STED super-resolution microscope based on first-order Bessel beams and adjustment method thereof

一种贝塞尔光束、超分辨的技术，应用在受激辐射损耗超分辨显微镜及其调节领域，达到提高成像信噪比、实验装置廉价、操作简单的效果

Active Publication Date: 2015-12-23
PEKING UNIV
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Problems solved by technology

[0005] Aiming at the above problems encountered by STED microscopes in deep imaging in the prior art, the present invention proposes a method that utilizes the anti-scattering and self-healing properties of Bessel beams to convert lost light into first-order Bessel beams, thereby having A certain aberration and scattering self-compensation ability to achieve stable super-resolution imaging inside the sample

Method used

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  • STED super-resolution microscope based on first-order Bessel beams and adjustment method thereof
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  • STED super-resolution microscope based on first-order Bessel beams and adjustment method thereof

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Embodiment 1

[0031] Such as figure 1As shown, the stimulated radiation loss super-resolution microscope based on the first-order Bessel beam of this embodiment includes: an excitation light source 1, a loss light source 2, an excitation light beam expander collimation system 3-1, a loss light beam expander collimator Straight system 3-2, spiral phase plate 4, half slide 5, Bessel beam generating system 6, lossy light focusing lens 14, beam combining system 8, objective lens 9, piezoelectric scanning system 10, filter 11. Signal collection system 12 and single photon detector 13; wherein, the excitation light output by excitation light source 1 fills the entrance pupil of objective lens 9 after passing through excitation light beam expander collimation system 3-1; the linear polarization output by loss light source 2 The lost light passes through the beam expander collimation system 3-2, the spiral phase plate 4, the half slide 5, the Bessel beam generating system 6 and the lost light focus...

Embodiment 2

[0037] Such as figure 2 As shown, in this embodiment, the Bessel beam generating system adopts an annular template 6-1 and a lens 6-2, by adjusting the distance between the lens 6-2 of the Bessel beam generating system and the lossy light focusing lens 14 The distance is such that the center of the point light formed by the excitation light after the objective lens is located at the axial center of the line light of the Bessel beam, and the lateral direction coincides precisely.

[0038] In this embodiment, the loss light is perpendicular to the excitation light, and the loss light is parallel to the signal light, the first dichroic mirror totally reflects the loss light, fully transmits the excitation light, and fully transmits the signal light; the second dichroic mirror In order to fully transmit the excitation light and fully reflect the signal light; thus, after the lost light is totally reflected by the first dichroic mirror, it combines with the excitation light after ...

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Abstract

The invention discloses an STED super-resolution microscope based on first-order Bessel beams and an adjustment method thereof. The STED microscope comprises an excitation light source, a loss light source, an excitation light beam expanding and collimation system, a loss light beam expanding and collimation system, a spiral phase plate, a Bessel beam generation system, a loss light focusing lens, a beam combining system, an object lens, a piezoelectric scanning system, a filter plate, a signal collection system and a single-photon detector. Loss light is the first-order Bessel beams and has scattering-resistant and self-healing characteristics, and great light spot form of the deep position of a sample can be maintained so that resolution of the deep area of the sample can be enhanced. Compared with a method of realizing STED super-resolution microscope deep imaging through adjusting an object lens correction ring, the method is relatively simple in experiment operation without active adjustment. Compared with a method of using a self-adaptive optical system, the experiment device is relatively simple and cheap.

Description

technical field [0001] The invention relates to microscope technology, in particular to a stimulated radiation loss super-resolution microscope based on first-order Bessel beams and an adjustment method thereof. Background technique [0002] Super-resolution microscopy has had a significant impact in the fields of biological imaging, material characterization, and laser fine processing. Among them, the stimulated radiation depletion STED microscope is a microscopic technique that realizes super-resolution imaging or excitation by directly adjusting the area of ​​the signal generation area on the basis of a confocal point scanning microscope. Compared with other types of super-resolution microscopy techniques , its principle is relatively simple, and the imaging speed is fast, and it can perform real-time super-resolution imaging in vivo. It has huge applications in the field of biomedicine, and provides a new feasible method in the fields of nanomaterial research, laser proc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G02B21/36G02B21/06
CPCG02B21/0076G02B27/58G01N21/6458G01N2021/6463G02B21/0032G02B21/0072G02B21/0036
Inventor 施可彬席鹏于文韬龚旗煌
Owner PEKING UNIV
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