Hi, I have been trying different methods of fixation to stain cells with lectins to detect glycans in the cell surface. I noticed that fixing the cells increases the MFI compared to live cells. Does anybody had experience this before?
Jonathan Cremer
It is not uncommon for fixation to cause autofluorescence to increase. That is why you should always include appropriate technical controls in your experiment.
However, you give very little information as with what and how you stain. Do you fix before the stain? If so, alcoholic fixation also permeabilizes the cells -more so than formaldehyde fixation-, you may very well be staining intracellularly.
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