High-density three-dimensional single-molecule positioning super-resolution microscopic imaging system and method

Abstract

The invention provides a high-density three-dimensional single-molecule positioning super-resolution microscopic imaging system and method. The high-density three-dimensional single-molecule positioning super-resolution microscopic imaging system comprises a first laser, a fluorescence signal generation unit, a beam splitting unit, a first signal channel unit, a second signal channel unit, a detector and a control terminal. The first signal channel unit comprises a first cylindrical lens, the second signal channel unit comprises a second cylindrical lens, and the orientations of the first cylindrical lens and the second cylindrical lens are mutually orthogonal. The beam splitting unit is used for splitting a fluorescence signal into two beams of fluorescence signals with mutually vertical propagation directions. The first signal channel unit and the second signal channel unit are used for projecting the two beams of fluorescence signals to the detector for imaging respectively. The control terminal is used for performing three-dimensional positioning and super-resolution imaging according to a forward and reverse astigmatic point spread function image pair. According to the system, the three-dimensional positioning precision of defocused fluorescent molecules is improved on the premise of not sacrificing the original imaging depth, the three-dimensional positioning accuracy is high, and the effective detection range is large.

Description

technical field [0001] The invention belongs to the field of optical microscopy technology, and in particular relates to a high-density three-dimensional single-molecule positioning super-resolution microscopic imaging system and method. Background technique [0002] In the past ten years, single-molecule localization super-resolution microscopy imaging technologies represented by Stochastic Optical Reconstruction Microscopy (STORM) and Photoactivated Localization Microscopy (PALM) have bypassed the optical diffraction limit for imaging Resolution limitation, realizing nanoscale spatial resolution, has been applied in life science research and promoted the development of related fields. Single-molecule localization technology separates the overlapping image spots (also called Point Spread Function, PSF) in space by controlling the blinking of fluorescent molecules, and then reconstructs them by detecting and positioning the sparse PSF. Therefore, obtaining a super-resolutio...

AI Technical Summary

Problems solved by technology

[0005] In view of the deficiencies in the prior art above, the purpose of the present invention is to provide a high-density three-dimensional single-molecule localization super-resolution microscopic imaging system and method, which overcomes the positioning accuracy of the existing high-density three-dimensional single-molecule localization super-resolution imaging technology. It decreases sharply with the increase of defocus degree, and the defects with small effective detection range

Images