Methods and Techniques Discussion => Immunocytochemistry (ICC) => Topic started by: igor on February 10, 2010, 12:13:00 PM

Title: ICC Bone Marrow Cytospins
Post by: igor on February 10, 2010, 12:13:00 PM
Dear All,

I am examining various bone marrow populations by IF, and wanted to check if there are any obvious flaws to my method:

1. Wash BM in PBS/1% Bovine albumin. Uses lysis buffer x2 to get rid of RBC's, platelets
2. Cyospin onto slide, allow air dry
3. Fix in paraformaldehyde 4%, wash 2x in PBS and air dry, store in fridge 4degreesC till next day for staining. ** Is this part ok?

I have had some very odd results with Acetone, in that the cells seem to become one amorphous clump, and their membranes are often disrupted. Hence the change to paraformaldehyde.

Many thanks