Well..... If it is truly GMA then you will not be able to remove it. What you describe is one of several methods to remove MMA. You will also lose the occasional section on exposure to the higher ( 70% +) grades of alcohol. If it is GMA I'd suggest you place your slides directly in PBS and then start your IHC procedure.
Robert Schoonhoven, HT/HTL (ASCP) ________________________________ From: Allyse Mazzarelli <allyse...@gmail.com> To: histonet@lists.utsouthwestern.edu Sent: Wednesday, February 13, 2013 1:19 PM Subject: [Histonet] Immuno Bed Plastic Question Hi histonetters! I have posted on this forum a while ago asking for opinions regardnig new types of plastics that work well with IHC. It was brought to my attention that "Immuno-Bed" (Polysciences) works rather well. I recently purchased the plastic, infiltrated/embedded well, and sectioned great, BUT, when I removed the excess GMA/rehydrated and labeled with my antibodies, NOTHING WORKED! Nothing showed up! Only DAPI!! The mechanism I used for removing GMA and rehydrating were as follows: 1.) 3 changes of 30 minutes each in 60 Celsius xylene. (Note: I recently let the slides sit in 60 Celsius xylene overnight too... but it did not work!!) 2.) 3 changes of 10 minutes each in 100% EtOH. 3.) 15 minutes in 95%, 75% & 50% EtOH 4.) 5 minutes in water 5.) 3 changes of 5 minutes each, PBS. I continued to block and apply my antibodies as normal. (Antibodies are all brand new, not duds!) Since this plastic was formulated specifically for IHC, I am at a loss. If anyone has used this plastic before, any insight directed my way would be extremely helpful!! Thanks, Allyse Mazzarelli Histologist NEUROTECH USA INC. _______________________________________________ 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