Spatiotemporal focusing-based widefield multiphoton microscopy for fast optical sectioning of thick tissues

Li Chung Cheng, Chia Yuan Chang, Wei Chung Yen, Shean Jen Chen

Research output: Chapter in Book/Report/Conference proceedingConference contribution

2 Citations (Scopus)


Conventional multiphoton microscopy employs beam scanning; however, in this study a microscope based on spatiotemporal focusing offering widefield multiphoton excitation has been developed to provide fast optical sectioning images. The microscope integrates a 10 kHz repetition rate ultrafast amplifier featuring strong instantaneous peak power (maximum 400 μJ/pulse at 90 fs pulse width) with a TE-cooled, ultra-sensitive photon detecting, electron multiplying charge-coupled device camera. This configuration can produce multiphoton excited images with an excitation area larger than 200 × 100 μm2 at a frame rate greater than 100 Hz. Brownian motions of fluorescent microbeads as small as 0.5 μm have been instantaneously observed with a lateral spatial resolution of less than 0.5 μm and an axial resolution of approximately 3.5 μm. Moreover, we combine the widefield multiphoton microscopy with structure illuminated technique named HiLo to reject the background scattering noise to get better quality for bioimaging.

Original languageEnglish
Title of host publicationUnconventional Imaging and Wavefront Sensing 2012
Publication statusPublished - 2012 Dec 1
EventUnconventional Imaging and Wavefront Sensing 2012 - San Diego, CA, United States
Duration: 2012 Aug 132012 Aug 14

Publication series

NameProceedings of SPIE - The International Society for Optical Engineering
ISSN (Print)0277-786X


ConferenceUnconventional Imaging and Wavefront Sensing 2012
Country/TerritoryUnited States
CitySan Diego, CA

All Science Journal Classification (ASJC) codes

  • Electronic, Optical and Magnetic Materials
  • Condensed Matter Physics
  • Computer Science Applications
  • Applied Mathematics
  • Electrical and Electronic Engineering


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