Airy Beam Light Sheet Illumination Microscopic Imaging Device Based on Binary Optical Components

A binary optical element and Airy beam technology, applied in the field of optical microscopy, can solve problems such as small field of view, unfavorable detection, complex system structure, etc., and achieve a large field of view, easy control of the position, and high imaging signal-to-noise ratio Effect

Active Publication Date: 2020-01-14
SHANGHAI INST OF OPTICS & FINE MECHANICS CHINESE ACAD OF SCI
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  • Abstract
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  • Claims
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Problems solved by technology

The above invention has certain advantages, but there are still some shortcomings: the invention patent "A device and method for label-free cell detection based on light-sheet illumination" adopts Gaussian beam light-sheet illumination, and the movement of the sample realizes the full-scale scanning of the sample For detection, because the thickness of the Gaussian beam is the thinnest only at the beam waist, and the thickness increases on both sides of the beam waist, the field of view of the system using Gaussian beam light sheet illumination is too small, which is not conducive to detection; the invention patent "Airy beam light sheet "And Airy beam light sheet microscope" is a microscope based on Airy beam light sheet illumination. This system is prone to aberrations, spherical aberrations, etc., and requires subsequent optical paths to compensate, making the system structure more complicated.

Method used

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  • Airy Beam Light Sheet Illumination Microscopic Imaging Device Based on Binary Optical Components
  • Airy Beam Light Sheet Illumination Microscopic Imaging Device Based on Binary Optical Components
  • Airy Beam Light Sheet Illumination Microscopic Imaging Device Based on Binary Optical Components

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Embodiment 1

[0030] The parameters of embodiment 1 are as follows:

[0031] Described laser 1 is used for emitting Gaussian beam, adopts 488nm laser, also can adopt He-Ne laser etc. in other example; Be 200mm, clear aperture is 30mm, certainly can use the beam expander collimating lens of other parameters in other embodiments; The size of described binary optical element is 20mm * 20mm, wherein there are 20 periods of phases on the binary optical element graphics, such as image 3 As shown, the cubic phase coefficient is 5, the tilt phase factor is 10, and the working wavelength of this binary optical element is 400nm-700nm; the focal length f=40mm of the cylindrical mirror, the cross-sectional size is 25, and the numerical aperture is 0.3; The field of view of the light sheet microscope of the Airy beam generated by the system reaches 0.3178mm;

[0032] The working process of embodiment 1:

[0033] The laser beam emitted by the laser 1 passes through the beam expander and collimator le...

Embodiment 2

[0038] Embodiment 2 specifically includes the following processes:

[0039] The only difference between embodiment 2 and embodiment 1 is that a baffle is used to block one of the light sheets when the light sheet irradiates the sample, and the other processes are exactly the same as in embodiment 1. Single Airy beam light sheet illumination can further reduce phototoxicity and photobleaching on the basis of double Airy beam light sheet illumination, and the operation of single Airy beam light sheet illumination is simpler.

[0040] Experiments show that the Airy beam light sheet illumination method adopted in the present invention can not only effectively limit the excitation area, reduce the influence of phototoxicity on the sample, but also improve the axial resolution and expand the effective field of view.

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Abstract

A binary optical element-based Airy beam light-sheet microscopy imaging device includes a laser device, a beam expanding and collimating lens group, a binary optical element, a cylindrical mirror and a sample to be tested are arranged along the laser output direction of the laser device, a microscope is arranged over the sample to be tested, and the output end of the microscope is connected with the input end of a computer. Airy beam light-sheet microscopy is adopted to effectively limit the excitation area, reduce the influences of light toxicity on the sample, improve the axial resolution and expand the effective view field range. The device has the advantages of simple structure, high axial resolution, low cost and wide view field range.

Description

technical field [0001] The invention relates to optical microscopy, in particular to an Airy beam light sheet illumination microscopic imaging device based on a binary optical element. [0002] technical background [0003] Important applications of light sheet illumination in biological microscopy. Light-sheet illumination fluorescence microscopy, as a non-invasive microscopic imaging method, is an indispensable technique in the detection of biomolecules. The idea of ​​light sheet fluorescence microscopy is to use a thin layer of light sheet to illuminate the sample, only excite a thin layer of the sample to make it emit fluorescence, and the detector collects the fluorescence from the vertical direction of the illuminated surface. Light sheet illumination fluorescence microscopy makes up for the shortcomings of confocal fluorescence microscopy and two-photon fluorescence microscopy. The number of samples is small, so the phototoxicity and photobleaching are weak; ③The ima...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64G02B21/00
CPCG01N21/6458G02B21/0032
Inventor 孙彦华范永涛
Owner SHANGHAI INST OF OPTICS & FINE MECHANICS CHINESE ACAD OF SCI
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