基于直接波前探测的深层动态自适应双光子荧光显微成像研究

Two-photon fluorescence microscopy has been widely used in the field of life sciences for three-dimensional imaging observation of deep tissues in vivo due to its advantages of large penetration depth, low phototoxicity and low photobleaching. Adaptive optics can overcome the influence of wavefront distortion caused by inhomogeneous refractive index distribution on imaging resolution. The current adaptive two-photon microscopy technology based on point scanning and isohalo exposure can solve the problem of low wavefront detection energy for fluorescent targets, but the imaging resolution for deeper biological tissues is still low. This project will intend to improve the accuracy and sensitivity of wavefront distortion detection by establishing a division mechanism of the isoplanatic region and improving the centroid detection algorithm of Hartmann wavefront detector for extended targets, and by automatically controlling the gain of EMCCD. The Optimized adaptive optics technology will make the resolution of two-photon microscopic imaging system close to the diffraction limit, and makes the imaging depth from the current micron level to the millimeter level. The research results of this project will have important scientific and applied value for basic research in the field of life science.

双光子荧光显微技术以其大穿透深度、低光毒性、低光漂白等优点而被广泛应用于生命科学领域进行活体的深层组织三维成像观察。自适应光学技术可以克服生物组织折射率分布不均匀导致的波前畸变对成像分辨率影响。当前的基于点扫描和等晕区曝光的自适应双光子显微成像技术很好的解决了荧光目标的波前探测能量低问题，但是对于深层生物组织的成像分辨率仍然较低。本项目拟通过建立等晕区划分机制和扩展目标的波前探测技术，以及通过探测器自动增益调整技术来提高波前畸变探测精度和灵敏度，从而提高自适应光学校正精度，使得双光子显微成像系统分辨率接近衍射极限的同时其成像深度由目前的微米级提升到毫米量级。本项目研究成果对于生命科学领域进行基础研究具有重要的科学及应用价值。

双光子荧光显微技术以其大穿透深度、低光毒性、低光漂白等优点而被广泛应用于生命科学领域进行活体的深层组织三维成像观察。自适应光学技术可以克服生物组织折射率分布不均匀导致的波前畸变对成像分辨率影响。当前的基于点扫描和等晕区曝光的自适应双光子显微成像技术很好的解决了荧光目标的波前探测能量低问题，但是对于深层生物组织的成像分辨率仍然较低。本项目通过组织像差模拟建立波前探测平台，进而建立等晕区划分机制和扩展目标的波前探测技术，提高波前畸变探测精度和灵敏度。本项目研究成果对于生命科学领域进行基础研究具有重要的科学及应用价值。