[Histonet] Humidity levels and IHC staining

Ross Benik ross <@t> premierlab.com
Fri Feb 17 13:32:25 CST 2012


We've experienced similar problems a few summers ago when the environmental conditions inside the Autostainers (dako) reach high temperatures and low humidity.  We keep thermometers inside the stainers to monitor these levels.  To remedy this problem, before every IHC  run, I add a few damp paper towels in the stainer sink.  As the stain progresses, liquid is added to the paper towels by the stainer so I never have to worry about it drying out.  Since we've started doing this, the humidity ranges anywhere from 35-55% and an ending temperature of no more than 28C.  With these conditions, we've yet to experience slides drying during a staining run.  Another factor to consider is volume of reagent added to each slide, we do 100 ul per drop zone (3 drop zones total).  The longest a reagent left on a slide is 1 hour max for a few of our primary antibodies.

Hope this helps,
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Jason McGough [jmcgough <@t> clinlab.com]
Sent: Friday, February 17, 2012 12:09 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Humidity levels and IHC staining

We are wondering what other labs are doing to control the humidity while IHC
stains are being performed. We currently place wet towels and a small weigh
boat with water in our Autostainer to help prevent our slides from drying
out but that seems to not be enough, they still tend to dry out and produce
background staining. What should the humidity level be at? Any help would be

Jason McGough HT(ASCP)
Account Representative - Anatomic Pathology
Clinical Laboratory of the Black Hills
2805 5th Street Suite 210
Rapid City, SD 57701
605-343-2267 Ext 127
605-718-3779 (Fax)
jmcgough <@t> clinlab.com

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