Super-resolution imaging method and system using square lattice structured light illumination

A technology of structured light illumination and super-resolution imaging, applied in the field of super-resolution imaging, can solve the problems of reducing system construction cost and construction difficulty, high construction cost, and high phototoxicity, reducing phototoxicity, reducing construction cost, and simplifying polarization control. Effect

Active Publication Date: 2021-06-22
XI AN JIAOTONG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Aiming at the problems of high phototoxicity, complex system, and high construction cost in the current super-resolution microscopy method of striped structured light illumination, the present invention proposes a super-resolution imaging method using square lattice structured light illumination (referred to as square lattice SIM method), its phototoxicity is doubled compared with the traditional method, and there is no need for rotation and polarization control during the imaging process, which can greatly reduce the construction cost and difficulty of the system

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  • Super-resolution imaging method and system using square lattice structured light illumination
  • Super-resolution imaging method and system using square lattice structured light illumination
  • Super-resolution imaging method and system using square lattice structured light illumination

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Embodiment 1

[0068] In this embodiment, the method for obtaining super-resolution images by the interferometric lattice illumination SIM super-resolution microscopy system based on spatial light modulator SLM modulation and laser illumination is specifically implemented by the following steps:

[0069] Step 1. Use figure 1The shown interferometric lattice SIM super-resolution microscopy system based on spatial light modulator SLM modulation and laser illumination, the collimated and expanded 488nm wavelength laser enters the polarization beam splitter 2, passes through the half-wave plate, and illuminates the space vertically The light modulator SLM4 returns to the original path, and generates a vertically linearly polarized diffracted beam in the reflected light path of the polarization beam splitter. The vertically polarized light beam is converted into circularly polarized light by a quarter-wave plate 5 and converged by a lens 6, and the zero-order and other high-order diffraction beam...

Embodiment 2

[0091] This embodiment is a method for obtaining a super-resolution image based on an interferometric lattice SIM super-resolution microscope system based on lattice diffraction grating and laser illumination, which is specifically implemented by the following steps:

[0092] Step 1. Use figure 2 The interferometric lattice SIM super-resolution microscopy system based on lattice diffraction gratings and laser illumination is shown. The collimated and expanded 488nm wavelength laser is incident on the two-dimensional lattice grating 2, and the grating divides the incident laser into four beams, and the vertically polarized beam is converted into circularly polarized light by the quarter-wave plate 3 and converged by the lens 4. The converged multi-order diffracted zero-order and other higher-order diffracted beams are custom-made spatial filters 5 (e.g. figure 2 As shown in the inset), only the ±1st-order diffracted beams can pass through, including four beams. Subsequently...

Embodiment 3

[0113] This embodiment is a projected lattice SIM super-resolution microscope system based on a digital micromirror device and LED lighting. In addition, this embodiment will illustrate the implementation of non-equally spaced phase shifts and the corresponding method for obtaining super-resolution images, which are specifically implemented by the following steps:

[0114] Step 1. Use image 3 In the projected lattice SIM super-resolution microscope system based on digital micromirror device DMD modulation and LED illumination, the light emitted by the LED is focused by the condenser lens 2, and then enters the total internal reflection beam splitter prism 3 and the digital micromirror device 4. The structured light generating module is collimated by the collimator lens 5 and filtered by the excitation light filter 6 , passes through the dichroic mirror 7 , and enters the rear pupil of the microscope objective lens 9 . The collimator lens 5 and the microscopic objective lens ...

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Abstract

The invention discloses a super-resolution imaging method and system using square lattice structure light illumination, which mainly includes the following steps: forming a lattice structure light field on a sample plane, and controlling the light field modulation device to perform phase shift; acquiring the lattice structure Fluorescence images illuminated by structured light; super-resolution images are obtained by using frequency domain reconstruction algorithm; compared with ordinary striped structured light, square lattice structured light has a lower light dose at the same peak intensity, which can greatly reduce the system's Phototoxicity enables longer observation of living cells. In addition, only phase shifting of the illumination pattern is required during imaging, and no rotation is required, which greatly simplifies polarization control during imaging, thus reducing system overhead. complexity, and effectively reduce the construction cost of the system.

Description

technical field [0001] The invention belongs to the field of optical technology, and in particular relates to a super-resolution imaging method and system using square lattice structured light illumination, which can be widely used in research in fields such as biology, medicine, microelectronics and material science. Background technique [0002] The spatial resolution of traditional optical microscopes is limited by the diffraction limit of light, which can only reach the order of half the wavelength of light, which greatly limits the application range of optical microscopes. How to achieve higher spatial resolution imaging has always been one of the important research topics in the field of optical microscopy. At the end of the 20th century, super-resolution fluorescence microscopy imaging technology broke the shackles of the optical diffraction limit, enabling humans to peek into the microscopic biological world at the nanometer level, and provided a set of unprecedented...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64G06T3/40
CPCG01N21/6456G01N2021/6463G06T3/4053G06T3/4084
Inventor 汪召军雷铭
Owner XI AN JIAOTONG UNIV
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