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> IS THERE ANYONE WITH EXPERIENCE OF USING A FACSCAN FOR QUANTITATING MALARIA
> INFECTED ERYTHROCYTES. OUR INITIAL EXPERIMENTS INDICATE THAT PI CAN BE USED
> AFTER GLUTARALDEHYDE FIXATION AND TWEEN PERMEABILIZATION. HOWEVER. I GUESS
> THERE MAY BE BETTER PROTOCOLS USING MEMBRANE PARMEABLE DYES?..
I don't know of a membrane permeable dye for use on a FACScan but we
have used Hoechst dyes on viable cels (Howard et al J.Histochem.Cytochem.
v27 p803-813 '79) or on fixed cells (Bianco et al Exp.Parasitol v62
p275-282 '86) on a sorter with UV laser. If you are restricted to a
FACScan, you may be interested in the information on fixation given in
the latter reference. Recently, we have been using the fixative
therein suggested (formaldehyde plus glucose) for FACScan analyses
using the stain Thiazole Orange. Unfortunately, the reason we used
that stain in preference to PI has been lost in the mists of time (I
think we were worried about possible background staining by PI) but we
have routinely measured parasitaemias of 0.3% to 25%. This question
may be academic only because I believe Thiazole Orange is no longer
produced (Polysciences is our last known supplier).
\ / < Flow Systems Laboratory
Frank Battye \__/ <<<<< The Walter & Eliza Hall Institute
Robyn Muir ----------!!<<<<<<<< Voice: 61_3_345 2540
Dora Constantinou /!!\ <<<<< Fax: 61_3_347 0852
o !! \ < IN:: "facs_copy@wehi.edu.au"
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