Hi everyone!
I was wondering if anyone knows which tissue preperation (frozen vs paraffin) is better for confocal microscopy. Also, I will be using antibodies against phospho-proteins and I have heard that these do no do so well when tissue has been embedded in paraffin.
Any insight would be greatly appreciated!
Thanks,
Carmen
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3 replies to this topic
#2
scolix
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Posted 09 February 2012 - 11:01 AM
WIth paraffin, you can get thin sections. and wIth frozen you get thicker sections. Thinner sections are good for staining.
I have no experience comparing phospho antibodies on sections.
I have no experience comparing phospho antibodies on sections.
#3
Tabaluga
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Posted 09 February 2012 - 11:49 AM
Have you seen this thread ? There's also something on anti-phospho antibodies...
http://www.protocol-...osts/16990.html
http://www.protocol-...osts/16990.html
#4
knuf
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Posted 13 February 2012 - 09:25 AM
Regarding the confocal, either frozen or paraffin-embedded sections should work as long as the antibody works. Because of how the confocal microscope works, it is able to see through the tissue section to a single plane, so the thickness of the frozen sections doesn't really affect the confocal microscope like it would in a normal fluorescent microscope. So if you find that you get better staining/morphology with the frozen sections then use the frozen sections.
