Hi World,

I'm having trouble finding a clear answer out there. We are trying to determine if FLIM-FRET can be performed using our widefield (or TIRF) microscope and an EMCCD camera. I generally see FLIM being performed on a single or multiphoton confocal, but have found some information suggesting that widefield FLIM is possible. Can a good EMCCD provide the acquisition speed to measure lifetime? Is any additional equipment required? Is there freeware available to crunch the numbers, or are specific software modules required? Thanks for your advice, oh quantitative imaging specialists!

Sara