Widefield multiphoton microscopy with image-based adaptive optics

C. Y. Chang*, L. C. Cheng, H. W. Su, W. C. Yen, Shean-Jen Chen

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review


Unlike conventional multiphoton microscopy according to pixel by pixel point scanning, a widefield multiphoton microscope based on spatiotemporal focusing has been developed to provide fast optical sectioning images at a frame rate over 100 Hz. In order to overcome the aberrations of the widefield multiphoton microscope and the wavefront distortion from turbid biospecimens, an image-based adaptive optics system (AOS) was integrated. The feedback control signal of the AOS was acquired according to locally maximize image intensity, which were provided by the widefield multiphoton excited microscope, by using a hill climbing algorithm. Then, the control signal was utilized to drive a deformable mirror in such a way as to eliminate the aberration and distortion. A R6G-doped PMMA thin film is also increased by 3.7-fold. Furthermore, the TPEF image quality of 1 μm fluorescent beads sealed in agarose gel at different depths is improved.

Original languageEnglish
Title of host publicationUnconventional Imaging and Wavefront Sensing 2012
StatePublished - 1 Dec 2012
EventUnconventional Imaging and Wavefront Sensing 2012 - San Diego, CA, United States
Duration: 13 Aug 201214 Aug 2012

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


  • Adaptive optics
  • Bioimaging
  • Spatiotemporal focusing
  • Widefield multiphoton microscopy


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