If I have cells on a glass slide and I put 4% PFA on it, are the cells going to be stuck on that particular position always? Will the cells move if I add PBS to it? What exactly does fixation do to the cells?
the cells fixed with PFH will not stay in the same position because you are fixing the cell not the surface plus the cells, so if you add PBS later they will move position but cells will not change biochemically or morphologically
Some type of cells such as macrophage tends to spread to the culture dish and bottom glass. Generally, PFA-fixed cell, which internal and membranous proteins are crosslinked, can not move. However, the process of the fixed cell can be partly removed from glass and plastic dish surfece by harsh washing. Therefore, it could be seen as if the fixed cells were moved even after fixation.
The trick is to handle the cells post-fixing very very gently (especially during the PBS-rinsing step). Try your best to minimize turbulence in your samples. Good luck.
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