Rapid three-dimensional (3D) super-resolution microscopic method and device

Abstract

The invention discloses a rapid three-dimensional super-resolution microscopic method. The method comprises that a laser beam is converted into linearly polarized light after being collimated, phase modulation is carried out on the linearly polarized light, the linearly polarized light is converted into circularly polarized light, the circularly polarized light is projected to a sample to be measured, and pilot light emitted by scanning points of the sample to be measured is collected; and 3D scanning is carried out on the sample. Phase modulation comprises primary phase modulation and secondary phase modulation, a spatial light modulator which modulates the phase of an s light component of the linearly polarized light is used in the primary phase modulation, a spatial light modulator which modulates the phase of a p light component of the linearly polarized light is used in the secondary phase modulation, and a 3D super-resolution image is obtained according to the effective pilot light intensity. The invention also discloses a rapid 3D super-resolution microscopic device. According to the invention, optical power is lower, the photobleaching effect is weakened, 3D imaging is rapid, the device is simple, and light splitting is not needed.

Description

technical field [0001] The invention belongs to the field of super-resolution, and in particular relates to a method and a device capable of quickly realizing three-dimensional super-diffraction limit resolution in the far field. Background technique [0002] Optical microscopic imaging is a commonly used and effective method for observing submicron microstructures, but the existence of the optical diffraction limit greatly limits the resolution of optical microscopic imaging. According to the principle of Abbe's diffraction limit, the ideal point light source is not an ideal image point after being focused by the microscope objective lens, but a diffraction spot, and its size is determined by the full width at half maximum of the intensity distribution curve of the diffraction spot In the formula, λ is the wavelength of the illumination light used by the microscope, and NA is the numerical aperture of the microscopic objective lens used. Since NA=nsinα, in the formula, n i...

AI Technical Summary

Problems solved by technology

[0006] Stimulated fluorescence microscopy has the advantages of low power consumption, weak bleaching, fast imaging, and simple optical path. However, when it is applied to three-dimensional super-resolution microscopy imaging, it is necessary to divide the excitation light into three channels and modulate them separately. The modulated three-way beams are combined, which undoubtedly greatly increases the space complexity of the optical microscope system

In addition, it is necessary to design a three-way shutter switch to control the switching of the excitation light mode, which also greatly increases the time complexity of the microscope system.

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