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Hyperspectral interference label-free imaging method and living cell quantitative tomography system

An imaging method and tomographic imaging technology, applied in the field of live cell imaging, can solve the problems of requiring precise operation, restricting the quality of reconstruction results, and being expensive, and achieving the effect of high sensitivity

Active Publication Date: 2020-06-02
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In summary, although interference phenomena can provide sub-nanometer resolution, current quantitative phase imaging techniques cannot provide nanometer-scale axial resolution
Usually the axial resolution of quantitative phase imaging instruments is greater than 1 μm, which severely restricts the quality of reconstruction results
In addition, common quantitative phase imaging instruments use the optical path structure of phase contrast or differential interference difference microscopes to generate coherent signals, and utilize spatial light modulators or liquid crystal color filters to modulate phase or wavelength. These components are very expensive and require delicate operation. , which limits the practical application of quantitative phase imaging techniques

Method used

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  • Hyperspectral interference label-free imaging method and living cell quantitative tomography system
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  • Hyperspectral interference label-free imaging method and living cell quantitative tomography system

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Embodiment 1

[0056] like figure 1 As shown, the living cell quantitative tomography system provided by the present invention includes a light source 1 , a reflective optical microscopic imaging system, a cell incubator 2 , a five-dimensional electric translation stage 3 , a spectrometer 4 and a computer 5 . The reflective optical microscopic imaging system includes a condenser collimating lens 6, a field lens 7, a first aperture stop 8, a second aperture stop 9, a first beam splitter 10, a second beam splitter 11, an objective lens 12, Imaging lens 13 , optical fiber 14 and camera 15 , the input end of spectrometer 4 is connected to one end of optical fiber 14 , the other end of optical fiber 14 is set at the focal point of imaging lens 13 , and the output end of spectrometer 4 is connected to computer 5 .

[0057] The cell incubator 2 is fixedly installed on the five-dimensional electric translation platform 3, and the five-dimensional electric translation platform 3 includes an XYZ three...

Embodiment 2

[0072] like figure 2 As shown, the present invention obtains hyperspectral interference information of live cell samples based on the interference principle, and realizes non-marked quantitative tomographic imaging of cells by establishing a hyperspectral interference model and a hyperspectral analysis algorithm based on Fourier transform. In this embodiment, single crystal silicon Taking this cell substrate as an example, the imaging process of the present invention is described in detail, which includes the following steps:

[0073] 1. Place the living cell sample on a high-reflectivity single-crystal silicon wafer substrate. The incident light is reflected by the cell membrane, scattered by organelles and biomacromolecules inside the cell, and reflected by the high-reflective substrate again; among them, reflected light and scattered light The coherent effect is produced, which is expressed in the form of hyperspectral white light interference signals; the basic structure ...

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Abstract

The invention relates to a hyperspectral interference non-labeled imaging method and a living cell quantitative tomographic imaging system. The imaging system comprises a light source; light emitted by the light source is collimated into parallel light, and the parallel light enters a first beam splitter; light transmitted by the first beam splitter is converged on a living cell sample in a cell culture box through an objective lens; light reflected or scattered by the living cells is collimated into parallel light by the objective lens, and the parallel light is returned to the first beam splitter; after the parallel light is reflected by the first beam splitter, the reflected light is vertically emitted to an imaging lens; light emitted by the imaging lens passes through a second beam splitter; a spectrograph receives the interference hyperspectral information of single pixels on the focal point of the imaging lens and sends the interference hyperspectral information to a computer, wherein the interference hyperspectral information is transmitted through the second beam splitter; and the computer controls an electric translation stage to complete the focusing of the cell sample,and the computer processes the interference hyperspectral signals of all pixel points so as to obtain the quantitative tomographic imaging result of the living cells; all tomographic images are spliced and combined, so that the three-dimensional structural image of the living cells is reconstructed.

Description

technical field [0001] The invention relates to a hyperspectral interference non-marking imaging method and a living cell quantitative tomographic imaging system, and relates to the technical field of living cell imaging. Background technique [0002] Long-term cell tomography is an indispensable tool in biomedical research and clinical diagnosis. Effective and continuous tomographic scanning of living cells can reveal natural, dynamic, and subcellular changes at the microscopic scale, which is crucial for grasping long-term cellular morphological information and metabolic conditions. To achieve efficient and continuous cell imaging, super-resolution fluorescence microscopy has become a much-needed imaging tool. However, with fluorescence imaging instruments, phototoxicity and photobleaching are unavoidable problems. These two phenomena limit the application of fluorescence microscopy for long-term stable imaging of live cells. Furthermore, sample preparation procedures f...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/45G01B11/06
CPCG01B11/0675G01N21/45
Inventor 黄国亮符荣鑫苏雅靳翔宇杨晗
Owner TSINGHUA UNIV