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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
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
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