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

A two-photon fluorescence, imaging system technology, applied in applications, analysis using fluorescence emission, medical imaging, etc., can solve the problem of inability to complete simultaneous observation requirements, avoid the use of acousto-optic deflection devices, solve the problem too slowly, The effect of improving the speed of 3D imaging

Inactive Publication Date: 2018-06-29
NYMPH TIANJIN TECH
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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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  • High-speed three-dimensional two-photon fluorescence imaging system and imaging method and application thereof
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  • High-speed three-dimensional two-photon fluorescence imaging system and imaging method and application thereof

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[0046] Implementation example: use the high-speed three-dimensional two-photon fluorescence imaging system of the present invention to record the changes in the calcium signal of the visual cortex nerve cell population of the living mouse brain, and the operation process is as follows:

[0047] (1) Mouse head surgery: Use C57BL / 6 28-40 day old mice, first anesthetize the mice, then place the mice on a constant temperature heating plate, monitor the respiration and body temperature of the mice to maintain Depth of anesthesia. The hair of the mouse head was clipped, and lidocaine hydrochloride was injected under the scalp for local anesthesia. The scalp and localized tissue over the skull are removed. A recording chamber was secured to the exposed skull using cyanoacrylate adhesive. The skull was resected with a high-speed micro electric drill and a steel needle, and the resection area was about 2mmx2.5mm. After perfusing the brain tissue with ACSF (artificial cerebrospinal f...

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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 ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61B5/00
CPCA61B5/0071A61B5/0042A61B2503/40A61B2503/42A61B2576/026
Inventor 徐强
Owner NYMPH TIANJIN TECH
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