210 BD FACSCanto II Instructions for Use
BD FACSCanto Software TBNK Analysis
QC Messages
Message
Possible Causes
Recommended Solutions
Sample quality
questionable
Donor-specific anomaly Adjust gates manually to include
required subsets.
Insufficient mixing
during sample
preparation: cell
populations elongated in
CD3 vs SSC and FITC vs
PE plots
Re-stain the sample. Vortex after
blood and reagent are added to
tube. Make sure no blood
remains on the side of the tube.
Sample not lysed
adequately: cell
populations in CD45 vs
SSC extend upward
Prepare sample again, ensuring
complete lysis.
Aged blood and/or
stained sample:
granulocytes have low
side scatter in CD45 vs
SSC plot; no distinct
monocyte population
Refer to the information
supplied with the reagent for
stability limitations.
Excessive mixing: debris
encroaching on
populations in CD45 vs
SSC plot
Re-stain the sample and run it
again.
Low voltage for SSC
detector
Increase the SSC voltage and run
the sample(s) again.
Various Look at the dot plots in
BD FACSCanto Software Four-
and Six-Color TBNK on
page 212. Re-run the sample.
Sample not mixed
properly
Vortex prior to loading the
sample onto the SIT.
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