High-speed three-dimensional two-photon fluorescence imaging system and imaging method and application thereof

Abstract

The invention provides a high-speed three-dimensional two-photon fluorescence imaging system and an imaging method and application thereof. The high-speed three-dimensional two-photon fluorescence imaging system comprises a two-photon microscope, a near-infrared femtosecond laser, an X-axis scanning device, a Y-axis scanning device, a Z-axis scanning device and an image recognition device; the X-axis scanning device is configured to include a resonance scanning mirror; the Y-axis scanning device is configured to include a galvanometer scanning mirror; the Z-axis scanning device is configured to include a high-speed piezoelectric scanning device, and the high-speed piezoelectric scanning device is used for achieving movement of high numerical aperture focused light spots in the Z-axis direction by vibrating an objective lens. The system is adopted and is simple and practical. Through mutual cooperation among the Z-axis high-speed scanning device, the X-axis resonant scanning mirror andthe Y-axis galvanometer scanning mirror and by means of a method for conducting three-dimensional imaging twice and optimizing a three-dimensional imaging space, the three-dimensional imaging speed isfurther increased, the problem of too low scanning speed is correspondingly solved, and the use of an acousto-optic deflection device is avoided.

Description

technical field [0001] The invention relates to the technical field of two-photon fluorescence imaging, in particular to a high-speed three-dimensional two-photon fluorescence imaging system and its imaging method and application. Background technique [0002] Nerve cells in the mammalian cerebral cortex form a neural network through a large number of dendrites, which form the material basis of the brain's computing function. Recent studies have shown that dendrites themselves can actively generate electrical spikes, so they are considered to have the ability to actively process external input information, which expands the brain's computing power. Therefore, the in-depth study of dendritic behavior will help to understand the operation mode and information processing mechanism of the brain's neural circuits. However, the existing research techniques on nerve cells have the following problems in the actual operation process: [0003] 1. At present, the research technology ...

AI Technical Summary

Problems solved by technology

Therefore, a commercial two-photon microscope is used to record the spike propagation of calcium ions in a single nerve cell. The nerve cells in the visual cortex of the brain are distributed in three dimensions. For information on the change of calcium ion fluorescence concentration over time at one site, if it is necessary to measure the calcium ion dynamics of different dendrites of a single nerve cell or the calcium ion dynamics of nerve cell groups, only time-sharing recording methods can be used, which cannot meet the simultaneous observation requirements

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