Automated spherical aberration correction in scanning confocal microscopy

Journal article (2014)
Department
Delft Center for Systems and Control (Mechanical, Maritime and Materials Engineering) (TU Delft)
Copyright: © 2014 AIP Publishing
DOI: https://doi.org/doi:10.1063/1.4904370
Medical imaging Entropy Mage sensors Spherical aberrations Confocal microscopes
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Published Date

18-12-2014

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Source:

Review of Scientific Instruments, 85 (12), 2014

ISSN:

0034-6748

Source:

http://scitation.aip.org/content/aip/journal/rsi/85/12/10.1063/1.4904370

Faculty

Mechanical, Maritime and Materials Engineering

Department

Delft Center for Systems and Control

Abstract

Mismatch between the refractive indexes of immersion media and glass coverslips introduces spherical aberrations in microscopes especially for high numerical aperture objectives. This contribution demonstrates an automated adjustment of the coverslip correction collar in scanning confocal microscopy to compensate for spherical aberrations due to coverslip thickness mismatch. With a motorized coverslip correction collar, the adjustment procedure consists of xz image scans, image processing, correction quality evaluation, the mismatch estimation, and eventually the optimal adjustment of the correction collar. For fast correction with less photodamage, coarse-fine Gaussian fitting algorithms are proposed and evaluated with various specimen for their estimation accuracy. The benefits of the proposed automated correction are demonstrated for various coverslips with biological specimens, showing the optimized resolution of the confocal microscope.

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