Supramolecular Assembly in Live Cells Mapped by Real-Time Phasor-Fluorescence Lifetime Imaging

• DOI: 10.1021/jacs.4c01279
• Publication Date: 2024-04-19T12:03:15Z
• Authors: Yong Ren; Zhixuan Zhou; Konrad Maxeiner; Anke Kaltbeitzel; Iain Harley; Jiaqi Xing; Yingke Wu; Manfred Wagner; Katharina Landfester; Ingo Lieberwirth; Tanja Weil; David Y. W. Ng
• Keywords: Live cell imaging; Fluorescence-lifetime imaging microscopy; Supramolecular assembly

The complex dynamics and transience of assembly pathways in living systems complicate the understanding these molecular to nanoscale processes. Current technologies are unable track events leading onset assembly, where real-time information is imperative correlate their rich biology. Using a chemically designed pro-assembling molecule, we map its transformation into nanofibers fusion with endosomes form hollow fiber clusters. Tracked by phasor-fluorescence lifetime imaging (phasor-FLIM) epithelial cells (L929, A549, MDA-MB 231) correlative light-electron microscopy tomography (CLEM), spatiotemporal splicing shows time-correlated metabolic dysfunction. biological impact begins assembly-induced endosomal disruption that reduces glucose transport cells, which, turn, stymies mitochondrial respiration.