Multi-focal structured illumination microscopy systems and methods

Abstract

Various embodiments (300, 400, 500) for a multi-focal selective illumination microscopy (SIM) system for generating multi-focal patterns of a sample are disclosed. The embodiments (300, 400, 500) of the multi-focal SIM system perform a focusing, scaling and summing operation on each generated multi-focal pattern in a sequence of multi-focal patterns that completely scan the sample to produce a high resolution composite image.

Description

technical field [0001] This document relates to multi-focal structured illumination microscopy, and in particular to multi-focal structured illumination microscopy systems for producing multiple multi-focal fluorescent emissions arising from a multi-focal pattern of a sample and methods. Background technique [0002] Classical fluorescence microscopy is limited in resolution by the wavelength of light, known as the "diffraction limit," which will laterally resolve a sample at typical excitation and emission wavelengths when it emits fluorescence that is detected by the microscope. The rate is limited to about 200 nm, and the axial resolution is limited to about 500 nm. Confocal microscopy is an optical imaging technique used to eliminate out-of-focus emission from specimens thicker than the focal plane by using point illumination and a spatial pinhole arrangement to Increasing the optical resolution beyond the diffraction limit delivers an image with 1.41 times the resolut...

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Problems solved by technology

However, such optical sectioning is performed computationally and thus suffers from shot (Poisson) noise

Therefore, SIM is not suitable for thick or highly stained samples when background fluorescence may cause this shot noise contribution to outweigh the in-focus signal

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