triple labeling

Gerald W. Gordon gordon at med.unc.edu
Tue Jan 13 12:42:56 EST 1998


Disclaimer: I'm familiar with neither your instrument nor
your application. But here are my 2 cents to get the ball
rolling.

A simple approach would be to collect more data so
that the precision would be adequate for color compensation.

Judging from the subject (triple label) I would guess that
you are using GFP to identify cells in which to look for
CD34 and CD38 which have longer wavelength dyes attached.
If this guess is correct you could use an inducible promoter
for the GFP to reduce the signal size and therefore the
crosstalk.

Jerry
gordon at med.unc.edu

Jordi Barquinero wrote:
> 
> Dear GFP netters,
> We are trying to analyze the simultaneous expression of two phenotypic
> markers (CD34 and CD38) in human hematopoietic cells transduced with the
> EGFP gene, but EGFP signal (FL1) is very bright and produces a level of
> background in the other channels (FL2, FL3 and FL4) that makes color
> compensation imposible. We are using a COULTER XL cytometer with an
> argon-ion laser (488 nm). Any hints to help us? Thanks.
> 
> Jordi Barquinero, M.D.
> Institut de Recerca Oncologica
> 08907 Hospitalet
> Barcelona, SPAIN
> Tel (93) 2633817
> Fax (93) 2632251
> jbarquinero at iro.es



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