Adaptive optics for confocal laser scanning microscopy with adjustable pinhole

Adaptive optics for confocal laser scanning microscopy with adjustable pinhole

Yoo, H.W. (Technische Universität Wien)
van Royen, M.E. (Erasmus MC)
van Cappellen, WA (Erasmus MC)
Houtsmuller, AB (Erasmus MC)
Verhaegen, M.H.G. (TU Delft Team Raf Van de Plas)
Schitter, G (Technische Universität Wien)

Popp, Jürgen (editor)
Tuchin, Valery V. (editor)
Matthews, Dennis L. (editor)
Pavone, Francesco S. (editor)

2016

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.

Adaptive Optics
Square Adjustable Pinhole
Confocal Laser Scanning Microscopy
Pinhole Intensity Ratio

http://resolver.tudelft.nl/uuid:9ddaee73-b3ec-496f-b332-f0cb3ae0c2bf

https://doi.org/10.1117/12.2227775

SPIE, Bellingham, WA, USA

978-1-510601321

Proceedings of SPIE: Biophotonics: Photonic Solutions for Better Health Care V, 9887

Biophotonics: Photonic Solutions for Better Health Care V, 2016-04-04 → 2016-04-07, Brussels, Belgium

Proceedings of SPIE, 1605-7422, 9887

Institutional Repository

conference paper

© 2016 H.W. Yoo, M.E. van Royen, WA van Cappellen, AB Houtsmuller, M.H.G. Verhaegen, G Schitter