Re: Mystery:Disappearing melanin!
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From: | "R.Wadley" <s9803537@pop3.unsw.edu.au> (by way of histonet) |
To: | histonet <histonet@magicnet.net> |
Reply-To: | |
Content-Type: | text/plain; charset="us-ascii" |
Dear Greg,
Sounds like the granules are not fixed, ie not fixed into place in the
cells & not cohesive within themselves. Have you tried drying the slides
after staining? My favorite xylene replacement fluid was not miscible with
my xylene mountant I got excellent results be drying the slides (on a hot
plate) then coverslipping.
In your case maybe using glycerine as the mountant (whether or not
you dry
the slide first) will fix your problem. When I was Curator of the Rodda
Pathology Museum, University of Tasmania, the manual recommended gelatin
solutions, glycerine and liquid paraffin as a mounting mediums for
melanomas because it prevented the melanin (& other colouring agents) from
leaking out of the specimen & discolouring the mounting fluid. A little
clear nail polish (or even your usual mounting medium) around the edge of
the coverslip prevents the mountant from drying out.
Hope this helps.
Regards
Rob W.
At 08:24 AM 2/19/99 -0400, you wrote:
...
>One of our graduate students is working on lobster hemolymph (blood).
>She is trying to use cytochemistry to demonstrate phenyloxidase
>activity in lobster hemocytes.
...
>She is obtaining nice brown-black staining of intracellular
>granules, BUT, the staining disappears when the mounting media is
>placed on the cells prior to coverslipping.
>
>The staining survives hematoxylin counterstaining and subsequent tap
>water wash, survives the alcohol dehydration, and xylene clearing,
>but disappears when mounting media added. She tried both solvent
>based and aqueous mounting medias. ANY IDEAS???
R. Wadley, B.App.Sc, M.L.S
Laboratory Manager
Cellular Analysis Facility
School of Microbiology & Immunology
UNSW, New South Wales, Australia, 2052
Ph (BH) +61 (2) 9385 3517
Ph (AH) +61 (2) 9555 1239
Fax +61 (2) 9385 1591
E-mail r.wadley@unsw.edu.au
www http://www.unsw.edu.au/clients/microbiology/CAF.html
(Under development)
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