I try to find a cell fixation method which preserves not only the morphology but the elastic parameters of cells, too. Paraformaldehyde and glutaraldehyde are out of the game.
Have you tried chilled methanol? That is another way of fixing the cells.For immunostaining purposes it works fine. However, it permablizes the cell membrane and I do not know if that would be a problem in your experiment.
All the best
Thank you for your answer. Actually, I would like to have cells fixed in situ within the AFM fluid cell, what I cannot chill. Anyway membrane pores and chilling has severe effect on cell stiffness. So I search for something what works at 37 and I can simply add to medium. I would like to examine the same cell before and after fixation, so I would prefer not to take them anywhere out from AFM.
Hi Attila, cell fixation implies that you are chemically crosslinking cell proteins, and anything with a primary amine together to preserve the overall cell shape and structure, I gather, that this is not what you want.
Instead of chemical fixation, you could look at treating the cells with a mixture of azide and digitonin:
http://scholar.google.com/scholar?q=azide+digitonin+afm&btnG=&hl=en&as_sdt=1%2C21
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