Open AccessAdaptive optics for confocal laser scanning microscopy with adjustable pinhole
Author(s) -
Han Woong Yoo,
Martin E. van Royen,
Wiggert A. van Cappellen,
Adriaan B. Houtsmuller,
Michel Verhaegen,
Georg Schitter
Publication year - 2016
Publication title -
proceedings of spie, the international society for optical engineering/proceedings of spie
Language(s) - English
Resource type - Conference proceedings
SCImago Journal Rank - 0.192
H-Index - 176
eISSN - 1996-756X
pISSN - 0277-786X
DOI - 10.1117/12.2227775
Subject(s) - pinhole (optics) , optics , adaptive optics , wavefront , distortion (music) , laser scanning , materials science , confocal , confocal microscopy , calibration , microscopy , resolution (logic) , spatial light modulator , image resolution , microscope , laser , physics , computer science , optoelectronics , artificial intelligence , amplifier , cmos , quantum mechanics
The pinhole plays an important role in confocal laser scanning microscopy (CLSM) for adaptive optics (AO) as well as in imaging, where the size of the pinhole denotes a trade-off between out-of-focus rejection and wavefront distortion. This contribution proposes an AO system for a commercial CLSM with an adjustable square pinhole to cope with such a trade-off. The proposed adjustable pinhole enables to calibrate the AO system and to evaluate the imaging performance. Experimental results with fluorescence beads on the coverslip and at a depth of 40 μm in the human hepatocellular carcinoma cell spheroid demonstrate that the proposed AO system can improve the image quality by the proposed calibration method. The proposed pinhole intensity ratio also indicates the image improvement by the AO correction in intensity as well as resolution
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