An adaptive optics 3D STED microscope for super- resolution imaging of thick samples with background noise suppression using digital image processing

摘要

The emerge of super-resolution (SR) microscopy enabled imaging below the diffraction barrier. One of the SR techniques, Stimulated Emission Depletion (STED) microscopy, has shown promise in super-resolution imaging of thick specimen. Imaging such structures is a non-trivial task due to the increased aberrations introduced by the sample. Adaptive optics provides the solution to this problem. AO can correct the aberrations by modulation of the phase. Although STED microscopy is theoretically a diffraction unlimited technique, the resolution limiting factor is noise. Modern filtering techniques, such as block matching and 3D filtering (BM3D), can increase the signal-to-noise ratio of the STED images. This work presents an AO 3D STED microscope with aberration correction and background noise filtering using BM3D algorithm. We show the super-resolution images of thick samples and emphasize the importance of image processing for recovering of object high spatial frequencies.