Cornelius Krasel wrote:
>> Steffen Dietzel <s.dietzel at pop.life.uiuc.edu> wrote:
> > I am looking at mammal cell culture cells expressing GFP. To facilitate
> > observation of cells in the fluorescent microscope, I would like to use an
> > Anti-Fade reagent that minimizes bleaching. Currently we have Propyl Galate
> > solved in 90% Glycerol in the lab, however it quenches the GFP-fluorescene in
> > formaldehyde fixed cells pretty much!
>> Are you sure that it is the Propyl Galate? When I mount GFP-expressing
> cells in Propyl Galate without fixing, I get a lot of fluorescence;
> however, when I first fix them in 4% paraformaldehyde, fluorescence is
> almost completely lost :-(
>
Yes, I am sure. When I do the same preparation (including
Formaldehydefixation) and mount the cells at the end in Glycerol only (without
Propyl Galate), fluorescence is o.k. Same if mounted in PBS. Yust that it
bleaches pretty fast. However, i have not tried unfixed cells with Propyl
Galate. Maybe this would work too, but not in the experiment I am planning :-(
Could it be that some GFPs work with a given Anti-Fade and others don't?
Steffen
