Abcam Ad

Author Topic: Splattered cells  (Read 3654 times)

0 Members and 1 Guest are viewing this topic.

Offline Maran

  • Newbie
  • *
  • Posts: 1
Splattered cells
« on: December 20, 2006, 05:05:50 PM »
I've been making cytospins of T cells and it worked fine for a while. Out of the blue, most cells now have a splattered appearance (nuclei are large and vague after staining with hematoxylin) and I can't figure out why this is.
I use 0.5-1 106 cells/ml cell suspensions, 50-100 ul per spin, spin 5 min at 450 rpm. The T cells have been isolated with MACS microbeads (does that make them more susceptible to splattering?).
I tried different speeds (250-450 rpm) and times (2-6 min) and different buffers, none of this seems to make a huge difference. I do get the impression though that the more cells I use (100 ul vs 50 ul of a 1 million cells/ml cell suspension), the more splattering.
Does anybody have any insights or suggestions?

Splattered cells
« on: December 20, 2006, 05:05:50 PM »