Adaptive optics for confocal laser scanning microscopy with adjustable pinhole

Han Woong Yoo; Martin E. van Royen; Wiggert A. van Cappellen; Adriaan B. Houtsmuller; Michel Verhaegen; Georg Schitter

doi:10.1117/12.2227775

27 April 2016 Adaptive optics for confocal laser scanning microscopy with adjustable pinhole

Han Woong Yoo, Martin E. van Royen, Wiggert A. van Cappellen, Adriaan B. Houtsmuller, Michel Verhaegen, Georg Schitter

Proceedings Volume 9887, Biophotonics: Photonic Solutions for Better Health Care V; 988739 (2016) https://doi.org/10.1117/12.2227775
Event: SPIE Photonics Europe, 2016, Brussels, Belgium

Abstract

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.

Citation Download Citation

Han Woong Yoo, Martin E. van Royen, Wiggert A. van Cappellen, Adriaan B. Houtsmuller, Michel Verhaegen, and Georg Schitter "Adaptive optics for confocal laser scanning microscopy with adjustable pinhole", Proc. SPIE 9887, Biophotonics: Photonic Solutions for Better Health Care V, 988739 (27 April 2016); https://doi.org/10.1117/12.2227775

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