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Fixation of GFP bacteria


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2 replies to this topic

#1 virusfan

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Posted 23 July 2009 - 06:23 AM

Hi all.

My live GFP expressing bacteria appeared as bright green under confocal microscope, but the Methanol fixed slides showed no green signal at all.

I'm wondering if the lost of GFP signal is due to the Methanol fixation? If yes, how can Methanol quench my GFP signal?

Thank you.

#2 Michaelro

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Posted 23 July 2009 - 06:38 AM

Hi
I know that GFP may escape during the methanol fixation.
The possible solution is to use formaldehyde fixation or mirystoylated GFP
Best
Michael

quote name='virusfan' date='Jul 23 2009, 05:23 PM' post='30198']
Hi all.

My live GFP expressing bacteria appeared as bright green under confocal microscope, but the Methanol fixed slides showed no green signal at all.

I'm wondering if the lost of GFP signal is due to the Methanol fixation? If yes, how can Methanol quench my GFP signal?

Thank you.
[/quote]

#3 little mouse

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Posted 23 July 2009 - 06:41 AM

I would use 0.5% formaldehyde to fix the bacteria




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