Widefiled multiphoton excited florescence microcopy for animal study in vivo

L. C. Cheng, C. Y. Chang, C. H. Lin, Y. D. Su, T. Y. Huang, S. J. Chen

研究成果: Conference contribution


Unlike conventional multiphoton excited microscopy according to pixel-by-pixel point scanning, a widefield multiphoton excited microscopy based on spatiotemporal focusing has been developed to construct three-dimensional (3D) multiphoton fluorescence images only with the need of an axial scanning. By implementing a 4.0 W 10 kHz femtosecond laser amplifier with an instant strong peak power and a fast TE-cooled EMCCD camera with an ultra-sensitive fluorescence detection, the multiphoton excited fluorescence images with the excitation area over 100 μm × 100 μm can be achieved at a frame rate up to 80 Hz. A mechanical shutter is utilized to control the exposure time of 1 ms, i.e. average ten laser pulses reach the fluorescent specimen, and hence an uniform enough multiphoton excited fluorescence image can be attained with less photobleaching. The Brownian motion of microbeads and 3D neuron cells of a rat cerebellum have been observed with a lateral spatial resolution of 0.24 μm and an axial resolution of 2.5 μm. Therefore, the developed widefield multiphoton microscopy can provide fast and high-resolution multiphoton excited fluorescence images for animal study in vivo.

主出版物子標題Processing, Characterization, and Applications III
出版狀態Published - 2010
事件Nanobiosystems: Processing, Characterization, and Applications III - San Diego, CA, United States
持續時間: 2010 八月 42010 八月 5


名字Proceedings of SPIE - The International Society for Optical Engineering


ConferenceNanobiosystems: Processing, Characterization, and Applications III
國家/地區United States
城市San Diego, CA

All Science Journal Classification (ASJC) codes

  • 電子、光磁材料
  • 凝聚態物理學
  • 電腦科學應用
  • 應用數學
  • 電氣與電子工程


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