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Collaborative Drug Discovery Inc ultrathin sections
ORP5 localize at ER-mitochondria contact sites near cristae junctions (CJ) (a) Electron micrographs of <t>ultrathin</t> cryosections of HeLa cells transfected with HA-ORP5 or Mitofilin-EGFP and immunogold stained with anti-HA or anti-GFP (10 or 15 nm gold), showing ORP5 localization at ER-mitochondria contacts in close proximity to CJ (arrow) and the localization of the MICOS complex (Mitofilin) at CJ (arrows). Scale bar 250 nm. (b) Quantification of the proximity of HA-ORP5 and EGFP-Sec61β gold particles to the CJ. Results are presented as the percentage of ORP5 or Sec61β gold particles (n = 23 cell profiles) at specific ranges of distance (in nm) from CJ. (c) Electron micrographs of ultrathin cryosections of HeLa cells transfected with EGFP-ORP5 or GFP-Sec61β and immunogold labeled with anti-GFP (15 nm gold) and anti-PDI (10nm gold). Note ORP5 localization at ER-mitochondria contacts near CJ (arrow) and Sec61β localization to ER membranes not in contact with the mitochondria membranes (arrowheads). Scale bar 250 nm.
Ultrathin Sections, supplied by Collaborative Drug Discovery Inc, used in various techniques. Bioz Stars score: 92/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "ORP5 Transfers Phosphatidylserine To Mitochondria And Regulates Mitochondrial Calcium Uptake At Endoplasmic Reticulum - Mitochondria Contact Sites"

Article Title: ORP5 Transfers Phosphatidylserine To Mitochondria And Regulates Mitochondrial Calcium Uptake At Endoplasmic Reticulum - Mitochondria Contact Sites

Journal: bioRxiv

doi: 10.1101/695577

ORP5 localize at ER-mitochondria contact sites near cristae junctions (CJ) (a) Electron micrographs of ultrathin cryosections of HeLa cells transfected with HA-ORP5 or Mitofilin-EGFP and immunogold stained with anti-HA or anti-GFP (10 or 15 nm gold), showing ORP5 localization at ER-mitochondria contacts in close proximity to CJ (arrow) and the localization of the MICOS complex (Mitofilin) at CJ (arrows). Scale bar 250 nm. (b) Quantification of the proximity of HA-ORP5 and EGFP-Sec61β gold particles to the CJ. Results are presented as the percentage of ORP5 or Sec61β gold particles (n = 23 cell profiles) at specific ranges of distance (in nm) from CJ. (c) Electron micrographs of ultrathin cryosections of HeLa cells transfected with EGFP-ORP5 or GFP-Sec61β and immunogold labeled with anti-GFP (15 nm gold) and anti-PDI (10nm gold). Note ORP5 localization at ER-mitochondria contacts near CJ (arrow) and Sec61β localization to ER membranes not in contact with the mitochondria membranes (arrowheads). Scale bar 250 nm.
Figure Legend Snippet: ORP5 localize at ER-mitochondria contact sites near cristae junctions (CJ) (a) Electron micrographs of ultrathin cryosections of HeLa cells transfected with HA-ORP5 or Mitofilin-EGFP and immunogold stained with anti-HA or anti-GFP (10 or 15 nm gold), showing ORP5 localization at ER-mitochondria contacts in close proximity to CJ (arrow) and the localization of the MICOS complex (Mitofilin) at CJ (arrows). Scale bar 250 nm. (b) Quantification of the proximity of HA-ORP5 and EGFP-Sec61β gold particles to the CJ. Results are presented as the percentage of ORP5 or Sec61β gold particles (n = 23 cell profiles) at specific ranges of distance (in nm) from CJ. (c) Electron micrographs of ultrathin cryosections of HeLa cells transfected with EGFP-ORP5 or GFP-Sec61β and immunogold labeled with anti-GFP (15 nm gold) and anti-PDI (10nm gold). Note ORP5 localization at ER-mitochondria contacts near CJ (arrow) and Sec61β localization to ER membranes not in contact with the mitochondria membranes (arrowheads). Scale bar 250 nm.

Techniques Used: Transfection, Staining, Labeling

2) Product Images from "C1 Catecholamine Neurons form Local Circuit Synaptic Connections Within the Rostroventrolateral Medulla of Rat"

Article Title: C1 Catecholamine Neurons form Local Circuit Synaptic Connections Within the Rostroventrolateral Medulla of Rat

Journal: Neuroscience

doi: 10.1016/j.neuroscience.2012.09.049

The experimental design incorporated in the TEM analysis and data typical of that collected for each case is illustrated. The relative position of the C1 cell group in the brainstem, as well as the four levels of analysis (L1, L2, L3 L4) selected for TEM sampling, are illustrated schematically in figure 1A. The approximate location of the C1 column is shown by the gray area in the sagittal schematic at the top of A; vertical lines extending through the rostrocaudal extent of the nucleus in that diagram illustrate the extent of coronal vibratome sections collected for analysis. The representation of the C1 cell column shown in the expansion at the bottom of A schematically illustrates the two projection-specific and phenotypically-defined populations of neurons known to populate this region. Filled red circles represent C1 reticulospinal neurons that are concentrated at rostral levels of the nucleus. The open red circles represent C1 neurons that colocalize neuropeptide Y, are concentrated caudally in cell column, and project to supraspinal targets. The filled grey circles represent non-C1 neurons that contribute to both pathways. In figure 1B neurons immunopositive for TH illustrate the position of the C1 column in the ventrolateral medulla and delineate the area cut from the vibratome section for TEM analysis. The lower portion of the schematic illustrates the sectioning strategy incorporated in the analysis to ensure precise localization of labeled profiles within the area of C1. Series of ultrathin sections bracketed by toluidine blue stained thick sections were collected on formvar-coated narrow slot copper grids. Landmarks ( e.g. ). AT = axon terminal, N = neuron cytoplasm, Nu = nucleus; asterisks identify blood vessels. Marker bars = 15 μm for A F, 3 μm for D G, and 0.5 μm for E H.
Figure Legend Snippet: The experimental design incorporated in the TEM analysis and data typical of that collected for each case is illustrated. The relative position of the C1 cell group in the brainstem, as well as the four levels of analysis (L1, L2, L3 L4) selected for TEM sampling, are illustrated schematically in figure 1A. The approximate location of the C1 column is shown by the gray area in the sagittal schematic at the top of A; vertical lines extending through the rostrocaudal extent of the nucleus in that diagram illustrate the extent of coronal vibratome sections collected for analysis. The representation of the C1 cell column shown in the expansion at the bottom of A schematically illustrates the two projection-specific and phenotypically-defined populations of neurons known to populate this region. Filled red circles represent C1 reticulospinal neurons that are concentrated at rostral levels of the nucleus. The open red circles represent C1 neurons that colocalize neuropeptide Y, are concentrated caudally in cell column, and project to supraspinal targets. The filled grey circles represent non-C1 neurons that contribute to both pathways. In figure 1B neurons immunopositive for TH illustrate the position of the C1 column in the ventrolateral medulla and delineate the area cut from the vibratome section for TEM analysis. The lower portion of the schematic illustrates the sectioning strategy incorporated in the analysis to ensure precise localization of labeled profiles within the area of C1. Series of ultrathin sections bracketed by toluidine blue stained thick sections were collected on formvar-coated narrow slot copper grids. Landmarks ( e.g. ). AT = axon terminal, N = neuron cytoplasm, Nu = nucleus; asterisks identify blood vessels. Marker bars = 15 μm for A F, 3 μm for D G, and 0.5 μm for E H.

Techniques Used: Transmission Electron Microscopy, Sampling, Labeling, Staining, Marker

3) Product Images from "Polarized Cell Migration during Cell-to-Cell Transmission of Herpes Simplex Virus in Human Skin Keratinocytes"

Article Title: Polarized Cell Migration during Cell-to-Cell Transmission of Herpes Simplex Virus in Human Skin Keratinocytes

Journal: Journal of Virology

doi: 10.1128/JVI.01172-13

Ultrastructural characterization of HSV plaques in HaCaT cells. Monolayers of HaCaT cells in 35-mm dishes were infected with 100 PFU of HSV-1[17]/well and processed at 24 h postinfection for TEM as described in Materials and Methods. (a) Low-power binocular image of the vertical face of the embedded resin block. This image gives a three-dimensional view along the horizontal surface of the plastic support. Arrows indicate the plaques of clustered piled-up cells, with the size of the arrow indicating the distance from the block front. (b) The block was then sectioned into semithin survey sections (500 to 1,000 nm) and stained with crystal violet. Images were collected on a Zeiss Axiovert 200M using a ×40 objective lens, and the panels were assembled using Adobe Photoshop. (c) Sequential ultrathin sections (50 to 70 nm) of the same sample were then obtained and collected onto slot grids, stained, and examined by TEM. Because the sections analyzed are immediately adjacent to the section stained with crystal violet, corresponding areas could be matched. The TEM images of the sections in panel c, sections 1 to 4, therefore correspond to the boxes in panel b, sections 1 to 4. Upper panels (lower magnification) and lower panels (higher magnification on inset area) show different features of infection within the plaque as discussed in the text. In the lower panels, the features include extracellular virions (black arrows with white outline), nucleocapsids (short black arrows), and cell tight junctions (white arrows). In the upper part of panels 3 and 4, cells with no distinct features of infection (e.g., no early chromatin marginalization) are indicated by long black arrows. Box 5 in panel b shows flat uninfected cells that correspond to the elongated cells (TEM data not shown).
Figure Legend Snippet: Ultrastructural characterization of HSV plaques in HaCaT cells. Monolayers of HaCaT cells in 35-mm dishes were infected with 100 PFU of HSV-1[17]/well and processed at 24 h postinfection for TEM as described in Materials and Methods. (a) Low-power binocular image of the vertical face of the embedded resin block. This image gives a three-dimensional view along the horizontal surface of the plastic support. Arrows indicate the plaques of clustered piled-up cells, with the size of the arrow indicating the distance from the block front. (b) The block was then sectioned into semithin survey sections (500 to 1,000 nm) and stained with crystal violet. Images were collected on a Zeiss Axiovert 200M using a ×40 objective lens, and the panels were assembled using Adobe Photoshop. (c) Sequential ultrathin sections (50 to 70 nm) of the same sample were then obtained and collected onto slot grids, stained, and examined by TEM. Because the sections analyzed are immediately adjacent to the section stained with crystal violet, corresponding areas could be matched. The TEM images of the sections in panel c, sections 1 to 4, therefore correspond to the boxes in panel b, sections 1 to 4. Upper panels (lower magnification) and lower panels (higher magnification on inset area) show different features of infection within the plaque as discussed in the text. In the lower panels, the features include extracellular virions (black arrows with white outline), nucleocapsids (short black arrows), and cell tight junctions (white arrows). In the upper part of panels 3 and 4, cells with no distinct features of infection (e.g., no early chromatin marginalization) are indicated by long black arrows. Box 5 in panel b shows flat uninfected cells that correspond to the elongated cells (TEM data not shown).

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

Related Articles

Staining:

Article Title: Polarized Cell Migration during Cell-to-Cell Transmission of Herpes Simplex Virus in Human Skin Keratinocytes
Article Snippet: .. The sample was sectioned again when appropriate transverse sections were observed, and ultrathin sections (typically 50 to 70 nm) were cut and collected onto slot grids, stained with Reynold's lead citrate examined in a FEI Tecnai electron microscope with CCD camera image acquisition. ..

Article Title: penner/lgl2 is required for the integrity of the photoreceptor layer in the zebrafish retina
Article Snippet: .. For TEM, ultrathin sections of 70 nm were stained with uranyl acetate and lead citrate, and imaged with a Morgagni TEM (80 kV) using a Morada CCD camera (EMSIS GmbH) and ITEM software (EMSIS GmbH). ..

Article Title: RNA-seq Analysis of Host and Viral Gene Expression Highlights Interaction between Varicella Zoster Virus and Keratinocyte Differentiation
Article Snippet: .. Ultrathin sections (typically 50–70 nm) were cut parallel to the dish stained with Reynold's lead citrate and examined in a FEI Tecnai electron microscope with CCD camera image acquisition. .. Plaque assays Plates were fixed at time points post infection with 4% paraformaldehyde and stained by immunohistochemistry using a mixed VZV mAb (Meridian Life Sciences, Memphis, US), followed by and biotin (Vector labs, Peterborough, UK) streptavidin (Jackson ImmunoResearch, PA, US) amplification.

Article Title: Megakaryocyte-derived microparticles: direct visualization and distinction from platelet-derived microparticles
Article Snippet: .. Ultrathin sections were stained with uranyl acetate and lead citrate, and examined with a Tecnai G2 Spirit BioTWIN transmission electron microscope (Hillsboro, OR) at an accelerating voltage of 80 kV, and images were recorded with an AMT 2k CCD camera (Danvers, MA). .. For analysis of murine platelet microparticles, blood was drawn from wild-type C57BL/6J mice by vena caval puncture and anticoagulated with acid citrate dextrose (ACD).

Software:

Article Title: penner/lgl2 is required for the integrity of the photoreceptor layer in the zebrafish retina
Article Snippet: .. For TEM, ultrathin sections of 70 nm were stained with uranyl acetate and lead citrate, and imaged with a Morgagni TEM (80 kV) using a Morada CCD camera (EMSIS GmbH) and ITEM software (EMSIS GmbH). ..

Article Title: TAT-Cx43266-283 impairs metabolic plasticity in glioma stem cells in vitro and in vivo
Article Snippet: .. Ultrathin sections were obtained with a Leica EM UC7 ultramicrotome, contrasted with uranyl acetate and lead citrate and analysed using a Tecnai Spirit Twin 120 kV electron microscope with a CCD Gatan Orius SC200D camera with DigitalMicrograph™ software. .. Procedures were performed at the Electron Microscopy Facilities-NUCLEUS of the University of Salamanca.

Microscopy:

Article Title: Polarized Cell Migration during Cell-to-Cell Transmission of Herpes Simplex Virus in Human Skin Keratinocytes
Article Snippet: .. The sample was sectioned again when appropriate transverse sections were observed, and ultrathin sections (typically 50 to 70 nm) were cut and collected onto slot grids, stained with Reynold's lead citrate examined in a FEI Tecnai electron microscope with CCD camera image acquisition. ..

Article Title: RNA-seq Analysis of Host and Viral Gene Expression Highlights Interaction between Varicella Zoster Virus and Keratinocyte Differentiation
Article Snippet: .. Ultrathin sections (typically 50–70 nm) were cut parallel to the dish stained with Reynold's lead citrate and examined in a FEI Tecnai electron microscope with CCD camera image acquisition. .. Plaque assays Plates were fixed at time points post infection with 4% paraformaldehyde and stained by immunohistochemistry using a mixed VZV mAb (Meridian Life Sciences, Memphis, US), followed by and biotin (Vector labs, Peterborough, UK) streptavidin (Jackson ImmunoResearch, PA, US) amplification.

Article Title: Megakaryocyte-derived microparticles: direct visualization and distinction from platelet-derived microparticles
Article Snippet: .. Ultrathin sections were stained with uranyl acetate and lead citrate, and examined with a Tecnai G2 Spirit BioTWIN transmission electron microscope (Hillsboro, OR) at an accelerating voltage of 80 kV, and images were recorded with an AMT 2k CCD camera (Danvers, MA). .. For analysis of murine platelet microparticles, blood was drawn from wild-type C57BL/6J mice by vena caval puncture and anticoagulated with acid citrate dextrose (ACD).

Article Title: ORP5 Transfers Phosphatidylserine To Mitochondria And Regulates Mitochondrial Calcium Uptake At Endoplasmic Reticulum - Mitochondria Contact Sites
Article Snippet: .. Ultrathin sections were counterstained with 2% uranyl acetate and observed under a FEI Tecnai 12 microscope equipped with a CCD (SiS 1kx1k keenView) camera. ..

Article Title: Herpes Simplex Virus 1 Enters Human Keratinocytes by a Nectin-1-Dependent, Rapid Plasma Membrane Fusion Pathway That Functions at Low Temperature
Article Snippet: .. Ultrathin sections (typically 50 to 70 nm) were cut parallel or perpendicular to the dish and examined in an FEI Tecnai electron microscope with charge-coupled-device (CCD) camera image acquisition. .. Protein samples were analyzed on 10% polyacrylamide gels and subjected to electrophoresis in Tris-glycine buffer.

Article Title: C1 Catecholamine Neurons form Local Circuit Synaptic Connections Within the Rostroventrolateral Medulla of Rat
Article Snippet: .. Ultrathin sections were analyzed unstained using a transmission electron microscope (TEM; Morgagni, FEI, Hillsboro, OR) equipped with a CCD camera (Advanced Microscopy Techniques, Danvers, MA). .. Insertion of objective apertures into the path of the electron beam provided the necessary contrast for identification of landmarks and labeled profiles.

Article Title: TAT-Cx43266-283 impairs metabolic plasticity in glioma stem cells in vitro and in vivo
Article Snippet: .. Ultrathin sections were obtained with a Leica EM UC7 ultramicrotome, contrasted with uranyl acetate and lead citrate and analysed using a Tecnai Spirit Twin 120 kV electron microscope with a CCD Gatan Orius SC200D camera with DigitalMicrograph™ software. .. Procedures were performed at the Electron Microscopy Facilities-NUCLEUS of the University of Salamanca.

Transmission Electron Microscopy:

Article Title: penner/lgl2 is required for the integrity of the photoreceptor layer in the zebrafish retina
Article Snippet: .. For TEM, ultrathin sections of 70 nm were stained with uranyl acetate and lead citrate, and imaged with a Morgagni TEM (80 kV) using a Morada CCD camera (EMSIS GmbH) and ITEM software (EMSIS GmbH). ..

Article Title: C1 Catecholamine Neurons form Local Circuit Synaptic Connections Within the Rostroventrolateral Medulla of Rat
Article Snippet: .. Ultrathin sections were analyzed unstained using a transmission electron microscope (TEM; Morgagni, FEI, Hillsboro, OR) equipped with a CCD camera (Advanced Microscopy Techniques, Danvers, MA). .. Insertion of objective apertures into the path of the electron beam provided the necessary contrast for identification of landmarks and labeled profiles.

Transmission Assay:

Article Title: Megakaryocyte-derived microparticles: direct visualization and distinction from platelet-derived microparticles
Article Snippet: .. Ultrathin sections were stained with uranyl acetate and lead citrate, and examined with a Tecnai G2 Spirit BioTWIN transmission electron microscope (Hillsboro, OR) at an accelerating voltage of 80 kV, and images were recorded with an AMT 2k CCD camera (Danvers, MA). .. For analysis of murine platelet microparticles, blood was drawn from wild-type C57BL/6J mice by vena caval puncture and anticoagulated with acid citrate dextrose (ACD).

Article Title: C1 Catecholamine Neurons form Local Circuit Synaptic Connections Within the Rostroventrolateral Medulla of Rat
Article Snippet: .. Ultrathin sections were analyzed unstained using a transmission electron microscope (TEM; Morgagni, FEI, Hillsboro, OR) equipped with a CCD camera (Advanced Microscopy Techniques, Danvers, MA). .. Insertion of objective apertures into the path of the electron beam provided the necessary contrast for identification of landmarks and labeled profiles.

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    Collaborative Drug Discovery Inc ultrathin sections
    ORP5 localize at ER-mitochondria contact sites near cristae junctions (CJ) (a) Electron micrographs of <t>ultrathin</t> cryosections of HeLa cells transfected with HA-ORP5 or Mitofilin-EGFP and immunogold stained with anti-HA or anti-GFP (10 or 15 nm gold), showing ORP5 localization at ER-mitochondria contacts in close proximity to CJ (arrow) and the localization of the MICOS complex (Mitofilin) at CJ (arrows). Scale bar 250 nm. (b) Quantification of the proximity of HA-ORP5 and EGFP-Sec61β gold particles to the CJ. Results are presented as the percentage of ORP5 or Sec61β gold particles (n = 23 cell profiles) at specific ranges of distance (in nm) from CJ. (c) Electron micrographs of ultrathin cryosections of HeLa cells transfected with EGFP-ORP5 or GFP-Sec61β and immunogold labeled with anti-GFP (15 nm gold) and anti-PDI (10nm gold). Note ORP5 localization at ER-mitochondria contacts near CJ (arrow) and Sec61β localization to ER membranes not in contact with the mitochondria membranes (arrowheads). Scale bar 250 nm.
    Ultrathin Sections, supplied by Collaborative Drug Discovery Inc, used in various techniques. Bioz Stars score: 92/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ultrathin sections/product/Collaborative Drug Discovery Inc
    Average 92 stars, based on 21 article reviews
    Price from $9.99 to $1999.99
    ultrathin sections - by Bioz Stars, 2020-08
    92/100 stars
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    ORP5 localize at ER-mitochondria contact sites near cristae junctions (CJ) (a) Electron micrographs of ultrathin cryosections of HeLa cells transfected with HA-ORP5 or Mitofilin-EGFP and immunogold stained with anti-HA or anti-GFP (10 or 15 nm gold), showing ORP5 localization at ER-mitochondria contacts in close proximity to CJ (arrow) and the localization of the MICOS complex (Mitofilin) at CJ (arrows). Scale bar 250 nm. (b) Quantification of the proximity of HA-ORP5 and EGFP-Sec61β gold particles to the CJ. Results are presented as the percentage of ORP5 or Sec61β gold particles (n = 23 cell profiles) at specific ranges of distance (in nm) from CJ. (c) Electron micrographs of ultrathin cryosections of HeLa cells transfected with EGFP-ORP5 or GFP-Sec61β and immunogold labeled with anti-GFP (15 nm gold) and anti-PDI (10nm gold). Note ORP5 localization at ER-mitochondria contacts near CJ (arrow) and Sec61β localization to ER membranes not in contact with the mitochondria membranes (arrowheads). Scale bar 250 nm.

    Journal: bioRxiv

    Article Title: ORP5 Transfers Phosphatidylserine To Mitochondria And Regulates Mitochondrial Calcium Uptake At Endoplasmic Reticulum - Mitochondria Contact Sites

    doi: 10.1101/695577

    Figure Lengend Snippet: ORP5 localize at ER-mitochondria contact sites near cristae junctions (CJ) (a) Electron micrographs of ultrathin cryosections of HeLa cells transfected with HA-ORP5 or Mitofilin-EGFP and immunogold stained with anti-HA or anti-GFP (10 or 15 nm gold), showing ORP5 localization at ER-mitochondria contacts in close proximity to CJ (arrow) and the localization of the MICOS complex (Mitofilin) at CJ (arrows). Scale bar 250 nm. (b) Quantification of the proximity of HA-ORP5 and EGFP-Sec61β gold particles to the CJ. Results are presented as the percentage of ORP5 or Sec61β gold particles (n = 23 cell profiles) at specific ranges of distance (in nm) from CJ. (c) Electron micrographs of ultrathin cryosections of HeLa cells transfected with EGFP-ORP5 or GFP-Sec61β and immunogold labeled with anti-GFP (15 nm gold) and anti-PDI (10nm gold). Note ORP5 localization at ER-mitochondria contacts near CJ (arrow) and Sec61β localization to ER membranes not in contact with the mitochondria membranes (arrowheads). Scale bar 250 nm.

    Article Snippet: Ultrathin sections were counterstained with 2% uranyl acetate and observed under a FEI Tecnai 12 microscope equipped with a CCD (SiS 1kx1k keenView) camera.

    Techniques: Transfection, Staining, Labeling

    The experimental design incorporated in the TEM analysis and data typical of that collected for each case is illustrated. The relative position of the C1 cell group in the brainstem, as well as the four levels of analysis (L1, L2, L3 L4) selected for TEM sampling, are illustrated schematically in figure 1A. The approximate location of the C1 column is shown by the gray area in the sagittal schematic at the top of A; vertical lines extending through the rostrocaudal extent of the nucleus in that diagram illustrate the extent of coronal vibratome sections collected for analysis. The representation of the C1 cell column shown in the expansion at the bottom of A schematically illustrates the two projection-specific and phenotypically-defined populations of neurons known to populate this region. Filled red circles represent C1 reticulospinal neurons that are concentrated at rostral levels of the nucleus. The open red circles represent C1 neurons that colocalize neuropeptide Y, are concentrated caudally in cell column, and project to supraspinal targets. The filled grey circles represent non-C1 neurons that contribute to both pathways. In figure 1B neurons immunopositive for TH illustrate the position of the C1 column in the ventrolateral medulla and delineate the area cut from the vibratome section for TEM analysis. The lower portion of the schematic illustrates the sectioning strategy incorporated in the analysis to ensure precise localization of labeled profiles within the area of C1. Series of ultrathin sections bracketed by toluidine blue stained thick sections were collected on formvar-coated narrow slot copper grids. Landmarks ( e.g. ). AT = axon terminal, N = neuron cytoplasm, Nu = nucleus; asterisks identify blood vessels. Marker bars = 15 μm for A F, 3 μm for D G, and 0.5 μm for E H.

    Journal: Neuroscience

    Article Title: C1 Catecholamine Neurons form Local Circuit Synaptic Connections Within the Rostroventrolateral Medulla of Rat

    doi: 10.1016/j.neuroscience.2012.09.049

    Figure Lengend Snippet: The experimental design incorporated in the TEM analysis and data typical of that collected for each case is illustrated. The relative position of the C1 cell group in the brainstem, as well as the four levels of analysis (L1, L2, L3 L4) selected for TEM sampling, are illustrated schematically in figure 1A. The approximate location of the C1 column is shown by the gray area in the sagittal schematic at the top of A; vertical lines extending through the rostrocaudal extent of the nucleus in that diagram illustrate the extent of coronal vibratome sections collected for analysis. The representation of the C1 cell column shown in the expansion at the bottom of A schematically illustrates the two projection-specific and phenotypically-defined populations of neurons known to populate this region. Filled red circles represent C1 reticulospinal neurons that are concentrated at rostral levels of the nucleus. The open red circles represent C1 neurons that colocalize neuropeptide Y, are concentrated caudally in cell column, and project to supraspinal targets. The filled grey circles represent non-C1 neurons that contribute to both pathways. In figure 1B neurons immunopositive for TH illustrate the position of the C1 column in the ventrolateral medulla and delineate the area cut from the vibratome section for TEM analysis. The lower portion of the schematic illustrates the sectioning strategy incorporated in the analysis to ensure precise localization of labeled profiles within the area of C1. Series of ultrathin sections bracketed by toluidine blue stained thick sections were collected on formvar-coated narrow slot copper grids. Landmarks ( e.g. ). AT = axon terminal, N = neuron cytoplasm, Nu = nucleus; asterisks identify blood vessels. Marker bars = 15 μm for A F, 3 μm for D G, and 0.5 μm for E H.

    Article Snippet: Ultrathin sections were analyzed unstained using a transmission electron microscope (TEM; Morgagni, FEI, Hillsboro, OR) equipped with a CCD camera (Advanced Microscopy Techniques, Danvers, MA).

    Techniques: Transmission Electron Microscopy, Sampling, Labeling, Staining, Marker

    Ultrastructural characterization of HSV plaques in HaCaT cells. Monolayers of HaCaT cells in 35-mm dishes were infected with 100 PFU of HSV-1[17]/well and processed at 24 h postinfection for TEM as described in Materials and Methods. (a) Low-power binocular image of the vertical face of the embedded resin block. This image gives a three-dimensional view along the horizontal surface of the plastic support. Arrows indicate the plaques of clustered piled-up cells, with the size of the arrow indicating the distance from the block front. (b) The block was then sectioned into semithin survey sections (500 to 1,000 nm) and stained with crystal violet. Images were collected on a Zeiss Axiovert 200M using a ×40 objective lens, and the panels were assembled using Adobe Photoshop. (c) Sequential ultrathin sections (50 to 70 nm) of the same sample were then obtained and collected onto slot grids, stained, and examined by TEM. Because the sections analyzed are immediately adjacent to the section stained with crystal violet, corresponding areas could be matched. The TEM images of the sections in panel c, sections 1 to 4, therefore correspond to the boxes in panel b, sections 1 to 4. Upper panels (lower magnification) and lower panels (higher magnification on inset area) show different features of infection within the plaque as discussed in the text. In the lower panels, the features include extracellular virions (black arrows with white outline), nucleocapsids (short black arrows), and cell tight junctions (white arrows). In the upper part of panels 3 and 4, cells with no distinct features of infection (e.g., no early chromatin marginalization) are indicated by long black arrows. Box 5 in panel b shows flat uninfected cells that correspond to the elongated cells (TEM data not shown).

    Journal: Journal of Virology

    Article Title: Polarized Cell Migration during Cell-to-Cell Transmission of Herpes Simplex Virus in Human Skin Keratinocytes

    doi: 10.1128/JVI.01172-13

    Figure Lengend Snippet: Ultrastructural characterization of HSV plaques in HaCaT cells. Monolayers of HaCaT cells in 35-mm dishes were infected with 100 PFU of HSV-1[17]/well and processed at 24 h postinfection for TEM as described in Materials and Methods. (a) Low-power binocular image of the vertical face of the embedded resin block. This image gives a three-dimensional view along the horizontal surface of the plastic support. Arrows indicate the plaques of clustered piled-up cells, with the size of the arrow indicating the distance from the block front. (b) The block was then sectioned into semithin survey sections (500 to 1,000 nm) and stained with crystal violet. Images were collected on a Zeiss Axiovert 200M using a ×40 objective lens, and the panels were assembled using Adobe Photoshop. (c) Sequential ultrathin sections (50 to 70 nm) of the same sample were then obtained and collected onto slot grids, stained, and examined by TEM. Because the sections analyzed are immediately adjacent to the section stained with crystal violet, corresponding areas could be matched. The TEM images of the sections in panel c, sections 1 to 4, therefore correspond to the boxes in panel b, sections 1 to 4. Upper panels (lower magnification) and lower panels (higher magnification on inset area) show different features of infection within the plaque as discussed in the text. In the lower panels, the features include extracellular virions (black arrows with white outline), nucleocapsids (short black arrows), and cell tight junctions (white arrows). In the upper part of panels 3 and 4, cells with no distinct features of infection (e.g., no early chromatin marginalization) are indicated by long black arrows. Box 5 in panel b shows flat uninfected cells that correspond to the elongated cells (TEM data not shown).

    Article Snippet: The sample was sectioned again when appropriate transverse sections were observed, and ultrathin sections (typically 50 to 70 nm) were cut and collected onto slot grids, stained with Reynold's lead citrate examined in a FEI Tecnai electron microscope with CCD camera image acquisition.

    Techniques: Infection, Transmission Electron Microscopy, Blocking Assay, Staining