Tem ccd camera

Manufactured by Ametek

The TEM CCD camera is a core imaging component for transmission electron microscopes (TEMs). It captures high-quality digital images of samples under electron beam examination. The camera is designed to provide reliable and accurate data collection for various TEM applications.

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Lab products found in correlation

Cm10 tem, by Ametek (2 mentions) Lr white resin, by Merck Group (1 mentions) Tecnai 12bt, by Thermo Fisher Scientific (1 mentions) Cm10 electron microscope, by Ametek (1 mentions)

9 protocols using tem ccd camera

Ultrastructural Analysis of Drosophila Imaginal Discs

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Imaginal discs from 3rd instar larvae were dissected and fixed in 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer pH 7.2 for 1 h at RT and post-fixed with 1% osmium tetroxide for 1 h at RT. Samples were in block stained with a 1% uranyl acetate aqueous solution (w/v) at 6 °C overnight, dehydrated in ethanol, and infiltrated first with two changes of 100% propylene oxide and then with a 50%/50% propylene oxide/SPI-Pon812 resin mixture, and finally polymerized at 68 °C in flat pre-filled embedding molds. The samples were then reoriented, and ultra-thin sections (~70 nm) were placed on copper support grids and contrasted with lead citrate and uranyl acetate. Sections were examined using the CM10 TEM with 100 kV accelerating voltage, and images were captured using a Gatan TEM CCD camera.

Diwanji N, & Bergmann A. (2020). Basement membrane damage by ROS- and JNK-mediated Mmp2 activation drives macrophage recruitment to overgrown tissue. Nature Communications, 11, 3631.

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Visualizing EGFP-tagged Proteins in HEK293 Cells

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HEK293 cells expressing EGFP-C1 and EGFP-(GR)₈0 plasmids were fixed with 4% PFA (v/v) in 0.75 M sodium phosphate buffer (pH 7.2) solution for 1 h, pelleted and dehydrated with ethanol, embedded in LR white resin (Sigma-Aldrich) and cut into ultrathin sections (80 nm) in gold support grids. The grids were incubated first in blocking solution (10% normal goat serum in PBS) for 1 h and then with rabbit polyclonal anti-(GR)₈ antibody (Covance; 1:100) overnight at room temperature in blocking solution. The grids were washed 5 times with PBS 1× and incubated with 1 μg/μl rabbit anti-IgG (1:20) conjugated with 12-nm gold particles in blocking buffer. The samples were examined on a Philips CM 10 transmission electron microscope using 80 kV accelerating voltage. Images were captured with a Gatan TEM CCD camera.

Choi S.Y., Lopez-Gonzalez R., Krishnan G., Phillips H.L., Li A.N., Seeley W.W., Yao W.D., Almeida S, & Gao F.B. (2019). C9ORF72-ALS/FTD-associated poly(GR) binds Atp5a1 and compromises mitochondrial function in vivo. Nature neuroscience, 22(6), 851-862.

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Ultrastructural Analysis of Fly Brain

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Three-week-old fly brains were dissected in PBS and immediately immersion fixed in 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer, pH 7.2. Samples were processed and analyzed by standard procedures at the Electron Microscopy Facility at the University of Massachusetts Medical School. Briefly, fixed samples were incubated overnight in fresh 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer at 4 °C, rinsed twice in the same fixation buffer, and postfixed with 1% osmium tetroxide for 1 h at room temperature. Samples were then washed twice for 5 min each with distilled water and dehydrated through a graded ethanol series (20% increments), before two changes in 100% ethanol. Samples were infiltrated first with two changes of 100% propylene oxide and then with a 50:50 mixture of propylene oxide and SPI-Pon 812 resin. The next day, after five changes of fresh 100% SPI-Pon 812 resin, the samples were polymerized at 68 °C in flat embedding molds. The samples were reoriented, cut into ~70-nm sections ~100 μm from the tip of the ventral ganglion or central brain (frontal sections), placed on copper support grids, stained with lead citrate and uranyl acetate, and examined with a FEI Tecnai 12 BT electron microscope at 100 Kv accelerating voltage. Images were captured with a Gatan TEM CCD camera.

Yuva-Aydemir Y., Almeida S., Krishnan G., Gendron T.F, & Gao F.B. (2019). Transcription elongation factor AFF2/FMR2 regulates expression of expanded GGGGCC repeat-containing C9ORF72 allele in ALS/FTD. Nature Communications, 10, 5466.

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Ultrastructural Analysis of Islet Cells

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Animals were deeply anesthetized and perfused with PBS until the blood was removed followed by 15 min perfusion with 2.5% glutaraldehyde, 2% paraformaldehyde in 100 mM cacodylate pH 7.2. Tissue was then removed and cut in 1 mm cubes or rings and fixed overnight in the same solution before osmication with 1% osmium tetroxide for 1 hr, at 22°C. Samples were then rinsed and then dehydrated through a graded ethanol series of 20% increments, before two changes in 100% ethanol. Samples were then infiltrated first with two changes of 100% Propylene Oxide and then with a 50%/50% propylene oxide / SPI-Pon 812 resin mixture. The following day three changes of fresh 100% SPI-Pon 812 resin were done before the samples were polymerized at 68^°C in plastic capsules. The samples were then sectioned (1 μm) and stained with toluidine blue to locate the islets. The samples were then re-trimmed with the islets in the center, and thin sections of approximately 70 nm were placed on copper support grids and contrasted with Lead citrate and Uranyl acetate. Sections were examined using the FEI Tecani 12 BT with 80Kv accelerating voltage, and images were captured using a Gatan TEM CCD camera.

Pearring J.N., San Agustin J.T., Lobanova E.S., Gabriel C.J., Lieu E.C., Monis W.J., Stuck M.W., Strittmatter L., Jaber S.M., Arshavsky V.Y, & Pazour G.J. (2017). Loss of Arf4 causes severe degeneration of the exocrine pancreas but not cystic kidney disease or retinal degeneration. PLoS Genetics, 13(4), e1006740.

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Ultrastructural Analysis of Ferroptosis

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Adherent siControl and siProminin2 cells were treated with DMSO or RSL3 (5μM) for two hr. ECM-detached siControl and siProminin2 cells were maintained for 2 hrs in low-adherence plates. All cells were fixed by adding 2.5% glutaraldehyde (v/v) in 0.1 M Na cacodylate buffer (pH 7.2) and processed for TEM. The samples were examined on a FEI Tecnai 12 BT transmission electron microscope using 100Kv accelerating voltage. Images were captured using a Gatan TEM CCD camera.

Brown C.W., Amante J.J., Chhoy P., Elaimy A.L., Liu H., Zhu L.J., Baer C.E., Dixon S.J, & Mercurio A.M. (2019). Prominin2 Drives Ferroptosis Resistance by Stimulating Multivesicular Body/Exosome-Mediated Iron Export. Developmental cell, 51(5), 575-586.e4.

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Ultrastructural Analysis of Fly Ovaries

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Fly ovaries were quickly dissected in dissecting solution and transferred to 1.7 ml microfuge tubes on ice. Dissecting solution was removed and ovaries fixed in 2.5% glutaraldehyde (w/v) in 0.1 M sodium cacodylate buffer, pH 7.2, overnight at 4°C. Samples were processed and analyzed at the University of Massachusetts Medical School Electron Microscopy Core Facility using standard procedures. Briefly, intact ovaries were rinsed three times in the fixation buffer and post-fixed with 1% osmium tetroxide (w/v) for 1 h at room temperature. Samples were then washed three times with water for 10 min each and dehydrated using a graded series of 10%, 30%, 50%, 70%, 85%, and 95% ethanol, finishing with three changes in 100% ethanol. Samples were then infiltrated first with two changes 100% propylene oxide and then with a mixture of 50% propylene oxide/50%SPI-Pon 812 resin. The sample was incubated in seven successive changes of fresh 100% SPI-Pon 812 resin over 2 days, then polymerized at 68°C in flat molds. The samples were then reoriente d for horizontal sections of the center of individual ovaries. Thin sections (~70 nm) were placed on gold support grids, and stained with lead citrate and uranyl acetate to increase contrast. Sections were examined using a CM10 with 80 KV accelerating voltage, and images captured using a Gatan TEM CCD camera.

Ge D.T., Wang W., Tipping C., Gainetdinov I., Weng Z, & Zamore P.D. (2019). The RNA-binding ATPase, Armitage, Couples piRNA Amplification in Nuage to Phased piRNA Production on Mitochondria. Molecular cell, 74(5), 982-995.e6.

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Ultrastructural Analysis of Frontal Cortex

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Frontal cortex tissues from CamKII and CamKII;(GR)go mice were cut into ~1-mm³ pieces, fixed in glutaraldehyde-containing phosphate buffer, pH 7.2, at room temperature and postfixed in 1% OsO4 solution for 1 h at 4 °C. The specimens were then dehydrated, embedded in araldite, cut into semi-thin and thin sections, stained with uranyl acetate and lead citrate, examined with a Philips CM10 electron microscope and photographed with a Gatan TEM CCD camera.

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Ultrastructural Analysis of C. elegans

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L4 larvae were transferred to 2.5% glutaraldehyde in 0.1 M Sodium Cacodylate buffer pH 7.2 for 10 min. The tail and head of each worm were dissected out and the main body was transferred to fresh 2.5% glutaraldehyde in 0.1 M Sodium Cacodylate buffer and kept at 4 °C overnight. Samples were processed and analyzed at the University of Massachusetts Medical School Electron Microscopy core facility according to standard procedures. Briefly, the samples were rinsed three times in the same fixation buffer and post-fixed with 1% osmium tetroxide for 1 h at room temperature. Samples were then washed three times with ddH₂O for 10 min and then dehydrated through a graded ethanol series of 20% increments, before two changes in 100% ethanol. Samples were then infiltrated first with two changes of 100% Propylene Oxide and then with a 50%/50% propylene oxide/SPI-Pon 812 resin mixture. The following day, five changes of fresh 100% SPI-Pon 812 resin were performed before the samples were polymerized at 68 °C in flat pre-filled embedding molds. The samples were then reoriented, and thin sections (~70 nm) were placed on copper support grids and contrasted with Lead citrate and Uranyl acetate. Sections were examined using a CM10 TEM with 100 Kv accelerating voltage and images were captured using a Gatan TEM CCD camera.

Shpilka T., Du Y., Yang Q., Melber A., Uma Naresh N., Lavelle J., Kim S., Liu P., Weidberg H., Li R., Yu J., Zhu L.J., Strittmatter L, & Haynes C.M. (2021). UPRmt scales mitochondrial network expansion with protein synthesis via mitochondrial import in Caenorhabditis elegans. Nature Communications, 12, 479.

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Electron Microscopic Analysis of C. elegans

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L4 larvae were transferred to 2.5% glutaraldehyde in 0.1 M Sodium Cacodylate buffer pH 7.2. for 10 min. The tail and head of each worm were dissected out and the main body was transferred to fresh 2.5% glutaraldehyde in 0.1 M Sodium Cacodylate buffer and kept at 4 o C overnight. Samples were processed and analyzed at the University of Massachusetts Medical School Electron Microscopy core facility according to standard procedures. Briefly, the samples were rinsed three times in the same fixation buffer and post-fixed with 1% osmium tetroxide for 1h at room temperature. Samples were then washed three times with ddH2O for 10 minutes and then dehydrated through a graded ethanol series of 20% increments, before two changes in 100% ethanol. Samples were then infiltrated first with two changes of 100% Propylene Oxide and then with a 50%/50%
propylene oxide/SPI-Pon 812 resin mixture. The following day, five changes of fresh 100% SPI-Pon 812 resin were performed before the samples were polymerized at 68 o C in flat pre-filled embedding molds. The samples were then reoriented, and thin sections (approx. 70nm) were placed on copper support grids and contrasted with Lead citrate and Uranyl acetate. Sections were examined using a CM10 TEM with 100Kv accelerating voltage, and images were captured using a Gatan TEM CCD camera.

Shpilka T., Du Y., Qi Y., Melber A., Naresh N.U., Lavelle J., Liu P., Weidberg H., Li J., Yu J., Zhu L.J., Strittmatter L., & Haynes C.M. (2020). UPR^mt scales mitochondrial network expansion with protein synthesis via mitochondrial import.

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