re: Sperm sorting

Michael Fox (MFox@vines.ColoState.EDU)
Tue, 14 Jan 97 10:34:19 MST

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 35 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

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