Blood clot samples were fixed in 2% glutaraldehyde in 0.05 M phosphate buffer and 100 mM sucrose. Then they were post-fixed overnight in 1% osmium tetroxide in 0.1 M phosphate buffer, followed by dehydration and embedment in a mixture of EM-bed/Araldite (Electron Microscopy Sciences). One μm-thick sections were then stained with Richardson’s stain for observation by light microscopy. One hundred ηM sections were then cut on a Leica Ultracut-R ultramicrotome and stained with uranyl acetate and lead citrate. Grids were viewed at 80 kV in a JEOL JEM-1400 transmission electron microscope. Images were captured by an AMT BioSprint digital camera.
Em bed araldite
Manufactured by Electron Microscopy Sciences
Sourced in United States
EM-bed/Araldite is an epoxy resin used for embedding biological and material science specimens for electron microscopy. It is a two-component resin system that provides a hard, durable embedding medium for ultrathin sectioning and analysis.
Automatically generated - may contain errors
5 protocols using em bed araldite
1
Ultrastructural Analysis of Blood Clots
2
Ultrastructural analysis of fetal cardiomyocytes
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Tissue was fixed in 2% glutaraldehyde in 0.05 M phosphate buffer and 100mM sucrose, post‐fixed overnight in 1% osmium tetroxide in 0.1 M phosphate buffer, dehydrated through a series of cold graded ethanols, and embedded in a mixture of EM‐bed/Araldite (Electron Microscopy Sciences). 1 μm thick sections were stained with Richardson’s stain for observation under a light microscope. 100 nmol/L sections were cut on a Leica Ultracut‐R ultramicrotome and stained with uranyl acetate and lead citrate. The grids were viewed at 80 kV in a Philips cardiomyocyte‐10 transmission electron microscope and images captured by a Gatan ES1000W digital camera. N=2 samples were examined per each group at 17.5 d of gestation.
3
Transmission Electron Microscopy Sample Preparation
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Samples were fixed in 2% glutaraldehyde in 0.05 m phosphate buffer and 100 mm sucrose, postfixed overnight in 1% osmium tetroxide in 0.1 m phosphate buffer, dehydrated through a series of graded ethanols, and embedded in a mixture of EM-bed/Araldite (Electron Microscopy Sciences). The 1-μm-thick sections were stained with Richardson’s stain for observation under a light microscope. One hundred nanometer sections were cut using a Leica Ultracut-R ultramicrotome and were stained with uranyl acetate and lead citrate. The grids were viewed at 80 kV on a transmission electron microscope (model JEM-1400, JEOL), and images were captured with a digital camera (BioSprint, AMT).
4
Ultrastructural Analysis of Tissue Samples
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Individual punches were fixed in 2% glutaraldehyde in 0.05 M phosphate buffer and 100 mM sucrose, post-fixed overnight in 1% osmium tetroxide in 0.1 M phosphate buffer, dehydrated through a series of cold graded ethanol, and embedded in a mixture of EM-bed/Araldite (Electron Microscopy Sciences). One-μm-thick sections were stained with Richardson’s stain for observation under a light microscope. A total of 100 nM sections were cut on a Leica UC7 ultramicrotome and stained with uranyl acetate and lead citrate. The grids were viewed t 80 kV in a JEOL JEM-1400 transmission electron microscope and images captured by an AMT BioSprint 12 digital camera.
5
Transmission Electron Microscopy of Autopsy and Biopsy Samples
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Four of the six autopsy patients and the one live testis biopsy patient had tissue that underwent TEM imaging. Samples were fixed in 2% glutaraldehyde in 0.05 M phosphate buffer and 100 mM sucrose, post-fixed overnight in 1% osmium tetroxide in 0.1 M phosphate buffer, dehydrated through a series of graded ethanols, and embedded in a mixture of EM-bed/Araldite (Electron Microscopy Sciences, Hatfield, PA, USA). A total of 1 µm thick sections were stained with Richardson's stain for observation under a light microscope. A total of 100 nM sections were cut on a Leica Ultracut-R ultramicrotome (Leica, Wetzlar, Germany) and stained with uranyl acetate and lead citrate. The grids were viewed at 80 kV in a JEOL JEM-1400 transmission electron microscope (JEOL, Tokyo, Japan) and images captured by an AMT BioSprint digital camera (AMT, Woburn, MA, USA).
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