Hi from Down Under I was wondering if anyone had a method for preparing buffy coats for immuno.
What we have done so far 1) is spun down whole blood, extracted the buffy coat and made a thrombin/plasma clot of the extracted buffy coat (minus RBCs). The clot was post fixed overnight in 10% formalin and subsequently paraffin processed. 5um Sections were cut and immuno done. The clumping cells did not stain, but the single ones on their own did. 2) To improve on fixation (assuming this was the problem with the unstained clumping cells), the buffy coat was collected and fixed in 10% formalin. The fixed buffy coat was washed in 0.9% NaCl twice, and then embedded in both a thrombin/plasma clot (unsuccessful), and an agar block. The agar block was paraffin processed. The subsequent immuno showed good staining in single WBC, and in one clump of cells. The other clump showed no staining and had a bit of cell damage. Thanks Stephen Pathology Sydney University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet