Cell carryover on FacsCalibur with loader

From: Richard McFarland PhD, MD (mcfarland.richard@pathology.swmed.edu)
Date: Mon Sep 20 1999 - 14:21:09 EST


>

>        Intermittently, we experience what appears to be cell carryover
>from one tube to the next when acquiring on our FacsCalibur with the
>automated sample loader attachment.  These cells
>form small populations which appear to express bizarre antigen combinations
>until it is recognized that these are cells from the immediately previous
>tube, stained with different antibodies.  When a re-stain is performed,
>using a new aliquot of the specimen, these cells disappear.  When the
>contaminated tube is later re-run, however, the cells are still there,
>indicating that a portion of the cells from the first tube actually entered
>the second tube.  We initially suspected that the tube-tube contamination
>occurred before they were put on the cytometer, but that is apparently not
>the case, as in one case data from all tubes looked normal, with no
>contamination, but when the case had to be re-run for an unrelated reason,
>one tube was now contaminated with cells from the immediately preceeding
>tube, and when it was run a third time, the cells were still there.  We
>thought there might be a problem with the pump mechanism, but it has been
>replaced (by a B-D FSE) without correcting the problem.  Has anyone else
>ever experienced
>this problem?  Any ideas what might cause it?


Thanks,


Richard McFarland
UT Southwestern at Dallas



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