Active PSF Shaping and Adaptive Optics Enable Volumetric Single Molecule Super-Resolution Microscopy through Brain Sections

Abstract

images with high temporal and spacial resolution. We combine this method with Multifocus microscopy (Abrahamsson et al 2015 Opt. Express 23, 7734-7754), which allows us to image 25 focal planes simultaneously for fast volumetric imaging. Multifocus Microscopy(MFM) has provided the opportunity to 3D image biological events at speeds limited by the camera readout of a single image. High speed MFM has been made possible by utilizing diffractive fourier optics to multiplex light from different focal planes on a single camera sensor to produce an instantaneous 3D image from the resulting 2D focal stack. When designing imaging systems for MFM applications, there is a strong desire to prevent distortion and reduce chromatic and spherical aberration in the 2D focal stack of images. In addition to improving image quality, imaging small singular biomolecules requires maximization of photon collection efficiency. It is the goal of this project to determine if large acromatic doublet lenses could be replaced with camera objectives to improve MFM system performance.