Line-scan confocal scanning light-field microscopic imaging apparatus and method

Through the linear scanning confocal scanning light field microscopy device, the linear scanning hardware optical path and camera separate the sample fluorescence signal and background fluorescence signal, solving the problem of background fluorescence interference in the scanning light field microscopy system, and achieving high-resolution three-dimensional imaging.

WO2025152202A1 Publication Date: 2025-07-24TSINGHUA UNIVERSITY
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Patent Information

Application Number
PCT/CN2024/073893
Authority / Receiving Office
WO · WO
Patent Type
Applications
Current Assignee / Owner
Priority Date
2024-01-18
Filing Date
2024-01-24
Publication Date
2025-07-24

AI Technical Summary

Technical Problem

In the scanning light field microscopy imaging system, the background fluorescent signal interferes with the sample structure signal, affecting the accuracy and reliability of observation, and the confocal light field microscopy system cannot reach the diffraction limit resolution.

Method used

The linear scanning confocal scanning light field microscopy imaging device is used to separate the sample fluorescence signal and the background fluorescence signal through the linear scanning hardware optical path, camera and control system. The light field modulation is performed using a microlens array and a two-dimensional galvanometer to generate a three-dimensional microscope image of background fluorescence removal.

Benefits of technology

It effectively alleviates the impact of background fluorescence on three-dimensional microscopy imaging, improves imaging results in strong background environments in living bodies, improves the accuracy and resolution of observation, and is suitable for rapid imaging of real scenes.

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Abstract

A line-scan confocal scanning light-field microscopic imaging apparatus (10) and method. Said apparatus comprises: a microscope (100), a target biological sample being placed on a stage of the microscope (100); an excitation optical path (200), which is used for outputting a line-shaped light source for exciting fluorescence of the target biological sample; a line-scan hardware optical path (300), which is used for performing multi-dimensional scanning of the line-shaped light source to obtain a scanned light-field image; a microlens array (400), which is used for light-field modulation of the line-shaped light source to obtain a line-shaped light field; a camera (500), which is used for performing three-dimensional imaging on the basis of the line-shaped light field to obtain a three-dimensional image; and a control system (600), which is used for controlling the line-scan hardware optical path (300) and the camera (500) to execute a line-scan confocal action, so that the number of pixel rows exposed simultaneously at a same moment is consistent with a sample region illuminated by the line-shaped light source, so as to separate a sample fluorescence signal and a background fluorescence signal of the target biological sample, and, on the basis of the scanned light-field image and the three-dimensional image, generate a three-dimensional microscopic image with background fluorescence removed for the target biological sample. Therefore, the problem of background fluorescence in a scanning light-field microscopy system is solved.
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