Hello everyone,

I am building a Prism TIRF microscope with 2 lasers 532/637 nm. Although I have already defined the plane using Tetraspeck beads (0.1 um), I find it hard to find the signals from labeled proteins and DNAs. I tried to check with different dyes conjugated with proteins and DNAs and with varying concentrations.

I checked those samples under the objective TIRF, they seem to have sufficient densities and the signals are good. Could you please share with me some tips so that I can visualize the samples more easily?

The camera we are using for the Prism TIRF is Prime BSI Express sCMOS, and the one for objective TIRF is iXon Ultra 888 EMCCD. The samples visualized under 60X objectives in both cases.

Thank you so much.