Fixing and Preserving GFP Expressing Cells

darkwng · #1 01-29-2009, 04:24 AM

Fixing and Preserving GFP Expressing Cells:
Hi, i would like to fix and section GFP expressing cells. What is the best way to do it in order to maintain GFP fluorescence?

admin · #2 01-29-2009, 04:24 AM

Hi, one can fix the GFP cells with 1:1 Methanol:Acetone quenches GFP but not Paraformaldehyde

jiji324 · #3 01-29-2009, 04:25 AM

4% PFA (ph 7.4) for 30min at RT did the trick for us....

admin · #4 01-29-2009, 04:25 AM

Hi jiji thats a good point, both PFA and Methanol are good to preserve GFP signal. However when using methanol, fixation at low temperature is critical. When using PFA, correct pH (7.4) is important.

prashmi · #5 01-21-2010, 04:50 PM

I do 2% PFA with 4% sucrose at room temp for 15 mins. Seems to work well in my case.

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Fixing and Preserving GFP Expressing Cells

Fixing and Preserving GFP Expressing Cells - Immunofluorescence Forum

Fixing and Preserving GFP Expressing Cells - Immunofluorescence Forum. Talk, discuss and post questions on immunofluorescence techniques including paraffin, frozen sections or cultured cells.