DMD (digital micromirror device) computing holographic scanning-based fully automated TCSPC-FLIM (time-correlated single photon counting-fluorescence lifetime imaging microscopy) system and time detection method

Abstract

The present invention discloses a DMD (digital micromirror device) computing holographic scanning-based fully automated TCSPC-FLIM (time-correlated single photon counting-fluorescence lifetime imagingmicroscopy) system and a time detection method. According to the DMD (digital micromirror device) computing holographic scanning-based fully automated TCSPC-FLIM system and the time detection method,a continuous laser excites a sample to generate fluorescence under a wide field condition; an SCMOS camera transmits obtained sample fluorescence information to a computer; the computer performs calculation and processing so as to obtain the position and intensity information of a fluorescence image, selectively generates DMD two-dimensional hologram distribution corresponding to a region of interest, and at the same time controls a DMD lens group to achieve selective optical excitation; under the control of the computer, a DMD can transmit laser light from a first laser to the sample througha light microscope group, a chromatic dispersion elimination system, the DMD and the like, so that the laser light can excite the sample to realize FLIM. According to the system of the invention, theDMD is applied to a fully-automated TCSPC-FLIM system; the DMD is adopted as a scanner, and is added to an automatic selection optical path system; fully automated fluorescence service life microscopic imaging can be realized according to the definition and selection of a user. With the optical path system of the present invention adopted, an imaging scanning speed can be greatly increased, and selective optical excitation is realized, and the fluorescence imaging of a weak fluorescent region can be effectively improved.

Description

technical field [0001] The present application relates to the field of biomedical imaging, in particular to a fully automatic TCSPC-FLIM system based on DMD computational holographic scanning and a time detection method. Background technique [0002] Fluorescence lifetime imaging is an extremely important direction in the field of biomedical imaging. When a substance is excited by excitation light, the molecules in the substance transition from the ground state to the first excited state after absorbing energy, and return from the excited state to the ground state. Fluorescence is emitted by way of radiative transitions. When the excitation stops, the fluorescence intensity of the fluorescent molecules will decrease, and the time taken when the intensity decreases to 1 / e of the maximum light intensity when excited is defined as the fluorescence lifetime. Two-photon is based on nonlinear optics, and its most notable features in imaging are its strong tissue penetration abili...

AI Technical Summary

Problems solved by technology

Using the fluorescence intensity imaging method is relatively straightforward, and can observe different degrees of biological imaging, but its disadvantage is that the concentration of fluorophores, the intensity of excitation light, photobleaching and the environment of fluorophores will affect the measured fluorescence intensity, which is not conducive to quantification analyze

Currently, two-photon FLIM (Fluorescence Life-time imaging Microcopy) based on TCSPC (time-correlated single photon counting) can easily visualize the fluorescence decay curve and Poisson Statistical distribution, but its imaging speed is slow and time-consuming, which is not conducive to the rapid measurement of fluorescence lifetime

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