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

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...

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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.

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