PFA fixation and dead cells

Ian Titley Ian.Titley at icr.ac.uk
Mon Oct 24 10:27:58 EST 2005


Hi Paolo

Never used them but Invitrogen/Molecular probes have a series of "LIVE/DEAD® Fixable"
dyes in different colours, address for the green variant below:


https://catalog.invitrogen.com/index.cfm?fuseaction=viewCatalog.viewProductDetails&produc
tDescription=14695&CMP=LEC-GCMSSEARCH&HQS=L23101 


They claim on the website that the differential (50x) is maintained between "live and
dead" cells after formaldehyde fixation.

Might be worth a peek.

Best wishes

Ian


Ian Titley PhD
Section of Haemato-oncology
Institute of Cancer Research
237 Fulham Road
London UK SW3 6JB

Tel +44 (0)20 7352 8133
Fax +44 (0)20 7352 3299

>>> paolo <paolo+ at pitt.edu> 21/10/2005 13:41:25 >>>
Dear Fellow Flowers,

I work with HIV infected cultures and I need to determine apoptotic vs live vs dead
cells. I use an Coulter XL with a single laser (488nm) and I have to fix cells with 1%
PFA as they are loaded with HIV. If I use PI or 7AAD all cells will turn dead after
fixation. Can anyone suggest alternatives to these dyes? I already tried EMA, but perhaps
there something else out there that does not require the extra step with a light
source...

Thanks in advance for all your help.
-- 
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