Expression of GFP fusion protein - Improving fluorescence visualization (Nov/20/2006 )
Hi everyone! I´m studing the localization of my bacterial protein and I have done a GFP fusion. My problem is that I can only see a tiny amount of fluorescence and would like to improve this. Right now I just look at the samples of the culture the bacteria is growing in but I´m reading other papers where cells are fixed for visualization (paraformaldehyde, etc). Coulld anyone give any advice or tell me the advantage of fixing the cell?
suitable for immuncytochemistry; but if you see only faint GFP-signals in living cells you won´t improve intensity by fixation;
Thanks, but I'm still wondering if the way I'm preparing the slides makes the image aquisiton hard? I only put a bit of bacterial culture on a slide and I see papers where the bacteria are placed on the coverslip?