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 a corner reflector in the system, the propagation optical path ofa beam of Gaussian pulse laser in a first optical path is prolonged or shortened, and the beam of Gaussian pulse laser is converted into annular pulse laser to focus and irradiate a sample, the otherbeam of Gaussian pulse laser propagates along a second light path and focuses to irradiate the sample, the pulse interval between the annular pulse laser for irradiating the sample and the Gaussian pulse laser is greater than the fluorescence lifetime of the fluorescent dye, and an exciting light pulse signal and a fluorescence signal are respectively acquired by two detectors. The first image and the second image are separated from the fluorescence signal, and the first image and the second image are analyzed and processed according to an image processing rule to obtain a target super-resolution image of which the resolution is further improved. Through the method, the damage to a biological sample is reduced by adopting the low-power pulse laser, and the problem that the quality of a super-resolution image is reduced due to pixel mismatch during traditional fluorescent differential super-resolution imaging is solved.

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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