[Cytometry] Quenching Autofluorescence
kprovost at buffalo.edu
Fri Apr 17 10:07:58 EDT 2009
I am looking for any suggestions on quenching red (and green) autofluorescence in
unfixed alveolar macrophages for multicolor flow cytometry. These cells
fluoresce bright green under a standard microscope and very brightly in all
channels of our cytometer (fitc, pe, pecy7, percp cy 5.5, apc, apc750).
We have tried the following protocols, with the outcomes listed.
B-D galactopyranoside (20% reduction but still very bright)
Crystal violet (decreased FITC/PE but increased in all red channels)
combinations of all of the above
Any thoughts on pH alterations, Janus dyes, light bleaching or other options?
Karin Provost, DO
Research Assistant Professor
SUNY at Buffalo
School of Medicine and Biomedical Sciences
VA Western NY Healthcare System
3495 Bailey Avenue
Buffalo, NY 14215
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