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Wide-field multispectral fluorescence microscopic imaging method and system based on dynamic speckle illumination

A dynamic speckle and microscopic imaging technology, applied in the field of biomedical imaging research, can solve the problems of low excitation efficiency, high cost and complex structure

Pending Publication Date: 2022-01-28
GUILIN UNIV OF ELECTRONIC TECH
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Problems solved by technology

However, the laser scanning confocal fluorescence microscopy system adopts point-by-point scanning imaging method, which leads to long imaging time, low excitation efficiency, complex structure and high cost.
In addition, phototoxicity, photodamage and photobleaching are also unavoidable problems.

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  • Wide-field multispectral fluorescence microscopic imaging method and system based on dynamic speckle illumination
  • Wide-field multispectral fluorescence microscopic imaging method and system based on dynamic speckle illumination
  • Wide-field multispectral fluorescence microscopic imaging method and system based on dynamic speckle illumination

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Embodiment Construction

[0019] The present invention will be further described in detail below in conjunction with examples, so that those skilled in the art can implement it with reference to the description.

[0020] It should be understood that terms such as "having", "comprising" and "including" used herein do not exclude the presence or addition of one or more other elements or combinations thereof.

[0021]A wide-field multi-spectral fluorescence microscopic imaging method and system based on dynamic speckle illumination, characterized in that the system is composed of a multi-spectrum dynamic speckle illumination system and a fluorescence spectrum signal collection and imaging system. The system mainly consists of mode-locked Ti:Sapphire femtosecond pulse laser 1; doublet lenses 2, 3, 6, 7; scatterer 4; micro-displacement stage 5; CCD camera 8; imaging lens 9; Group 10; dichromatic mirror 11; apochromatic microscope objective lens 12; stage 13; cell culture dish 14; computer terminal 15; In t...

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Abstract

The invention provides a wide-field multispectral fluorescence microscopic imaging method and system based on dynamic speckle illumination. The system is characterized in that a mode-locked titanium sapphire femtosecond pulse laser is used as a light source. A femtosecond laser pulse with a wide spectral range forms a speckle pattern after passing through a scatterer, full-field illumination is formed in the field of view of the microscope objective, and different types of fluorophores in a biological sample to be detected are excited at the same time. Illumination speckle patterns are sequentially changed, a CCD camera detects and receives fluorescence images obtained through illumination of the dynamically-changed speckle patterns, and the wide-field multispectral fluorescence microscopic imaging method based on dynamic speckle illumination is achieved through a tomography extraction algorithm. The system constructed by the invention can simultaneously and quickly acquire three-dimensional space distribution images of different types of fluorophores in a biological sample, has the characteristics of high temporal and spatial resolution, simple structure, easiness in operation and the like, and has wide application prospects in the research fields of biology, medicine, life science and the like.

Description

(1) Technical field [0001] The present invention relates to a wide-field multi-spectral fluorescence microscopy imaging method and system based on dynamic speckle illumination, which can be used for multi-wavelength excitation conditions, high temporal resolution, high spatial resolution and multiple Spectral fluorescence imaging belongs to the field of biomedical imaging research. (2) Background technology [0002] A cell is an independent, orderly, self-regulating structural and functional system. Every cell has a complete set of equipment to meet its own metabolic needs. Unicellular organisms can carry out all life activities independently. In multicellular organisms, although the function of each cell is coordinated and controlled by the whole, each cell is an independent, highly ordered, self-controlled metabolic system. They have relatively independent life activities, and various tissues use cells as the basic unit to perform specific functions. As the basic unit o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N15/10G01N15/14G01N21/64G01N21/01
CPCG01N15/10G01N15/1436G01N21/6486G01N21/6458G01N21/01G01N2015/1006G01N2015/1445G01N2021/0112
Inventor 尹君王少飞于凌尧陈宏宇胡徐锦贾源苑立波
Owner GUILIN UNIV OF ELECTRONIC TECH
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