Fluorescence differential super-resolution imaging method and imaging system

Abstract

The invention discloses a fluorescent differential super-resolution imaging method and an imaging system. By adjusting the position of the corner reflector in the system, the optical path of a beam of Gaussian pulsed laser propagating in the first optical path is extended or shortened and converted into a ring pulsed laser to focus on the sample, and another beam of Gaussian pulsed laser propagates along the second optical path to focus on the irradiation The sample, the pulse interval between the ring pulse laser and the Gaussian pulse laser irradiating the sample is greater than the fluorescence lifetime of the fluorescent dye, and the excitation light pulse signal and the fluorescence signal are collected by two detectors respectively. The first image and the second image are separated from the fluorescent signal, and the target super-resolution image with further improved resolution is obtained after analyzing and processing the first image and the second image according to image processing rules. Through the above method, the use of low-power pulsed laser reduces the damage to biological samples, and solves the problem of the degradation of super-resolution image quality caused by pixel mismatch in traditional fluorescence differential super-resolution imaging.

Description

technical field [0001] The invention relates to the technical field of super-resolution optical microscopic imaging, in particular to a fluorescence differential super-resolution imaging method and an imaging system. Background technique [0002] Imaging of living cells and tissues is crucial to the research in the field of biomedicine. In addition to correct cell culture conditions and sample preparation methods, advanced imaging methods and imaging systems can maximize the protection of biological characteristics of the observed samples on the basis of maintaining their own biological characteristics. The authenticity and validity of the obtained information. Optical microscopy has the advantages of non-contact, non-injury and specificity, and is an important symbol of the beginning of the development of modern natural science. It can be well applied to the imaging of living cells and tissues. However, the diffraction of light limits the resolution capability of the optic...

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Problems solved by technology

[0004] The embodiment of the present invention provides a fluorescence differential super-resolution imaging method and an imaging system, aiming to solve the problem of inability to observe biological samples for a long time and obtain high-quality super-resolution images due to pixel mismatch in the existing fluorescence differential super-resolution imaging method The problem

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