marking cells

sue at murphy.prestel.co.uk sue at murphy.prestel.co.uk
Mon Oct 7 16:09:49 EST 1996


whc at mole.bio.cam.ac.uk (W.H. Colledge) wrote:

>Does anyone have a method for labelling (non-radioactive) or marking
>non-dividing cells that will persist for the lifetime of the cell?

>-- 
>Dr.W.H.Colledge
>Physiological Laboratory
>University of Cambridge
>Cambridge CB2 3EG

Dear Dr Colledge,
	You could perhaps try CFSE (carboxyfluorescein succinimidyl ester)
from Molecular Probes Inc USA.  Its a precursor fluorochrome which
only becomes active once acted on by esterases within the cell, and is
also a viability stain.  Also its non-membrane permeable after its
been converted thus.  I used it to label 3T3 fibroblasts and it was
also passed on to daughter cells, although that's apparently not an
issue with your cell type.  Cells were incubated in tubes of HBSS
(should work with whatever your favoured salt solution is) with the
stain for 15 min at 37C and then washed and pelleted twice prior to
replating.  I tried final concentrations from 500 microgrammes/ml down
to 50ng/ml and assessed the cell cultures for fluorescence under
normal FITC filters the next day.  I got best results with 500ng/ml,
but this varies with cell type.  Its been used by someone else for
looking at cell fates in embryogenesis, so presumably can hang around
for quite a while- depends how long your cells live for I guess!  Hope
this is some help,
 	Sue Murphy.
--
Sue Murphy PhD         ><          
St Thomas'Hosp IVF Unit><          
London UK              ><      
Above views are my own, not necessarily my organization's!!!




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