RE: Small cytologic specimen handling recommendations

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From:Penelope Marr <MarrP@sesahs.nsw.GOV.AU> (by way of histonet)
To:histonet <>
Content-Type:text/plain; charset="us-ascii"

May I add a word of caution about colouring small specimens.  If
immunohistochemistry is to be performed on any of these tissues or cell
blocks remaining colour can mask positive staining, particularly if it
is weak.  Both pink and blue compete with the brown colour of the DAB
deposit.  Too much of either will mask weak staining thus making an
accurate assessment of the result very difficult if not impossible.
Also, colouring a specimen can make orientation at embedding more

Where ever I have worked we have always returned to the use of thrombin
clots / ager pellets for cytological material and the use of biopsy pads
/ paper for small biopsy material.

Penny Marr
> -----Original Message-----
> From:	R.Wadley []
> Sent:	Wednesday, 24 February 1999 12:32
> To:
> Subject:	Re: Small cytologic specimen handling recommendations
> 	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
> www
> 	(Under development)
South Eastern Sydney Area Health Service

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