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Proceedings Paper

Sensorless adaptive optics for isoSTED nanoscopy
Author(s): Jacopo Antonello; Xiang Hao; Edward S. Allgeyer; Joerg Bewersdorf; Jens Rittscher; Martin J. Booth
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Paper Abstract

The presence of aberrations is a major concern when using fluorescence microscopy to image deep inside tissue. Aberrations due to refractive index mismatch and heterogeneity of the specimen under investigation cause severe reduction in the amount of fluorescence emission that is collected by the microscope. Furthermore, aberrations adversely affect the resolution, leading to loss of fine detail in the acquired images. These phenomena are particularly troublesome for super-resolution microscopy techniques such as isotropic stimulated-emission-depletion microscopy (isoSTED), which relies on accurate control of the shape and co-alignment of multiple excitation and depletion foci to operate as expected and to achieve the super-resolution effect.

Aberrations can be suppressed by implementing sensorless adaptive optics techniques, whereby aberration correction is achieved by maximising a certain image quality metric. In confocal microscopy for example, one can employ the total image brightness as an image quality metric. Aberration correction is subsequently achieved by iteratively changing the settings of a wavefront corrector device until the metric is maximised. This simplistic approach has limited applicability to isoSTED microscopy where, due to the complex interplay between the excitation and depletion foci, maximising the total image brightness can lead to introducing aberrations in the depletion foci. In this work we first consider the effects that different aberration modes have on isoSTED microscopes. We then propose an iterative, wavelet-based aberration correction algorithm and evaluate its benefits.

Paper Details

Date Published: 23 February 2018
PDF: 6 pages
Proc. SPIE 10502, Adaptive Optics and Wavefront Control for Biological Systems IV, 1050206 (23 February 2018); doi: 10.1117/12.2288973
Show Author Affiliations
Jacopo Antonello, Univ. of Oxford (United Kingdom)
Xiang Hao, Yale School of Medicine (United States)
Edward S. Allgeyer, Univ. of Cambridge (United Kingdom)
Joerg Bewersdorf, Yale School of Medicine (United States)
Yale Univ. (United States)
Jens Rittscher, Univ. of Oxford (United Kingdom)
Martin J. Booth, Univ. of Oxford (United Kingdom)


Published in SPIE Proceedings Vol. 10502:
Adaptive Optics and Wavefront Control for Biological Systems IV
Thomas G. Bifano; Joel Kubby; Sylvain Gigan, Editor(s)

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