I believe that you can identify SP cell populations with a low power UV laser but your optical allignment and choice of dichroic filters, PMT's, and LP filters is very important. The red signal collected through a 675LP filter is very important. My suggestion is to acheive as much red signal as possible by looking at this fluorescence while exciting molecular probes UV beads. Allignment and optimization of signal is critical. On the other hand having 100-150MW of UV power is very helpfull. I have found I can obtain much better separation of SP population using a higher power UV laser. But ot is also very important to optimize allignment for best signal. The amount of Hoechst used for dye loading and incubation time are also critical. David M. Dombkowski dombkowski@helix.mgh.harvard.edu Flow Cytometry-Pathology-CNY rm7017 149 13th Street Massachusetts General Hospital-East Boston, MA 02129 Tel. (617)726-1683 Fax.(617)724-3164
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:01:08 EST