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I am isolating single plasma cells by FACS sorting into 384-well plates, with the intent to assay the supernatant and clone H/L chains from positive wells. The efficiency of the PCR is however low, because the medium must be removed, yet many plasma cells get sucked up during medium removal. Is there any way to make plasma cells adhere to the bottom, and therefore withstand sucking? I am thinking of poly-L-lysine, gelatine, or similar things - but I do not know whether these would be effective, or even tolerated by the plasma cells.

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    $\begingroup$ Have you tried spinning the plate down (say, 400xg for 5 minutes) just before removing the sup, then using a robot to slowly aspirate 1/2 or 2/3 of the well? $\endgroup$ – MattDMo Mar 5 '16 at 17:35
  • $\begingroup$ We use a PE Janus to suck the medium, and may be moving to a Biotek robot. The problem is that >95% of the medium must be sucked off, or else the PCR won't work. Spinning will not help since microscopy shows that the cells are already at the bottom of the plate. I still wonder whether some kind of gluing may help. $\endgroup$ – aag Mar 6 '16 at 9:41
  • $\begingroup$ Oh, I didn't realize the PCR wouldn't work with excess liquid. How long are you culturing the cells before assaying for Ig (I assume) in the sup and deciding to PCR? If it's a relatively short amount of time (maybe 24-36 hours or less?), I can't see great harm in trying poly-D-lysine or gelatin or something. Have you tried Corning's Cell-Bind plates? I've had mixed results with them increasing adherence of suspension cells. Unfortunately, I haven't every tried adhering plasma cells, as I've always had microfluidics devices available should I need to do single-cell cloning. $\endgroup$ – MattDMo Mar 6 '16 at 18:41

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