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Super-resolution structure illumination obvious micro-imaging method based on space-frequency domain hybrid reconstruction

A structured light illumination and microscopic imaging technology, applied in the field of optics, can solve the problems of low spatial resolution, inability to image, and not taking into account the out-of-focus background, etc., and achieve the effect of simple reconstruction steps, reduced calculation, and high parallelism

Active Publication Date: 2021-11-19
XI AN JIAOTONG UNIV
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Problems solved by technology

This workflow has achieved a 7-fold improvement in reconstruction speed, but there are problems: 1) Since the SDR reconstruction method lacks an OTF compensation step, its spatial resolution is lower than that of the frequency domain reconstruction method, such as Figure 4 (b)
2) The SDR reconstruction method does not take into account the out-of-focus background, so it is only suitable for two-dimensional super-resolution imaging of thin samples with a thickness of microns, but cannot image thicker samples

Method used

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  • Super-resolution structure illumination obvious micro-imaging method based on space-frequency domain hybrid reconstruction
  • Super-resolution structure illumination obvious micro-imaging method based on space-frequency domain hybrid reconstruction
  • Super-resolution structure illumination obvious micro-imaging method based on space-frequency domain hybrid reconstruction

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Embodiment 1

[0125] The present embodiment is based on the spatial light modulator SLM modulating laser illumination and the interferometric fringe lighting on the SIM super-resolution microscope COS7 cells mitochondria imaging, since the distribution of mitochondria COS7 cells are more dispersed, in accordance with the local area may no defocus background processing, the specific implementation of the following steps:

[0126] Step 1, figure 1 Based on the spatial light modulator SLM modulating laser illumination and the interferometric fringes SIM super-resolution microscopy system shown, to collimate the laser wavelength of 488nm spread the incident polarization beam splitter 2, through the half-wave plate 3, the vertical irradiation space and an optical modulator SLM4 backtrack, diffracted light beams vertically linear polarization in the reflected light path of the polarization beam splitter 2. Diffracted beam is converted into vertically polarized the quarter wave plate 5 and is circular...

Embodiment 2

[0154] This embodiment is a system using the same COS7 cells were imaged tubulin, tubulin has a continuous three-dimensional structure, it is necessary to collect data scanned layer by layer, and then there is performed according to a fluorescence image from the original focus background deal with. Specific implementation of the following steps:

[0155] Step 1, figure 1 Based on the spatial light modulator SLM modulating laser illumination and the interferometric fringes SIM super-resolution microscopy system shown, to collimate the laser wavelength of 488nm spread the incident polarization beam splitter 2, through the half-wave plate, the vertical light irradiation space SLM4 modulator and backtrack, diffracted light beams linearly polarized in the vertical path of the reflected light of the polarization beam splitter. Vertically polarized beam is converted into quarter wave plate 5 and is circularly polarized converging lens 6, a multi-stage aggregation diffracted zero-order di...

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Abstract

The invention discloses a super-resolution structure illumination obvious micro-imaging method based on space-frequency domain hybrid reconstruction. The method comprises the following steps: 1) generating a stripe-shaped illumination light field; 2) illuminating and exciting a to-be-detected sample to generate a fluorescence signal by utilizing the stripe-shaped illumination light field under the conditions of different stripe directions and different phase shifting steps; 3) collecting fluorescence signals to obtain original fluorescence images excited by different stripe-shaped illumination light fields; and 4) if an original fluorescence image does not comprise a defocus background, processing the original fluorescence image which does not comprise the defocus background by using a space-frequency domain hybrid reconstruction algorithm to obtain a super-resolution image; and if an original fluorescence image comprises the defocus background, processing the original fluorescence image comprising the defocus background by using the space-frequency domain hybrid reconstruction algorithm to obtain a super-resolution image with a chromatography effect. According to the method, the calculation amount required in the reconstruction process is reduced by more than two times, and the reconstruction speed can be increased to more than 80 times of that of an existing super-resolution reconstruction algorithm.

Description

Technical field [0001] The present invention belongs to the field of optical technology, relates to a method of forming a micro light significantly, in particular, relates to a widely used in biology, medicine, science, microelectronics and materials based on space-frequency domain of the hybrid reconstruction of Super micro structure clearly distinguish light imaging method. Background technique [0002] Spatial resolution of conventional optical microscope is limited to the diffraction limit of light can only reach the order of half the wavelength of light, which greatly limits the application of an optical microscope. How to achieve higher spatial resolution imaging has been one of the important research topics in the field of optical microscopy. Essence overcome diffraction limit that allows the system to distinguish between the fluorescent molecules in the region of the diffraction limit, can generally be accomplished by two methods. The first class method by randomly activa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6458G01N21/6402G01N21/6428G01N21/645G01N2021/6439G01N2021/6471G01N2021/6478
Inventor 汪召军雷铭梁言生
Owner XI AN JIAOTONG UNIV
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