i am measuring superoxides in chlorella cells with dihydroethidium (DHE). Using a final concentration of 30 micromolar DHE (sigma) working stock in MeOH for 100000 cells per ml washed and suspended in PBS, I incubated for 15 min in dark at 37 C with shaking.
I am using a Perkin Elmer LS 55 to measure/read fluoresence at ex 392 nm and em 410 nm. Wat would be the suitable slit width of both monochromators?
am i doing anything wrong in the above protocol? there was fluoresence intensity above 100 using slit widths 10nm/5nm shown for the concentration of DHE i used. still can it be improved upon....???
Edited by belladonna22, 17 July 2010 - 12:38 PM.