Super-resolution microscopy method based on dual-mode competition stimulation and super-resolution microscopy device based on dual-mode competition stimulation

Abstract

The invention discloses a super-resolution microscopy method based on dual-mode competition stimulation and a super-resolution microscopy device based on dual-mode competition stimulation. The method comprises the following steps of: irradiating the surface of a fluorescent sample by use of two types of modulated light beams emitted from a same light source, wherein one light beam is focused to form high-energy hollow light spot on the sample after passing through a spatial light modulator, so as to generate a saturation effect, and the other light beam is modulated into pulsed light by an acousto-optic modulator so as to be focused to form low-energy solid light spots on the sample; and collecting signal lights emitted from each scanning point of the sample, and extracting the signal lights which have same frequency with the pulsed light, wherein the extracted signal lights serve as effective signal lights for acquiring super-resolution images. The device is simple, and convenient to operate; by use of the competitive mechanism of the solid light spots and the hollow light spots during a process of stimulating sample fluorescent, the resolution ratio for saturated fluorescence to stimulate an ultra-diffraction limit is realized by only using one laser light source.

Description

technical field [0001] The invention belongs to the field of super-resolution, and in particular relates to a super-resolution microscopic method and device for realizing super-diffraction-limited resolution in the far field by utilizing the principle of fluorescence saturation. Background technique [0002] Due to the effect of diffraction from the optical system, there is a limit to the resolution achievable by conventional far-field optical microscopy methods. According to Abbe's diffraction limit theory, the size of the spot formed by the beam focused by the microscope objective lens is expressed as Where λ is the operating wavelength of the microscope, and NA is the numerical aperture of the microscope objective used. Therefore, the limiting resolution of conventional far-field optical microscopes is generally limited to about half a wavelength. [0003] In order to break through the limitation of the optical diffraction limit and improve the resolution of the micros...

AI Technical Summary

Problems solved by technology

Among them, the resolution of STED microscopy is determined by the optical power of the added loss light. Therefore, when achieving high resolution, the required optical power is very strong, which easily leads to the bleaching of fluorescent molecules.

SIM microscopy does not require high optical power, but due to the need for raster scanning, the imaging speed is slow and the imaging system is relatively complicated.

The imaging speed of STORM microscopy is also very slow, and it is currently difficult to apply it to the real-time detection of living cells

Images