I am looking for a good protocol to stain B cells for cell cycle analysis. One of our antibodies is not really compatible with PFA. I would like a method that would be apt for using in confocal microscopy since using methanol has not helped us much. Any suggestions?
Dorothea Dijkstra
You could look for a buffer from BD, they have several kind of "balanced" fixation buffers. We use isolated human B cells for flow cytometry, stain them first with antibodies and then fix the cells with Cytofix from BD. Then we stain with DAPI and Pyronin Y. This works well in flow cytometry. I hope this helps.
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