2004).Īpoptosis is a form of regulated cell death( D’Arcy 2019 D. The resulting MP-IVM data stream is a complex and invaluable source of information, contributing to enhance our understanding of several fundamental processes( Beltman et al. In particular, multiphoton intravital microscopy (MP-IVM) generates in-depth 3D data that encompass multiple channels for up to several hours of acquisition (x,y,z + t)( Secklehner, Celso, and Carlin 2017 Helmchen and Denk 2005 Rocheleau and Piston 2003), thus providing unprecedented insights into cellular dynamics and interactions(Pizzagalli et al., n.d.). In the last two decades, Intravital microscopy (IVM) has revolutionized live-cell imaging by enabling microscopy acquisitions in situ across different organs, making it one of the most accurate model to describe cellular activities within a living host( Sumen et al. Our findings suggest that ADeS is a valuable tool for the accurate detection and quantification of apoptosis in live-cell imaging and, in particular, intravital microscopy data, providing insights into the complex spatial-temporal regulation of this process. Finally, we employed ADeS to quantify cell survival in vitro and tissue damage in vivo, demonstrating its potential application in toxicity assays, treatment evaluation, and inflammatory dynamics. We demonstrated the effectiveness and robustness of ADeS across various imaging modalities, cell types, and staining techniques. ADeS is the first method capable of detecting the location and duration of multiple apoptotic events in full microscopy time-lapses, surpassing human performance in the same task. We trained ADeS on extensive datasets containing more than 10,000 apoptotic instances collected both in vitro and in vivo, achieving a classification accuracy above 98% and outperforming state-of-the-art solutions. To overcome this limitation, we developed ADeS, a deep learning-based apoptosis detection system that employs the principle of activity recognition. However, at present, no computational method can deliver label-free detection of apoptosis in microscopy time-lapses. Live-cell imaging enabled the study of apoptosis at the cellular level, enhancing our understanding of its spatial-temporal regulation. Amongst them, apoptosis is a crucial form of regulated cell death involved in tissue homeostasis and host defense. However, the complexity of the data generated by this technology has limited the development of effective computational tools to identify and quantify cell processes. Intravital microscopy has revolutionized live cell imaging by allowing the study of spatial-temporal cell dynamics in living animals.
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