Performance of the holographic system.

a: Normalized fluorescence intensity of a focal spot as a function of number of spots, Nb: Normalized uncaging power per spot as a function of number of stimulation spots. Number of foci is kept constant and superfluous foci are moved outside of the holographic FOV in transverse direction. c: Measured PSF of two-photon excitation in the axial (⚬) and transverse (●) directions for 60× and 20× objective lenses. d: Normalized power as a function of displacement along the x for 60× for FOV centered at (0, 0) (⚬), (−100, −100) μm (△) and (100, 100) μm (□). e: Normalized power as a function of displacement along the x for 20× for FOV centered at (0, 0) (⚬), (−300, −300) μm (△) and (300, 300) μm (□). Filled circles with dashed line denote power distribution after amplitude weighting for FOV centered at (0, 0). f: Normalized power as a function of displacement along the z for 60× (red markers in red area) and 20× (black markers in gray area) for FOV centered at different coordinates specified in d and e. Solid lines denote empirical Gaussian fit used for amplitude weighting. Filled circles with dashed line denote power distribution for 20× after amplitude weighting for FOV centered at (0, 0).