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Surface plasmon nonlinear structured light illumination super-resolution microscopy imaging method and device

A surface plasmon and structured light illumination technology, applied in microscopes, measuring devices, material analysis by optical means, etc., can solve the problems of coating roughness, thickness and groove cycle error, resolution bottleneck, etc., and achieve imaging resolution. The effect of improving the imaging rate, fast imaging speed, and high imaging resolution

Active Publication Date: 2020-05-19
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

On the other hand, in order to obtain a fine structured light illumination pattern, the requirements for the parameters are very high, not only the high matching between the dielectric constant of the metal and the medium is required, but also the thickness of the film needs to reach the nanometer level. There are errors in coating roughness, thickness and groove cycle
The above factors make PSIM encounter a resolution bottleneck

Method used

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  • Surface plasmon nonlinear structured light illumination super-resolution microscopy imaging method and device
  • Surface plasmon nonlinear structured light illumination super-resolution microscopy imaging method and device
  • Surface plasmon nonlinear structured light illumination super-resolution microscopy imaging method and device

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Embodiment 1

[0042] Such as figure 2 The shown surface plasmon nonlinear structured light illumination super-resolution microscopy imaging device includes: a laser group 201 as an illumination light source, a 1 / 4 wave plate 202 and a rotating polarizer 205 for polarization adjustment of the laser light source; optical path modulation Unit, including the vibrating mirror 203 and its host computer 204 for adjusting the light path direction, mirror 207, optical filter 211; PSIM chip 209; imaging unit, including imaging lens 206, imaging lens 208, microscope objective lens 210 and camera 212; and image Reconfiguration unit 213 .

[0043] The laser group 201 is a multi-wavelength short-pulse light source with adjustable light intensity, and its light pulse width must be smaller than the fluorescence lifetime of the selected fluorescent particles. In this embodiment, the laser group 201 includes a plurality of lasers emitting pulsed laser light with different wavelengths.

[0044] The vibrati...

Embodiment 2

[0048] According to the above-mentioned imaging device, the surface plasmon nonlinear structured light super-resolution microscopic imaging steps in this embodiment are as follows:

[0049] (1) Sample preparation, placing the biological sample stained with fluorescent particles on the PSIM chip.

[0050] (2) A beam of low-intensity pulsed laser turns into circularly polarized light through the corresponding 1 / 4 wave plate, and the rotating polarizer modulates the circularly polarized light into linearly polarized light perpendicular to the grooves in one of the directions, and the incident angle is perpendicular to the PSIM chip , using a camera (CCD) to collect an image;

[0051] (3) Change the angle of the incident light by the galvanometer to deviate from the vertical direction θ and -θ degrees, while keeping the polarization direction unchanged, so that the phase of the surface plasmon interference fringe shifts by 2π / 3, and collect the other two images;

[0052] (4) rota...

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Abstract

The invention discloses a super-resolution microscopic imaging device of surface plasma nonlinear structured light illumination. The device comprises a laser source emits pulse laser of different wavelengths, a polarization adjusting unit carrying out polarization adjustment on the laser, a vibrating mirror adjusting the laser angle to realize phase modulation of interference light, a PSIM chip accommodating a fluorescent sample, an imaging collecting a fluorescence image and an image reconstruction unit; the input laser generates surface plasma interference fringes and further excites fluorescence; and the image reconstruction unit controls the vibrating mirror to adjust angle of laser and recover the collected image. The invention also discloses a super-resolution microscopic imaging method of surface plasma nonlinear structured light illumination. Compared with a present PSIM technology, the frequency shift is deeper, the resolution of sub-ten nanometer can be reached theoretically;and compared with a present point scanning super-resolution microscopic method, the imaging speed and resolution are higher.

Description

technical field [0001] The invention relates to the field of optical super-resolution microscopic imaging, in particular to a surface plasmon nonlinear structured light illumination super-resolution microscopic imaging method and a device thereof. Background technique [0002] In the post-genome era, the focus of life science research has shifted from revealing all the genetic information of life to dynamically studying the structure and function of all proteins in biological systems. Optical microscopy has become an important tool for studying life sciences due to its low biological damage compared to other imaging methods. [0003] However, the resolution of the traditional optical microscope is limited by the Abbe diffraction limit, and its resolution cannot exceed λ / (2NA), (λ: wavelength of incident light, NA: numerical aperture of the optical system). At present, the super-resolution microscopy methods that break the diffraction limit mainly include: stimulated emissio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G02B21/06G02B21/36G02B27/58G01N21/552G01N21/64
CPCG01N21/553G01N21/648G02B21/06G02B21/365G02B27/58
Inventor 杨青汤明炜庞陈雷刘小威孟超刘旭
Owner ZHEJIANG UNIV