Hi, Steve: I haven't worked with human sperm, only bull and some other mammals, but staining with Hoechst 33342 works just fine. We are sorting bull sperm based on X-Y separation, but since you don't need to do that, it should be very easy to sort individual sperm based on Ho342 staining. We stain 5x10^^7 sperm/ml with 10 ul Ho342 (1 mg/ml stock) and incubate for 45 min at 35ø C. That would probably work well for you also. I don't think you will have any trouble with doing PCR on the sorted sperm. However, I wonder whether you couldn't just sort based on light scatter and avoid staining altogether. Normally you get a pretty decent light scatter signal. Mike Fox Colorado State University
This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:49:17 EST