A wide-field super-resolution microscopy imaging method and system

A microscopic imaging and super-resolution technology, applied in microscopes, analytical materials, material excitation analysis, etc., can solve problems such as damage, large light, and inapplicability to live cell imaging, and achieve the effect of avoiding damage and improving resolution.

Active Publication Date: 2022-07-05
SHENZHEN UNIV
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Problems solved by technology

At present, in SIM technology, only SSIM can achieve higher resolution than linear SIM. However, since SSIM requires high optical power excitation light to perform fluorescence saturation excitation on the sample, resulting in large optical damage, this method is not feasible. Suitable for live cell imaging, unable to take advantage of SIM

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  • A wide-field super-resolution microscopy imaging method and system
  • A wide-field super-resolution microscopy imaging method and system
  • A wide-field super-resolution microscopy imaging method and system

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Embodiment Construction

[0040] In order to make the objectives, technical solutions and advantages of the present invention clearer and clearer, the present invention will be further described in detail below with reference to the accompanying drawings and examples. It should be understood that the specific embodiments described herein are only used to explain the present invention, but not to limit the present invention.

[0041] see also Figure 1-Figure 5 , the present invention provides some embodiments of a wide-field super-resolution microscopy imaging system.

[0042] like figure 1As shown in the figure, a wide-field super-resolution microscopic imaging system of the present invention includes: a light source 1, a beam expanding and collimating device, a half-wave plate 4, a spatial light modulator 6, a Fourier lens 7, and a tube mirror, which are arranged in sequence. 8. The excitation filter 9, the beam splitter 10; the objective lens 11 located in the reflection direction of the beam spli...

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Abstract

The invention discloses a wide-field super-resolution microscopic imaging method and a system thereof. The method comprises the steps of: performing phase modulation on laser light emitted by a light source through a spatial light modulator, and obtaining intensity-modulated light through a Fourier lens; The intensity-modulated light is sequentially passed through the tube mirror, excitation filter, beam splitter, and objective lens to excite the sample to emit signal light; the signal light is collected through the objective lens, and transmitted to the detector by the beam splitter above; a super-resolution image is obtained by reconstructing the signal light structure patterns with different phases in at least three orientations. The spatial light modulator modulates the laser light into an excitation light pattern whose intensity satisfies the sine power distribution, and excites the fluorescence pattern, thereby generating multi-level spectral aliasing. The higher frequency sample information is obtained by de-frequency and resetting of the aliased spectrum. Under the condition of non-saturated excitation, the super-resolution structured light imaging is realized, and the spatial resolution is determined by the harmonic order of the striped structured light.

Description

technical field [0001] The invention relates to the technical field of optical microscopic imaging, in particular to a wide-field super-resolution microscopic imaging method and a system thereof. Background technique [0002] Fluorescence microscopy has been widely used in life science research because of its advantages of non-destructive, non-invasive, specific labeling and real-time dynamic imaging of living cells. However, due to the limitation of optical diffraction limit, the highest resolution of general imaging can only reach about 200nm. In order to break through the diffraction limit of fluorescence microscopy resolution, a series of novel super-resolution microscopy imaging methods have been proposed. [0003] Hell's group proposed the stimulated emission depletion (STED) technology, which can greatly improve the resolution to levels far higher than the diffraction limit resolution (tens of nanometers or even higher). However, high-power STED light causes great d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64G02B21/00G02B21/06
CPCG01N21/6458G02B21/0076G02B21/06G01N2021/6463
Inventor 邵永红郑晓敏汪磊王美婷屈军乐
Owner SHENZHEN UNIV
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