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2. Decrease of H2O2 production in the presence of
thiazolylcephalosporins, See Labro et.al. J. Immunol 1994 Vol 152
(5) 244-55.
Your combo includes Mycostatin (a molecule that increases membrane
permeability and leakage) and Penicillin V/Streptomcin. Pen V is a
major amplifier of chemiluminescence in the youngster's assay,
apparently the beta lactam alters the route and efficiency of
peroxide radical transfer, by a mechanism unknown. I think it is
perfecly feasible that the same process that enhances the luminol
chemiluminescence may inhibit peroxidation of Dihydrorhodamine.
Just a speculation, but it is worth testing.
J. Peter Bercz
USEPA/EMSL/Cincinnati, OH
HI, everyone!
A problem has come up with one of our grad students using the ACAS
570 confocal laser scanning microscope. He is looking for the
oxidative burst in live cultured macrophages using:
dihydrorhodamine --H2O2, enzymes-->rhodamine 123
He used to get good rhodamine 123 fluorescence. Then he changed the
cell culture conditions - mainly added Pen/Strep and Nystatin to hold
down contamination. Now he gets little or no rhodamine 123
flourescence. We're wondering if the addition of antibiotics to the
culture medium could be the cause, or if there are other pitfalls in
this method. If any of you have any experience with this technique,
we would appreciate your input.
TIA
* * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * *
* Richard K. Meister Email: Meister.1@osu.edu *
* The Ohio State University Voice: (614) 292-9716 *
* Dept. of Veterinary Biosciences FAX: (614) 292-6473 *
* Cytometry Instrumentation Lab *
* 1925 Coffey Road *
* Columbus, OH 43210 U.S.A. *
* * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * *
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