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Measuring the kinetics of the violaxanthin cycle imposes an experimental challenge. Traditionally, carotenoid analysis was carried out laboriously with high-performance liquid chromatography. In this work, we present the first in vivo approach to directly measure the kinetics of the violaxanthin cycle, using resonance Raman spectroscopy in combination with baseline correction and principal component analysis. Applying the new approach allows measuring thousands of data points as opposed to the few possible with chemical analysis over the course of a violaxanthin cycle kinetics experiment. In vivo analysis of the violaxanthin cycle is necessary to fully understand adaptation kinetics to varying light conditions, the knowledge of which is especially important for assessing the stress tolerance of plants in the wake of the increasing climate change. Three experiments on the green alga Dunaliella salina were performed, featuring both the light-to-dark and dark-to-light transitions response of the algae. This is the peer reviewed version of the following article: Koch, M. et al.: Violaxanthin cycle kinetics analysed in vivo with resonance Raman spectroscopy. In: Journal of Raman Spectroscopy (2017), which has been published in final form at http://onlinelibrary.wiley.com/doi/10.1002/jrs.5102/abstract. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.