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Methods for improving the performance of structured illumination microscopy with adaptive optics

Creative Commons 'BY' version 4.0 license
Abstract

While the light microscope offers biologists the advantage in vivo imaging, it suffers from a reduced resolution due to constraints imposed in the optical domain. The hard limit on resolving power, known as the diffraction limit of resolution, is the distance of roughly one-half wavelength of the excitation light of an imaging system. Super-resolution imaging can relax the constraint on resolution.

Further constraints on resolving power are imposed by light wavefront aberrations. Sample-induced aberrations are inherent to biological samples. Other aberrations come from the microscope system itself. The canonical solution for wavefront aberration in a microscope is known as adaptive optics (AO).

Structured Illumination Microscopy (SIM) is an optical transfer function-based super-resolution technique that increases the support region in Fourier space via spatial frequency mixing. In particular, the optical imaging system that acts as a low-pass filter on the image of the biological sample, favors low spatial frequencies. By optically mixing the biological sample with SIM fringes, high spatial frequency information from the sample is mixed into the pass band of the microscope.

This work contains the results of a SIM microscope implementation with an adaptive optics system (AOSIM). The results of this microscope are presented. In addition, further improvements for AO SIM microscopes are given, backed by simulations based on the mathematics of SIM and both Fourier and geometric optics. The simulations show the utility of the improvements in terms of both aberration mitigation and resolution improvement.

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