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We are currently working with alveolar macrophages, which express a huge
amount of autofluorescence (nice diagonal background on Fl-1 v Fl-2). Upon
staining eg. with a CD14PE or CD11cPE it becomes almost impossible to
qualify the stain, due to this high background. My question is: Is there any
possible way to quench this fluorescence in alveolar macs (is the background
carbon particles phagocytosed in the alveoli by the macs...maybe, but how
can I reduce it...is it possible to quench intracellular particles, pre
stain) I have attempted to use compensation, with reduction of PMT volts,
but I have had as much luck as a fish out of water breathing. Is this
background going to be as high if I stain with non PE or FITC (does the
background bleed into higher wavelengths eg texas red or rhodamine ~630 nm).
Saturation with antibody has very little effect in our case. So many
questions.... I would love to hear from other frustrated macists and their
reslove to this problem (if there is one).
Fanx for your time
Geza Paukovics - Flow Laboratory - AIDS Pathogenesis Research Unit
Macfarlane Burnet Centre for Medical Research .***. .***. .**
PO Box 254 _--_|\ * | | | * * | |
Fairfield, VIC 3078 / \ * * | | | * * | | |
AUSTRALIA. \_.--._/ * * | | | * | | | *
ph. (+61 3) 9282 2132, O '***' '***'
FAX (+61 3) 9282 2100
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web