Human RBC are also quite brittle, and my 4-5% FA fixation used to destroy quite a few of them. I found that the following helps, and it may be good for mouse cells too: 1. step up the FA concentration slowly, first 0.5, then 1,2 and 4 over a day or so. Start in the fridge, later steps at RT. 2. make sure the FA is in the proper salt concentration (e.g.PBS). For a time we made the mistake to mix a 10% FA solution in H2O with cells in PBS at 1:1 to make 5% FA. The nucleated cells can handle it, but many of the RBCs blow up. Hope that helps Ralph Bohmer, New England Medical Center, Boston
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