Metabolic Nanoscopy for Studying Aging and Diseases

Understanding how metabolism functions in multicellular organisms is essential for revealing the fundamental mechanisms of numerous biological processes. Our research has developed multimodal microscopy which integrate deuterium-probed stimulated Raman scattering (DO-SRS), multiphoton fluorescence (MPF), and second harmonic generation (SHG) into a unified nanoscopy. We have converted the metabolic imaging platform into a super resolution multiplex nanoscopy by developing A-PoD and PRM algorithms, for delivering 59 nm resolution volumetric imaging capabilities. By using various deuterated molecules—including glucose, amino acids, fatty acids, and water molecules—as bioorthogonal metabolic probes. The process of enzymatic incorporation of deuterium generates carbon-deuterium (C-D) bonds in newly formed molecules, detectable by DO-SRS within the spectral cell-silence region of the Raman spectrum, which identifies them apart from older molecules. This nanoscopy provides new biological insights into the metabolic heterogeneity of various cells and organ tissues under physiological and pathological conditions. One of the significant findings is that the over-expressed tau proteins significantly altered the lipid metabolism in aged or Alzheimer's brains, and an excessive amount of newly formed lipid droplets were accumulated in glia cells, and this phenomenon can be rescued by the activation of AMPK. This nanoscopy imaging platform can be used for disease detection, diagnosis, drug discovery, and assessing drug efficacy or resistance, as well as for mechanistic understanding of scientific fundamentals in aging processes and disease development.