Mike, I free float all of my rat brain sections (40-60um) and transfer them with a paintbrush. It takes a delicate touch to do this without causing damage to the tissue. I do know people who use pipettes to change the solutions. Either way works, it's a matter of personal preference. There are plastic vessels with mesh screens that can be purchased commercially, that allow you to transfer tissue without touching them. The only problem with that method is it requires additional amounts of antibody. For example: I dispense 500ul of antibody+diluent into the well when using the paintbrush transfer method (this is done under constant agitation on a shaker table). The plastic tissue vessel method requires 2 ml. of antibody+diluent (to cover tissue with enough fluid to account for the size of the vessel within the well itself). The link below offers commercially made tissue transfer vessels (look under circular staining nets and dishes).
http://www.brainresearchlab.com/productinform.html Good luck, Tina Montina J. Van Meter, HT (ASCP) Lab Manager Autonomic Neuroscience Pennington Biomedical Research Center 6400 Perkins Rd. Baton Rouge, LA 70791 225-763-2564 -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mitchell Jean A Sent: Friday, July 31, 2009 8:30 AM To: Michael Patrick; histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: IHC free floating sections Michael: I use 1.5mm nichrome transfer loops for IHC staining of 50um free floating sections and I move them into clean culture wells for each solution. I purchase mine from Ted Pella. It does takes some practice but the loops work quite well with minimal damage to the tissue. Jean Mitchell, BS, HT (ASCP) University of Wisconsin Hospital & Clinics Neuromuscular Laboratory 600 Highland Avenue Madison, WI 53792-5132 -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Michael Patrick Sent: Friday, July 31, 2009 2:14 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC free floating sections I would like to stain free floating sections (fixed rat brain; 50um). Is it necessary to transfer sections to "clean" culture wells with a paint brush every time I wash or incubate with antibodies? I find this causes significant damage to my tissue (especially since I do a lot of washes etc.). Could I simply use a pipette to draw out the old solution and then replace with the appropriate solution? What do most people do in these situations? Much thanks Mike _________________________________________________________________ Stay in the loop and chat with friends, right from your inbox! http://go.microsoft.com/?linkid=9671354_________________________________ ______________ 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet