what does the best ratio of Formaldehyde and Glutaraldehyde for cell Fixation to image by TEM? and if I want to make Combining fixatives what does the best way?How long will my cells last once fixed? and which one is good to make fixation in plate or tube?
2.5% glut works for most (all?) cell cultures beautifully. Formaldehyde has faster penetration rate, but it is irrelevant for thin cell cultures. For pure love of formaldehyde some researchers add it to their fixatives, but why not, if it please them aesthetically? For those who just needs good results formaldehyde is not needed at all.
As answered by Vladimir, glutaraldehyde alone is sufficient for TEM preparation.
For cell monolayer, a fixation of 30 minutes is enough. You can fix for longer time by keeping the sample at 4°C for example over the night or the week-end (or even longer). It is never over-fixed!
Instead of formaldehyde fixation, glutaraldehyde fixation is not reversible.
I would also like to recomment using a good buffer for fixation (not PBS). Sörensens phosphate buffer with 4% PFA and 2.5% GA usually does the job.
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