Lipfectamine fluorescent

Wolfgang Resch wresch at email.unc.edu
Mon Aug 31 20:24:27 EST 1998



On Mon, 31 Aug 1998, Tam Anh Le wrote:

> Hi all
>     I've just started using lipofectamine from Gibco (life technologies) to
> transfect mouse Rho0 cells, however in the initial experiments I used a
> pGreen plasmid (  which codes a for a fluorescent protein). I found in the
> control of Rho0 cells and lipofectamine only that fluorescent activity was
> quiet intense in comparison to my autofluorescent control ( ie Rho0 cells
> only). My question  is that although the intensity of the combination of
> pGreen plasmid with lipofectamine was higher than all controls :
>  1. has anyone notice the same thing with this particular transfecting
> reagent?
> 2.  is there a way of eliminating this problem of fluorescent caused by the
> lipofectamine.
> 
> -with regards
> t.l
> --
> Tam Anh Le
> Clinical Neuroscience
> St Vincent Hospital Melbourne.
> 
hi there,

i don't have much experience in GFP fluorescence, but my question would be
: do you  look at living cells in medium ? if so, it's probably the phenol
red indicator in the medium which shows autofluorescence. at least for
FACS analysis, there cannot be any phenol red and with the scope i
observed much less background without the indicator.
hope this helps.
wolfgang




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