>Date: Tue, 27 Apr 1999 15:58:43 -0500 (CDT) >From: Jeffrey M Scott <jeffreys@bcm.tmc.edu> >To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> >Subject: High pressure cell survival > > > > We're just figuring out sorting with a Coulter Altra over here, >and the high-pressure unit was recently installed. Given that the cells >are gonna be put through a gauntlet of ~50 PSI, has anybody checked >viability and/or proliferative capacity after such treatment? We're >afraid it will produce some aberrant cell behaviors. > > Cheers, > > Jeff Hi Jeff, I don't have an Altra, but I do have a MoFlo that I typically run at 60psi. I have had no problem with viability after a sort if the samples are handled properly. I sort mainly mouse cells, from bone marrow, spleen, etc.,. or cell culture all with good viability. However I have heard that large delicate cells such as insect cells can fragment at high pressures. I reckon that this will occur at the nozzle tip owing to the rapid decrease in pressure there. I would recommend that you carefully prepare your samples, ensuring that there are no clumps. Try to minimise the debris found in samples as much as possible by filtering your reagents. Lots of debris can lead to blocked tubing and nozzle at these kinds of pressures. Small particulate matter can collect and foul the tubing leading to sheath core instability and ruin your laser alignment. I have found that small particles can also effect droplet formation, perhaps the debris causes local changes in viscosity of the stream? (Any ideas welcome?) Hope this helps. Andy Andy Riddell PNAC MRC-LMB Hills Road Cambridge UK tel (0) 1223 402218 fax (0) 1223 412178 email ar3@mrc-lmb.cam.ac.uk
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