Embryos were dissected in PBS (pH 7.2), temporarily placed in 3% PFA in 0.2 M Hepes (pH 7.2), and then fixed in 4% PFA + 1.5% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.4) on ice. Embryos were then processed and imaged on a Philips CM100 transmission electron microscope equipped with an Olympus Megaview III CCD camera as previously described (Gokhin et al., 2010 (link)).
Cm100 transmission electron microscope
Manufactured by Olympus
The CM100 is a transmission electron microscope (TEM) manufactured by Olympus. It is designed to produce high-resolution images of thin samples by transmitting an electron beam through the specimen. The CM100 TEM is capable of magnifying samples up to 1,000,000x and can provide detailed information about the internal structure and composition of materials at the nanoscale level.
Automatically generated - may contain errors
3 protocols using cm100 transmission electron microscope
1
Embryo Fixation and Electron Microscopy
2
TEM Imaging of Microbial Cells
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Cells were prepared and pre-grown as previously described. The culture was grown in the presence of 1 mM IPTG for 6 h and harvested by centrifugation. Samples were prepared as previously described (Andersen et al. 2021 (link)). Semi-thin sections (2 μm) were cut using a glass knife and ultramicrotome (Leica Ultracut, Leica Microsystems, Wetzlar, Germany) and stained with 1% toluidine blue (Millipore) and 1% Borax (LabChem). Ultra-thin sections (50–70 nm) were cut using a diamond knife (Jumdi, 2 mm) and the ultramicrotome, contrasted in 2% uranyl acetate (Polyscience) and lead citrate (Merck). The samples were examined in a Philips CM100 transmission electron microscope, and images were obtained using an Olympus Morada camera and iTEM software (Olympus).
3
TEM Imaging of Microbial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were prepared and pre-grown as previously described. The culture was grown in the presence of 1 mM IPTG for 6 h and harvested by centrifugation. Samples were prepared as previously described (Andersen et al. 2021 (link)). Semi-thin sections (2 μm) were cut using a glass knife and ultramicrotome (Leica Ultracut, Leica Microsystems, Wetzlar, Germany) and stained with 1% toluidine blue (Millipore) and 1% Borax (LabChem). Ultra-thin sections (50–70 nm) were cut using a diamond knife (Jumdi, 2 mm) and the ultramicrotome, contrasted in 2% uranyl acetate (Polyscience) and lead citrate (Merck). The samples were examined in a Philips CM100 transmission electron microscope, and images were obtained using an Olympus Morada camera and iTEM software (Olympus).
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