Spatiotemporal focusing-based widefield multiphoton microscopy for fast optical sectioning

Li Chung Cheng, Chia Yuan Chang, Chun Yu Lin, Keng Chi Cho, Wei Chung Yen, Nan Shan Chang, Chris Xu, Chen Yuan Dong, Shean Jen Chen

Research output: Contribution to journalArticlepeer-review

72 Citations (Scopus)


In this study, a microscope based on spatiotemporal focusing offering widefield multiphoton excitation has been developed to provide fast optical sectioning images. Key features of this microscope are the integrations of a 10 kHz repetition rate ultrafast amplifier featuring high instantaneous peak power (maximum 400 μ J/pulse at a 90 fs pulse width) and a TE-cooled, ultra-sensitive photon detecting, electron multiplying charge-coupled camera into a spatiotemporal focusing microscope. This configuration can produce multiphoton images with an excitation area larger than 200 × 100 μ m2 at a frame rate greater than 100 Hz (current maximum of 200 Hz). Brownian motions of fluorescent microbeads as small as 0.5 μ m were observed in real-time with a lateral spatial resolution of less than 0.5 μ m and an axial resolution of approximately 3.5 μ m. Furthermore, second harmonic images of chicken tendons demonstrate that the developed widefield multiphoton microscope can provide high resolution z-sectioning for bioimaging.

Original languageEnglish
Pages (from-to)8939-8948
Number of pages10
JournalOptics Express
Issue number8
Publication statusPublished - 2012 Apr 9

All Science Journal Classification (ASJC) codes

  • Atomic and Molecular Physics, and Optics


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