Is there a way to fix cell membranes for SEM?
I have features on the cells membrane which collapse when prepping the cell for SEM. The features don’t contain actin, so I need to find a way of fixing the membrane without cryofixation.
Cheers
depends what you want to see. But generally you should get reasonable results if you fix in fixative for EM (4% paraformaldehyde, 2% glutaraldehyde in some suitable buffer). then osmificate to increase stability of cell membranes. Careful and slow dehydration into acetone for critical point drying. The most important thing will be the quality of your critical point drying. Then sputter coat and have a look whether it worked.
You need to keep all solutions really clean and you may want to consider putting them through a membrane filter.
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September 29th, 2009 at 4:25 pm
depends what you want to see. But generally you should get reasonable results if you fix in fixative for EM (4% paraformaldehyde, 2% glutaraldehyde in some suitable buffer). then osmificate to increase stability of cell membranes. Careful and slow dehydration into acetone for critical point drying. The most important thing will be the quality of your critical point drying. Then sputter coat and have a look whether it worked.
You need to keep all solutions really clean and you may want to consider putting them through a membrane filter.
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