Re: Calcium determination by flow cytometry

From: PeterG (gogy@freemail.hu)
Date: Fri Jul 19 2002 - 06:40:32 EST


Hello Alfredo,

It is depend on your cytometer.
You should have UV laser.
FURA-2 is used mainly for microscopy. Indo-1 is used in
flow cytometry instead.
If you have 488nm blue laser, you  can use eg. Fluo-3 as
Ca++ indicator. But in this case you cannot do ratiometric
measurement directly.
You can find the reasons and quiet good descriptions about
different Ca++ indicators and methods here:

http://www.probes.com/handbook/sections/2000.html

Best regards,

Gogy

Peter Gogolak, Ph.D.
Institute of Immunology
University of Debrecen, Hungary
Phone/Fax: +36 52 417159

Your original message:
Dear flowers,
I am interested in studying calcium response in human blood
leukocytes after a specific stimulation.
I used to analyse intracellular calcium in many cell types
by a spectrofluorimetric method using FURA 2AM.
May I use the same fluorochrome in flow cytometry?
Do you have any suggestions on the matter?
Any hint is welcome!

Thank you in advance,

Alfredo Vannacci


------------------------------------------------------------
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Alfredo Vannacci, MD

Department of Preclinical and Clinical Pharmacology
Medical Toxicology Unit
University of Florence
Viale Gaetano Pieraccini, 6
Firenze
50139, Italy

tel: +39(0)554271236
fax: +39(0)554271280

email: alfredo.vannacci@unifi.it



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