Multicolor structured light illumination super-resolution microscopic imaging method and device based on DMD

Abstract

The invention discloses a multi-color structured light illumination super-resolution microscopic imaging method based on a DMD (Digital Micromirror Device), which is characterized in that the DMD is used as a digital blazed grating to diffract laser with different wavelengths, and positive and negative first-order diffraction light generated by Fourier filtering is subjected to accurate polarization modulation and then interferes on the surface of a fluorescent sample to be detected to generate a high-contrast stripe illumination pattern; fringe patterns in different directions and phases are loaded through the DMD to quickly perform direction rotation and phase movement on interference fringes, and a plurality of obtained original fluorescence intensity images are reconstructed to obtain a super-resolution microscopic image. The invention further discloses a multi-color structured light illumination super-resolution microscopic imaging device based on the DMD, different laser wavelengths are rapidly switched by a galvanometer to be gated or cut off to enter different square-core multimode optical fibers, and the incident angles of all the wavelengths are aligned by adjusting the output ends of the square-core multimode optical fibers. Therefore, the blaze level corresponding to each wavelength is ensured to be located at the same position so as to facilitate Fourier filtering. The device is low in complexity and high in flexibility, stability and expandability, and meanwhile, a modular design is adopted, so that the system can be extensively applied to imaging research of a three-dimensional interference type structure illumination obvious microtechnology and a projection type structure illumination obvious microtechnology.

Description

technical field [0001] The invention belongs to the field of optical super-resolution fluorescence microscopic imaging, in particular to a DMD-based multicolor structured light illumination super-resolution microscopic imaging method and device. Background technique [0002] In the field of super-resolution fluorescence microscopy imaging, the three mainstream technologies are stimulated radiation depletion technology, single-molecule localization technology and structured light illumination technology. Stimulated radiation depletion technology belongs to the point scanning technology. It realizes super-resolution by superimposing the point spread function of excitation light and loss light. At present, the highest resolution can be obtained at 20nm, but it requires a high density of fluorescent labels and high optical power. It will cause serious photobleaching of the sample. Both single-molecule localization technology and structured light illumination technology belong t...

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