Miniature dual-photon micro-imaging device and method, and living sample behavior imaging system

Abstract

The invention discloses a miniature dual-photon micro-imaging device and method, and a living sample behavior imaging system. The miniature dual-photon micro-imaging device comprises a femtosecond pulse laser, a femtosecond pulse laser modulator, a miniature probe and a fluorescent output optical fiber, wherein the femtosecond pulse laser is used for generating laser with a wavelength of 920 nanometers; the femtosecond pulse laser modulator is used for receiving the laser outputted by the femtosecond pulse laser, pre-chirping the pulse width of the compensated laser to a preset value and outputting; the miniature probe comprises a scanning imaging part, the scanning imaging part is used for receiving the laser outputted by the femtosecond pulse laser modulator, and the laser is used for scanning tissues inside a living sample to excite the live sample to generate a fluorescent signal; the fluorescent output optical fiber is used for receiving and outputting the fluorescent signal outputted by the scanning imaging part. The miniature dual-photon micro-imaging device disclosed by the invention can stably observe the activities of dendrites and dendrite spines of an unrestrained animal in a natural physiological environment.

Description

technical field [0001] The invention relates to the technical field of optical imaging, in particular to a miniature two-photon microscopic imaging device and method, and a living sample behavioral imaging system. Background technique [0002] One of the ultimate goals of neuroscience is to understand the fundamentals of subcellular, cellular, circuit, and higher-level neuronal information processing in freely moving animals. Combined with fluorescent indicators, light microscopy has become a fundamental research tool in this task, as it allows direct observation of neuronal activity at multiple temporal and spatial scales. A single synapse is the fundamental unit of information transmission, processing and storage, and is critical for understanding brain function and disease mechanisms. Postsynaptic structures -- dendritic spines are submicron structures buried deep in the brain and active on the order of milliseconds. Due to its inherent optical sectioning and deep tissu...

AI Technical Summary

Problems solved by technology

However, none of these mTPMs has been used for subsequent biological applications, mainly due to two major limitations: First, no one has been able to image the most widely used fluorescent probes, such as GCaMPs, due to the lack of suitable optical fibers for delivery 920nm femtosecond laser pulses for undistorted pulse to sample

Second, mTPMs often show lower than their theoretical resolution in vivo experiments, which may be due to the low sampling rate, miniature graded-index lenses (English full name "Graded-Index Lenses", hereinafter referred to as "GRIN") aberrations, imperfections in micro-optics and imaging noise caused by motion, etc.

However, current noninvasive miniature widefield microscopes can only achieve cellular resolution, and image contrast is compromised by accumulated out-of-focus background signals

So far, novel mTPMs that offer good imaging capabilities and experimental protocols, and are powerful enough for neuroscientists' daily experiments are still to be completed.

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