[Histonet] RE: brain sections falling off slides during IHC

[Watson, Linda]

Just to add to Christina,
Try Gold Plus Slides-Thermo Shandon (previously Erie Scientific).

Linda

>-----Original Message-----
>From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-
>bounces <@t> lists.utsouthwestern.edu] On Behalf Of Linke, Noelle
>Sent: Wednesday, February 11, 2009 11:12 AM
>To: Thurby, Christina; histonet <@t> lists.utsouthwestern.edu
>Subject: [Histonet] RE: brain sections falling off slides during IHC
>
>Hi Christina,
>
>In my experience, there are a few things to try.  First, don't soak your
>blocks too long before cutting, and try not to let too much water get
>underneath the sections as you pick them up from the waterbath.  I
>wouldn't put them in an oven at all, just let them air dry as long as
>you can (overnight, a day if you can spare the time) before staining.
>
>If you're having troubles, I wouldn't use the autostainer at all (it
>rinses too aggressively for some tissues), I would do them manually and
>never let the stream from your wash bottle touch the tissue, gently
>rinse after each step.
>
>I hope this helps.
>
>Noelle
>
>Noёlle Linke M.S., HTL(ASCP)QIHC
>Manager, Histology Services
>Department of Pathology & Laboratory Medicine
>David Geffen School of Medicine at UCLA
>Phone: 310-825-7397
>Pager: 97471
>nlinke <@t> mednet.ucla.edu
>
>-----Original Message-----
>From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-
>bounces <@t> lists.utsouthwestern.edu] On Behalf Of Thurby, Christina
>Sent: Wednesday, February 11, 2009 7:38 AM
>To: histonet <@t> lists.utsouthwestern.edu
>Subject: [Histonet] brain sections falling off slides during IHC
>
>Hello all,
>Can any of you all with experience immunostaining brain sections please
>help.  My sections are almost completely washing off the slides.  I am
>using 10% NBF fixed canine brain sections.  Embedded in paraffin and
>sectioned at 4 microns on plus slides with distilled water in the water
>bath.  The slides are dried at 65 C for 15 minutes and allowed to stand
>overnight.  I am using a Dako Autostainer for staining.  There is no
>antigen retrieval step.  My staining procedure is approximately 3.5
>hours long.  At the completion of the procedure, the tissue on my slides
>is 50-80% washed off.
>
>Any thoughts??
>Christina Thurby
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>IMPORTANT WARNING:  This email (and any attachments) is only intended
>for the use of the person or entity to which it is addressed, and may
>contain information that is privileged and confidential.  You, the
>recipient, are obligated to maintain it in a safe, secure and
>confidential manner.  Unauthorized redisclosure or failure to maintain
>confidentiality may subject you to federal and state penalties. If you
>are not the intended recipient, please immediately notify us by return
>email, and delete this message from your computer.
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet