[Cytometry] Sorting microparticles from Tcells

Monica DeLay Monica.DeLay at cchmc.org
Tue Feb 3 13:22:40 EST 2009


Hello all,
First I want to say that I appreciate all of the feedback that I get when I submit questions to this list.  What a great community!

For the question, I have a client that wants to sort microparticles from Tcells.  We have FACSAria II SORPs in our facility.  The smallest nozzle we have is 70 um.  My client supplied me with a very nice paper from Journal of Virology (Vol 82 No. 15 p7700 see supplementary data), where they analyzed MPs on an LSRII.  FSC and SSC are in biexponential and they use beads to set up the gate as the MPs are 0.1-0.5 um in size.  The beads were 0.1, 0.2, 0.5 and 1.0 um from Molecular Probes and were diluted 1:100,000.  They ran buffer alone as a control for background debris.  
My client would like to stain MPs with  Annexin-FITC and CD3 (not sure what label).

Does anyone have experience with sorting MPs?  I'm concerned about the nozzle size and whether the MPs will survive the sort as we usually use 70 um nozzle with 70 psi.  What about doublet discrimination?  Any other information or details to consider would be of great help.

Thanks!



Monica DeLay, M.S.
Manager, Research Flow Cytometry Core
Cincinnati Children's Hospital Medical Center
Department of Rheumatology
TCHRF 5543
3333 Burnet Ave
Cincinnati, Ohio 45229
Phone (513) 636-3575
Fax (513) 636-3328


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