摘要
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。
Multiphoton fluorescence microscopy is a powerful tool for studying biology. However, the current scanning method is not fast enough to fulfill the requirement for fast functional event detection in the neuroimaging field. A random-access scanning multiphoton fluorescence microscope system is reported. Two-dimensional acoustooptic deflectors （AODs） are used to scan rapidly the femtosecond excitation laser. The system is capable of achiering the scanning speed of 10 μs per scanning spot in the regin of thterest, and increases the effective scanning speed greatly, A single prism is used to compensate the severe dispersion of the excitation light introduced by the AODs. Using a 170 nm fluorescent bead, the lateral resolution of the system is measured to be 0. 3 μm and the axial resolution is about 1.3 μm, Multiphoton fluorescence image of a fluorophore-labelled cell is acquired by using the random-access scanning system and a commercial multiphoton fluorescence microscope, respectively. The result proves the multiphoton fluorescence collection efficiency of the system.
出处
《光学学报》
EI
CAS
CSCD
北大核心
2006年第12期1823-1828,共6页
Acta Optica Sinica
基金
国家自然科学(30370463
60278017
30328014)
国家重点基础研究发展规划项目(2004CB520804)资助课题
关键词
显微成像
随机扫描
声光偏转器
飞秒激光
多光子荧光成像
microscopy
random-access scanning
acousto-optic deflector
femtosecond laser
multiphoton fluorescence imaging
