[Histonet] LR White and immunofluorescence

Va Paula Sicurello vapatpxs <@t> yahoo.com
Fri Jun 19 12:35:03 CDT 2009

Happy Friday Everybody,

I have a user who has a precious sample and wants to embed in LR White, maybe.  I'm having him practice on something not precious first.  My questions are:

1.  He wants to do immunofluorescence and morphology on these nerve samples.  He is planning on perfusing in 2% PFA, 0.1% glut in 0.1M sodium cacodylate.  Then he will dissect out the nerve, cut it in half and place half in 2% PFA, 0.1% glut (embed this in LR White) and the other half in 4% PFA, 2.5% glut (embed this in Epon) to finish the fixation.  Will that work? Or will the perfusion fixation with the weaker fix cause the stronger submersion fix to be moot?

2.  He wants to do immunofluorescence on the LR white sample.  I figure, primary antibody, then secondary with glow is no different that secondary with gold ball, am I correct in this?

Thanks and have a great weekend.

All aglow waiting for your answer,

Paula :-)

Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core Research Imaging Center (CRIC)
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397

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