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Sub-millisecond real-time three-dimensional super-resolution microscopic imaging system

A real-time three-dimensional, microscopic imaging technology, applied in microscopes, material analysis, material excitation analysis, etc., can solve the problems of PSF damage, inapplicability, and unsuitability for rapid molecular motion process monitoring, etc., and achieve the effect of easy operation and simple structure

Active Publication Date: 2019-09-13
FUDAN UNIV
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  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the above methods are not suitable for the monitoring of fast molecular motion process.
First, due to the high speed of molecular motion, the PSF of the molecule will be destroyed, and all PSF-based techniques will not be applicable

Method used

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  • Sub-millisecond real-time three-dimensional super-resolution microscopic imaging system

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Embodiment Construction

[0018] refer to figure 1 As shown, the sample is excited by the laser light source of the inverted fluorescence microscope to generate fluorescence signals. The first objective lens M1 of the inverted fluorescence microscope adopts 40× and the numerical aperture is 0.9 to collect the original single-molecule fluorescence signal S1 on the lower hemisphere for imaging. The sample plane of the inverted microscope is called the first plane. On the first plane 2, the second plane is formed by connecting the optical plate 4 with the bracket 3. The second plane must be firmly fixed with the microscope and independent of the external system to achieve the purpose of reducing vibration. Place the first plane mirror 6, the concave mirror M37, the second plane 8, the triangle mirror frame 9, and the L-shaped mirror frame 10 on the second plane 4. S1 is the original fluorescent signal, S2 is the image point of S1 after passing through the second objective lens M25, S3 is the imaging poin...

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Abstract

The invention belongs to the technical field of single molecule imaging and particularly relates to a sub-millisecond real-time three-dimensional super-resolution microscopic imaging system. The system comprises a first objective lens, a second objective lens, a first plane, a second plane, a first plane mirror, a concave mirror, a second plane mirror and an EMCCD. According to the system in the invention, on the basis of a basic inverted fluorescence microscope high-speed two-dimensional detection system, the second objective lens is built, a fluorescence signal of the upper hemisphere surface is collected to the second plane, and then the fluorescence signal is returned to a sample plane by the concave mirror on the second plane; after being amplified by the second objective lens, the distance between the single-molecule signal and the central axis of the concave mirror is close to the magnitude of the focal length of the concave mirror, so as to reach the optimal measurement precision in the vertical direction.

Description

technical field [0001] The invention belongs to the technical field of single-molecule imaging, and in particular relates to a submillisecond-level real-time three-dimensional super-resolution microscopic imaging system. Background technique [0002] Super-resolution microscopy brings optical microscopy into the nanometer dimension, enabling the visualization of individual molecular pathways in living cells. High temporal resolution and high three-dimensional spatial resolution microscopy are widely used to further study the mechanism of nucleoporin regulation of genes, millisecond or even faster molecular transport in cilia, intercellular molecular exchange, and transmembrane channel switching and other biological issues. Therefore, it is very important to require the super-resolution microscope experimental system to have both high temporal resolution and high three-dimensional spatial resolution of the photoelectric signal detection function. [0003] Today, super-resolu...

Claims

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Application Information

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IPC IPC(8): G01N21/64G02B21/00G02B21/02
CPCG01N21/64G01N21/6486G01N2021/6423G02B21/0088G02B21/02
Inventor 马炯刘晓兰
Owner FUDAN UNIV
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