Re: Small cytologic specimen handling recommendations

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From:"R.Wadley" <> (by way of histonet)
To:histonet <>
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	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
	(Under development)

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