Understanding water dynamics in different subcellular environments by combining multiplexing imaging and dimethylamino naphthalene fluorescent derivatives

Water dynamics in different subcellular locations are crucial to modulate several cellular processes such as enzyme function, membrane dynamics, and phase separation. Methods to study water dynamics at subcellular level in vivo are scarce and technically challenging. The environmentally sensitive fluorescent probes derived from dimethylaminonaphthalene (DAN) were designed to sense polarity and dipolar relaxation around its proximity and address macromolecules' structural dynamics. In this work, we used four different DAN probes: LAURDAN, PRODAN, ADCAN, and DANCA, with a differential affinity for different subcellular locations due to its functional group included in the carbonyl group.