Sensitivity in Diabetic Cardiac Muscle

Journal: Diabetes

doi: 10.2337/db14-1107

Differential OGT and OGA subcellular localization and myofilament interactions in control and diabetic myocardium. Representative transmission electron microscopy images of control ( A ) and STZ diabetic rat myocardium ( B ). Ultrathin sections were examined with immunoelectron microscopy with primary antibodies (anti-OGT AL-28 and anti-OGA), and secondary antibodies gold-labeled (anti-rabbit, 12 nm; anti-chicken, 6 nm). Z-line, green arrowhead; OGT, purple circles; OGA, red circles; OGT and OGA in close vicinity, pink circles; OGT, purple arrowhead; and OGA, red arrowhead. Quantification of OGT ( C ) and OGA ( D ) number of particles/field in nine fields shows an increase for OGT (2 ± 0.6 vs. 8.6 ± 2.6, * P ≤ 0.028) and OGA (9.4 ± 2.9 vs 37.6 ± 12.6, * P ≤ 0.05) immunoelectron microscopy in STZ diabetic myocardium. Images were analyzed in ImageJ software (NIH). Representative coimmunoprecipitations (co-IP) of OGT ( E ) and OGA ( F ), followed by Western blots (WB) for α-sarcomeric actin, Tm, and MLC 1. A fraction of the inputs from controls (C1, C2), STZ (S1, S2), and agarose beads with no antibody (M) or isotype-specific normal antibody + agarose beads (1°) were included. G : Analysis of integrated signal density of myofilament immunoreactivity normalized to total IP OGT displayed a trend toward increased interactions with Tm and MLC 1. H : OGA co-IP shows that diabetic STZ rats display several fold increased associations with α-actin, α-Tm, and MLC 1. * P ≤ 0.05.
Figure Legend Snippet: Differential OGT and OGA subcellular localization and myofilament interactions in control and diabetic myocardium. Representative transmission electron microscopy images of control ( A ) and STZ diabetic rat myocardium ( B ). Ultrathin sections were examined with immunoelectron microscopy with primary antibodies (anti-OGT AL-28 and anti-OGA), and secondary antibodies gold-labeled (anti-rabbit, 12 nm; anti-chicken, 6 nm). Z-line, green arrowhead; OGT, purple circles; OGA, red circles; OGT and OGA in close vicinity, pink circles; OGT, purple arrowhead; and OGA, red arrowhead. Quantification of OGT ( C ) and OGA ( D ) number of particles/field in nine fields shows an increase for OGT (2 ± 0.6 vs. 8.6 ± 2.6, * P ≤ 0.028) and OGA (9.4 ± 2.9 vs 37.6 ± 12.6, * P ≤ 0.05) immunoelectron microscopy in STZ diabetic myocardium. Images were analyzed in ImageJ software (NIH). Representative coimmunoprecipitations (co-IP) of OGT ( E ) and OGA ( F ), followed by Western blots (WB) for α-sarcomeric actin, Tm, and MLC 1. A fraction of the inputs from controls (C1, C2), STZ (S1, S2), and agarose beads with no antibody (M) or isotype-specific normal antibody + agarose beads (1°) were included. G : Analysis of integrated signal density of myofilament immunoreactivity normalized to total IP OGT displayed a trend toward increased interactions with Tm and MLC 1. H : OGA co-IP shows that diabetic STZ rats display several fold increased associations with α-actin, α-Tm, and MLC 1. * P ≤ 0.05.

Techniques Used: Transmission Assay, Electron Microscopy, Immuno-Electron Microscopy, Labeling, Software, Co-Immunoprecipitation Assay, Western Blot

16) Product Images from "The Adherent/Invasive Escherichia coli Strain LF82 Invades and Persists in Human Prostate Cell Line RWPE-1, Activating a Strong Inflammatory Response"

Article Title: The Adherent/Invasive Escherichia coli Strain LF82 Invades and Persists in Human Prostate Cell Line RWPE-1, Activating a Strong Inflammatory Response

Journal: Infection and Immunity

doi: 10.1128/IAI.00438-16

Light and electron microscopy of Epon-embedded RWPE-1 cells infected with LF82 (C and D) or EC73 (A and B) for 24 h. Groups of bacteria are frequently observed to occupy cytoplasmic vacuoles, suggesting active proliferation and/or clustering. (A and C) Light micrograph of toluidine blue-stained thin sections. Arrows indicate cells with groups of bacteria within cytoplasmic compartments. (B and D) Electron micrographs of ultrathin sections. Bars: A and C, 8 μm; B and D, 0.9 μm.
Figure Legend Snippet: Light and electron microscopy of Epon-embedded RWPE-1 cells infected with LF82 (C and D) or EC73 (A and B) for 24 h. Groups of bacteria are frequently observed to occupy cytoplasmic vacuoles, suggesting active proliferation and/or clustering. (A and C) Light micrograph of toluidine blue-stained thin sections. Arrows indicate cells with groups of bacteria within cytoplasmic compartments. (B and D) Electron micrographs of ultrathin sections. Bars: A and C, 8 μm; B and D, 0.9 μm.

Techniques Used: Electron Microscopy, Infection, Staining

17) Product Images from "Morphological Features of the Anther Development in Tomato Plants with Non-Specific Male Sterility"

Article Title: Morphological Features of the Anther Development in Tomato Plants with Non-Specific Male Sterility

Journal: Biology

doi: 10.3390/biology9020032

Cross-sections of anthers with various types of developmental abnormalities ( a – d ). Pollen sacs ( а ’– d ’) and microspore ultrastructure ( a’ ’– d’’ ) from these anthers. black asterisks—plastids, white asterisks—mitochondria. Bar—30 μm (for semi-thin sections: a–d’) and 3 μm (for ultrathin sections: a”–d”).
Figure Legend Snippet: Cross-sections of anthers with various types of developmental abnormalities ( a – d ). Pollen sacs ( а ’– d ’) and microspore ultrastructure ( a’ ’– d’’ ) from these anthers. black asterisks—plastids, white asterisks—mitochondria. Bar—30 μm (for semi-thin sections: a–d’) and 3 μm (for ultrathin sections: a”–d”).

Techniques Used:

Semi-thin cross sections of anther from the control ( а , a’ , a’’ ) and transgenic tomato plants without abnormalities ( b , b ’, b ’’) at the microspore stage. Microspore ultrastructure at some asymmetric mitosis stages ( с – g ), and mature pollen grain ultrastructure ( h ). Symbols: er–endoplasmic reticulum; gс–generative cell; vc–vegetative cell; black asterisks–plastids, white asterisks–mitochondria. Bar—100 μm (for semi-thin sections: a–b’) and 10 μm (for ultrathin sections: a”; b”; c–h).
Figure Legend Snippet: Semi-thin cross sections of anther from the control ( а , a’ , a’’ ) and transgenic tomato plants without abnormalities ( b , b ’, b ’’) at the microspore stage. Microspore ultrastructure at some asymmetric mitosis stages ( с – g ), and mature pollen grain ultrastructure ( h ). Symbols: er–endoplasmic reticulum; gс–generative cell; vc–vegetative cell; black asterisks–plastids, white asterisks–mitochondria. Bar—100 μm (for semi-thin sections: a–b’) and 10 μm (for ultrathin sections: a”; b”; c–h).

Techniques Used: Transgenic Assay

18) Product Images from "Co-transmission of acetylcholine and GABA regulates hippocampal states"

Article Title: Co-transmission of acetylcholine and GABA regulates hippocampal states

Journal: Nature Communications

doi: 10.1038/s41467-018-05136-1

Cholinergic cells express the molecular machinery required for GABA release. a , b The cholinergic neurons of the MS are GABAergic. White box in a contains area enlarged in b . Images show neurons stained for ChAT in red, while the green labelling marks the vGAT-expressing neurons in vGAT-ZsGreen reporter mouse. c , d AAV-eYFP virus-traced septo-hippocampal fibres express GAD65 ( c ). AAV-eYFP virus-traced septo-hippocampal fibres express vGAT and vAChT ( d ). Insets show xz and yz projections of the terminal labelled with an arrow. Arrowhead points to another terminal. Green line marks the fibre outline. (Scale bar on d is 210, 14, 2 and 1 μm for a , b , c and d , respectively.) e , f Hippocampal cholinergic terminals contain GABA. Three consecutive EM sections of a vAChT-positive terminal ( e , red pseudocolor) are shown. vAChT was visualised by pre-embedding immunogold method (the first panel of e , silver-intensified gold particles, large arrows), whereas on the next ultrathin sections (the second and third panels of e ) postembedding GABA immunostaining was performed (smaller gold particles, thin arrows, some GABA molecules penetrate into mitochondria during fixation). vAChT signal is absent in postembedding images, because of the etching procedure. Scale bar is 200 nm for all EM images. f Cholinergic terminals contained significantly higher immunogold signal than glutamatergic ones ( p
Figure Legend Snippet: Cholinergic cells express the molecular machinery required for GABA release. a , b The cholinergic neurons of the MS are GABAergic. White box in a contains area enlarged in b . Images show neurons stained for ChAT in red, while the green labelling marks the vGAT-expressing neurons in vGAT-ZsGreen reporter mouse. c , d AAV-eYFP virus-traced septo-hippocampal fibres express GAD65 ( c ). AAV-eYFP virus-traced septo-hippocampal fibres express vGAT and vAChT ( d ). Insets show xz and yz projections of the terminal labelled with an arrow. Arrowhead points to another terminal. Green line marks the fibre outline. (Scale bar on d is 210, 14, 2 and 1 μm for a , b , c and d , respectively.) e , f Hippocampal cholinergic terminals contain GABA. Three consecutive EM sections of a vAChT-positive terminal ( e , red pseudocolor) are shown. vAChT was visualised by pre-embedding immunogold method (the first panel of e , silver-intensified gold particles, large arrows), whereas on the next ultrathin sections (the second and third panels of e ) postembedding GABA immunostaining was performed (smaller gold particles, thin arrows, some GABA molecules penetrate into mitochondria during fixation). vAChT signal is absent in postembedding images, because of the etching procedure. Scale bar is 200 nm for all EM images. f Cholinergic terminals contained significantly higher immunogold signal than glutamatergic ones ( p

Techniques Used: Mass Spectrometry, Staining, Expressing, Immunostaining

19) Product Images from "Overexpression of α-Internexin Causes Abnormal Neurofilamentous Accumulations and Motor Coordination Deficits in Transgenic Mice"

Article Title: Overexpression of α-Internexin Causes Abnormal Neurofilamentous Accumulations and Motor Coordination Deficits in Transgenic Mice

Journal: The Journal of Neuroscience

doi: 10.1523/JNEUROSCI.19-08-02974.1999

Ultrastructural analysis of Purkinje cells and neocortical pyramidal neurons from 4.5-month-old transgenic mice. A , Ultrathin sections of cerebellum from an α16k-T5 homozygous mouse examined by electron microscopy revealed accumulation of high levels of intermediate filaments in the soma of a Purkinje cell. Lying within the neurofilamentous masses are aggregates of mitochondria and sER, some of which appear to associate with each other ( arrows ). Although the external membranes of these mitochondria appear to be continuous with sER, some other mitochondria look normal. B , Electron micrograph showed massive accumulation of intermediate filaments in the soma of a neocortical pyramidal neuron from a 4.5-month-old α16k-T5 homozygous mouse. Note that the nucleus is eccentrically localized, and many cellular organelles such as mitochondria are peripherally displaced. Some mitochondria trapped within the masses of neuronal intermediate filaments often aggregate and appear abnormal. The neuronal intermediate filaments in A and B are highly packed and randomly oriented. m , Mitochondria; sER , smooth endoplasmic reticulum; n , nucleus. Scale bars: A , 0.75 μm; B , 2.0 μm.
Figure Legend Snippet: Ultrastructural analysis of Purkinje cells and neocortical pyramidal neurons from 4.5-month-old transgenic mice. A , Ultrathin sections of cerebellum from an α16k-T5 homozygous mouse examined by electron microscopy revealed accumulation of high levels of intermediate filaments in the soma of a Purkinje cell. Lying within the neurofilamentous masses are aggregates of mitochondria and sER, some of which appear to associate with each other ( arrows ). Although the external membranes of these mitochondria appear to be continuous with sER, some other mitochondria look normal. B , Electron micrograph showed massive accumulation of intermediate filaments in the soma of a neocortical pyramidal neuron from a 4.5-month-old α16k-T5 homozygous mouse. Note that the nucleus is eccentrically localized, and many cellular organelles such as mitochondria are peripherally displaced. Some mitochondria trapped within the masses of neuronal intermediate filaments often aggregate and appear abnormal. The neuronal intermediate filaments in A and B are highly packed and randomly oriented. m , Mitochondria; sER , smooth endoplasmic reticulum; n , nucleus. Scale bars: A , 0.75 μm; B , 2.0 μm.

Techniques Used: Transgenic Assay, Mouse Assay, Electron Microscopy

20) Product Images from "Adipose-derived mesenchymal stem cells accelerate nerve regeneration and functional recovery in a rat model of recurrent laryngeal nerve injury"

Article Title: Adipose-derived mesenchymal stem cells accelerate nerve regeneration and functional recovery in a rat model of recurrent laryngeal nerve injury

Journal: Neural Regeneration Research

doi: 10.4103/1673-5374.215267

Effects of adipose-derived mesenchymal stem cells (ADSCs), differentiated Schwann-like adipose-derived mesenchymal stem cells (dADSCs) and extracellular matrix (ECM) transplantation on the morphological changes of axons following recurrent laryngeal nerve (RLN) injury. (A–C) RLN in the ADSCs group, the dADSCs group and the ECM group at 2 weeks post-surgery, respectively (toluidine blue staining, light microscopy). (D) The ultrathin transverse section of the RLN as visualized by transmission electron microscopy (ADSCs group). Black arrows point to myelinated axons, while white arrows point to unmyelinated axons. (E, F) For stereological analysis, the density of myelinated fibers (E) and the myelin thickness (F) were quantitatively evaluated and compared by statistical analyses. Data are expressed as the mean ± SEM ( n = 5) and analyzed by one-way analysis of variance followed by Scheffe's post hoc test at each time point. * P
Figure Legend Snippet: Effects of adipose-derived mesenchymal stem cells (ADSCs), differentiated Schwann-like adipose-derived mesenchymal stem cells (dADSCs) and extracellular matrix (ECM) transplantation on the morphological changes of axons following recurrent laryngeal nerve (RLN) injury. (A–C) RLN in the ADSCs group, the dADSCs group and the ECM group at 2 weeks post-surgery, respectively (toluidine blue staining, light microscopy). (D) The ultrathin transverse section of the RLN as visualized by transmission electron microscopy (ADSCs group). Black arrows point to myelinated axons, while white arrows point to unmyelinated axons. (E, F) For stereological analysis, the density of myelinated fibers (E) and the myelin thickness (F) were quantitatively evaluated and compared by statistical analyses. Data are expressed as the mean ± SEM ( n = 5) and analyzed by one-way analysis of variance followed by Scheffe's post hoc test at each time point. * P

Techniques Used: Derivative Assay, Transplantation Assay, Staining, Light Microscopy, Transmission Assay, Electron Microscopy

21) Product Images from "Overexpression of α-Internexin Causes Abnormal Neurofilamentous Accumulations and Motor Coordination Deficits in Transgenic Mice"

Article Title: Overexpression of α-Internexin Causes Abnormal Neurofilamentous Accumulations and Motor Coordination Deficits in Transgenic Mice

Journal: The Journal of Neuroscience

doi: 10.1523/JNEUROSCI.19-08-02974.1999

Ultrastructural analysis of Purkinje cells and neocortical pyramidal neurons from 4.5-month-old transgenic mice. A , Ultrathin sections of cerebellum from an α16k-T5 homozygous mouse examined by electron microscopy revealed accumulation of high levels of intermediate filaments in the soma of a Purkinje cell. Lying within the neurofilamentous masses are aggregates of mitochondria and sER, some of which appear to associate with each other ( arrows ). Although the external membranes of these mitochondria appear to be continuous with sER, some other mitochondria look normal. B , Electron micrograph showed massive accumulation of intermediate filaments in the soma of a neocortical pyramidal neuron from a 4.5-month-old α16k-T5 homozygous mouse. Note that the nucleus is eccentrically localized, and many cellular organelles such as mitochondria are peripherally displaced. Some mitochondria trapped within the masses of neuronal intermediate filaments often aggregate and appear abnormal. The neuronal intermediate filaments in A and B are highly packed and randomly oriented. m , Mitochondria; sER , smooth endoplasmic reticulum; n , nucleus. Scale bars: A , 0.75 μm; B , 2.0 μm.
Figure Legend Snippet: Ultrastructural analysis of Purkinje cells and neocortical pyramidal neurons from 4.5-month-old transgenic mice. A , Ultrathin sections of cerebellum from an α16k-T5 homozygous mouse examined by electron microscopy revealed accumulation of high levels of intermediate filaments in the soma of a Purkinje cell. Lying within the neurofilamentous masses are aggregates of mitochondria and sER, some of which appear to associate with each other ( arrows ). Although the external membranes of these mitochondria appear to be continuous with sER, some other mitochondria look normal. B , Electron micrograph showed massive accumulation of intermediate filaments in the soma of a neocortical pyramidal neuron from a 4.5-month-old α16k-T5 homozygous mouse. Note that the nucleus is eccentrically localized, and many cellular organelles such as mitochondria are peripherally displaced. Some mitochondria trapped within the masses of neuronal intermediate filaments often aggregate and appear abnormal. The neuronal intermediate filaments in A and B are highly packed and randomly oriented. m , Mitochondria; sER , smooth endoplasmic reticulum; n , nucleus. Scale bars: A , 0.75 μm; B , 2.0 μm.

Techniques Used: Transgenic Assay, Mouse Assay, Electron Microscopy

22) Product Images from "Co-transmission of acetylcholine and GABA regulates hippocampal states"

Article Title: Co-transmission of acetylcholine and GABA regulates hippocampal states

Journal: Nature Communications

doi: 10.1038/s41467-018-05136-1

Cholinergic cells express the molecular machinery required for GABA release. a , b The cholinergic neurons of the MS are GABAergic. White box in a contains area enlarged in b . Images show neurons stained for ChAT in red, while the green labelling marks the vGAT-expressing neurons in vGAT-ZsGreen reporter mouse. c , d AAV-eYFP virus-traced septo-hippocampal fibres express GAD65 ( c ). AAV-eYFP virus-traced septo-hippocampal fibres express vGAT and vAChT ( d ). Insets show xz and yz projections of the terminal labelled with an arrow. Arrowhead points to another terminal. Green line marks the fibre outline. (Scale bar on d is 210, 14, 2 and 1 μm for a , b , c and d , respectively.) e , f Hippocampal cholinergic terminals contain GABA. Three consecutive EM sections of a vAChT-positive terminal ( e , red pseudocolor) are shown. vAChT was visualised by pre-embedding immunogold method (the first panel of e , silver-intensified gold particles, large arrows), whereas on the next ultrathin sections (the second and third panels of e ) postembedding GABA immunostaining was performed (smaller gold particles, thin arrows, some GABA molecules penetrate into mitochondria during fixation). vAChT signal is absent in postembedding images, because of the etching procedure. Scale bar is 200 nm for all EM images. f Cholinergic terminals contained significantly higher immunogold signal than glutamatergic ones ( p )
Figure Legend Snippet: Cholinergic cells express the molecular machinery required for GABA release. a , b The cholinergic neurons of the MS are GABAergic. White box in a contains area enlarged in b . Images show neurons stained for ChAT in red, while the green labelling marks the vGAT-expressing neurons in vGAT-ZsGreen reporter mouse. c , d AAV-eYFP virus-traced septo-hippocampal fibres express GAD65 ( c ). AAV-eYFP virus-traced septo-hippocampal fibres express vGAT and vAChT ( d ). Insets show xz and yz projections of the terminal labelled with an arrow. Arrowhead points to another terminal. Green line marks the fibre outline. (Scale bar on d is 210, 14, 2 and 1 μm for a , b , c and d , respectively.) e , f Hippocampal cholinergic terminals contain GABA. Three consecutive EM sections of a vAChT-positive terminal ( e , red pseudocolor) are shown. vAChT was visualised by pre-embedding immunogold method (the first panel of e , silver-intensified gold particles, large arrows), whereas on the next ultrathin sections (the second and third panels of e ) postembedding GABA immunostaining was performed (smaller gold particles, thin arrows, some GABA molecules penetrate into mitochondria during fixation). vAChT signal is absent in postembedding images, because of the etching procedure. Scale bar is 200 nm for all EM images. f Cholinergic terminals contained significantly higher immunogold signal than glutamatergic ones ( p )

Techniques Used: Mass Spectrometry, Staining, Expressing, Immunostaining

23) Product Images from "Impairment of vesicular ATP release affects glucose metabolism and increases insulin sensitivity"

Article Title: Impairment of vesicular ATP release affects glucose metabolism and increases insulin sensitivity

Journal: Scientific Reports

doi: 10.1038/srep06689

VNUT acted as a negative regulator of insulin secretion in pancreatic islets. (A) Indirect immunofluorescence microscopy revealed that VNUT was expressed in mouse pancreatic islets (WT). No VNUT immunoreactivity was seen in pancreatic islets isolated from Vnut −/− mice (KO). L, islet of Langerhans; a, acinar cell. Scale bar = 20 μm. (B) Localization of VNUT in islets. Sections of pancreas were double immunostained with antibodies against VNUT and insulin, VNUT and glucagon (marker of α−cells) and VNUT and somatostatin (marker of δ-cells), and observed by confocal laser microscopy. Merged pictures are also shown to the right. Areas surrounded by dotted line were enlarged in Supplementary Fig. S7 . Bar = 20 μm. (C) Immunoelectron microscopy indicated that VNUT was localized at dense core granules of β-cells. Arrows indicate VNUT-positive signals (gold nanoparticles). The inset shows the control sample for which normal serum instead of mouse VNUT antibody was used. Scale bar = 500 nm. (D) Electron micrographs of ultrathin-sections of islets from WT and KO mice. The number and morphology of secretory granules in islet β-cells of KO mice were normal. Scale bar = 1 μm. (E) High glucose concentration-dependent release of ATP from pancreatic islets of WT (open bars, n = 10, 19) and KO mice (filled bars, n = 15, 14). (F) High glucose concentration-dependent secretion of insulin from pancreatic islets of WT (n = 8, 14, 8) and KO mice (n = 8, 14, 6). Where indicated, insulin secretion was measured in the presence of 0.1 mM ATP. (G) The amount of insulin in pancreatic islets of WT (n = 3) and KO mice (n = 4). (H) The ratio of proinsulin/insulin + proinsulin in pancreatic islets of WT (n = 16) and KO mice (n = 13). (I) The amount of ATP content in pancreatic islets of WT (n = 4) and KO mice (n = 4). Error bars represent mean ± S.E. *: p
Figure Legend Snippet: VNUT acted as a negative regulator of insulin secretion in pancreatic islets. (A) Indirect immunofluorescence microscopy revealed that VNUT was expressed in mouse pancreatic islets (WT). No VNUT immunoreactivity was seen in pancreatic islets isolated from Vnut −/− mice (KO). L, islet of Langerhans; a, acinar cell. Scale bar = 20 μm. (B) Localization of VNUT in islets. Sections of pancreas were double immunostained with antibodies against VNUT and insulin, VNUT and glucagon (marker of α−cells) and VNUT and somatostatin (marker of δ-cells), and observed by confocal laser microscopy. Merged pictures are also shown to the right. Areas surrounded by dotted line were enlarged in Supplementary Fig. S7 . Bar = 20 μm. (C) Immunoelectron microscopy indicated that VNUT was localized at dense core granules of β-cells. Arrows indicate VNUT-positive signals (gold nanoparticles). The inset shows the control sample for which normal serum instead of mouse VNUT antibody was used. Scale bar = 500 nm. (D) Electron micrographs of ultrathin-sections of islets from WT and KO mice. The number and morphology of secretory granules in islet β-cells of KO mice were normal. Scale bar = 1 μm. (E) High glucose concentration-dependent release of ATP from pancreatic islets of WT (open bars, n = 10, 19) and KO mice (filled bars, n = 15, 14). (F) High glucose concentration-dependent secretion of insulin from pancreatic islets of WT (n = 8, 14, 8) and KO mice (n = 8, 14, 6). Where indicated, insulin secretion was measured in the presence of 0.1 mM ATP. (G) The amount of insulin in pancreatic islets of WT (n = 3) and KO mice (n = 4). (H) The ratio of proinsulin/insulin + proinsulin in pancreatic islets of WT (n = 16) and KO mice (n = 13). (I) The amount of ATP content in pancreatic islets of WT (n = 4) and KO mice (n = 4). Error bars represent mean ± S.E. *: p

Techniques Used: Immunofluorescence, Microscopy, Isolation, Mouse Assay, Marker, Immuno-Electron Microscopy, Concentration Assay

24) Product Images from "Removal of Abnormal Myofilament O-GlcNAcylation Restores Ca2+ Sensitivity in Diabetic Cardiac Muscle"

Article Title: Removal of Abnormal Myofilament O-GlcNAcylation Restores Ca2+ Sensitivity in Diabetic Cardiac Muscle

Journal: Diabetes

doi: 10.2337/db14-1107

Differential OGT and OGA subcellular localization and myofilament interactions in control and diabetic myocardium. Representative transmission electron microscopy images of control ( A ) and STZ diabetic rat myocardium ( B ). Ultrathin sections were examined with immunoelectron microscopy with primary antibodies (anti-OGT AL-28 and anti-OGA), and secondary antibodies gold-labeled (anti-rabbit, 12 nm; anti-chicken, 6 nm). Z-line, green arrowhead; OGT, purple circles; OGA, red circles; OGT and OGA in close vicinity, pink circles; OGT, purple arrowhead; and OGA, red arrowhead. Quantification of OGT ( C ) and OGA ( D ) number of particles/field in nine fields shows an increase for OGT (2 ± 0.6 vs. 8.6 ± 2.6, * P ≤ 0.028) and OGA (9.4 ± 2.9 vs 37.6 ± 12.6, * P ≤ 0.05) immunoelectron microscopy in STZ diabetic myocardium. Images were analyzed in ImageJ software (NIH). Representative coimmunoprecipitations (co-IP) of OGT ( E ) and OGA ( F ), followed by Western blots (WB) for α-sarcomeric actin, Tm, and MLC 1. A fraction of the inputs from controls (C1, C2), STZ (S1, S2), and agarose beads with no antibody (M) or isotype-specific normal antibody + agarose beads (1°) were included. G : Analysis of integrated signal density of myofilament immunoreactivity normalized to total IP OGT displayed a trend toward increased interactions with Tm and MLC 1. H : OGA co-IP shows that diabetic STZ rats display several fold increased associations with α-actin, α-Tm, and MLC 1. * P ≤ 0.05.
Figure Legend Snippet: Differential OGT and OGA subcellular localization and myofilament interactions in control and diabetic myocardium. Representative transmission electron microscopy images of control ( A ) and STZ diabetic rat myocardium ( B ). Ultrathin sections were examined with immunoelectron microscopy with primary antibodies (anti-OGT AL-28 and anti-OGA), and secondary antibodies gold-labeled (anti-rabbit, 12 nm; anti-chicken, 6 nm). Z-line, green arrowhead; OGT, purple circles; OGA, red circles; OGT and OGA in close vicinity, pink circles; OGT, purple arrowhead; and OGA, red arrowhead. Quantification of OGT ( C ) and OGA ( D ) number of particles/field in nine fields shows an increase for OGT (2 ± 0.6 vs. 8.6 ± 2.6, * P ≤ 0.028) and OGA (9.4 ± 2.9 vs 37.6 ± 12.6, * P ≤ 0.05) immunoelectron microscopy in STZ diabetic myocardium. Images were analyzed in ImageJ software (NIH). Representative coimmunoprecipitations (co-IP) of OGT ( E ) and OGA ( F ), followed by Western blots (WB) for α-sarcomeric actin, Tm, and MLC 1. A fraction of the inputs from controls (C1, C2), STZ (S1, S2), and agarose beads with no antibody (M) or isotype-specific normal antibody + agarose beads (1°) were included. G : Analysis of integrated signal density of myofilament immunoreactivity normalized to total IP OGT displayed a trend toward increased interactions with Tm and MLC 1. H : OGA co-IP shows that diabetic STZ rats display several fold increased associations with α-actin, α-Tm, and MLC 1. * P ≤ 0.05.

Techniques Used: Transmission Assay, Electron Microscopy, Immuno-Electron Microscopy, Labeling, Software, Co-Immunoprecipitation Assay, Western Blot

25) Product Images from "Synaptic characteristics of dentate gyrus axonal boutons and their relationships with aging, menopause, and memory in female rhesus monkeys"

Article Title: Synaptic characteristics of dentate gyrus axonal boutons and their relationships with aging, menopause, and memory in female rhesus monkeys

Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

doi: 10.1523/JNEUROSCI.0822-11.2011

Electron micrographs of 15 serial ultrathin sections illustrating the methods involved in obtaining axonal bouton measures. The middle section (8th section) was used as a reference section. All presynaptic boutons containing at least 3 synaptic vesicles
Figure Legend Snippet: Electron micrographs of 15 serial ultrathin sections illustrating the methods involved in obtaining axonal bouton measures. The middle section (8th section) was used as a reference section. All presynaptic boutons containing at least 3 synaptic vesicles

Techniques Used:

26) Product Images from "Microglia control the spread of neurotropic virus infection via P2Y12 signalling and recruit monocytes through P2Y12-independent mechanisms"

Article Title: Microglia control the spread of neurotropic virus infection via P2Y12 signalling and recruit monocytes through P2Y12-independent mechanisms

Journal: Acta Neuropathologica

doi: 10.1007/s00401-018-1885-0

Microglia rapidly isolate virus-infected neurons in the brain. a Microglia (Iba1, yellow) are recruited to neurons infected with PRV expressing GFP with immediate-early kinetics (BDG) in the hypothalamic paraventricular nucleus (PVN). Note that microglia recruitment starts after the expression of the immediate-early marker GFP (phase I), when low levels of viral structural proteins become detectable (phase II). High levels of viral structural proteins indicate a late stage of viral infection (phase III), which is associated with higher number of microglia recruited to infected cells. b Microglial numbers increase significantly around virus-infected neurons in parallel with the propagation of infection c In vivo two-photon imaging reveals the recruitment of microglia (green) to virus-infected neurons (red) in real-time. Retrograde transsynaptic infection was induced by the virus BDR in Cx3Cr1 +/GFP microglia reporter mice 7 days prior to imaging to visualize infected cells in the cerebral cortex based on the immediate-early DSRed expression. d Merged Z -planes of microglial cells around a PRV-DSRed-positive neuron (arrows indicate recruited microglia, arrowheads indicate microglial processes contacting the infected cell). e Microglial process velocity increases significantly in response to viral infection compared to that seen in control mice. f – i Correlated CLSM, electron microscopy and electron tomography confirms direct microglia–neuron contact with intercellular molecular links in early phase of viral infection. f , 3D-reconstruction from deconvolved confocal stack of a recruited microglia (green) engulfing a PRV-positive neuronal cell body (cyan) with CD68-positive phagolysosomes (magenta) within the microglia, arranged around infected neuron. Upper insert: single image plane of the same confocal stack. Middle insert: 3D-reconstruction of the same microglial cell, rotated 180° around the vertical axis. Asterisk labels the bay engulfing the soma of the infected neuron. Lower insert: 3D-reconstruction of the infected neuronal cell body with surrounding phagolysosomes located within the microglia. g Transmission electron micrograph shows the same cells on an ultrathin section matching the confocal image plane in the upper insert in f . Note that microglial processes surround the apical dendrites of the infected neuron. h Part of g (in white box) enlarged. Note that the nucleus is void of viral capsids and the membrane of the neuron is intact (white arrowheads), confirming the early phase of infection. i 3 nm thick electron tomographic section and corresponding 3D-reconstruction shows the very close cell–cell contact between the same microglia (green pseudocolor) and infected neuron (cyan pseudocolor, mitochondria are in yellow). White arrows point to putative contact sites, where the distance between the membranes is the smallest, and several filament-like structures (magenta) can be observed connecting the two cells. b *** p
Figure Legend Snippet: Microglia rapidly isolate virus-infected neurons in the brain. a Microglia (Iba1, yellow) are recruited to neurons infected with PRV expressing GFP with immediate-early kinetics (BDG) in the hypothalamic paraventricular nucleus (PVN). Note that microglia recruitment starts after the expression of the immediate-early marker GFP (phase I), when low levels of viral structural proteins become detectable (phase II). High levels of viral structural proteins indicate a late stage of viral infection (phase III), which is associated with higher number of microglia recruited to infected cells. b Microglial numbers increase significantly around virus-infected neurons in parallel with the propagation of infection c In vivo two-photon imaging reveals the recruitment of microglia (green) to virus-infected neurons (red) in real-time. Retrograde transsynaptic infection was induced by the virus BDR in Cx3Cr1 +/GFP microglia reporter mice 7 days prior to imaging to visualize infected cells in the cerebral cortex based on the immediate-early DSRed expression. d Merged Z -planes of microglial cells around a PRV-DSRed-positive neuron (arrows indicate recruited microglia, arrowheads indicate microglial processes contacting the infected cell). e Microglial process velocity increases significantly in response to viral infection compared to that seen in control mice. f – i Correlated CLSM, electron microscopy and electron tomography confirms direct microglia–neuron contact with intercellular molecular links in early phase of viral infection. f , 3D-reconstruction from deconvolved confocal stack of a recruited microglia (green) engulfing a PRV-positive neuronal cell body (cyan) with CD68-positive phagolysosomes (magenta) within the microglia, arranged around infected neuron. Upper insert: single image plane of the same confocal stack. Middle insert: 3D-reconstruction of the same microglial cell, rotated 180° around the vertical axis. Asterisk labels the bay engulfing the soma of the infected neuron. Lower insert: 3D-reconstruction of the infected neuronal cell body with surrounding phagolysosomes located within the microglia. g Transmission electron micrograph shows the same cells on an ultrathin section matching the confocal image plane in the upper insert in f . Note that microglial processes surround the apical dendrites of the infected neuron. h Part of g (in white box) enlarged. Note that the nucleus is void of viral capsids and the membrane of the neuron is intact (white arrowheads), confirming the early phase of infection. i 3 nm thick electron tomographic section and corresponding 3D-reconstruction shows the very close cell–cell contact between the same microglia (green pseudocolor) and infected neuron (cyan pseudocolor, mitochondria are in yellow). White arrows point to putative contact sites, where the distance between the membranes is the smallest, and several filament-like structures (magenta) can be observed connecting the two cells. b *** p

Techniques Used: Infection, Expressing, Marker, In Vivo, Imaging, Mouse Assay, Confocal Laser Scanning Microscopy, Electron Microscopy, Transmission Assay

27) Product Images from "Rab8b Regulates Transport of West Nile Virus Particles from Recycling Endosomes *"

Article Title: Rab8b Regulates Transport of West Nile Virus Particles from Recycling Endosomes *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M115.712760

Ultrastructural analysis of WNV-infected WT or Rab8b KO MEFs. Ultrathin sections of WNV-infected, Epon-embedded WT or Rab8b MEFs fixed at 48 hpi are shown in ( A–C ) or ( D–F ), respectively. The red and white-boxed areas in A are shown at
Figure Legend Snippet: Ultrastructural analysis of WNV-infected WT or Rab8b KO MEFs. Ultrathin sections of WNV-infected, Epon-embedded WT or Rab8b MEFs fixed at 48 hpi are shown in ( A–C ) or ( D–F ), respectively. The red and white-boxed areas in A are shown at

Techniques Used: Infection

28) Product Images from "The ultrastructural characteristics of porcine hepatocytes donated after cardiac death and preserved with warm machine perfusion preservation"

Article Title: The ultrastructural characteristics of porcine hepatocytes donated after cardiac death and preserved with warm machine perfusion preservation

Journal: PLoS ONE

doi: 10.1371/journal.pone.0186352

The ultrastructure of the endomembrane structure in the hepatocytes of the control liver. (A and C) Typical hepatocytes were identified in the ultrathin sections of the Epon 812-embedded control liver tissue (A). Nucleus was colored blue. The partial area indicated in A was further photographed at a higher magnification (C). Nucleus was colored blue and mitochondria were colored green. Bars = 1 μm. (B and D) Similar typical hepatocytes in osmium-macerated control porcine liver tissues were viewed with a scanning electron microscope (B). Nucleus was colored blue. The p area indicated in B was further photographed at a higher magnification (D). Nucleus was colored blue and mitochondria were colored green. Open arrows indicated cristae of mitochondria. Bars = 1 μm.
Figure Legend Snippet: The ultrastructure of the endomembrane structure in the hepatocytes of the control liver. (A and C) Typical hepatocytes were identified in the ultrathin sections of the Epon 812-embedded control liver tissue (A). Nucleus was colored blue. The partial area indicated in A was further photographed at a higher magnification (C). Nucleus was colored blue and mitochondria were colored green. Bars = 1 μm. (B and D) Similar typical hepatocytes in osmium-macerated control porcine liver tissues were viewed with a scanning electron microscope (B). Nucleus was colored blue. The p area indicated in B was further photographed at a higher magnification (D). Nucleus was colored blue and mitochondria were colored green. Open arrows indicated cristae of mitochondria. Bars = 1 μm.

Techniques Used: Microscopy

29) Product Images from "Optimal concentration and time window for proliferation and differentiation of neural stem cells from embryonic cerebral cortex: 5% oxygen preconditioning for 72 hours"

Article Title: Optimal concentration and time window for proliferation and differentiation of neural stem cells from embryonic cerebral cortex: 5% oxygen preconditioning for 72 hours

Journal: Neural Regeneration Research

doi: 10.4103/1673-5374.165526

Influence of hypoxic preconditioning on the ultrastructure of neural stem cells from embryonic rat cerebral cortex (uranyl acetate and lead citrate double staining, transmission electron microscope). After 7 days of culture in serum-free medium, neurospheres from each group were collected and treated for ultrathin sectioning. (A) In the normal control group (20% O 2 , 7 days), only modest changes in the morphological signs of apoptosis were detected (× 6,000). (B) Pyknosis and condensed chromatin near the nuclear membrane were detected in some neural stem cells in the 5% O 2 120-hour group (× 3,000).
Figure Legend Snippet: Influence of hypoxic preconditioning on the ultrastructure of neural stem cells from embryonic rat cerebral cortex (uranyl acetate and lead citrate double staining, transmission electron microscope). After 7 days of culture in serum-free medium, neurospheres from each group were collected and treated for ultrathin sectioning. (A) In the normal control group (20% O 2 , 7 days), only modest changes in the morphological signs of apoptosis were detected (× 6,000). (B) Pyknosis and condensed chromatin near the nuclear membrane were detected in some neural stem cells in the 5% O 2 120-hour group (× 3,000).

Techniques Used: Double Staining, Transmission Assay, Microscopy

30) Product Images from "Pharmacological, behavioural and mechanistic analysis of HIV-1 gp120 induced painful neuropathy"

Article Title: Pharmacological, behavioural and mechanistic analysis of HIV-1 gp120 induced painful neuropathy

Journal: Pain

doi: 10.1016/j.pain.2007.02.015

Effects of perineural treatment with RSA or HIV-1 gp120 on the integrity of axons in the sciatic nerve. (A–B) Toluidine blue-stained resin sections (1 μm) from the site of application via an oxycellulose wrap of (i) RSA and (ii) HIV-1 gp120 to the sciatic nerve on day 14 post treatment. (A) Examples of the sciatic nerve highlighting small areas of damage which occurred in the perineural window of the nerve. The broken line demarcates the area of morphologically normal nerve from the damaged area. Scale bar = 20 μm. (B) Examples of nerve in which axons appear undisturbed. (C) Examples of ultrathin sections of nerve within 1cm distal to the site of application of (i) RSA and (ii) HIV-1 gp120 to the sciatic nerve on day 14 post treatment assessed by EM representing the majority of the nerve where myelinated and unmyelinated axons appear normal. Scale bar = 10 μm.
Figure Legend Snippet: Effects of perineural treatment with RSA or HIV-1 gp120 on the integrity of axons in the sciatic nerve. (A–B) Toluidine blue-stained resin sections (1 μm) from the site of application via an oxycellulose wrap of (i) RSA and (ii) HIV-1 gp120 to the sciatic nerve on day 14 post treatment. (A) Examples of the sciatic nerve highlighting small areas of damage which occurred in the perineural window of the nerve. The broken line demarcates the area of morphologically normal nerve from the damaged area. Scale bar = 20 μm. (B) Examples of nerve in which axons appear undisturbed. (C) Examples of ultrathin sections of nerve within 1cm distal to the site of application of (i) RSA and (ii) HIV-1 gp120 to the sciatic nerve on day 14 post treatment assessed by EM representing the majority of the nerve where myelinated and unmyelinated axons appear normal. Scale bar = 10 μm.

Techniques Used: Staining

31) Product Images from "Direct Labeling of Polyphosphate at the Ultrastructural Level in Saccharomyces cerevisiae by Using the Affinity of the Polyphosphate Binding Domain of Escherichia coli Exopolyphosphatase"

Article Title: Direct Labeling of Polyphosphate at the Ultrastructural Level in Saccharomyces cerevisiae by Using the Affinity of the Polyphosphate Binding Domain of Escherichia coli Exopolyphosphatase

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.71.10.5692-5701.2005

(A) SDS-PAGE analysis of purified PPBD. Standards and samples were fractionated by 10% SDS-PAGE. Lane 1, size standards (in kilodaltons); lane 2, purified PPBD. (B) Schematic illustration of polyP labeling for transmission electron microscopic observation. polyP in ultrathin sections was treated with PPBD linked with an Xpress epitope tag. The epitope tag was detected by indirect immunogold labeling using anti-Xpress epitope antibody and secondary antibody conjugated with colloidal gold. For laser scanning confocal microscopic observation, the secondary antibody was conjugated with Alexa 488 instead of colloidal gold.
Figure Legend Snippet: (A) SDS-PAGE analysis of purified PPBD. Standards and samples were fractionated by 10% SDS-PAGE. Lane 1, size standards (in kilodaltons); lane 2, purified PPBD. (B) Schematic illustration of polyP labeling for transmission electron microscopic observation. polyP in ultrathin sections was treated with PPBD linked with an Xpress epitope tag. The epitope tag was detected by indirect immunogold labeling using anti-Xpress epitope antibody and secondary antibody conjugated with colloidal gold. For laser scanning confocal microscopic observation, the secondary antibody was conjugated with Alexa 488 instead of colloidal gold.

Techniques Used: SDS Page, Purification, Labeling, Transmission Assay

32) Product Images from "Calcium-dependent proteasome activation is required for axonal neurofilament degradation"

Article Title: Calcium-dependent proteasome activation is required for axonal neurofilament degradation

Journal: Neural Regeneration Research

doi: 10.3969/j.issn.1673-5374.2013.36.005

Calcium and the proteasome are late effectors in axonal degeneration. (A) Representative ultrathin longitudinal cross-sections showing protection against axonal degeneration by MG132 (20 μmol/L) and calpeptin (50 μmol/L) in sciatic nerve explant cultures. Arrowheads: Microtubules. Scale bars: 0.2 μm. (B) Mean diameters (μm) of mitochondria in degenerating axons. (C) Mean length index (length/diameter) of mitochondria in degenerating axons. (D) The decrease of NAD and ATP levels in sciatic nerve explants cultured for 1 day could not be rescued by MG132. (B–D) Data were expressed as mean ± SD ( n = 3). At least three independent experiments were performed for each condition. a P
Figure Legend Snippet: Calcium and the proteasome are late effectors in axonal degeneration. (A) Representative ultrathin longitudinal cross-sections showing protection against axonal degeneration by MG132 (20 μmol/L) and calpeptin (50 μmol/L) in sciatic nerve explant cultures. Arrowheads: Microtubules. Scale bars: 0.2 μm. (B) Mean diameters (μm) of mitochondria in degenerating axons. (C) Mean length index (length/diameter) of mitochondria in degenerating axons. (D) The decrease of NAD and ATP levels in sciatic nerve explants cultured for 1 day could not be rescued by MG132. (B–D) Data were expressed as mean ± SD ( n = 3). At least three independent experiments were performed for each condition. a P

Techniques Used: Cell Culture

33) Product Images from "Multivesicular bodies in developing tobacco seed and mung bean are functionally equivalent"

Article Title: Multivesicular bodies in developing tobacco seed and mung bean are functionally equivalent

Journal: Plant Signaling & Behavior

doi: 10.4161/psb.19524

Figure 2. Immunogold EM and ultrastructural analysis of MVB in developing tobacco seeds. (A and B) Developing tobacco seeds were fixed by the indicated methods and embedded in HM20 resin. Ultrathin sections were prepared and labeled with indicated
Figure Legend Snippet: Figure 2. Immunogold EM and ultrastructural analysis of MVB in developing tobacco seeds. (A and B) Developing tobacco seeds were fixed by the indicated methods and embedded in HM20 resin. Ultrathin sections were prepared and labeled with indicated

Techniques Used: Labeling

34) Product Images from "Origin of fundus hyperautofluorescent spots and their role in retinal degeneration in a mouse model of Goldmann-Favre syndrome"

Article Title: Origin of fundus hyperautofluorescent spots and their role in retinal degeneration in a mouse model of Goldmann-Favre syndrome

Journal: Disease Models & Mechanisms

doi: 10.1242/dmm.012112

Histology and electron microscopy of P60 rd7 mouse retinas compared with P60 wild-type mouse retinas. (A) Semi-thin section of an rd7 mouse retina showing outer nuclear layer foldings. (B) Control: semi-thin section of a WT mouse retina. INL, inner nuclear layer; ONL, outer nuclear layer. (C,D) Ultrathin section of retinas of rd7 and WT mice, showing the junction between the outer segment (OS) and RPE. (E,F) The ultrastructural analysis of the retina of rd7 mice revealed the presence of microglial cells between the OS and the RPE. Asterisks indicate microglia. (G) Higher magnification of the boxed area from panel F, showing the accumulation of lysosomes inside a microglial cell (arrow).
Figure Legend Snippet: Histology and electron microscopy of P60 rd7 mouse retinas compared with P60 wild-type mouse retinas. (A) Semi-thin section of an rd7 mouse retina showing outer nuclear layer foldings. (B) Control: semi-thin section of a WT mouse retina. INL, inner nuclear layer; ONL, outer nuclear layer. (C,D) Ultrathin section of retinas of rd7 and WT mice, showing the junction between the outer segment (OS) and RPE. (E,F) The ultrastructural analysis of the retina of rd7 mice revealed the presence of microglial cells between the OS and the RPE. Asterisks indicate microglia. (G) Higher magnification of the boxed area from panel F, showing the accumulation of lysosomes inside a microglial cell (arrow).

Techniques Used: Electron Microscopy, Mouse Assay

35) Product Images from "Virus-Infection or 5?ppp-RNA Activates Antiviral Signal through Redistribution of IPS-1 Mediated by MFN1"

Article Title: Virus-Infection or 5?ppp-RNA Activates Antiviral Signal through Redistribution of IPS-1 Mediated by MFN1

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1001012

Localization of IPS-1 and mitochondria. A , IPS-1-HeLa cells infected with NDV for 9 h were fixed, stained with anti-NP antibody, and subjected to ultrathin sectioning as shown in the Methods . The area enclosed by a red rectangle is enlarged. NP: NP foci stained with the anti-NP antibody were visualized using gold particles. B , IPS-1-HeLa cells infected with NDV for 9 h were fixed, stained with anti-FLAG antibody, and subjected to ultra thin sectioning. The area enclosed by a red rectangle is enlarged. NP: morphologically similar structures are in A . IPS-1 was visualized using gold particles. The arrowheads indicate boundaries between IPS-1 and NP foci.
Figure Legend Snippet: Localization of IPS-1 and mitochondria. A , IPS-1-HeLa cells infected with NDV for 9 h were fixed, stained with anti-NP antibody, and subjected to ultrathin sectioning as shown in the Methods . The area enclosed by a red rectangle is enlarged. NP: NP foci stained with the anti-NP antibody were visualized using gold particles. B , IPS-1-HeLa cells infected with NDV for 9 h were fixed, stained with anti-FLAG antibody, and subjected to ultra thin sectioning. The area enclosed by a red rectangle is enlarged. NP: morphologically similar structures are in A . IPS-1 was visualized using gold particles. The arrowheads indicate boundaries between IPS-1 and NP foci.

Techniques Used: Infection, Staining, Thin Sectioning

36) Product Images from "A novel ADOA-associated OPA1 mutation alters the mitochondrial function, membrane potential, ROS production and apoptosis"

Article Title: A novel ADOA-associated OPA1 mutation alters the mitochondrial function, membrane potential, ROS production and apoptosis

Journal: Scientific Reports

doi: 10.1038/s41598-017-05571-y

The OPA1 mutation altered the fragmentation of the mitochondrial network. ( a ) Mitochondrial morphology was visualized by immunofluorescent labeling with MitoTracker (red) and was analyzed by confocal microscopy using lymphoblastoid cell lines. DAPI-stained nuclei were identified by their blue fluorescence. Internal control cell (III-12) lacking the OPA1 mutation exhibited a balanced mitochondrial network of the filamentous and fragmented states. Cells carrying the OPA1 mutation (III-11) displayed a distinct punctate pattern of mitochodnria. ( b ) Mitochondrial networks from patient and control cells were examined by transmission electron microscopy. Ultrathin sections were stained with uranyl acetate and alkaline lead citrate. 25,000X and 40,000X magnifications were used. Control lymphoblastoid cell lines (III-12) have tubular distribution of mitochondria. Patient-derived lymphoblastoid cell lines (III-11) have spotty distribution of mitochondria.
Figure Legend Snippet: The OPA1 mutation altered the fragmentation of the mitochondrial network. ( a ) Mitochondrial morphology was visualized by immunofluorescent labeling with MitoTracker (red) and was analyzed by confocal microscopy using lymphoblastoid cell lines. DAPI-stained nuclei were identified by their blue fluorescence. Internal control cell (III-12) lacking the OPA1 mutation exhibited a balanced mitochondrial network of the filamentous and fragmented states. Cells carrying the OPA1 mutation (III-11) displayed a distinct punctate pattern of mitochodnria. ( b ) Mitochondrial networks from patient and control cells were examined by transmission electron microscopy. Ultrathin sections were stained with uranyl acetate and alkaline lead citrate. 25,000X and 40,000X magnifications were used. Control lymphoblastoid cell lines (III-12) have tubular distribution of mitochondria. Patient-derived lymphoblastoid cell lines (III-11) have spotty distribution of mitochondria.

Techniques Used: Mutagenesis, Labeling, Confocal Microscopy, Staining, Fluorescence, Transmission Assay, Electron Microscopy, Derivative Assay

37) Product Images from "Testicular regulation of seasonal change in apocrine glands in the back skin of the brown bear (Ursus arctos)"

Article Title: Testicular regulation of seasonal change in apocrine glands in the back skin of the brown bear (Ursus arctos)

Journal: The Journal of Veterinary Medical Science

doi: 10.1292/jvms.17-0689

Localization of VVA reaction in apocrine cells of castrated males during the breeding season. Semi-thin sections with VVA staining (A). Square (A) corresponds to panel (B). Arrowheads, positive VVA staining. Transmission electron microscopy (TEM) image of ultrathin section (B) adjacent to panel (A). TEM image without VVA staining from same individual (C). Square (B) and (C) correspond to panel (D) and (E), respectively. Asterisk, secretory granule. G, Golgi apparatus. Scale bar: 5 µ m.
Figure Legend Snippet: Localization of VVA reaction in apocrine cells of castrated males during the breeding season. Semi-thin sections with VVA staining (A). Square (A) corresponds to panel (B). Arrowheads, positive VVA staining. Transmission electron microscopy (TEM) image of ultrathin section (B) adjacent to panel (A). TEM image without VVA staining from same individual (C). Square (B) and (C) correspond to panel (D) and (E), respectively. Asterisk, secretory granule. G, Golgi apparatus. Scale bar: 5 µ m.

Techniques Used: Staining, Transmission Assay, Electron Microscopy, Transmission Electron Microscopy

38) Product Images from "Stylet Morphometrics and Citrus Leaf Vein Structure in Relation to Feeding Behavior of the Asian Citrus Psyllid Diaphorina citri, Vector of Citrus Huanglongbing Bacterium"

Article Title: Stylet Morphometrics and Citrus Leaf Vein Structure in Relation to Feeding Behavior of the Asian Citrus Psyllid Diaphorina citri, Vector of Citrus Huanglongbing Bacterium

Journal: PLoS ONE

doi: 10.1371/journal.pone.0059914

Electron micrographs of ultrathin cross sections in the stylets of a psyllid adult (A), stylets of 1 st instar nymph (B), and in the phloem of a LAS-infected citrus leaf (C). Unlabeled arrows (in C) indicate bacterial structures found in the phloem of infected leaves. Abbreviations: d, dendrites; fc, food canal; md1 2, mandibular stylets 1 2; mx1 2, maxillary stylets 1 2; sc, salivary canal.
Figure Legend Snippet: Electron micrographs of ultrathin cross sections in the stylets of a psyllid adult (A), stylets of 1 st instar nymph (B), and in the phloem of a LAS-infected citrus leaf (C). Unlabeled arrows (in C) indicate bacterial structures found in the phloem of infected leaves. Abbreviations: d, dendrites; fc, food canal; md1 2, mandibular stylets 1 2; mx1 2, maxillary stylets 1 2; sc, salivary canal.

Techniques Used: Infection

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Electron Microscopy:

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Microscopy:

Article Title: Activated N
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Article Title: HS1 Is Involved in Hygromycin Resistance Through Facilitating Hygromycin Phosphotransferase Transportation From Cytosol to Chloroplast
Article Snippet: .. Immunized ultrathin sections were washed, post-stained, and examined with a transmission electron microscope (Hitachi H-7650, Japan). .. Identification of Hygromycin B-Sensitive Mutants We previously developed a rolling-circle amplification-based method for constructing libraries of long hpRNA gene-silencing constructs targeting all expressed genes , and obtained over 5,000 hpRNA transgenic Arabidopsis lines using HyB as the selective agent.

Article Title: Co-transmission of acetylcholine and GABA regulates hippocampal states
Article Snippet: .. Ultrathin sections were examined in a Hitachi 7100 electron microscope equipped with a Veleta CCD camera (Olympus Soft Imaging Solutions, Germany). .. For CLSM analysis coverslips were washed in PB and Tris-buffered saline (TBS).

Imaging:

Article Title: Co-transmission of acetylcholine and GABA regulates hippocampal states
Article Snippet: .. Ultrathin sections were examined in a Hitachi 7100 electron microscope equipped with a Veleta CCD camera (Olympus Soft Imaging Solutions, Germany). .. For CLSM analysis coverslips were washed in PB and Tris-buffered saline (TBS).

Transmission Assay:

Article Title: Activated N
Article Snippet: .. Ultrathin sections were placed on copper grids, stained with uranyl acetate and lead citrate, and images were acquired using a Hitachi 600 transmission electron microscope ( ). .. Electron microscopy (EM) negatives were digitized using an ArtixScan1100 scanner (Microtek, Carson, CA).

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Article Title: Three-dimensional reconstruction of electron micrographs reveals intrabulbar circuit differences between accessory and main olfactory bulbs
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Staining:

Article Title: Activated N
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Article Title: Three-dimensional reconstruction of electron micrographs reveals intrabulbar circuit differences between accessory and main olfactory bulbs
Article Snippet: .. After staining with uranyl acetate and lead citrate, ultrathin sections were observed under a transmission electron microscope (H-7650, HITACHI). .. Electron micrographs were consecutively aligned using Photoshop (Adobe) to identify the synaptic types on labeled dendrites.

Transmission Electron Microscopy:

Article Title: Derlin-1 Regulates Mutant VCP-Linked Pathogenesis and Endoplasmic Reticulum Stress-Induced Apoptosis
Article Snippet: .. For transmission electron microscopy, samples and ultrathin sections were prepared as previously described , and imaged with HT7700 TEM (Hitachi). .. Supporting Information Knockdown of endogenous Sip3 and Ufd1 levels with the expression of RNAi constructs. (A) RT-PCR and (B) Western blotting results show the knockdown efficacy of (A) sip3 and (B) ufd1 RNAi lines used in .

Article Title: Vaccinia Virus Mutations in the L4R Gene Encoding a Virion Structural Protein Produce Abnormal Mature Particles Lacking a Nucleocapsid
Article Snippet: .. Ultrathin sections (70 to 80 nm) were poststained with 2% uranyl acetate and lead citrate and examined with an H-7000 transmission electron microscope (TEM; Hitachi High Technologies America, Inc., Schaumburg, IL) operated at 100 kV. ..

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    Hitachi Ltd ultrathin section
    Changes in the ultrastructure of the bile canaliculi in porcine hepatocytes after warm ischemia. (A and B) Representative hepatocytes and bile canaliculi were observed by SEM in osmium-macerated porcine liver graft samples after warm ischemia for 60 minutes. The partial area indicated in A was further photographed at a higher magnification (B). Bile canaliculi are colored green, nuclei are colored blue, huge vacuoles are colored red. Asterisks indicate vacant space without any other endomembrane organelles around the bile canaliculi. (C) Typical bile canaliculi were identified in the <t>ultrathin</t> sections of Epon 812-embedded tissues from liver graft samples after warm ischemia for 60 minutes. Bile canaliculi are colored green and asterisks indicate the vacant space without any other endomembrane organelles around the bile canaliculi. Bars = 1 μm.
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    Changes in the ultrastructure of the bile canaliculi in porcine hepatocytes after warm ischemia. (A and B) Representative hepatocytes and bile canaliculi were observed by SEM in osmium-macerated porcine liver graft samples after warm ischemia for 60 minutes. The partial area indicated in A was further photographed at a higher magnification (B). Bile canaliculi are colored green, nuclei are colored blue, huge vacuoles are colored red. Asterisks indicate vacant space without any other endomembrane organelles around the bile canaliculi. (C) Typical bile canaliculi were identified in the ultrathin sections of Epon 812-embedded tissues from liver graft samples after warm ischemia for 60 minutes. Bile canaliculi are colored green and asterisks indicate the vacant space without any other endomembrane organelles around the bile canaliculi. Bars = 1 μm.

    Journal: PLoS ONE

    Article Title: The ultrastructural characteristics of bile canaliculus in porcine liver donated after cardiac death and machine perfusion preservation

    doi: 10.1371/journal.pone.0233917

    Figure Lengend Snippet: Changes in the ultrastructure of the bile canaliculi in porcine hepatocytes after warm ischemia. (A and B) Representative hepatocytes and bile canaliculi were observed by SEM in osmium-macerated porcine liver graft samples after warm ischemia for 60 minutes. The partial area indicated in A was further photographed at a higher magnification (B). Bile canaliculi are colored green, nuclei are colored blue, huge vacuoles are colored red. Asterisks indicate vacant space without any other endomembrane organelles around the bile canaliculi. (C) Typical bile canaliculi were identified in the ultrathin sections of Epon 812-embedded tissues from liver graft samples after warm ischemia for 60 minutes. Bile canaliculi are colored green and asterisks indicate the vacant space without any other endomembrane organelles around the bile canaliculi. Bars = 1 μm.

    Article Snippet: Ultrathin section (80 nm thick) were cut, stained with uranyl acetate and lead citrate, and observed using an HT7700 transmission electron microscope (Hitachi High Technologies, Tokyo, Japan).

    Techniques:

    The ultrastructure of the bile canaliculi in porcine hepatocytes of the control liver. (A and B) Representative hepatocytes and bile canaliculi were observed by SEM in osmium-macerated control porcine liver graft samples. The partial area indicated in A was further photographed at a higher magnification (B). (C) Typical bile canaliculi were identified in the ultrathin sections of the Epon 812-embedded control liver tissue. Bile canaliculi are colored green. Arrows indicate the Golgi apparatus, and asterisks indicate vacant spaces without any other endomembrane organelles around the bile canaliculi. Bars = 1 μm.

    Journal: PLoS ONE

    Article Title: The ultrastructural characteristics of bile canaliculus in porcine liver donated after cardiac death and machine perfusion preservation

    doi: 10.1371/journal.pone.0233917

    Figure Lengend Snippet: The ultrastructure of the bile canaliculi in porcine hepatocytes of the control liver. (A and B) Representative hepatocytes and bile canaliculi were observed by SEM in osmium-macerated control porcine liver graft samples. The partial area indicated in A was further photographed at a higher magnification (B). (C) Typical bile canaliculi were identified in the ultrathin sections of the Epon 812-embedded control liver tissue. Bile canaliculi are colored green. Arrows indicate the Golgi apparatus, and asterisks indicate vacant spaces without any other endomembrane organelles around the bile canaliculi. Bars = 1 μm.

    Article Snippet: Ultrathin section (80 nm thick) were cut, stained with uranyl acetate and lead citrate, and observed using an HT7700 transmission electron microscope (Hitachi High Technologies, Tokyo, Japan).

    Techniques:

    The ultrastructural changes of the bile canaliculi in porcine liver grafts preserved by HMP. (A and B) Representative hepatocytes and bile canaliculi were observed by SEM in osmium-macerated porcine liver graft samples preserved by HMP for 4 h after 60 minutes of warm ischemia. The partial area indicated in A was further photographed under higher magnification (B). Bile canaliculi are colored green, nuclei are colored blue. Asterisks indicate vacant space without any other endomembrane organelles around the bile canaliculi. (C) Typical bile canaliculi were identified in the ultrathin sections of Epon 812-embedded tissues from liver graft samples preserved by HMP for 4 h after 60 minutes of warm ischemia. Bile canaliculi are colored green. Asterisks indicate the vacant space without any other endomembrane organelles around the bile canaliculi. Bars = 1 μm.

    Journal: PLoS ONE

    Article Title: The ultrastructural characteristics of bile canaliculus in porcine liver donated after cardiac death and machine perfusion preservation

    doi: 10.1371/journal.pone.0233917

    Figure Lengend Snippet: The ultrastructural changes of the bile canaliculi in porcine liver grafts preserved by HMP. (A and B) Representative hepatocytes and bile canaliculi were observed by SEM in osmium-macerated porcine liver graft samples preserved by HMP for 4 h after 60 minutes of warm ischemia. The partial area indicated in A was further photographed under higher magnification (B). Bile canaliculi are colored green, nuclei are colored blue. Asterisks indicate vacant space without any other endomembrane organelles around the bile canaliculi. (C) Typical bile canaliculi were identified in the ultrathin sections of Epon 812-embedded tissues from liver graft samples preserved by HMP for 4 h after 60 minutes of warm ischemia. Bile canaliculi are colored green. Asterisks indicate the vacant space without any other endomembrane organelles around the bile canaliculi. Bars = 1 μm.

    Article Snippet: Ultrathin section (80 nm thick) were cut, stained with uranyl acetate and lead citrate, and observed using an HT7700 transmission electron microscope (Hitachi High Technologies, Tokyo, Japan).

    Techniques:

    The ultrastructural changes of the bile canaliculi in porcine liver grafts preserved by MMP. (A and B) Representative hepatocytes and bile canaliculi were observed by SEM in osmium-macerated porcine liver graft samples preserved by MMP for 4 h after 60 minutes of warm ischemia. The partial area indicated in A was further photographed under higher magnification (B). Bile canaliculi are colored green, nuclei are colored blue, huge vacuoles are colored red. Asterisks indicate vacant space without any other endomembrane organelles around the bile canaliculi. (C) Typical bile canaliculi were identified in the ultrathin sections of the Epon 812-embedded tissues from liver graft samples preserved by MMP for 4 h after 60 minutes of warm ischemia. Bile canaliculi are colored green. Asterisks indicate vacant space without any other endomembrane organelles around the bile canaliculi. Bars = 1 μm.

    Journal: PLoS ONE

    Article Title: The ultrastructural characteristics of bile canaliculus in porcine liver donated after cardiac death and machine perfusion preservation

    doi: 10.1371/journal.pone.0233917

    Figure Lengend Snippet: The ultrastructural changes of the bile canaliculi in porcine liver grafts preserved by MMP. (A and B) Representative hepatocytes and bile canaliculi were observed by SEM in osmium-macerated porcine liver graft samples preserved by MMP for 4 h after 60 minutes of warm ischemia. The partial area indicated in A was further photographed under higher magnification (B). Bile canaliculi are colored green, nuclei are colored blue, huge vacuoles are colored red. Asterisks indicate vacant space without any other endomembrane organelles around the bile canaliculi. (C) Typical bile canaliculi were identified in the ultrathin sections of the Epon 812-embedded tissues from liver graft samples preserved by MMP for 4 h after 60 minutes of warm ischemia. Bile canaliculi are colored green. Asterisks indicate vacant space without any other endomembrane organelles around the bile canaliculi. Bars = 1 μm.

    Article Snippet: Ultrathin section (80 nm thick) were cut, stained with uranyl acetate and lead citrate, and observed using an HT7700 transmission electron microscope (Hitachi High Technologies, Tokyo, Japan).

    Techniques:

    HS1 is involved in cytosol-to-chloroplast transport of HPT. Subcellular localization of HPT-GFP in the WT (A) and hs1-1 (B) backgrounds in Arabidopsis. HPT-GFP constructs were transformed transiently into plant cells. Arrowheads indicate higher chloroplast GFP fluorescence intensity in a mesophyll protoplast of WT compared to hs1-1 . Bars = 10 μm. (C , D) Western blot of HPT in WT/ HPT (empty vector transgenic lines), HS1 RNAi/ HPT , and hs1-1 / HPT (empty vector transformed into hs1-1 ) transgenic plants. HPT accumulation was reduced significantly or undetectable in HS1 RNAi/ HPT and hs1-1/HPT chloroplasts compared with the control (C) , while total HPT varied slightly (D) . The large subunit of RUBISCO (RBCL) was probed as a protein loading control. Approximately 20 μg (C) and 10 μg (D) of protein were loaded in each lane. Similar results were obtained in two additional independent experiments. (E–H) Transmission electron microscopic images of immunogold localization of HPT in transgenic Arabidopsis . HPT in ultrathin leaf sections reacted with anti-HPT antibody and a gold-conjugated secondary antibody. Gold particles were detected by transmission electron microscopy. Immunogold labeling of HPT in leaves from WT/ HPT (E) , hs1-1 / HPT (F) , hs1-1 / HPT / HS1 (G) , negative control (H) transgenic Arabidopsis plants, and negative control without anti-HPT antibody in blocking buffer showed no gold particles. Arrowheads show typical gold particles (10 nm, black dots). HPT present inside of chloroplasts (chl, the dark area) in WT / HPT , hs1-1 / HPT / HS1 , and outside of chloroplasts in hs1-1 / HPT . Three different leaf samples and more than 30 immunogold-labeled positive cells were observed with similar results. Images represent typical observations in different leaf samples. chl, chloroplast; cyt, cytosol; va, vacuole; cw, cell wall. Bars = 1 μm.

    Journal: Frontiers in Plant Science

    Article Title: HS1 Is Involved in Hygromycin Resistance Through Facilitating Hygromycin Phosphotransferase Transportation From Cytosol to Chloroplast

    doi: 10.3389/fpls.2020.00613

    Figure Lengend Snippet: HS1 is involved in cytosol-to-chloroplast transport of HPT. Subcellular localization of HPT-GFP in the WT (A) and hs1-1 (B) backgrounds in Arabidopsis. HPT-GFP constructs were transformed transiently into plant cells. Arrowheads indicate higher chloroplast GFP fluorescence intensity in a mesophyll protoplast of WT compared to hs1-1 . Bars = 10 μm. (C , D) Western blot of HPT in WT/ HPT (empty vector transgenic lines), HS1 RNAi/ HPT , and hs1-1 / HPT (empty vector transformed into hs1-1 ) transgenic plants. HPT accumulation was reduced significantly or undetectable in HS1 RNAi/ HPT and hs1-1/HPT chloroplasts compared with the control (C) , while total HPT varied slightly (D) . The large subunit of RUBISCO (RBCL) was probed as a protein loading control. Approximately 20 μg (C) and 10 μg (D) of protein were loaded in each lane. Similar results were obtained in two additional independent experiments. (E–H) Transmission electron microscopic images of immunogold localization of HPT in transgenic Arabidopsis . HPT in ultrathin leaf sections reacted with anti-HPT antibody and a gold-conjugated secondary antibody. Gold particles were detected by transmission electron microscopy. Immunogold labeling of HPT in leaves from WT/ HPT (E) , hs1-1 / HPT (F) , hs1-1 / HPT / HS1 (G) , negative control (H) transgenic Arabidopsis plants, and negative control without anti-HPT antibody in blocking buffer showed no gold particles. Arrowheads show typical gold particles (10 nm, black dots). HPT present inside of chloroplasts (chl, the dark area) in WT / HPT , hs1-1 / HPT / HS1 , and outside of chloroplasts in hs1-1 / HPT . Three different leaf samples and more than 30 immunogold-labeled positive cells were observed with similar results. Images represent typical observations in different leaf samples. chl, chloroplast; cyt, cytosol; va, vacuole; cw, cell wall. Bars = 1 μm.

    Article Snippet: Immunized ultrathin sections were washed, post-stained, and examined with a transmission electron microscope (Hitachi H-7650, Japan).

    Techniques: Construct, Transformation Assay, Fluorescence, Western Blot, Plasmid Preparation, Transgenic Assay, Transmission Assay, Electron Microscopy, Labeling, Negative Control, Blocking Assay

    Immunogold labeling of core proteins. Cells were infected with WR or Ets85 at an MOI of 10 and incubated at 31°C or 39.7°C. At 24 h postinfection, cells were processed for immunoelectron microscopy. Ultrathin sections were probed with

    Journal: Journal of Virology

    Article Title: Vaccinia Virus Mutations in the L4R Gene Encoding a Virion Structural Protein Produce Abnormal Mature Particles Lacking a Nucleocapsid

    doi: 10.1128/JVI.02126-14

    Figure Lengend Snippet: Immunogold labeling of core proteins. Cells were infected with WR or Ets85 at an MOI of 10 and incubated at 31°C or 39.7°C. At 24 h postinfection, cells were processed for immunoelectron microscopy. Ultrathin sections were probed with

    Article Snippet: Ultrathin sections (70 to 80 nm) were poststained with 2% uranyl acetate and lead citrate and examined with an H-7000 transmission electron microscope (TEM; Hitachi High Technologies America, Inc., Schaumburg, IL) operated at 100 kV.

    Techniques: Labeling, Infection, Incubation, Immuno-Electron Microscopy

    A 3D image of an AOB Gr cell. (A) A drawing of a prepared rat AOB slice (upper panel) and the position of a biocytin-injected Gr cell (red cross, upper panel), and a joint image of differential interference and fluorescence of the slice reacted with avidin-FITC-gold (lower panel). Scale bar = 200 μm. (B) A confocal fluorescent image of the injected cell. The red dotted line indicates the border between the MTL, the olfactory tract layer (OTL), and the GRL. The red box indicates the area observed with electron microscopy corresponding to the 3D image of C . Scale bar = 50 μm. (C) 3D reconstruction of the apical dendrite in B by serial electron micrograph. The green is the injected dendrite, and the yellow structures indicate the neuron's dendritic spines. The white structures are filopodia-like structures. Arrows indicate synaptic transmission direction. iSSs (mainly excitatory synapses) are labeled in red, oSSs (all inhibitory synapses) are labeled in blue, and bidirectional arrows indicate RSs. Orange boxes indicate the areas shown in D,E . Scale bar = 10 μm. (D and E) An example of 3D morphology of the spine containing the iSS (iSS3 in C ) and the gemmule containing the RS (RS10 in C ). The 3D image (left panels) was reconstructed with the serial electron micrograph of a labeled spine by silver-enhances gold particles (right panels). Numbers on the right panels indicate ultrathin section sequence. The asterisk indicates the same section of the low-magnification panel (upper). Arrows indicate synaptic transmission direction. Red and blue areas indicate the locations and sizes of synaptic sites (red, PSD of the labeled cell; blue, PSD of a paired MT cell). Ax: axon-like structure. White scale bar = 500 nm (in D ) and 1 μm (in E ). Black scale bar = 500 nm (in D ), 1 μm (low-magnification photo in E ), and 500 nm (high-magnification photos in E ).

    Journal: Frontiers in Neuroanatomy

    Article Title: Three-dimensional reconstruction of electron micrographs reveals intrabulbar circuit differences between accessory and main olfactory bulbs

    doi: 10.3389/fnana.2013.00005

    Figure Lengend Snippet: A 3D image of an AOB Gr cell. (A) A drawing of a prepared rat AOB slice (upper panel) and the position of a biocytin-injected Gr cell (red cross, upper panel), and a joint image of differential interference and fluorescence of the slice reacted with avidin-FITC-gold (lower panel). Scale bar = 200 μm. (B) A confocal fluorescent image of the injected cell. The red dotted line indicates the border between the MTL, the olfactory tract layer (OTL), and the GRL. The red box indicates the area observed with electron microscopy corresponding to the 3D image of C . Scale bar = 50 μm. (C) 3D reconstruction of the apical dendrite in B by serial electron micrograph. The green is the injected dendrite, and the yellow structures indicate the neuron's dendritic spines. The white structures are filopodia-like structures. Arrows indicate synaptic transmission direction. iSSs (mainly excitatory synapses) are labeled in red, oSSs (all inhibitory synapses) are labeled in blue, and bidirectional arrows indicate RSs. Orange boxes indicate the areas shown in D,E . Scale bar = 10 μm. (D and E) An example of 3D morphology of the spine containing the iSS (iSS3 in C ) and the gemmule containing the RS (RS10 in C ). The 3D image (left panels) was reconstructed with the serial electron micrograph of a labeled spine by silver-enhances gold particles (right panels). Numbers on the right panels indicate ultrathin section sequence. The asterisk indicates the same section of the low-magnification panel (upper). Arrows indicate synaptic transmission direction. Red and blue areas indicate the locations and sizes of synaptic sites (red, PSD of the labeled cell; blue, PSD of a paired MT cell). Ax: axon-like structure. White scale bar = 500 nm (in D ) and 1 μm (in E ). Black scale bar = 500 nm (in D ), 1 μm (low-magnification photo in E ), and 500 nm (high-magnification photos in E ).

    Article Snippet: After staining with uranyl acetate and lead citrate, ultrathin sections were observed under a transmission electron microscope (H-7650, HITACHI).

    Techniques: Injection, Fluorescence, Avidin-Biotin Assay, Electron Microscopy, Transmission Assay, Labeling, Sequencing