Abstract
Stimulated emission depletion (STED) microscopes, like all super-resolution methods, are sensitive to aberrations. Of particular importance are aberrations that affect the quality of the depletion focus, which requires a point of near-zero intensity surrounded by strong illumination. We present analysis, modeling, and experimental measurements that show the effects of coma aberrations on depletion patterns of two-dimensional (2D) and three-dimensional (3D) STED configurations. Specifically, we find that identical coma aberrations create focal shifts in opposite directions in 2D and 3D STED. This phenomenon could affect the precision of microscopic measurements and has ramifications for the efficacy of combined 2D/3D STED systems.
Highlights
Stimulated emission depletion (STED) microscopes, like all super-resolution methods, are sensitive to aberrations
The excitation and emission paths together form, in effect, a confocal laser scanning microscope; the depletion path is co-linear with the excitation path and contains a phase mask that generates a ring-shaped depletion focus
Equation (10) shows that, within the validity range of the approximation in Eq (6), the zero of the 2D depletion focus is translated along the x axis by a distance Δx2, which is proportional to the amplitude c of the coma aberration defined in Eq (4)
Summary
Stimulated emission depletion (STED) microscopes, like all super-resolution methods, are sensitive to aberrations. [5], it was shown how misalignment of the phase mask can be mistakenly attributed to coma aberrations for the 2D or 3D STED modes. In this Letter we show that, to first approximation, small amounts of coma do not fill in the zero of the depletion focus but shift its position laterally by amounts that can be much larger than the resolution of the microscope.
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