Re: Small cytologic specimen handling recommendations

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From:WAYNE HOLLAND <tissueman@home.com> (by way of histonet)
To:histonet <histonet@magicnet.net>
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Have you used the histoscreen cassette? It is agreat time saver, good
processing cassette with no artifact caused to the tissue. If you need
even more security you can use them with the Histogel thay have out now.
Its great for many time consuming tasks, it is some sort of gel that
melts when it is heated and gels up at room temp, or guick gel up when
it is cooled down.
Let me know if you need the info.
"R.Wadley" wrote:
>
>         Dear Gary,
>
>         When I was at the Royal Hobart Hospital the main way we handled
>very small
> biopsies, by putting them a small folded parcel of cigarette paper.  The
> paper was held closed between 2 pieces of cassette foam.  We found that
> using this paper on our fine needle biopsies prevented artifacts caused by
> the rough surface of the sponge imprinting on the biopsy.  We did not
> colour our biopsies at that time.  Other techniques included using thrombin
> clots, & agar pellets.  With Haematology down one end of the corridor &
> Cytology at the other end it was easy to get some aggregation of cells
> happening.  The cigarette paper sounds fiddly, but the technique is quickly
> mastered.  I used it exclusively with biopolymer specimens that tend to
> curl during dehydration to keep them flat.  Wrapping tiny specimens for TEM
> & SEM in this manner works just as well as for histo specimens.
>
>         While at the CSIRO I had a technician who coloured her samples with
> celestine blue.  I was not convinced that the stain actually made it
> through to the embedded block. But my 70% ethanol was very blue!  I agree
> with the comments by other histonetters that 1% eosin in ethanol (50%, 70%
> or 95%) with a few drops of acetic acid to make it colour fast is probably
> the way to go.  My prepference would by 70% ethanolic eosin, only because
> that is what I used in my H&E run.
>
> At 10:10 PM 2/22/99 -0500, you wrote:
> >Can anyone recommend materials and methods or products for processing small
> >specimens of loosely aggregated cells?  Is there, for example, material that
> >when added to formalin will coalesce/solidify/harden etc. so that the cells
> >will remain together and the bonding agent will remain intact through
> >embedment in paraffin?
> >
> >Specifically, these are specimens such as fine needle aspirates, endometrial
> >biopsies and endocervical curettages collected in formalin.  They are sent
> >to the histo lab where they are individually wrapped [in tissue], dehydrated
> >in alcohol, cleared in xylene, and embedded in paraffin.  Even if stained,
> >these can be very difficult for the histotechs to see the specimens, which
> >are small and blend in with the paraffin, and therefore difficult to
> >determine how far to section into the block.  Also, the specimen
> >disaggregates during processing so the cells are widely separated (or lost)
> >and less useful than they could be otherwise.
> >
> >Thanks for any and all suggestions.
> >
> >Gary Gill
> >
> >
> >
> >
> >
> 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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