[Histonet] Help

[Rene J Buesa]

As to your issue of tissue not adhering to the slides, you could try to check the expiration date of your (+) slides. Perhaps it is just an issue with the slide.
As to controlling the "concentration" of an antibody by changing the incubation time, that is somewhat unorthodox to say the least. You modify a concentration with dilution, not with time. Perhaps you could modify the HIER step, or try to dilute the antibody.
René J.

--- On Fri, 5/18/12, Cloughley-Gray, Nancy <CloughleyN <@t> rvh.on.ca> wrote:


From: Cloughley-Gray, Nancy <CloughleyN <@t> rvh.on.ca>
Subject: [Histonet] Help
To: "'histonet <@t> lists.utsouthwestern.edu'" <histonet <@t> lists.utsouthwestern.edu>
Cc: "Callan, Lisa" <CallanL <@t> rvh.on.ca>
Date: Friday, May 18, 2012, 4:02 PM


I'm a Histotechnologist working in the Regional Hospital in Barrie, ON Canada. We are using the Ventana Ultra for our Immunohistochemistry (IHC). Since the end of February, we have been having issues with some tissues lifting off our positive (marked with +) charged slides. It seems to be mostly with the fatty and/or larger sections. We now dry our slides for one hour at room temperature (R.T.) and an additional hour at 60 degrees C. We cut our IHC sections at 4 um. Since we have tried 2 different types of + slides and will be trying another type of charged slide (from Newcomer this time) I was wondering if anyone has any other suggestions?
I also have another question regarding a QC (quality control) issue. We use a multi-tissue control that is applied to the top of all our test slides for IHC. One of our paths commented that there is some positive staining in the smooth muscle nuclei of the normal bowel when we are testing for Progesterone (PR). We are using a Heat Induce Epitope Retrieval (HEIR) of 36 minutes with CC1 (Ventana's proprietary buffer @ pH of 8.0-8.5) and a primary antibody incubation time of 16 minutes with PR clone 1E2. (Ventana instrumentation provides pre-diluted antibodies and the user adjusts the concentration of the antibody by adjusting the time the primary antibody is incubated with the tissue).
I am concerned about the implications of this staining and I have not been able to find a reference to this kind of unusual staining pattern. The bowel tissue that we are using as QC is from a 62 year old female patient. I was wondering if anyone has had any experience with this kind of staining and /or any references that I could use.

Thanking you in advance,
I look forward to your input,
Nancy Cloughley-Gray MLT

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet