Wavefront sensorless adaptive optics temporal focusing-based multiphoton microscopy

Chia Yuan Chang, Li Chung Cheng, Hung Wei Su, Yvonne Yuling Hu, Keng Chi Cho, Wei Chung Yen, Chris Xu, Chen Yuan Dong, Shean Jen Chen

Research output: Contribution to journalArticlepeer-review

24 Citations (Scopus)


Temporal profile distortions reduce excitation efficiency and image quality in temporal focusing-based multiphoton microscopy. In order to compensate the distortions, a wavefront sensorless adaptive optics system (AOS) was integrated into the microscope. The feedback control signal of the AOS was acquired from local image intensity maximization via a hill-climbing algorithm. The control signal was then utilized to drive a deformable mirror in such a way as to eliminate the distortions. With the AOS correction, not only is the axial excitation symmetrically refocused, but the axial resolution with full two-photon excited fluorescence (TPEF) intensity is also maintained. Hence, the contrast of the TPEF image of a R6G-doped PMMA thin film is enhanced along with a 3.7-fold increase in intensity. Furthermore, the TPEF image quality of 1μm fluorescent beads sealed in agarose gel at different depths is improved.

Original languageEnglish
Article numberA1768
Pages (from-to)1768-1777
Number of pages10
JournalBiomedical Optics Express
Issue number6
Publication statusPublished - 2014 Jun 1

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Atomic and Molecular Physics, and Optics


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