[Histonet] RE: Hematoxylin issue

[Rittman, Barry R]

Difficult to say without seeing but there could be many reasons for this.
The hematoxylin could have either been removed by traces of acid before mountiung or if transferred to the mounting medium.

Hematoxylin could have been further oxidized to a colorless oxyhematein. This can occur with some mounting media , exposiure to light etc.

Too alkaline mountiung medioum unlikely or else you would have also lost the eosin.

Not sure about xylene substitute.

It is possible to retain hematoxylin - I have some slide stained in 1957 and they still have excellent hematoxylin and eosin staining but were mounted in Canada balsam in xylene.
Barry



From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of SimmsColon, Janine [Janine.SimmsColon <@t> jmmc.com]
Sent: Tuesday, December 18, 2012 1:45 PM
To: Histonet
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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