[Histonet] RE: Frozen sections for Immunoflouresence

[Tony Henwood (SCHN)]

Hi Jim,

You mention that the OCT seems a little softer.
Could it be that the sections are thicker than you usually produce. Check the Frozen section H&E to check this.
If sections are thicker they will tend to detach from the slides.

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager & Senior 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: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Vickroy, Jim
Sent: Wednesday, 3 September 2014 12:51 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Frozen sections for Immunoflouresence

Lately we are experiencing an issue where the tissue sections are coming off of the slide during the staining process.   We have always used positive charged slides and have not experienced this problem on a routine case.   We do not fix the slides but allow them to air dry for Immunology.  Some have suggested a quick fixation in acetone or 95% ETOH.   This doesn't seem to correct the problem and we don't find the IF stains work as well.   We have tried a different batch of slides and are looking into a different kind of coating for the slides.   Our regular frozen sections stay on the slides.  We are using OCT.   The only other thing I have noticed is that maybe it's my imagination but the OCT in the block surrounding the kidney biopsy seems a little softer than normal.   I did try a different mounting medium and maybe the softness was my imagination.   Has anyone else experienced this problem and if so how did you correct it?   The lab is air conditioned so I don't think the humidity is the main factor.  The Immunology staff changed all of their reagents so we have reduced that variable hopefully.   We are using Leica positive charged slides.  (I have found that the Apex control box slides from Leica are a little better than the regular Apex slides.   Any ideas?

Jim

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