Two-photon multi-focus microscopic imaging system and method based on spatial light modulator

Abstract

The invention provides a two-photon multi-focus microscopic imaging system and method based on a spatial light modulator. The two-photon multi-focus microscopic imaging system comprises a laser lightsource, a spatial light modulator, an objective lens, a detector and a control terminal; the spatial light modulator is used for carrying out phase modulation on a laser beam according to the importedsuperimposed phase diagram which is switched at equal intervals to generate a multi-focus array; the superimposed phase diagram is formed by superposing a multi-focus array phase diagram and a linearphase grating phase diagram; the objective lens is used for projecting the multi-focus array to a sample surface to generate a multi-focus fluorescence signal; the detector is used for acquiring a multi-focus fluorescence signal to obtain a plurality of images; and the control terminal is used for processing the plurality of images to obtain a super-resolution image. According to the invention, the multi-focus array phase diagram and the linear phase grating phase diagram are loaded on the spatial light modulator at the same time, so the generation of the multi-focus array and the high-precision parallel digital random addressing scanning and excitation imaging of the sample surface are carried out, and a problem of mechanical inertia of a two-photon multi-focus microimaging system is solved; the complexity of the system is reduced, and the flexibility is improved.

Description

technical field [0001] The invention belongs to the technical field of optical imaging, and in particular relates to a two-photon multi-focus microscopic imaging system and method based on a spatial light modulator. Background technique [0002] Optical microscopic imaging system is one of the important tools for biologists to study biological phenomena and life activities. The traditional optical microscopic imaging system uses wide-field illumination, which limits its imaging depth and imaging resolution. Although laser confocal microscopy can achieve super-resolution imaging, its resolution is affected by the size of the pinhole. Although the image scanning microscope can improve the signal-to-noise ratio of the image, the imaging speed is slow. The multi-focus scanning microscope adopts a parallel scanning method, which can increase the imaging speed while improving the imaging depth. [0003] Existing multi-focus scanning microscopes are divided into single-photon mul...

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Problems solved by technology

[0005] In view of the deficiencies in the above-mentioned prior art, the purpose of the present invention is to provide a two-photon multi-focus microscopic imaging system and method based on a spatial light modulator, which overcomes the complex structure of the existing two-photon multi-focus scanning microscopic imaging system , the defect of poor flexibility due to the influence of mechanical inertia

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