I am not sure if Fluoromount G is a good media to perform TIRF because his refractive index. Has anyone tried?
By my calculation, with your low refractive index media (Fluoromount G seems to be 1.40 from what I find) and high refractive index media (glass goverslip 1.515) give a required angle of incidence of 67 degrees to achieve critical angle. With water and coverslip it would only need to be 62 degrees -which standard TIRF optics can do. Depends on your wavelength and lens NA as to whether you can hit critical angle. All the calculations can be found here: https://www.microscopyu.com/techniques/fluorescence/total-internal-reflection-fluorescence-tirf-microscopy
Thank you Christopher. I know that theoretically it should work, but I read that the refactive index change when it stars to dry, so is hard to perform TIRF. I have the same problem, with Fluoromont my images do not look like a tirf image, so I wonder if this works for someone.
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