自适应光学在生物荧光显微成像技术中的应用

Application of adaptive optics in biological fluorescent microscopy

显微成像是开展微观生命科学研究的重要手段,其中荧光显微成像以其可特异性标记、暗场对比度高、动态示踪性能出色等优势在生物成像领域占据重要地位.由于活体生物组织通常是非透明、非均匀、各向异性的复杂三维结构,激发光和发射荧光在生物组织内传播时均会由于折射、散射、吸收等作用使得光波波前发生明显畸变,造成激发光点扩散函数以及荧光成像点扩散函数的性能显著降低.借助自适应光学技术可对上述经由生物组织传播产生的波前畸变进行实时探测和精确校正,从而提升激发光照明和发射荧光成像的空间分辨率和深度.本文归纳了生物荧光显微成像中的不同像差来源,介绍了自适应光学波前传感和校正的方法,概括了不同成像方式下波前校正的要求,重点讨论了自适应光学技术在不同类型荧光显微成像中的运用和实现方法,并对其今后的应用目标和发展方向提出展望.

Microscopy is an essential means for the Life Sciences research under the sub-micron scale. Because of the advantage in specific marking, dynamic indication, high contrast and non-destructive style, the biological fluorescence microscopy plays an important role in in-vivo imaging. When propagating in the non-transparent, non-homogeneous, anisotropic biological tissue, the excitation and emission light both will be distorted by refraction, scattering, absorption and other unknown effects and the point spread function （PSF） in each path will be consequently degraded. In order to improve the spatial resolution of fluorescence microscopy and increase the imaging depth, the adaptive optical technology has been employed in the microscope system to detect and correct the above aberrations in real-time. In this paper, aberrations in fluorescence microscope have been specified in different sources, both aberration sensing and correction methods have been introduced, and the characteristics of aberration correction in different imaging modes of fluorescence microscopy have been summarized. Then the latest applications of adaptive optics in different types of fluorescence microscopy have been discussed here and the further development trend is concluded.