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Super-resolution microscopy method and system based on saturated pumping-stimulated emission detection

A technology of stimulated radiation and microscopic system, which is applied in the super-resolution microscopic method and system field based on saturated pump-stimulated radiation detection, can solve the problem of signal strength weakening, achieve signal strength improvement, improve imaging speed, The effect of high imaging resolution

Active Publication Date: 2020-02-25
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The laser power required by this method is significantly lower than that of STED, and the system structure is relatively simple. However, because the signal strength decreases with the increase of the harmonic signal order, the existing methods can only detect the second order of the sample. (few to third-order) signal, which limits higher-resolution microscopic imaging by this method

Method used

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  • Super-resolution microscopy method and system based on saturated pumping-stimulated emission detection
  • Super-resolution microscopy method and system based on saturated pumping-stimulated emission detection
  • Super-resolution microscopy method and system based on saturated pumping-stimulated emission detection

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Embodiment 1

[0037] Such as figure 1 As shown, in this embodiment, a super-resolution microscopy system based on saturated pump-stimulated radiation detection for time-domain modulation by an electro-optical modulator includes a laser (wavelength λ 1 ) 1, laser 2 (wavelength λ 2 ), the first collimating lens 3, the second collimating lens 4, the first polarizing beam splitting prism 5, the second polarizing beam splitting prism 6, beam reducing mirror 7, electro-optic modulator 8, beam expanding mirror 9, 1 / 2 wave plate 10, 1 / 4 wave plate 11, 1 / 4 wave plate 12, mirror 13, dichroic mirror 14, scanning galvanometer 15, scanning mirror 16, field lens 17, illumination objective 18, sample 19, collection objective 20, filter Light sheet 21, photodetector 22, signal generator 23, lock-in amplifier 24, acquisition card and computer 25.

[0038] use figure 1 The system shown realizes super-resolution imaging based on saturated pump-stimulated radiation detection, and the process is as follows: ...

Embodiment 2

[0050] Such as image 3 As shown, in this embodiment, the super-resolution microscopy system based on saturated pump-stimulated radiation detection for time-domain modulation through an acousto-optic modulator, its structural composition is the same as that of the electro-optic modulator 28 except that the modulator is set as the Example 1 is the same. I won't repeat them here.

[0051] use image 3 The system shown realizes super-resolution imaging based on saturated pump-stimulated radiation detection, and the process is as follows:

[0052] (1) The wavelength emitted by laser 1 is λ 1 The excitation light is then collimated by the first collimating lens 3, and then converted into linearly polarized light by the first polarization beam splitter prism 5;

[0053] (2) The exciting light is narrowed by the beam shrinker 7, then the multi-level diffraction spot is obtained after the acousto-optic modulator 28, and the first-order diffracted light (time frequency is 10kHz), t...

Embodiment 3

[0063] This embodiment provides a super-resolution microscopy method based on saturated pump-stimulated radiation detection, including the following steps:

[0064] S1 The first light source emits a wavelength of λ 1 The excitation light is modulated in the time domain to make it loaded with a time frequency of 50kHz; the modulation device is an electro-optic modulator or an acousto-optic modulator.

[0065] S2 The second light source emits a wavelength of λ 2 the reading light.

[0066] S3 combining the excitation light with the time frequency and the reading light to form a combined beam.

[0067] S4 uses the combined beam to scan the sample, so that the sample emits stimulated radiation signal light.

[0068] The S5 stimulated emission signal light enters the photodetector after being collected and converted into an electrical signal. The method of backward detection is adopted, and the stimulated emission signal light is collected by the collection objective lens.

[...

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Abstract

The invention discloses a super-resolution microscopy method and system based on saturated pumping-stimulated emission detection and belongs to fields of super-resolution microscopy imaging and nonlinear optics. The system comprises a light source module, a detection module and a processing module, wherein a sample is put between the light source module and the detection module, the light source module comprises a first light source emitting excitation light, a second light source emitting reading light, a modulation device, a dichroscope and a galvanometer scanner, the detection module comprises a photo detector, the photo detector is connected with a lock-in amplifier, and the lock-in amplifier is in communicated connection with the processing module. On the basis of confocal microscopy,saturated excitation light is introduced and is limited in time domain, so that the sample can be entangled with a fluorescent high-order harmonic signal; in addition, another beam of reading laser is introduced, so that to-be-emitted fluorescence is emitted in a form of stimulated emission light, finally, a high-order harmonic signal of the stimulated emission light is demodulated through phase-locked detection, so as to obtain a final imaging result.

Description

technical field [0001] The invention relates to the fields of super-resolution microscopic imaging and nonlinear optics, in particular to a super-resolution microscopic method and system based on saturated pump-stimulated radiation detection. Background technique [0002] Light microscopy enables live imaging of biological tissues and cells, an advantage that some other methods, such as electron microscopy, cannot match. Although the imaging resolution cannot reach the level of electron microscopy, optical microscopy can observe living tissues and cells without causing serious damage to biological samples, thus visually presenting a variety of biological Phenomenon. Among various optical microscopy methods, confocal fluorescence microscopy can improve the axial slice ability and lateral resolution of traditional far-field optical microscopy. At the same time, with the help of various specific fluorescent probes, tissue And specific imaging of various components or structur...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6402G01N21/645
Inventor 匡翠方王文生徐良丁志华李海峰刘旭
Owner ZHEJIANG UNIV
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