Supramolecular Assembly in Live Cells Mapped by Real-Time Phasor-Fluorescence Lifetime Imaging.
Yong RenZhixuan ZhouKonrad MaxeinerAnke KaltbeitzelIain HarleyJiaqi XingYingke WuManfred WagnerKatharina LandfesterIngo LieberwirthTanja WeilDavid Yuen Wah NgPublished in: Journal of the American Chemical Society (2024)
The complex dynamics and transience of assembly pathways in living systems complicate the understanding of these molecular to nanoscale processes. Current technologies are unable to track the molecular events leading to the onset of assembly, where real-time information is imperative to correlate their rich biology. Using a chemically designed pro-assembling molecule, we map its transformation into nanofibers and their fusion with endosomes to form hollow fiber clusters. Tracked by phasor-fluorescence lifetime imaging (phasor-FLIM) in epithelial cells (L929, A549, MDA-MB 231) and correlative light-electron microscopy and tomography (CLEM), spatiotemporal splicing of the assembly events shows time-correlated metabolic dysfunction. The biological impact begins with assembly-induced endosomal disruption that reduces glucose transport into the cells, which, in turn, stymies mitochondrial respiration.
Keyphrases
- electron microscopy
- induced apoptosis
- cell cycle arrest
- high resolution
- single molecule
- oxidative stress
- cell death
- healthcare
- energy transfer
- endoplasmic reticulum stress
- high glucose
- metabolic syndrome
- adipose tissue
- fluorescence imaging
- mass spectrometry
- health information
- insulin resistance
- drug induced
- photodynamic therapy
- endothelial cells
- social media
- liquid chromatography