An efficient algorithm for measurement and correction of chromatic aberrations in fluorescence microscopy

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Authors

KOZUBEK Michal MATULA Petr

Year of publication 2000
Type Article in Periodical
Magazine / Source Journal of Microscopy
MU Faculty or unit

Faculty of Informatics

Citation
Web URL
Field Use of computers, robotics and its application
Keywords chromatic aberration; fluorescence microscopy; fluorescent bead; image analysis
Description Even the best optical microscopes available on the market exhibit chromatic aberrations to some extent. In some types of study, chromatic aberrations of current optics cannot be neglected and a software correction is highly desirable. This paper describes a novel method of chromatic aberration measurement and software correction using sub-resolution bead imaging and computer image analysis. The method is quick, precise and enables the determination of both longitudinal and lateral chromatic aberrations. Correction function can be computed in about half an hour, including image acquisition. Using this approach, chromatic aberrations can be reduced to 10-20 nm laterally and 10-60 nm axially depending on the type of optical set-up. The method is especially suitable for fluorescence microscopy, where a limited number of wavelengths are observed.
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