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Three-dimensional super-resolution microscopy imaging method and device based on variable-angle total internal reflection structured light illumination

A technology of structured light illumination and total internal reflection, which is applied in the field of optical super-resolution microscopy and can solve the problems of limited lateral resolution and diffraction of total internal reflection fluorescence microscopy.

Active Publication Date: 2019-09-20
ZHEJIANG UNIV
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

However, the lateral resolution of total internal reflection fluorescence microscopy is still limited by diffraction, and its axial resolution needs to be further improved.

Method used

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  • Three-dimensional super-resolution microscopy imaging method and device based on variable-angle total internal reflection structured light illumination
  • Three-dimensional super-resolution microscopy imaging method and device based on variable-angle total internal reflection structured light illumination
  • Three-dimensional super-resolution microscopy imaging method and device based on variable-angle total internal reflection structured light illumination

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Experimental program
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Embodiment

[0061] see figure 2 The three-dimensional super-resolution microscopic imaging device of this embodiment includes a laser 1 and a single-mode optical fiber 2 arranged in sequence along the optical path of the light emitted by the laser 1, a first convex lens 3, a second convex lens 4, a first half-wave plate 5 and The first beam splitter 6. The first beam splitter 6 divides the light path into a transmission light path and a reflection light path, and a second half-wave plate 7, a transmission path vibrating mirror system 8, a second reflection mirror 9 and a first scanning lens 10 are sequentially arranged on the transmission light path. A light barrier 12, a first reflector 13, a reflective galvanometer system 15 and a second scanning lens 16 are arranged in turn on the reflected light path, and piezoelectric ceramics for driving the first reflector to change the reflection angle are mounted on the first reflector 13. 14. A second beam splitter is provided at the confluen...

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Abstract

The invention discloses a three-dimensional super resolution microscopic imaging method and a device based on variable angle total internal reflection structured light illumination. An illumination beam is split to two paths of incident light with the consistent polarization directions and with total reflection happening, generated evanescent waves interfere to form a total internal reflection structured light illumination sample, fluorescence signals emitted by the sample are gathered to obtain a total internal reflection structured light illumination original image, and a transversal super resolution image is reconstructed. Total internal reflection of a single path of incident light on the surface of the imaging sample is used, the incidence angle and the azimuth angle of the incident light are changed one by one to scan the sample, the fluorescence signals emitted by the sample are gathered to obtain a variable-incidence angle variable-azimuth angle total internal reflection structured light illumination original image; and the variable-incidence angle variable-azimuth angle total internal reflection structured light illumination original image is preprocessed to reconstruct anaxial super resolution image of the sample, and in combination of the transversal super resolution image, a three-dimensional super resolution image is reconstructed.

Description

technical field [0001] The invention relates to the field of optical super-resolution microscopic imaging, in particular to a three-dimensional super-resolution microscopic imaging method and device based on variable-angle total internal reflection structured light illumination. Background technique [0002] Observing and studying subcellular structure and function is an urgent need in the biological field. However, for a long time, limited by the optical diffraction limit, traditional optical microscopes cannot distinguish the detailed information of objects in the lateral range of 200nm and the axial range of 600nm, which seriously limits the development and application of optical microscopes in the fields of life science, medicine and material science. application. [0003] In order to solve this problem, scientists have proposed a series of super-resolution microscopy imaging techniques that break the diffraction limit, thus realizing the nanoscale observation of tissue...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G02B21/36G02B21/06G02B21/00
Inventor 匡翠方刘文杰陈友华郑程朱大钊刘旭李海峰刘向东张克奇毛磊
Owner ZHEJIANG UNIV
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