Hi microscopist,
I am trying observe single fluorophore blinking by an andor EMCCD (e2v cd97, 512*512, 16um) and nikon TIRF (60*, 1.49). Cy3-oligonucleotides and GelGreen stained DNA were used. To scavenge O2 and slow the blinking time down to 10ms scale, glucose, glucose oxidase, catalase and DTT were added to buffer.
But no large eat and flow of fluo intensity was observed within 20s.
Is there any mistake in my system? Does EMCCD suit for blinking observing?
Any suggesttion will be welcome.
Thank you!
Tian Tian,
State key lab of bioelectronics,
Southeast University, China,
titi_wudi@163.com