First, the very best of holidays to everyone. Now for the histology part. Our lab's focus is on the early stages of Alzheimer's Disease in the Brainstem using celloidin processing & embedding for IHC staining. This year, our lab will be receiving 6 post-mortem whole human brains (1 every other month). After fixation, processing & celloidin embedding, the whole brain will be serially cut at 100um thick. Each brain section will be 5 inches x 4.5 inches in size.
I will given 250 of these whole brain sections to stain for tau IHC...that's 1500 whole brain sections/year!!! 1) Does anyone have experience doing manual IHC staining of large free-floating brain sections? 2) What type of staining tools, dishes or other essential equipment can anyone recommend? 3) What's the most efficient way to stain 250 sections for batch IHC staining - such as transferring batch sections (maybe 5-10) from reagent to reagent? 4) What type of batch apparatus to use? As for the antibody & ABC steps, I was thinking of placing each section inside a large glass cigar tube (yep, people use large glass tubes with fitted cap to store cigars), with 5ml of antibody or ABC reagent & gently agitate on a shaker/rotator at room temp during the incubation. Does anyone have ideas on this? Please, any ideas, suggestions or recommendation anyone can provide will be most greatly appreciated. Best regards Maria Mejia UCSF Department of Neurology San Francisco, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet