Super resolution fluorescence lifetime imaging method and system

Abstract

The invention is applied to the fields of optics, biology, chemistry and the like and provides a super resolution fluorescence lifetime imaging method. The method comprises the following steps of: sparsely activating an optical switch dye molecule marked in a sample; exciting the activated optical switch dye molecule in the sample; collecting photons transmitted by the activated optical switch dye molecule and recording a fluorescent image of the optical switch dye molecule; carrying out the centroid positioning on the optical switch dye molecule in the fluorescent image; counting the photons received at the centroid positioning site and determining a fluorescence lifetime of the activated optical switch dye molecule; and constructing the super resolution fluorescence lifetime image by combining the centroid positioning result with the fluorescence lifetime of the obtained optical switch dye molecule. By combining the super resolution fluorescence microtechnique based on unimolecule positioning with the fluorescence lifetime imaging based on time relevant single photon counting, the invention realizes the super resolution fluorescence lifetime imaging, breaks through the traditional optical diffraction limit and has higher scientific significance and application value.

Description

technical field [0001] The invention belongs to the fields of optics, biology and chemistry, and in particular relates to a super-resolution fluorescence lifetime imaging method and an imaging system. Background technique [0002] Fluorescence microscopy has outstanding advantages such as non-destructive, non-contact, high specificity, high sensitivity, high in vivo friendliness, and ability to provide functional information. It has always been an important tool for life sciences, especially cell biology research. In recent years, with the development of life sciences, higher and higher requirements have been put forward for fluorescence microscopy. The continuous development of laser technology, fluorescent probe labeling technology, new fluorescence detection technology and imaging methods has greatly promoted the development of fluorescence microscopy. The development of microscopic technology has become an important driving force for the development of life sciences. In...

AI Technical Summary

Problems solved by technology

[0004] However, limited by the optical diffraction limit, the spatial resolution of fluorescence microscopy can only reach about 200 nanometers, which is difficult to meet the needs of life science research

For fluorescence lifetime imaging, the spatial resolution is particularly severely affected by the diffraction limit.

The fluorescence lifetime value of each pixel in the traditional diffraction-limited FLIM image cannot reflect the fluorescence lifetime of the corresponding molecule, and cannot achieve molecular-scale fluorescence lifetime imaging

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