[Histonet] Strange circles in IHC slides

[Kim Merriam]

I use this system quite often, when doing manual IHC.  I believe that these are air bubbles that were in the buffer solution that you used when you first put the slides into the cytoclips; I usually let the buffer settle a little bit in the beaker before I load my slides (to allow time for any bubbles to pop).  Then I put about 1-2 mls of buffer (with detergent) at the bottom of the clip and slowly put the slide onto the clip (just like coverslipping); I always make sure that there are no visible bubbles when loading the slides (again, just like coverslipping).  Then I use an eppendorf repeat-pipettor (set at 2-mls) for my washes (again, with buffer + detergent).
 Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA 




________________________________
From: V. Neubert <histonet.nospam <@t> vneubert.com>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Thursday, April 2, 2009 12:12:18 PM
Subject: [Histonet] Strange circles in IHC slides

Hello,

I really have some real links to real pictures of real relevance of the real histonet list. Really promised.

http://img12.imageshack.us/img12/8513/ts0402162049.jpg
http://img13.imageshack.us/img13/6514/ts0402162104.jpg

So, has ever anyone experienced sth. like this?
My conjugate control (every step except the antibody) was fine, nothing to be seen about DAB and no circles at all.

I used Shandon single-use coverplates, sterile buffer, fresh antibody aliquots. Any idea?

Thanks,

V. Neubert

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