Coaxial three-dimensional stimulated radiation loss super-resolution microscopic imaging method and apparatus thereof

Abstract

The invention discloses a coaxial three-dimensional stimulated radiation loss super-resolution microscopic imaging method. The method comprises the following steps: 1) exciting light and loss light are subjected to beam combination, then modulated to linearly polarized light and adjusted to a line polarization direction; 2) a spatial light modulator is used for loading a 0-2 Pi vortex phase plateand a 0-Pi vortex phase plate to simultaneously perform twice modulation on the exciting light and the loss light; a part of the loss light is modulated to transverse hollow light spot, and the otherpart is modulated to axial hollow light spot; 3) the exciting light polarization is adjusted to circular polarization, and the direction of rotation is opposite to the direction of rotation of the vortex phase plate, and the loss light polarization is converted to the circular polarization, and the direction of rotation is same with the direction of rotation of the vortex phase plate; 4) the exciting light and the loss light are focused to a sample, the exciting light is the solid light spot, the loss light is the hollow light spot, and signal light emitted by a sample can be respectively excited and loosed; and 5) the signal light is collected to obtain the microscopic images corresponding to a scanning spot of the sample. The invention also discloses a coaxial three-dimensional stimulated radiation loss super-resolution microscopic imaging apparatus.

Description

technical field [0001] The invention belongs to the field of optical super-resolution microscopic imaging, in particular to a coaxial three-dimensional stimulated radiation loss super-resolution microscopic imaging method and device. Background technique [0002] Super-resolution fluorescence microscopy imaging technology has made remarkable progress in the past ten years and has been widely used in research fields such as life sciences. Super-resolution fluorescence microscopy is often used to obtain high-resolution structures of a single sample, such as single cells, single fluorescent particles, etc., and is also used to observe the relative spatial distribution and interaction process between different samples. But at the same time, the application of super-resolution fluorescence imaging in the field of biomedical research also faces many experimental challenges, because the fluorescent labeling process and imaging beam may change the structure of the imaged sample. ...

AI Technical Summary

Problems solved by technology

Although this method is simpler than the isoSTED system, the adjustment of the three-dimensional hollow spot is realized in two optical paths, which is more likely to generate additional aberrations and deteriorate the imaging quality. Difficulty is still relatively high

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