Wavefront sensorless adaptive optics temporal focusing-based multiphoton microscopy

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 ac...

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Veröffentlicht in:Biomedical optics express 2014-06, Vol.5 (6), p.1768-1777
Hauptverfasser: Chang, Chia-Yuan, Cheng, Li-Chung, Su, Hung-Wei, Hu, Yvonne Yuling, Cho, Keng-Chi, Yen, Wei-Chung, Xu, Chris, Dong, Chen Yuan, Chen, Shean-Jen
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Sprache:eng
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Zusammenfassung: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.
ISSN:2156-7085
2156-7085
DOI:10.1364/BOE.5.001768