Method and device of stimulated emission depletion (STED) microscopy based on tangential polarized light

Abstract

The invention discloses a stimulated emission depletion (STED) microscopy method based on tangential polarized light. The stimulated emission depletion microscopy method comprises the following steps of: respectively converting a first laser beam and a second laser beam into first tangential polarized light and second tangential polarized light; projecting the first tangential polarized light onto a fluorescence sample to form an exciting light spot after phase modulation; decomposing the second tangential polarized light into a first working beam and a second working beam, and carrying out phase modulation on the first working beam and the second working beam to form a first STED beam and a second STED beam; projecting the first STED beam and the second STED beam onto the fluorescence sample to form a corresponding STED light spot through incoherence addition, wherein the central point of the STED light spot coincide with the central point of the exciting light spot to obtain a focusing light spot; and collecting fluorescence emitted by the fluorescence sample under the action of the focusing light spot, and obtaining a microscopic image after processing. The invention also discloses an STED microscopy device based on the STED microscopy method. According to the STED microscopy method and the STED microscopy device, a higher resolution can be realized under the same working light intensity.

Description

technical field [0001] The invention relates to the field of super-resolution microscopy, in particular to a stimulated emission loss microscopy method and device based on tangentially polarized light. Background technique [0002] According to Abbe's diffraction limit theory, the size of the spot formed by the beam after being focused by a general lens can be expressed as Where λ is the wavelength of the incident beam, and NA is the numerical aperture of the lens used, so the limiting resolution of conventional far-field fluorescence microscopes is generally limited to about half a wavelength. [0003] In order to break through the limitation of Abbe's diffraction limit and enable fluorescence microscopy to achieve super-resolution microscopy in the far field, researchers have proposed a variety of super-resolution microscopy methods. Among them, stimulated emission depletion microscopy (STED: Stimulated Emission Depletion Microscopy) is currently the most mature and wide...

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Problems solved by technology

[0002] According to Abbe's diffraction limit theory, the size of the spot formed by the beam after being focused by a general lens can be expressed as Where λ is the wavelength of the incident beam, and NA is the numerical aperture of the lens used, so the limiting resolution of conventional far-field fluorescence microscopes is generally limited to about half a wavelength

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