Adaptive optics for confocal laser scanning microscopy with adjustable pinhole

Conference paper (2016)

Authors

H.W. Yoo Technische Universität Wien
M.E. van Royen Erasmus MC
WA van Cappellen Erasmus MC
AB Houtsmuller Erasmus MC
M.H.G. Verhaegen Team Raf Van de Plas - Mechanical, Maritime and Materials Engineering
G Schitter Technische Universität Wien
Research Group
Team Raf Van de Plas (Mechanical, Maritime and Materials Engineering) (TU Delft)
Copyright: © 2016 H.W. Yoo, M.E. van Royen, WA van Cappellen, AB Houtsmuller, M.H.G. Verhaegen, G Schitter
DOI
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https://doi.org/10.1117/12.2227775
Adaptive Optics Confocal Laser Scanning Microscopy Square Adjustable Pinhole Pinhole Intensity Ratio
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Published Date

2016

Language

English

Reuse Rights

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Faculty

Mechanical, Maritime and Materials Engineering

Department

Delft Center for Systems and Control

Research Group

Team Raf Van de Plas

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.

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