Re: Cell carryover on FacsCalibur with loader

From: Keith Bahjat (kbahjat@ufl.edu)
Date: Fri Sep 24 1999 - 16:05:15 EST


As I understand it, with no tube on the SIP, sheath fluid flow backwards
out the SIP, where it would normally drip out, but is then caught by the
DCM. Giving the instrument a couple of seconds with the DCM running should
get enough sheath fluid backflushing to clear cells from the previous
sample.

Clinical labs utilizing cytometers are supposed to verify that
insignificant carryover between samples occurs. To test your instrument,
run a tube of PBS at various times after removing a tube contaiing stained
cells. We found that the typical 3 to 5 second delay implemented by the
loader was enough to flush sample from the SIP, but that sometimes users
manually placing tubes on the SIP would not wait long enough for this
backflush to complete.

Additionally, when initially placing a sample tube on the SIP, a small
amount of sheath fluid drips back into the tube. If you do not allow time
for the SIP to flush fully, this drop will contain cells from the previous
sample.

I use a 10 second mix between tubes, and see no carryover. During intense
investigation of this matter while running thiazole orange, I was unable to
detect rbc in subsequent PBS samples. I had suspect RBC's would cling to
the outer metal of the SIP, but they apparently did not.

Keith Bahjat
kbahjat@ufl.edu

Keith Bahjat
Graduate Assistant
University of Florida
College of Medicine
Gainesville, Florida
kbahjat@ufl.edu



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