Re: GFP tissue preparation for confocal microscopy

Posted by Caroline Bass on
URL: http://confocal-microscopy-list.275.s1.nabble.com/GFP-tissue-preparation-for-confocal-microscopy-tp2680696p2684173.html

I concur. I regularly perfuse rats and mice with formaldehyde, and I don’t have any problem getting a signal (viral vector delivered EGFP in brain, spleen, liver, muscle, etc.). I have seen severe loss with any sort of dehydration or methanol fixation in fresh tissue.

Can someone recommend a good PFA? (Product number and company please!) The ampules sound interesting...


On 4/23/09 11:19 AM, "Stephen Bunnell" <Stephen.Bunnell@...> wrote:

PFA does not kill GFP fluorescence.

BAD PFA kills GFP fluorescence.

We routinely use 1% PFA with GFP, YFP, and CFP, and have never had a problem so long as the PFA is fresh and pH’ed to between 7.0 and 7.4.

    -Steve



On 4/22/09 10:34 PM, "Young Jik Kwon" <kwonyj@...> wrote:

All,

We are trying to take confocal micrographs of at tumor tissues that express GFP. What would be the sample preparation procedures? We tried cryosectioned samples but the cell morphology seems weird. We already know paraformaldehyde fixation kills GFP fluorescence. Any expert's advice?

Best,

Young

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