High-speed volumetric imaging by combining multifocus multiphoton microscopy with a fast tunable lens
Wei-Kuan Lin1*, Yousuke Sakai2, Kuo-Jen Hsu1, Katsumasa Fujita2, Yen-Yin Lin3, Shi-Wei Chu1,4
1physics, National Taiwan Univeristy, Taipei, Taiwan
2Applied Physics, Osaka University, Osaka, Japan
3Institute of Photonics Technologies, National Tsing Hua University, Hsinchu, Taiwan
4Molecular Imaging Center, National Taiwan University, Taipei, Taiwan
* presenting author:Wei-Kuan Lin, email:mic200501@gmail.com
Optical microscopy plays an important role in brain study because it provides non-invasive and high-resolution features to observe the morphology of neurons. However, for monitoring brain functional dynamics in a three-dimensional volume, the imaging speed of traditional optical microscope is not enough. To achieve high-speed whole-brain functional imaging, we designed a novel system of multifocus two-photon microscopy with a fast tunable lens. The multifocus system provides millisecond sampling on a two-dimensional plane, while the tunable lens offers axial focus movement on a microsecond scale. The technique allows complete volumetric mapping with a speed comparable to the action potential, and paves the way for in vivo functional connectome study.

Keywords: optical microscopy , multifocus microscopy, multiphoton microscopy, fast tunable lens