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

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 d esirable. This paper describes a novel method of chromatic aberration measu rement and software correction using sub-resolution bead imaging and comput er image analysis. The method is quick, precise and enables the determinati on of both longitudinal and lateral chromatic aberrations. Correction funct ion can be computed in about half an hour, including image acquisition. Usi ng this approach, chromatic aberrations can be reduced to 10-20 nm laterall y 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.