The laser power is about 0.2 mW above the coverglass, is that able to obtain clear single molecule fluorescence spots?
If your system alignment is perfect you may get good image even in much lower power than 0.2mW.
It depends on the technique you are using and several factors of your optics. What counts is rather the power density (the laser power per illuminated area.)
In our widefield microscopes and TIRF we usually use more then 1mW, but u can also increase the exposure time of your camera.
In confocal microscopy, 0.2mW are definitely enough to detect single molecules in a carefully aligned setup. Looking in my PCCP article I used 0.6kW/cm2
So u have to find out/tell us the diameter of the beam at the coverglass interface and the NA of your objective to give u a betterestimate.
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