Re: Immunostaining in Neuropathology

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From:"Joyce Kotzuk" <JKotzuk@salud.unm.edu>
To:<histonet@pathology.swmed.edu>, <wheelock@zippy.mclean.org>
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Timothy, I also run a Neuropathology lab, and my primary experimental technique is immunostaining. We send the weekly autopsy brain cutting slides out to a clinical lab for H&E and any other special stains or immuno that the pathologists want for diagnosis.  I  do mainly research work, concerning Huntington's disease, ALS, Alzheimer's disease, and some other small projects. There are many, many good antibodies to all different types of neuronal components, receptor types, cell types, proteins that are components of pathological entities such as nuclear inclusion bodies associated with HD, Lewy bodies, amyloid etc.  I do my immuno by hand as opposed to using an automated stainer, and I use Vector ABC kits (an avidan/biotin labeling method) and substrate kits (DAB or NovaRed usually).  So yes, I would say immunostaining is a commonly used method in neuropathology these days.  The list of antibodies you would need would depend on the type of research you would be doing, but as I say, there are antibodies to  everything these days. I use one good web site when trying to locate new antibodies, as it is a free source of information and getting more complete all the time. The web site is http://www.antibodyresource.com/  From here you can search about six different databases to locate your antibodies. Hope this answers some of your questions, good luck.
Joyce Kotzuk, Neuropathology lab manager, Univ. of New Mexico Health Sciences Center

>>> "Timothy R. Wheelock" <wheelock@zippy.mclean.org> 02/03/00 01:02PM >>>
        I manage and single-handedly work a neuropathology lab at a
Brain Bank, and I am considering adding an immuno-staining capability to
my classical staining routine. I have some general questions to ask
before making such a move.
    (1) Has immuno-staining become a  standard routine practice in
neuropathology these days?
          We concentrate on degenerative and psychiatric disorders, and
rarely see tumors, so we don't  have
            to worry about tumor identification. Or is it only used
occasionally, for example,  to distinguish Diffuse
            Lewy Body disease from Pick's disease using Alpha B
crystalline antibodies? I get different answers
            from different pathologists.
    (2) If it is used routinely, what sort of immuno-stains are used?
                From one pathologist I get a list of Tau, Ubiquitin,
Amyloid, and Alpha B crystalline. From another
                I'm told that he rarely uses immuno-staining at all
except occasionally and that my Bielschowsky
                stains and LHE's show more than 99% of what's needed to
make a complete and detailed
                diagnosis.
    (3) Whether routine, frequent or occasional, does the neuropathology
lab tech do the immuno themselves, or simply out-source the immuno work
?
            One pathologist says that his histo-tech sends the unstained
slides to another lab that specializes in
            immuno and gets the slides back in one day,  rather than his
tech having to re-invent the wheel, at
            considerable expense and time, in adding an immuno routine
to her schedule.
    (4) If the immuno function is out-sourced what sort of fees can we
be expected to encounter for such a
          service?
    (5) Is there a review paper or protocols book that I could consult
that reviews the immuno-stains used in neuropath (other than tumors)?

        Thank you very much for any information and advice that you
could give me,

         Tim Wheelock






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