Multi-focus scanning three-dimensional imaging method and system based on double-helix-point spread function

Abstract

The invention discloses a multi-focus scanning three-dimensional imaging method and system based on a double-helix-point spread function. The method comprises the following steps: irradiating a laserbeam on a digital micromirror element at a preset angle; reflecting the laser beam by the digital micromirror element, and projecting onto the sample surface; switching the illumination mode of the digital micromirror element at equal intervals, exciting the generation of periodic arrays on the sample surface to move along with the switching of the illumination mode; performing phase modulation onfluorescent light generated by excitation of the sample surface, converting a fluorescence signal of Gaussian distribution into a fluorescence signal of a double-helix form, acquiring a fluorescencesignal of the double-helix form by a detector, and acquiring a plurality of image data; locating and intercepting all double helix points on each image according to the acquired image data to obtain aplurality of sub-regions, and performing wavefront reconstruction processing on all sub-regions to obtain a three-dimensional restructuring graph of a sample. The sample is excited simultaneously through multiple focus points, so that the sample acquisition time is reduced, and the time resolution of a three-dimensional image scanning microscopy system is greatly improved.

Description

technical field [0001] The invention relates to the field of optical microscopy technology, in particular to a multi-focus scanning three-dimensional imaging method and system based on a double helix point spread function. Background technique [0002] Laser scanning confocal microscopy is an effective technical means to study biological microstructure, and it is widely used in the field of biomedicine. In the confocal microscope system, through a pair of conjugated precision pinholes and a single focal point razor scanning method, the system can suppress the stray light from the non-focus plane, filter out the information outside the focal plane, and obtain high image contrast. Although confocal microscopy can achieve super-resolution imaging, its resolution is affected by the size of the pinhole. The smaller the pinhole, the higher the resolution. However, the signal light that can be collected is weaker, which directly leads to the improvement of resolution Reduce the s...

AI Technical Summary

Problems solved by technology

[0003] Although the image scanning microscope has many advantages, it can improve the resolution and obtain a higher signal-to-noise ratio at the same time. However, due to the weak signal ability received by the CCD detector itself, the reading time is long, which leads to the failure of the image scanning microscope system. The imaging speed is slow, and its scanning It takes 60s for a large sample area, and it takes a lot of time to image the three-dimensional structure of the sample

Images