Female worms recovered from 2–3 jirds per group were fixed in 2.5% glutaraldehyde, 2% paraformaldehyde in 0.1 M sodium cacodylate buffer, pH 7.4 (EMS, USA). Worms were cut into 2–3 mm long pieces in the fixative and incubated for 3 h at room temperature and at 4 °C overnight. Samples were washed thoroughly in the buffer and post-fixed in 1% osmium tetroxide in 0.1 M sodium cacodylate buffer for 1 h. Samples were then washed in the buffer, in distilled water and a series of ethanol dilutions: 30%, 50% (with 5% uranyl acetate), 70%, 95%, for 10 min each, and twice with 100% ethanol for 20 min each. Samples were infiltrated with a gradient of acetone-Embed 812 resin and embedded in 100% resin. After sectioning the solidified blocks, ~70 nm sections were stained with UranyLess (EMS, USA) and lead citrate followed by observation using a FEI Tecnai 12 Spirit transmission electron microscope (Microscopy core facility, NYBC).
Tecnai 12 spirit transmission electron microscope
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Tecnai 12 Spirit transmission electron microscope is a high-performance instrument designed for advanced imaging and analysis of microscopic samples. The core function of this equipment is to provide users with the ability to obtain detailed, magnified images of materials at the nanoscale level, enabling the study of the structural and compositional properties of a wide range of specimens.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Eagle 4k 4k ccd camera, by Thermo Fisher Scientific (1 mentions)
G5882, by Merck Group (1 mentions)
Embed 812 resin, by Electron Microscopy Sciences (1 mentions)
2330p xa, by SPI Supplies (1 mentions)
Eponate 12, by Ted Pella (1 mentions)
Glutaraldehyde, by Electron Microscopy Sciences (1 mentions)
5 protocols using tecnai 12 spirit transmission electron microscope
1
Ultrastructural Analysis of Parasitic Worms
2
Ultrastructural Analysis of HUVECs
3
Ultrastructural Analysis of Centriole Morphology
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Cells grown on glass coverslips were fixed in 2.5% glutaraldehyde (G5882; Sigma-Aldrich) and 0.25% formaldehyde in 1× PBS (pH 7.4) for 1 h at RT. Centriole position within the target cells was recorded, and the position of the cells was marked on the coverslip. Cells were prestained with 1% osmium tetroxide (19100; Electron Microscopy Sciences) and 1% uranyl acetate (22400; Electron Microscopy Sciences), dehydrated in ethanol, and embedded in EMbed-812 resin (13940; Electron Microscopy Sciences). 80-nm-thick serial sections were sectioned, transferred to the formvar-coated copper grids (2330P-XA; SPI Supplies), and stained with uranyl acetate and lead citrate. Samples were imaged using an FEI Tecnai 12 Spirit transmission electron microscope operating at 80 kV. Image analysis was performed in Photoshop and Fiji. Centriole length was determined from one of the sections of centrioles sectioned longitudinally or near-longitudinally. Centriole width was determined from centrioles in various favorable orientations.
4
Ultrastructural Analysis of Ebolavirus Proteins
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Huh7 cells were seeded at a density of 2.5x105 cells per well in a 6-well plate. Twenty-four hours after seeding, cells were transfected 1ug NP, 1ug VP35, and 1ug VP24. As a control, empty pCAGGS vectors was used instead of VP24. Cells were transfected with Lipofectamine2000 (Thermofisher Scientific, Waltham, MA, USA) at a ratio of 1ug DNA to 1ul reagent. Twenty-four hours post transfection cells were fixed in 2% glutaraldehyde in 0.1 M sodium cacodylate buffer (pH 7.2) 2 mM CaCl2 for over 1 hour at room temperature and post-fixed in 1% osmium/0.8% potassium ferricyanide in 0.1 M cacodylate buffer. This was followed by post-staining in 1% uranyl acetate in water, dehydration in an ethanol series, and embedding in Eponate 12 (Ted Pella, Inc). Ultrathin sections (60–65 nm) were stained with uranyl acetate and lead citrate, and images were acquired using a Tecnai 12 Spirit transmission electron microscope (FEI, Hillsboro, Oregon, USA) operated at 120 kV, equipped with an AMT BioSprint29 digital camera.
5
Megakaryocyte Ultrastructure Imaging
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On day 11 of culture megakaryocytes treated with latrunculin and control cells were fixed with 2.5% Glutaraldehyde (Electron Microscopy Sciences, Hatfield, PA) and prepared for electron microscopy analysis. Ultrathin sections were sectioned on RMC MTX ultramicrotome (Boeckeler Instruments, Tucson, AZ) and imaged on an FEI Tecnai 12 spirit transmission electron microscope (FEI, Morristown, NJ) with an AMT camera (Advanced Microscopy Techniques, Woburn, MA).
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