RE: Processing Carnoy's Fixed Tissue

From:"Marshall Terry Dr, Consultant Histopathologist"

Teri Johnson suggests that the cause of the black/brown nuclei Nicola is having trouble with may be due to cornflake artifact.
This sounds the only likely explanation to me too.
Slip one in an envelope and push it over the Pennines to me Nicola.


Terry L Marshall B.A.(Law), M.B.Ch.B., F.R.C.Path
Consultant Histopathologist
Rotherham General Hospital, 
Moorgate Rd
Oakwood
Rotherham
S60 2UD
Yorkshire
terry.marshall@rgh-tr.trent.nhs.uk

-----Original Message-----
From: Johnson, Teri [mailto:TJJ@Stowers-Institute.org]
Sent: 18 September 2002 15:48
To: Histonet (E-mail)
Subject: RE: Processing Carnoy's Fixed Tissue


Hi Nicola,

Is it possible to post a photomicrograph onto the www.histonet.org website?  I'd really like to see what your nuclei look like.  It could be the "cornflaking" artifact that you get when you have sections that dry before coverslipping.

Initially my first thought is that you do not need to transfer to 70% alcohol prior to processing.  It's an anhydrous fixative, and therefore should probably stay anhydrous through holding and processing.  Otherwise, more information about your processing schedule and reagents would also be helpful.

Teri Johnson
Managing Director Histology Core Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, Missouri  64110
tjj@stowers-institute.org


-----Original Message-----
From: ncragg [mailto:n.cragg@epistem.co.uk]
Sent: Wednesday, September 18, 2002 3:24 AM
To: 'HistoNet Server'
Subject: Processing Carnoy's Fixed Tissue


Hello,

I'm emailing from Manchester in the UK and just starting my histology 
career.  Please can anyone help?

Just read some archived mail on Histonet regarding processing carnoy's 
fixed tissue - I really need some help with this, as we are getting strange 
black/brown nuclei throughout the tissue.  It has been suggested that this 
is the acid remaining in the nuclei.  We have a Leica carousel tissue 
processor without a vacuum, however, I have used an overnight 18 hours 
program and still can't get rid of this. We also use other fixatives which 
aren't a problem, but we would like to resolve the problem with carnoy's 
fixed tissue, as we use this fixative a lot in our lab and it is necessary 
for soem of our work.  Tissues are fixed for approx. 1 hour in Carnoy's and 
then transferred to 70% prior to processing.

I have also read on Histonet that there are 2 references regarding the 
processing of carnoy's fixed tissue, however, there aren't any abstracts on 
PubMed and I'm not sure whether our library holds this journal.  Does 
anyone know how I can get hold of them?  They are:-
Puchtler H et al  Histochemie 16:361-371, 1968
Puchtler H et al  Histochemie 21:97-116,	1970

If anyone has any other advice for dealing with these tissues, I would be 
very grateful if you could pass it on.

Kind regards,

Nicola Cragg
Epistem Ltd
Manchester, UK







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