Re: GFP tissue preparation for confocal microscopy

Posted by anh2006 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/GFP-tissue-preparation-for-confocal-microscopy-tp2680696p2686595.html

Your understanding is correct. It's a misnomer, extremely common and although incorrect, it's generally accepted.

At 4:04 PM -0400 4/23/09, David Knecht wrote:

 I have always used the comparable Polysciences ampules of 16% formaldehyde.  I don't understand the Alfa product. My understanding was that paraformaldehyde was a solid, and dissolving it led to it being converted to formaldehyde.  They are selling it as 16% paraformaldehyde.    Dave

On Apr 23, 2009, at 12:15 PM, Andrea Hooper wrote:

Dear Caroline,

What do you mean by dehydration precisely? Do you dry your slides after sectioning?

Here is our ampoules info:

(1) http://www.alfa.com, Alfa Aesar, Item #43368

(2) http://www.emsdiasum.com, Electron Microscopy Sciences, Item # 15710

Thanks,

Andrea

At 12:05 PM -0400 4/23/09, Caroline Bass wrote:

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...

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Dr. David Knecht  

Department of Molecular and Cell Biology

Co-head Flow Cytometry and Confocal Microscopy Facility

U-3125

91 N. Eagleville Rd.

University of Connecticut

Storrs, CT 06269

860-486-2200

860-486-4331 (fax)

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