I have been using histopaque very unsuccessfully to isolate granulocytes and PBMCs from human whole blood. I tried layering histo-1077 onto histo-1119 followed by the blood. I did not get a clear distinction of layers (all at room temp). Second I tried doing an initial layering on histo-1119 to remove RBCs (worked lovely) followed by an additional layering on 1077. Although the leukocyte layer on the 1077 looked good, after spinning the 1077 seemed diffuse throughout the sample and there were no clear layers.
Any advice or thoughts would be greatly appreciated.