Coaxial three-dimensional stimulated radiation loss super-resolution microscopic imaging method and apparatus thereof

A stimulated radiation loss and microscopic imaging technology, applied in the field of optical super-resolution microscopic imaging, can solve the problems of difficult adjustment, complex imaging system, and poor imaging quality, and achieve simple system, high imaging quality, and low aberration small effect

Active Publication Date: 2018-04-20
ZHEJIANG UNIV
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Problems solved by technology

Although this method is simpler than the isoSTED system, the adjustment of the three-dimensional hollow spot is realized in two optical

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  • Coaxial three-dimensional stimulated radiation loss super-resolution microscopic imaging method and apparatus thereof
  • Coaxial three-dimensional stimulated radiation loss super-resolution microscopic imaging method and apparatus thereof
  • Coaxial three-dimensional stimulated radiation loss super-resolution microscopic imaging method and apparatus thereof

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Embodiment Construction

[0048] The present invention will be described in detail below in conjunction with the embodiments and accompanying drawings, but the present invention is not limited thereto.

[0049] Such as figure 1 As shown, a schematic diagram of a coaxial three-dimensional stimulated emission depletion super-resolution microscopic imaging system device based on the use of lenses and mirrors, including: excitation light laser 1, STED light laser 2, dichromatic mirror 3, converging lens 4a, converging Lens 4b and converging lens 4c, single-mode polarization-maintaining fiber 5, collimator lens 6, polarizer 7, reflector 8a, reflector 8b and reflector 8c, achromatic 1 / 2 wave plate 9, D-type reflector 10 , spatial light modulator (SLM) 11, achromatic 1 / 4 wave plate 12, excitation light wavelength 1 / 4 wave plate 13, STED light wavelength 1 / 2 wave plate 14, STED light wavelength 1 / 4 wave plate 15, double Galvanometer 4f scanning system 16, scanning mirror 17, field lens 18, high numerical aper...

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Abstract

The invention discloses a coaxial three-dimensional stimulated radiation loss super-resolution microscopic imaging method. The method comprises the following steps: 1) exciting light and loss light are subjected to beam combination, then modulated to linearly polarized light and adjusted to a line polarization direction; 2) a spatial light modulator is used for loading a 0-2 Pi vortex phase plateand a 0-Pi vortex phase plate to simultaneously perform twice modulation on the exciting light and the loss light; a part of the loss light is modulated to transverse hollow light spot, and the otherpart is modulated to axial hollow light spot; 3) the exciting light polarization is adjusted to circular polarization, and the direction of rotation is opposite to the direction of rotation of the vortex phase plate, and the loss light polarization is converted to the circular polarization, and the direction of rotation is same with the direction of rotation of the vortex phase plate; 4) the exciting light and the loss light are focused to a sample, the exciting light is the solid light spot, the loss light is the hollow light spot, and signal light emitted by a sample can be respectively excited and loosed; and 5) the signal light is collected to obtain the microscopic images corresponding to a scanning spot of the sample. The invention also discloses a coaxial three-dimensional stimulated radiation loss super-resolution microscopic imaging apparatus.

Description

technical field [0001] The invention belongs to the field of optical super-resolution microscopic imaging, in particular to a coaxial three-dimensional stimulated radiation loss super-resolution microscopic imaging method and device. Background technique [0002] Super-resolution fluorescence microscopy imaging technology has made remarkable progress in the past ten years and has been widely used in research fields such as life sciences. Super-resolution fluorescence microscopy is often used to obtain high-resolution structures of a single sample, such as single cells, single fluorescent particles, etc., and is also used to observe the relative spatial distribution and interaction process between different samples. But at the same time, the application of super-resolution fluorescence imaging in the field of biomedical research also faces many experimental challenges, because the fluorescent labeling process and imaging beam may change the structure of the imaged sample. ...

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6458
Inventor 匡翠方刘少聪刘文杰陈友华朱大钊刘旭李海峰张克奇毛磊
Owner ZHEJIANG UNIV
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