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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.


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  #1  
Old 01-29-2009, 05:24 AM
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Question Fixing and Preserving GFP Expressing Cells



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?
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Old 01-29-2009, 05:24 AM
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Default Re: Fixing and Preserving GFP Expressing Cells

Hi, one can fix the GFP cells with 1:1 Methanol:Acetone quenches GFP but not Paraformaldehyde
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Old 01-29-2009, 05:25 AM
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Default Re: Fixing and Preserving GFP Expressing Cells

4% PFA (ph 7.4) for 30min at RT did the trick for us....
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Old 01-29-2009, 05:25 AM
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Default Re: Fixing and Preserving GFP Expressing Cells

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.
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Old 01-21-2010, 05:50 PM
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Default Re: Fixing and Preserving GFP Expressing Cells

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