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Multi-image reconstruction microscopic imaging method and device based on parallel detection

A microscopic imaging and multi-image technology, applied in the field of optical super-resolution microscopy, can solve problems such as indistinguishability and achieve high-resolution effects

Active Publication Date: 2020-04-17
ZHEJIANG UNIV
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Problems solved by technology

However, the resolution limit of image scanning microscopy is twice that of the diffraction limit, so in microscopic imaging, it is impossible to distinguish the finer (less than one hundred nanometers) structures and physiological activities

Method used

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  • Multi-image reconstruction microscopic imaging method and device based on parallel detection
  • Multi-image reconstruction microscopic imaging method and device based on parallel detection
  • Multi-image reconstruction microscopic imaging method and device based on parallel detection

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Embodiment

[0050] see figure 1 In this embodiment, the multi-image reconstruction microscopic imaging device based on parallel detection includes: a laser 1, a single-mode fiber 2, a collimator lens 3, a 1 / 2 wave plate 4, a first beam splitter 5, and a first optical shutter 6 , the first 1 / 4 wave plate 7, the first mirror 8, the second optical shutter 9, the vortex phase plate 10, the second 1 / 4 wave plate 11, the second mirror 12, the second beam splitter 13, Dichroic mirror 14, two-dimensional scanning galvanometer system 15, scanning lens 16, field lens 17, third mirror 18, objective lens 19, sample stage 20, optical filter 21, focusing lens 22, pinhole 23, fiber array 24 , detector array 25, computer 26.

[0051] The above device is mainly divided into five parts: an illumination system for generating laser beams, a solid beam optical path, a hollow beam optical path, a microscopic system for generating fluorescence, a detection system for collecting fluorescent signals, and a proce...

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Abstract

The invention discloses a multi-image reconstruction microscopic imaging method and device based on parallel detection, and belongs to the technical field of optical super-resolution microscopy, and the method comprises the steps: projecting one laser beam onto a to-be-detected sample under modulation of a two-dimensional scanning galvanometer system, and carrying out the two-dimensional scanning;using a detector array for collecting fluorescence signals emitted by the to-be-detected sample in the two-dimensional scanning process, moving signals collected by peripheral detectors back to the center, carrying out normalization after all images are added, and obtaining parallel detection fluorescence signal light intensity I1 (x, y) with x and y representing two-dimensional coordinates of scanning points on the sample; carrying out phase modulation on the other laser beam; obtaining parallel detection fluorescence signal light intensity I2 (x, y); and performing iterative computation onthe light intensity of the parallel detection fluorescence signals obtained by scanning on the two sides to obtain effective signal light intensity I (x, y). The effective spatial frequency bandwidthof an imaging system is expanded by using the nonlinear effect of fluorescence excitation, so that higher resolution is realized.

Description

technical field [0001] The invention relates to the technical field of optical super-resolution microscopy, in particular to a multi-image reconstruction microscopic imaging method and device based on parallel detection. Background technique [0002] Confocal microscopy is a high-tech research technology that has developed rapidly in the past ten years. Its current application fields have expanded to the research of cytology, microbiology, developmental biology, genetics, neurobiology, physiology and pathology. In the work, it has become an important tool for the microscopic research of modern biology. [0003] Confocal microscopy is divided into laser scanning confocal and digital confocal microscopy according to the principle of microscope structure. Confocal technology has the advantages of clear imaging, three-dimensional images, multi-label observation, real-time observation and recording of dynamic physiological responses in living cells, qualitative and quantitative ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6452G01N21/6458G01N2021/6463G01N2021/6484
Inventor 匡翠方陈宇宸徐良刘旭李海峰毛磊张克奇
Owner ZHEJIANG UNIV
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