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Sleep science is entering a new era, thanks to new data-driven analysis approaches that, combined with mouse gene–editing technologies, show a promise in functional genomics and translational research. However, the investigation of sleep is time consuming and not suitable for large-scale phenotypic datasets, mainly due to the need for subjective manual annotations of electrophysiological states. Moreover, the heterogeneous nature of sleep, with all its physiological aspects, is not fully accounted for by the current system of sleep stage classification. In this study, we present a new data-driven analysis approach offering a plethora of novel features for the characterization of sleep. This novel approach allowed for identifying several substages of sleep that were hidden to standard analysis. For each of these substages, we report an independent set of homeostatic responses following sleep deprivation. By using our new substages classification, we have identified novel differences among various genetic backgrounds. Moreover, in a specific experiment with the Zfhx3 mouse line, a recent circadian mutant expressing both shortening of the circadian period and abnormal sleep architecture, we identified specific sleep states that account for genotypic differences at specific times of the day. These results add a further level of interaction between circadian clock and sleep homeostasis and indicate that dissecting sleep in multiple states is physiologically relevant and can lead to the discovery of new links between sleep phenotypes and genetic determinants. Therefore, our approach has the potential to significantly enhance the understanding of sleep physiology through the study of single mutations. Moreover, this study paves the way to systematic high-throughput analyses of sleep.

Extracting a statistically significant result from video of natural phenomenon can be difficult for two reasons: (i) there can be considerable natural variation in the observed behaviour and (ii) computer vision algorithms applied to natural phenomena may not perform correctly on a significant number of samples. This study presents one approach to clean a large noisy visual tracking dataset to allow extracting statistically sound results from the image data. In particular, analyses of 3.6 million underwater trajectories of a fish with the water temperature at the time of acquisition are presented. Although there are many false detections and incorrect trajectory assignments, by a combination of data binning and robust estimation methods, reliable evidence for an increase in fish speed as water temperature increases are demonstrated. Then, a method for data cleaning which removes outliers arising from false detections and incorrect trajectory assignments using a deep learning-based clustering algorithm is proposed. The corresponding results show a rise in fish speed as temperature goes up. Several statistical tests applied to both cleaned and not-cleaned data confirm that both results are statistically significant and show an increasing trend. However, the latter approach also generates a cleaner dataset suitable for other analysis.

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