Supplementary Figure 3: Estimating system resolution at coverslip surface in AO-2P ISIM.
From: Adaptive optics improves multiphoton super-resolution imaging
a) Exemplary images of 100 nm yellow-green fluorescence beads on coverslip surface, as viewed in lateral (top) and axial (bottom) cuts through the image stacks. From left to right: two-photon wide-field (non-rescan) image, two-photon wide field image after deconvolution, two-photon ISIM image (i.e. after rescanning), two-photon ISIM image after deconvolution. System aberrations were corrected by measuring the wavefront distortion using a Shack-Hartmann sensor, and subsequently compensating for the aberrated wavefront by using a deformable mirror. b) Modulation transfer functions corresponding to a). c) Comparative line-profiles through the center of each lateral (left) and axial (right) bead image further emphasize improvement after deconvolution. (d) Statistical comparison of estimated lateral (left) and axial (right) resolution (FWHM of bead image), derived from > 30 beads. Means and standard deviations are shown. Scale bars: 0.5 μm.
