3D multi-focal plane structured light fast microscopic imaging system and method

Abstract

The invention discloses a 3D multi-focal-plane structured illumination quick microscopic imaging system and method. The system is composed of an objective lens OL, a delay lens group, a diffraction multi-focal grating MFG, an aberration correction grating CCG, a digital microlens array DMD, a first 4f group, a second 4f group, and an imaging module. The delay lens group is used for enabling lightwith different depths to arrive at a light-sensitive surface of a camera after passing through a multi-focal grating. The aberration correction grating CCG is used for eliminating an aberration of a corresponding sub graph of a sample at a same depth. The digital microlens array is used for modulating illumination patterns of the sample at different depths. The first 4f group is used for realizingmatching of the size of a laser beam with the size of the digital microlens array DMD; and the second 4f module is used for matching of the size of the pattern on the digital microlens array DMD withthe size of the diffraction multi-focal grating MFG. And the imaging module is used for collecting data, carrying out structured illumination super-resolution microscopic reconstruction on the sampleat different depths, and stacking the reconstruction results of all layers to form a final 3D super-resolution imaging result. Therefore, the speed of microscopic imaging is increased effectively andthe stability of the system is improved.

Description

technical field [0001] The invention relates to the technical field of microscopic super-resolution imaging and microscopic body imaging, in particular to a 3D multi-focal plane structured light fast microscopic imaging system and method. Background technique [0002] SIM (structured illumination microscopy, structured illumination microscopy) plays a very important role in biological microscopic imaging, and this method can bring about a two-fold increase in spatial resolution. Compared with other super-resolution microscopy methods, structured light microscopy requires fewer shots, so it has obvious speed advantages and can be better applied to the study of live cell imaging. [0003] However, when performing three-dimensional imaging of an object, structured light microscopy usually needs to move the sample and scan the sample in the axial direction. The mechanical movement brought about by scanning will reduce the imaging speed on the one hand, and will affect the stabil...

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Problems solved by technology

[0003] However, when performing three-dimensional imaging of an object, structured light microscopy usually needs to move the sample and scan the sample in the axial direction. The mechanical movement brought about by scanning will reduce the imaging speed on the one hand, and will affect the stability of the system on the other hand. negative effect

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