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多光子荧光显微成像是生物学研究的有力手段,但目前的成像速度难以满足神经成像中快事件检测的需要。针对这一问题,提出了一套随机扫描快速多光子荧光显微成像系统。系统采用二维声光偏转器快速扫描飞秒激发光束,能够以每点10μs的速度对特定的感兴趣区域进行跳跃式扫描,即随机扫描,使得有效的扫描速度大为提高。引入单棱镜补偿方法解决应用声光偏转器带来的色散问题。以170 nm荧光小球为样品,测得系统的横向分辨力为0.3μm,纵向分辨力为1.3μm。给出了随机扫描系统和商品化多光子荧光显微镜对同一个荧光细胞的成像结果,证明了新系统的成像能力。
Multi-photon fluorescence microscopy is a powerful tool for biological research, but the current imaging speed is difficult to meet the needs of fast event detection in neuroimaging. In response to this problem, a set of random scanning fast multi-photon fluorescence microscopy imaging system is proposed. The system uses two-dimensional acousto-optic deflector to rapidly scan the femtosecond excitation beam, which can jump-scan a specific region of interest at a rate of 10 μs per point, ie, a random scan, so that the effective scanning speed is greatly improved. The introduction of single prism compensation method to solve the dispersion problem caused by the use of acousto-optic deflector. With 170 nm fluorescent beads as sample, the lateral resolution of the system was 0.3μm and the vertical resolution was 1.3μm. The imaging results of the same fluorescent cell by a randomized scanning system and a commercial multi-photon fluorescence microscope are given, demonstrating the imaging capabilities of the new system.