Hello, I suspect that 0.1% triton X-100 in my blocking solution may be causing my frozen (unfixed) sections of brain to come off the slide.
These are APTES slides and the sections were 20 and 40 micron thick. At first it looked to me that the entire section had come off but under the microscope I could see a thin layer (of single?) cells still stuck to the slide. There was also a chance in antibody staining. The antiserum I am using normally gives very defined intense staining of a surface antigen but now the staining was punctate. I suspect the triton is too strong and as well as washing off the tissue it is also disrupting the membranes and interfering with the staining. I have not seen this when I was using the same blocking solution without the triton X-100. For wholemounts I had used DMSO for permeabilization but also have NP-40 available. Does anyone have an opinion on the matter? -- Tyrone Genade Dept. Human Biology University of Cape Town _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet