[Histonet] Hematoxylin issue

[WILLIAM DESALVO]

I suggest you look at the reagents and protocol used 5 years ago and first determine if the same is used today. If you know all the reagents used, then start by check three specifics of the process: 1.) Removal of alum - make sure the slides are sufficiently rinsed. Suggested is 3-4 minutes to adequately remove the alum, you are using only 1 minute; 2.) Mounting Media - make sure the mounting media used had an antioxidant to prevent fading; 3.) Xylene - mixtures can cause fading, you made need to move to higher grade.
 
This is only a starting point, having the exact protocol and reagents used then and compare them to now is critical. I would pull samples for each year forward from the found problem and check to see if you have fading. I do not have an easy answere to fix and it may be necessary to recut slides, from the retained blocks, if the archived case is requested for review.
 
Most important is to find the cause, stop the problem and move forward.


William DeSalvo, B.S., HTL(ASCP)
Production Manager-Anatomic Pathology
Chair, NSH Quality Management Committee
Owner/Consultant, Collaborative Advantage Consulting

 

> Date: Tue, 18 Dec 2012 14:45:42 -0500
> From: Janine.SimmsColon <@t> jmmc.com
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Hematoxylin issue
> 
> Good afternoon all, 
> 
> 
> 
> I have been addressed with an issue which I would like some assistance
> with. My pathologist recently reviewed some skin slides from 2007 and
> noticed the hematoxylin was completely gone and only the eosin remained.
> We use Gill's II hematoxylin, and glass cover slips with xylene
> substitute mountant. The concern is that since we must retain slides for
> 10 years and the stain has "washed out" after only 5 this could be a
> pretty big problem. I do not believe this particular lab has experienced
> this problem before but I have only worked here for less than one year.
> We stain in hematoxylin for 3 minutes, use commercially available
> clarifier and bluing solution, each for one minute as well as one minute
> tap water rinses in between and dip in 95% alcohol before and after a 1%
> alcoholic eosin y solution. I searched the histonet archives for this
> problem as well which is why I am mentioning the staining process but I
> was curious if anyone out there had any suggestions or advice to avoid
> this issue for future slides. Thank you in advance. 
> 
> 
> 
> Janine Simms Colon, CPhT(PTCB), HT(ASCP)
> 
> Histology/Pathology
> 
> Johnson Memorial Hospital
> 
> 201 Chestnut Hill Road
> 
> Stafford Springs, CT 06076
> 
> Office: 860-684-8230 ext. 5197
> 
> janine.simmscolon <@t> jmmc.com
> 
> 
> 
> 
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