Intensity difference based three-dimensional super-resolution microscopic method and device

Abstract

The invention discloses a intensity difference based three-dimensional super-resolution microscopic method, which includes the following steps of: 1) starting a confocal imaging mode, converting the light beam sent out from a first light source into first linearly polarized light; 2) projecting the first linearly polarized light onto a sample to be tested; 3) collecting the signal light sent by the sample to be tested, thus obtaining first signal light intensity I1; 4) switching to a negative confocal imaging mode, converting the light beams sent out from a second light source and a third light source into second linearly polarized light and third linearly polarized light respectively; 5) carrying out phase modulation on the second linearly polarized light and the third linearly polarized light, and then converting them into a first modulated light beam and a second modulated light beam respectively; 6) projecting the first modulated light beam and the second modulated light beam onto the sample to be tested; 7) collecting the signal light sent by the sample to be tested, thus obtaining second signal light intensity I2; and 8) calculating effective signal light intensity I so as to obtain a super-resolution image. The invention also discloses an intensity difference based three-dimensional super-resolution microscopic device.

Description

technical field [0001] The invention belongs to the field of optical microscopy, in particular to a three-dimensional super-resolution microscopy method and device based on intensity difference. Background technique [0002] Due to the influence of the optical diffraction limit, the resolution of conventional optical microscopes is generally difficult to be less than 200nm, so it is impossible to observe nanoscale samples, which limits its application in nanotechnology and biotechnology. [0003] In recent years, in order to break through the limitations of the optical diffraction limit, researchers have proposed a variety of super-resolution microscopy methods, including: [0004] Stimulated Emission Depletion Microscopy (STED: Stimulated Emission Depletion Microscopy): Utilizes the nonlinear relationship between fluorescence saturation and excited-state fluorescence stimulated loss, and by limiting the area of ​​stimulated radiation attenuation, the size of the fluorescent...

AI Technical Summary

Problems solved by technology

STED and SIM have high requirements on system equipment, and the cost of the system is very expensive; the imaging speed of STORM and PALM is still relatively slow, which cannot meet the needs of real-time detection

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