[Histonet] Staining issues

[Tony Henwood (SCHN)]

Hi Charles,

Many issues could be in play.
Since the centre of the tissue is causing problems, then I would look at fixation; are the spaces around cells in the centre of the block wider than at the edges (conversely do the cells look like they are smaller than their counterparts at the edge)?

If part of this tissue is lightly fixed, they seem to be more prone to heating effects. If the haematoxylin is paler (washed-out?) in the centre then it is possible that the slides have been heated above 70oC before H&E staining. This will also affect the IPX's. Maybe the oven is over-heating.

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Principal Scientist, the Children's Hospital at Westmead
Adjunct Fellow, School of Medicine, University of Western Sydney 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 

-----Original Message-----
From: Charles Riley via Histonet [mailto:histonet at lists.utsouthwestern.edu] 
Sent: Thursday, 29 December 2016 12:19 AM
To: histonet at lists.utsouthwestern.edu
Subject: [Histonet] Staining issues

Our pathologists are complaining that the center of tissues are not staining properly in both our H&E's and IHC's. Can anyone provide some thoughts as to where this problem could be occurring?


As a separate situation we are experiencing the tissues folding or falling off during the staining process. Both this issue and the staining issue are recent occurrences and we have not changed our process in any way from previous years.


Does anyone think these problems are related? Why or why not?   I have
racked my brain with all the trouble shooting techniques I can think of so any help will greatly be appreciated

-- 

Charles Riley HT(ASCP)CM

Histopathology Coordinator/ Mohs
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