One-step rapid tracking and isolation of senescent cells in cellular systems, tissues, or animal models via GLF16
Sophia Magkouta, Dimitris Veroutis, Athanasios Pousias, Angelos Papaspyropoulos, Kety Giannetti, Νatassa Pippa, Nikolaos Lougiakis, Konstantinos Kambas, Nefeli Lаgopati, Aikaterini Polyzou, Μαρία Γεωργίου, Maria Chountoulesi, Stergios Pispas, Spyros Foutadakis, Efthymios Kyrodimos, Nicole Pouli, Panagiotis Marakos, Athanassios Kotsinas, Panayotis Verginis, Dimitrios Valakos, Giannis Vatsellas, Russell Petty, Dimitris Thanos, Marco Demaria, Konstantinos Evangelou, Raffaella Di Micco, Vassilis G. Gorgoulis
Abstract
Identification and isolation of senescent cells is challenging, rendering their detailed analysis an unmet need. We describe a precise one-step protocol to fluorescently label senescent cells, for flow cytometry and fluorescence microscopy, implementing a fluorophore-conjugated Sudan Black-B analog, GLF16. Also, a micelle-based approach allows identification of senescent cells in vivo and in vitro, enabling live-cell sorting for downstream analyses and live in vivo tracking. Our protocols are applicable to cellular systems, tissues, or animal models where senescence is present. For complete details on the use and execution of this protocol, please refer to Magkouta et al.1