[Cytometry] sorting sperm

Guzik, Lynda GuziLJ at ccm.upmc.edu
Mon Jan 26 14:24:34 EST 2009


Hello All,

I have an investigator who would like to sort non-human primate and mouse sperm.  So, I am looking for any information I can get.  In particular, info regarding nozzle tip size, sheath pressure, and drop frequency is needed.  We will be sorting gfp+ sperm, hopefully.  But, is there any other parameter we should be looking at - surface markers, dna dye, viability dye, etc?  The investigator hopes to recover functional sperm.  He believes the sperm head size should be about 4u, so I'm guessing a 50u tip would be best.  But, does the long tail interfere with drop formation or drop deflection with such a small tip?  Or does the tail break off?  And if it does break off, is the sperm still functional?  Does the shape of the sperm head have any affect on FSC or SSC?  Is the standard 1x PBS sheath fluid sufficient, or do sperm require a different pH or buffer?

I have a BDFACSVantage SE DiVa with 3 lasers:  488, 635, and UV.  Generally, I sort tumor cells based on surface markers and viability.

Any and all information will be appreciated.  Thank you!

Lynda Guzik
McGowan Institute - Lagasse Labs
Rm 520A - Bridgeside Point Bldg
100 Technology Dr - Suite 200
Pittsburgh, PA 15219
412-235-5213
guzilj at upmc.edu<mailto:lguzilj at upmc.edu> or lyg1 at pitt.edu<mailto:lyg1 at pitt.edu>




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