基于波前无传感器自适应光学时间聚焦的多光子显微镜
Wavefront sensorless adaptive optics temporal focusing-based multiphoton microscopy.
作者信息
Chang Chia-Yuan, Cheng Li-Chung, Su Hung-Wei, Hu Yvonne Yuling, Cho Keng-Chi, Yen Wei-Chung, Xu Chris, Dong Chen Yuan, Chen Shean-Jen
机构信息
Department of Photonics, National Cheng Kung University, Tainan 701, Taiwan.
Department of Engineering Science, National Cheng Kung University, Tainan 701, Taiwan.
出版信息
Biomed Opt Express. 2014 May 9;5(6):1768-77. doi: 10.1364/BOE.5.001768. eCollection 2014 Jun 1.
Temporal profile distortions reduce excitation efficiency and image quality in temporal focusing-based multiphoton microscopy. In order to compensate the distortions, a wavefront sensorless adaptive optics system (AOS) was integrated into the microscope. The feedback control signal of the AOS was acquired from local image intensity maximization via a hill-climbing algorithm. The control signal was then utilized to drive a deformable mirror in such a way as to eliminate the distortions. With the AOS correction, not only is the axial excitation symmetrically refocused, but the axial resolution with full two-photon excited fluorescence (TPEF) intensity is also maintained. Hence, the contrast of the TPEF image of a R6G-doped PMMA thin film is enhanced along with a 3.7-fold increase in intensity. Furthermore, the TPEF image quality of 1μm fluorescent beads sealed in agarose gel at different depths is improved.
时间轮廓畸变会降低基于时间聚焦的多光子显微镜中的激发效率和图像质量。为了补偿这些畸变，一个无波前传感器自适应光学系统（AOS）被集成到显微镜中。AOS的反馈控制信号通过爬山算法从局部图像强度最大化中获取。然后利用该控制信号以消除畸变的方式驱动可变形镜。通过AOS校正，不仅轴向激发被对称地重新聚焦，而且具有全双光子激发荧光（TPEF）强度的轴向分辨率也得以保持。因此，掺杂R6G的PMMA薄膜的TPEF图像的对比度得到增强，同时强度增加了3.7倍。此外，不同深度封装在琼脂糖凝胶中的1μm荧光珠的TPEF图像质量也得到了改善。
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