Systems and methods for three-dimensional structured illumination microscopy with isotropic spatial resolution improvement of axial resolution in 3d sim via optical and computational means
AssigneesThe U.S.A., as represented by the Secretary, Department of Health and Human Services, The University of Chicago
InventorsHari Peng SHROFF, Yicong WU, Patrick LA RIVIERE, Xuesong LI
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Abstract
Various embodiments for a three-dimensional structured illumination microscopy system having a mirror positioned in diametric opposition to the objective for producing additional illumination components by a fourth beam generated by the mirror and a computational means for accomplishing the same are disclosed herein.
Claims28
A three-dimensional structured illumination microscopy system comprising:
- at least one illumination source for transmitting a respective illumination beam through an acoustic-optic tunable filter for producing a combined laser beam;
- a pair of lenses in communication with a pinhole for generating a beam expander that expands and spatially filters the combined laser beam;
- a spatial light modulator for producing an illumination pattern of three illumination beams;
- an objective in communication with the spatial light modulator such that the three illumination beams are re-imaged to the back focal plane of the objective as off-axis illumination beams and an on-axis central illumination beam;
- and
- a mirror positioned in diametric opposition to the objective such that the on-axis central illumination beam is reflected back towards the objective as an on-axis reflected illumination beam to generate a four-beam interference pattern at the sample plane.
A method of improving spatial resolution in a three-dimensional structured illumination microscopy system (3D-SIM) comprising:
- generating an illumination pattern of three illumination beams by the 3D-SIM system;
- imaging the illumination pattern of three illumination beams to a back focal plane of an objective such that the three illumination beams comprise first and second off-axis illumination beams and an on-axis central illumination beam relative to a sample plane;
- reflecting the on-axis central illumination beam off a mirror positioned in diametric opposition to the objective such that an on-axis reflected illumination beam is reflected directly back towards the objective and sample plane;
- and
- generating a four-beam interference pattern by the first off-axis illumination beam, the second off-axis illumination beam, the on-axis central illumination beam, and the on-axis reflected illumination beam at the sample plane.
A system for training a neural network for improving axial resolution comprising:
- a computing system including a processor in communication with a memory, the memory including instructions, which, when executed, cause the processor to:
- obtain a 3D-SIM image;
- blur the 3D-SIM image along a first axial direction to suppress artifacts;
- blur the 3D-SIM image blurred along the first axial direction along a second lateral direction;
- downsample the 3D-SIM image blurred along the first and second directions;
- and
- upsample the 3D-SIM image blurred along the first and second directions to produce a low resolution 3D-SIM image.
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Dependencies are inferred from the claim text.