[Histonet] Problems with rhodanine

[Gayle Callis]

Katherine,

Are you even fixing the frozen sections AFTER you pick them up onto the 
slides and before staining? Sounds as though unfixed frozen sections are 
washing off the slides

At 01:25 PM 6/18/2007, you wrote:
>Hi all,
>
>I'm a graduate student in a neuropsychology lab, and we are having some
>problems with our histology. I was hoping someone might have some advice.
>
>My lab is doing staining for metals in fresh frozen mouse brain, sectioned
>at 10u or 20u. Some of our animals are fed high copper supplements and we
>see high copper in areas of our tissue with other methods, so we are trying
>to pilot rhodanine for copper. However, the protocols we have for rhodanine
>give conflicting instructions.
>
>We have unfixed, fresh frozen tissue, which is part of the problem, since
>ALL our protocols are for FFPE tissue. Two of the protocols call for
>microwave method, but we don't have an appropriate microwave so we are using
>the conventional oven method, which says to heat the slides for 18 hours at
>37 or 30 degrees centigrade.
>
>When we looked at the coverslipped slides, it looked as though the tissue
>had washed off in place. I have seen this happen before with immuno, and my
>histology professor said it was a problem with the slide subbing; however,
>we are using Erie Superfrost Plus slides, which I did not think would have
>that problem. We DID end up moving the slides between buildings, so maybe
>the tissue was jostled too much during transport and that's why it washed
>off?
>
>Does anyone have any experience with rhodanine on fresh frozen tissue? Do
>you need a special protocol?
>Does it just not work? Any input would be appreciated.
>
>Thanks in advance,
>Katherine
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610