Formvar coated single slot copper grids

Manufactured by Electron Microscopy Sciences

Sourced in Japan

Formvar-coated single-slot copper grids are a type of laboratory equipment used for sample preparation in electron microscopy. They provide a stable and uniform support for specimens during examination under an electron microscope.

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Lab products found in correlation

Uranyl acetate, by Electron Microscopy Sciences (4 mentions) Osmium tetroxide, by Electron Microscopy Sciences (4 mentions) Fei tecnai g2 spirit, by Thermo Fisher Scientific (3 mentions) Morada ccd digital camera, by Olympus (3 mentions) Ultracut uc7 ultramicrotome, by Leica (3 mentions) Durcupan acm epoxy resin, by Merck Group (1 mentions) Glutaraldehyde, by Electron Microscopy Sciences (1 mentions) Veleta, by Olympus (1 mentions) Ultracut uc6 ultramicrotome, by Leica (1 mentions)

5 protocols using formvar coated single slot copper grids

Ultrastructural Analysis of SVZ Samples

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Samples (n = 3 per group) were sectioned into 200 µm sections and post-fixed with 2% osmium tetroxide (Electron Microscopy Sciences) for 2 h. Sections were then washed in deionized water, and partially dehydrated in 70% ethanol. Afterwards, the samples were contrasted with 2% uranyl acetate (Electron Microscopy Sciences) in 70% ethanol for 2 h at 4 °C. The samples were further dehydrated and infiltrated in Durcupan ACM epoxy resin (Sigma) at room temperature overnight, and then at 60 °C for 72 h. Once the resin was cured, SVZ sections were selected and cut into ultrathin Sects. (60–80 nm) using an Ultracut UC7 ultramicrotome (Leica Biosystems). These sections were placed on Formvar-coated single-slot copper grids (Electron Microscopy Sciences) and stained with lead citrate.

Norton E.S., Whaley L.A., Ulloa-Navas M.J., García-Tárraga P., Meneses K.M., Lara-Velazquez M., Zarco N., Carrano A., Quiñones-Hinojosa A., García-Verdugo J.M, & Guerrero-Cázares H. (2022). Glioblastoma disrupts the ependymal wall and extracellular matrix structures of the subventricular zone. Fluids and Barriers of the CNS, 19, 58.

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Transmission Electron Microscopy Sample Preparation

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For TEM analysis, sections were embedded in epoxy resin. First, samples were post-fixed with 1% osmium tetroxide (Electron Microscopy Sciences), 7% glucose in 0.1 M PB for 30 min at room temperature, washed in deionized water, and partially dehydrated in 70% ethanol. Afterward, the samples were contrasted in 2% uranyl acetate (Electron Microscopy Sciences) in 70% ethanol for 2 h at 4°C. The samples were further dehydrated and embedded in Durcupan ACM epoxy resin at room temperature overnight, and then at 70°C for 72 h. Once the resin was polymerized, immunolabeled sections were selected and cut into semithin (1.5 μm) and then into ultrathin sections (60–80 nm) using an Ultracut UC7 ultramicrotome (Leica). Ultrathin sections were placed on formvar-coated single-slot copper grids (Electron Microscopy Sciences) stained with lead citrate and examined at 80 kV on a FEI Tecnai G² Spirit (FEI Company, Hillsboro, OR) transmission electron microscope equipped with a Morada CCD digital camera (Olympus, Tokyo, Japan).

Ulloa-Navas M.J., Pérez-Borredá P., Morales-Gallel R., Saurí-Tamarit A., García-Tárraga P., Gutiérrez-Martín A.J., Herranz-Pérez V, & García-Verdugo J.M. (2021). Ultrastructural Characterization of Human Oligodendrocytes and Their Progenitor Cells by Pre-embedding Immunogold. Frontiers in Neuroanatomy, 15, 696376.

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Detailed Protocol for Transmission Electron Microscopy

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For transmission electron microscopy, sections were embedded in epoxy resin. First, samples were post-fixed with 1% osmium tetroxide (Electron Microscopy Sciences), 7% glucose in 0.1 M PB for 30 min at room temperature, washed in deionized water, and partially dehydrated in 70% ethanol. Afterward, the samples were contrasted in 2% uranyl acetate (Electron Microscopy Sciences) in 70% ethanol for 2 h at 4°C. The samples were further dehydrated and embedded in Durcupan ACM epoxy resin at room temperature overnight, and then at 70°C for 72 h. Once the resin was polymerized, immunolabeled sections were selected and cut into serial semithin (1.5 mm) and then into serial ultrathin sections (60–80 nm) using an Ultracut UC7 ultramicrotome (Leica). We examined 20–25 serial ultrathin sections per cell. Ultrathin sections were placed on formvar-coated single-slot copper grids (Electron Microscopy Sciences) stained with lead citrate and examined at 80 kV on a FEI Tecnai G2 Spirit (FEI Company, Hillsboro, OR) transmission electron microscope equipped with a Morada CCD digital camera (Olympus, Tokyo, Japan).

Bönnemann C.G., Cox G.F., Shapiro F., Wu J.J., Feener C.A., Thompson T.G., Anthony D.C., Eyre D.R., Darras B.T, & Kunkel L.M. (2000). A mutation in the alpha 3 chain of type IX collagen causes autosomal dominant multiple epiphyseal dysplasia with mild myopathy. Proceedings of the National Academy of Sciences of the United States of America, 97(3), 1212-1217.

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Mouse Liver Ultrastructural Analysis

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The mouse liver was cut into 2-3 mm 3 cubes and immediately immersed in a fixation solution containing 2.5% glutaraldehyde (Electron Microscopy Science) in 0.1 M cacodylate buffer pH 7.4 for 1 hour at room temperature and then overnight at 4°C. After five washes in 0.1 M cacodylate buffer (pH 7.4), a post-fixation in 1% osmium tetroxide in 0.1 M cacodylate buffer (pH7.4) was done. After five washes in ddH2O and dehydration steps in graded alcohol series, the mouse liver pieces were embedded in Embed 812 epoxy resin hard (Electron Microscopy Science) for 12 h and polymerized at 60°C during 48 h. For TEM analysis, a region of interest was selected under bright field. After trimming, silver/gray thin sections (50 nm thickness) were collected on formvar-coated single-slot copper grids (Electron Microscopy Science). After post-staining with 1% uranyl acetate and lead citrate (6 min, each), images were recorded using a FEI Tecnai Spirit (FEI Company) operated at 120 keV using a side-mounted 2K × 2K CCD camera (Veleta, Olympus).

Saponara E., Penno C.A., Guzmán M.L.M., Brun V., Fischer B., Brousseau M.E., Wang Z., O’Donnell P., Turner J., Meyer A.G., Ansel-Bollepalli L., d’Ario G., Roma G., Carbone W., Orsini V., Annunziato S., Obrecht M., Beckmann N., Saravanan C., Osmont A., Tropberger P., Richards S.M., Genoud C., Aebi A., Ley S., Ksiazek I., Nigsch F., Terracciano L., Bouwmeester T., Tchorz J.S., & Ruffner H. (2021). Loss of hepatic Lgr4 and Lgr5 promotes nonalcoholic fatty liver disease.

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Transmission Electron Microscopy Sample Preparation

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Samples were embedded in resin as previously described. 20 (link) Briefly, samples were postfixed with 1% osmium tetroxide (Electron Microscopy Sciences), 7% glucose in 0.1 M PB for 30 min at room temperature, washed in deionized water, and partially dehydrated in 70% ethanol. Afterwards, the samples were contrasted in 2% uranyl acetate (Electron Microscopy Sciences) in 70% ethanol for 2 h at 4 ºC. The samples were further dehydrated and infiltrated in Durcupan ACM epoxy resin at room temperature overnight, and then at 60 ºC for 72 h. Once the resin was cured, immunolabeled sections were selected and cut into ultrathin sections (60-80 nm) using an Ultracut UC6 ultramicrotome (Leica Biosystems). These sections were placed on Formvarcoated single-slot copper grids (Electron Microscopy Sciences) stained with lead citrate and examined at 80 kV on a FEI Tecnai G 2 Spirit (FEI Company) transmission electron microscope equipped with a Morada CCD digital camera (Olympus).

Ulloa-Navas M.J., García-Tárraga P., García-Verdugo J.M, & Herranz-Pérez V. (2019). Immunogold Labeling to Detect Streptococcus pyogenes Cas9 in Cell Culture and Tissues by Electron Microscopy. The CRISPR journal, 2(6).

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