Method and device for improving multiphoton fluorescence microscope imaging resolution

Abstract

Relating to the field of optical technologies, the embodiment of the invention discloses a method for improving the multiphoton fluorescence microscope imaging resolution. The method includes: scanning a sample through a microscope repeatedly, acquiring the data of scanned multiple images, and acquiring the pixel point signal strength of sample target points in the multiple images according to the obtained multiple image data; and calculating the signal strength of sample target points in a reconstructed image obtained by processing the data of the multiple images according to the pixel point signal strength of the sample target points in the multiple images. The embodiment of the invention also discloses a device for improving the multiphoton fluorescence microscope imaging resolution. By means of the method and the device provided by the invention, and through specific mathematical calculation formulas, fusion of a multiphoton imaging technology and a structured illumination imaging technology is realized, while keeping the advantage of multiphoton original imaging depth, a super-resolution imaging ability is increased.

Description

technical field [0001] The invention relates to the field of optics, in particular to a method and device for improving the imaging resolution of a multiphoton fluorescence microscope. Background technique [0002] In the field of fluorescence imaging of living biological tissues, because the focus position of multiphoton fluorescence microscopy imaging can be accurately determined and controlled, multiphoton fluorescence is very suitable for exciting fluorescent probes below the surface of the specimen. This high-precision positioning method ensures It minimizes photobleaching of fluorescent dyes and reduces photodamage, thereby increasing the viability of cells and maintaining longer cell performance research experiments. Therefore, multiphoton microscopy imaging technology is widely used in the field of optical imaging. However, due to the existence of the optical diffraction limit, the lateral resolution and longitudinal resolution of multiphoton fluorescence microscopy ...

AI Technical Summary

Problems solved by technology

However, the confocal fluorescence microscope is a three-dimensional tomographic capability obtained by blocking the light signal outside the focal plane through the microhole in front of the detector. As the depth increases, a large amount of illumination light is scattered in the sample, and only a small part The light can reach the focal point, and the fluorescence generated by excitation outside the focal point will also have a certain amount of photons that can pass through the micropore and reach the detector after multiple scattering. These photons are imaging noise signals, which directly lead to a decrease in the signal-to-noise ratio of the imaging result , thus limiting the imaging depth of the laser scanning confocal microscope

Currently, laser scanning confocal microscopy combined with structured light illumination can only image single cell thickness samples

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