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Nanopore-micro lens scanning based super-resolution microscopic imaging system

A microscopic imaging and nanopore technology, applied in microscopes, analytical materials, material excitation analysis, etc., can solve problems such as difficulty in distinguishing different proteins, lack of specificity in the interaction force of cutting-edge samples, and inability to apply living cells

Active Publication Date: 2019-09-13
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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Problems solved by technology

Among them, scanning electron microscopy (SEM) can achieve ultra-high resolution of 0.1nm, but because biological samples cannot survive in a high vacuum environment, they cannot be applied to living cells
The resolution of atomic force microscope (AFM) can reach the atomic scale, but AFM can only obtain a few special structural or chemical information through topography
Furthermore, due to the lack of specificity of tip-sample interaction forces, it is difficult to distinguish different proteins co-existing in samples

Method used

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  • Nanopore-micro lens scanning based super-resolution microscopic imaging system

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specific Embodiment approach 1

[0027] Specific implementation mode 1. Combination Figure 1 to Figure 7 This embodiment is described, based on a nanopore-microlens scanning super-resolution microscopy imaging system, including a scanning optical system, a Z-axis scanning control system, an inverted fluorescence microscopy imaging system, and an automatic focusing and locking system,

[0028] The scanning optical system is used to generate a focused light spot after the laser passes through the microlens, and the focused light spot is used to excite the sample to generate a fluorescent signal. The scanning optical system includes a laser 4, a polarizer 5, a first focusing lens 6, a first fiber coupler 7, a scanning probe 8 and a three-dimensional piezoelectric ceramic stage 9;

[0029] It can be seen that the monochromatic excitation light is emitted by the laser 4 , passes through the polarizer 5 and the first focusing lens 6 successively to the fiber coupler 7 , and the laser light is transmitted to the sc...

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Abstract

The invention discloses a nanopore-micro lens scanning based super-resolution microscopic imaging system, related to the technical field of microscopic particle super-resolution imaging and used for solving the existing problem of imaging of a living cell under single molecule scale. The nanopore-micro lens scanning based super-resolution microscopic imaging system comprises a scanning optical system, a Z-axis scanning detection system, an inverted fluorescent microscopic imaging system and an automatic focus fixing and focus locking system. A super-diffraction limit focusing spot generated bya nanopore-micro lens stimulates a fluorescence signal of a sample, distance between a nanopore-micro lens focusing spot and the sample is determined according to the Z-axis scanning detection systemof the nanopore-micro lens, a three-dimensional piezoelectric ceramic displacement platform is controlled to realize three-dimensional direction scanning for the sample, the automatic focus fixing and focus locking system determines a focal plane of a microscope and the sample and locks the focal plane, and finally, super-resolution imaging is executed for the sample through an inverted fluorescent microscope. A nanopore-micro lens scanning based super-resolution microscope provided by the invention can be applied to single molecule imaging at the inside of the living cell.

Description

technical field [0001] The invention relates to the technical field of super-resolution imaging of microscopic particles, in particular to a method for realizing fast super-resolution imaging inside living cells by combining three-dimensional scanning of nanopore-microlens with fluorescence microscopy imaging technology. Background technique [0002] Due to the influence of the far-field diffraction limit of traditional optical microscopes, according to the Rayleigh criterion, the resolution generally does not exceed half of the wavelength. Light with wavelengths below 400 nm usually damages living cells and cannot be used for live cell observation. Therefore, the lateral resolution of traditional optical microscopes is generally only about 230 nm. For life sciences, the pursuit of dynamic imaging of single living cells at the small molecular scale has always been the focus of instrument development. Among them, scanning electron microscopy (SEM) can achieve ultra-high res...

Claims

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Application Information

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IPC IPC(8): G01N21/64G02B21/00
CPCG01N21/6458G01N2021/6463G01N2021/6478G02B21/0036G02B21/0076
Inventor 王宏达邵丽娜石岩初宏亮王慧利孙佳音
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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