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Author Topic: Ask help for ICC on RPFNA samples  (Read 4639 times)

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Offline fishguoguo

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Ask help for ICC on RPFNA samples
« on: July 23, 2009, 04:50:32 PM »
Hi everyone, thanks a lot first for any suggestions!

I am going to do phospho antibodies ICC on RPFNA samples. Acturally the collaborators will mail us the samples in CytoLyt solutions (modified with 10% buffered formalin) but not the slides. So, problems are what kind of slides I can make? I have to make ThinPrep slides or I can make normal cytospin as well? If they use fixation solution first before sent to us. Can I still do ICC on these cells? What do you prefer, ThinPrep or cytospin? I heart that Thinprep need more cells but clear background. Acturally we are going to have limited samples and the boss asked to do as many slides as possible. We have cytospin machine in our own lab. I checked literatures these days, but didn't get useful information. So I think experience is more necessary from your guys.

Another quick question. Do you always do antigen retrieval before performing staining? Can someone give me a ICC protocol on ThinPrep or Cytospin slides?

A long story there. Thank you very much for any suggesion again!!!

Looking forward to answers.

Ask help for ICC on RPFNA samples
« on: July 23, 2009, 04:50:32 PM »

Offline fishguoguo

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Re: Ask help for ICC on RPFNA samples
« Reply #1 on: August 10, 2009, 01:33:25 PM »
come on. No reply!

Offline richard03

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Re: Ask help for ICC on RPFNA samples
« Reply #2 on: August 10, 2009, 10:35:09 PM »
I have no experience with ThinPrep nor cytospin. Since you have a cytospin machine in your lab, it will be easy to go with cytospin. The links below are methods and sugguestions for making and storing cytospin slides for your reference.

http://www.ihcworld.com/_protocols/histology/cytospin.htm
http://www.ihcworld.com/smf/index.php?topic=1398.0

Antigen retrieval is mostly used for paraffin sections and occasionally used for frozen sections. It is rarely used on cultured cells. To enhance antibody penetration and binding to its antigen, Triton X-100 (0.1%-0.5%) is usually used.

Re: Ask help for ICC on RPFNA samples
« Reply #2 on: August 10, 2009, 10:35:09 PM »