Ultra-high-speed real-time super-resolution microscopic imaging method

Abstract

The invention discloses an ultra-high-speed real-time super-resolution microscopic imaging method, which solves the problems that a super-resolution imaging system in the prior art is low in shooting speed, long in later reconstruction time and poor in shooting result of living cells and the like. The ultra-high-speed real-time super-resolution microscopic imaging method comprises the following steps of S1, dividing an incident light beam emitted by a laser into two beams / three beams, and enabling the two beams / three beams to enter a sample through a microscope to generate interference so as to realize structured light illumination, S2, jointly controlling interference fringes to change at a high speed through a camera, a data acquisition card and a computer, and acquiring a plurality of original images of the sample at a high speed, and S3, performing real-time reconstruction on a plurality of collected original images through a designed real-time reconstruction framework to obtain a super-resolution image of the sample. After an original image is collected, the original image can be reconstructed immediately, real-time super-resolution imaging is achieved, and the method has great advantages in the aspect of observing living cells.

Description

technical field [0001] The invention relates to the technical field of super-resolution imaging, in particular to an ultra-high-speed real-time super-resolution microscopic imaging method. Background technique [0002] Structured light illumination microscopy extracts high spatial frequency information from multiple fluorescence images to improve resolution, and can break through the optical diffraction limit to observe intracellular structures. The current mainstream SIM microscopes mainly generate ±1st-order diffracted beams through physical gratings or spatial light modulators (SLMs). For a structure that uses a physical grating to generate a diffracted beam, it is necessary to use a mechanical platform to rotate and translate the grating to generate interference fringes of different phases and directions. The imaging speed is slow and the system is likely to be unstable due to mechanical vibration. For structures that use SLM to generate diffracted beams, fast and stabl...

AI Technical Summary

Problems solved by technology

Therefore, in order to find the initial area of ​​interest, time-consuming switching back and forth between measurement and post-processing steps is time-consuming and labor-intensive, and it is prone to premature bleaching of live cells, resulting in unsatisfactory shooting results

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