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.nos...@vneubert.com> To: histonet@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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
