Super-resolution microscopic method and device of time-gated wide-field stimulated emission

A technology of stimulated radiation and time gate, applied in the field of optical super-resolution microscopy, can solve problems such as imaging speed limitation, and achieve the effect of fast imaging speed and high energy utilization rate

Inactive Publication Date: 2014-01-01
ZHEJIANG UNIV
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  • Application Information

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Problems solved by technology

However, since the current STED microscopy is imaged by single-point scanning, its imaging speed is limited to a certain extent.

Method used

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  • Super-resolution microscopic method and device of time-gated wide-field stimulated emission
  • Super-resolution microscopic method and device of time-gated wide-field stimulated emission

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Embodiment Construction

[0038] 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.

[0039] Such as figure 1 As shown, a time-gated wide-field stimulated emission super-resolution microscopy device includes: a first lens 1, a beam modulation module 2, a second lens 3, a first dichromatic mirror 4, a second dichromatic mirror 5, A microscope objective lens 6, a nanoscale horizontal displacement stage 7, a field lens 8 and a photoelectric sensing device 9.

[0040] Wherein, the first lens 1 is located on the optical path of the excitation light, and is used for focusing the parallel incident excitation light beam.

[0041] The beam modulation module 2 is located on the loss light optical path, and is used to modulate the parallel incident loss light so that it forms a dark spot array-like illumination spot on the sample to be tested; generally two Wollaston prisms can be used to make tw...

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Abstract

The invention discloses a super-resolution microscopic method of time-gated wide-field stimulated emission. The method comprises the steps as follows: 1), exciting light is projected to a to-be-tested sample by a microscope objective, and the to-be-tested sample is subjected to wide-field excitation to generate fluorescence; 2), depletion light is similarly projected to the to-be-tested sample after modulated by a beam modulation module, so that illumination light spots in a dark-spot array are formed, and stimulated emission depletion is performed in a wide-field excitation area; 3), fluorescence emitted by the to-be-tested sample after subjected to the stimulated emission depletion is collected by the microscope objective and focused and projected to a photoelectric sensor, so that a fluorescence image of the to-be-tested sample is obtained; 4), the to-be-tested sample is moved horizontally, step 1) to step3) are repeated, and horizontal two-dimensional scanning is performed on the to-be-tested sample to obtain fluorescence images corresponding to scanning positions; and 5), all fluorescence images are moved horizontally and superimposed to recover a two-dimensional super-resolution image finally. The invention further discloses a super-resolution microscopic device of the time-gated wide-field stimulated emission.

Description

technical field [0001] The invention belongs to the field of optical super-resolution microscopy, in particular to a time-gated wide-field stimulated radiation super-resolution microscopy method and device. Background technique [0002] Due to the existence of the diffraction limit, the lateral resolution of conventional optical microscopes has been limited to about half a wavelength, and it is impossible to resolve sub-hundred nanometer-sized microscopic objects in the visible light band. Since the 1990s, researchers have successively proposed a variety of super-resolution microscopy imaging methods to achieve spatial resolutions smaller than the diffraction limit. Among them, Stimulated Emission Depletion Microscopy (STED) is the first technology that can realize super-resolution microscopy in the far field, and it is also the most widely used at present. [0003] STED microscope is based on the conventional confocal microscope. By introducing a loss of light, the excited...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 匡翠方李帅葛剑虹刘旭
Owner ZHEJIANG UNIV
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