Re: Brain sections - Advice.

From:Geoff McAuliffe
Dear Ian:

    Mount the sections on gelatin coated slides from dilute (0.05M or less) phosphate buffer without saline. Saline leaves lots of crystals when it evaporates. Mounting from distilled water may cause sections to curl, espcially folia of the cerebellum, at least in my hands. Dry overnight or longer, I use gentle heat but that is not essential. Rinse off any crystals, dehydrate, clear, mount. If the sections are free of precipitates (from the buffer) you can air dry and go straight to xylene. Definitely use DPX. Give serious consideration to treating the DAB with nickel, nickel+cobalt, osmium, gold chloride, something to prevent fading.
    If you dehydrate free-floating sections they will get brittle and you won't be able to flatten them on a slide without damage.

Geoff

Ian Montgomery wrote:
        50 micron free floating brain sections sitting in phosphate buffer after IHC, DAB final product. What is the preferred method for dehydration? Take them through free floating then mount from final xylene. Or, dry them onto a slide then dehydrate. If the latter, coated slides? and how long drying, air temperature or using heat?
Ian.

Dr. Ian Montgomery,
Histotechnology,
Graham Kerr Building,
Institute of Biomedical & Life Sciences,
University of Glasgow,
Glasgow,
G12 8QQ.
Tel: 0141 339 8855
Office: 4652
Lab: 6644.
Pager: 07625 702883
e-mail: ian.montgomery@bio.gla.ac.uk


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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029 
mcauliff@umdnj.edu
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