Immunohistochemistry - CD4 and CD8 cell (Sep/27/2005 )

I am working in a small lab where flow cytometry is not available and i have to perform the CD4 and CD8 counts of PBMCs.

I have gone through many literature but most of them mentions about a method which utilises enzymatic method for signaling (eg DAB and precipitating TMB) but i am not able to get a protocol where i can perform this enumeration by simple light or phase contrast microscope and immunochemistry.

can any body please help me by giving a protocol for same or a reference for same.

regards

nirav

could you try to fractionate the cells with something like dynabeads using the CD4 or CD8 antibody and then do trypan blue on a hemacytometer? You may also have to stain the cells and look under a scope to make sure your fraction is pure, but if you don't need to keep the cells positive or negative isolation with magnetic beads may work....