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However, resolving these biochemical features quantitatively and spatially at subcellular resolution remains challenging because no single imaging modality can capture molecular composition, redox state, and tissue architecture simultaneously without labeling. Here, we present\n                  <jats:bold>MANIFEST<\/jats:bold>\n                  (\n                  <jats:bold>M<\/jats:bold>\n                  ultimodal\n                  <jats:bold>A<\/jats:bold>\n                  nalysis of\n                  <jats:bold>N<\/jats:bold>\n                  onlinear\n                  <jats:bold>I<\/jats:bold>\n                  maging for\n                  <jats:bold>F<\/jats:bold>\n                  unctional\n                  <jats:bold>E<\/jats:bold>\n                  ndogenous\n                  <jats:bold>S<\/jats:bold>\n                  patial correla\n                  <jats:bold>T<\/jats:bold>\n                  ion), a label-free imaging platform that integrates stimulated Raman scattering (SRS), second harmonic generation (SHG), multiphoton fluorescence (MPF), and fluorescence lifetime imaging microscopy (FLIM) with their spatial correlation. MANIFEST combines chemical imaging of lipids with autofluorescence- and lifetime-based quantification of NADH and FAD metabolism, enabling spatially resolved analysis of metabolic heterogeneity at organelle and tissue-compartment levels. We apply this framework to four distinct aging or disease models: amyloid-beta-treated tri-cultured brain cells, high-fat diet mouse liver, human non-ischemic cardiomyopathy tissue, and aging mouse retina. Across these systems, MANIFEST reveals disease-associated lipid remodeling, redox imbalance, disrupted metabolic zonation, collagen reorganization, and layer-specific metabolic changes. By integrating complementary nonlinear optical modalities into a single label-free platform, MANIFEST provides an extensible, proof-of-concept platform for high-resolution metabolic phenotyping in complex biological systems and offers new opportunities for studying disease mechanisms, aging biology, and metabolism-driven tissue pathology.\n                <\/jats:p>","DOI":"10.64898\/2026.04.11.717872","type":"posted-content","created":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T00:05:47Z","timestamp":1776211547000},"source":"Crossref","is-referenced-by-count":0,"title":["Label free multimodal optical imaging of metabolic heterogeneity and correlation in aging by integrating SRS, MPF, FLIM, and SHG"],"prefix":"10.64898","author":[{"given":"Hongje","family":"Jang","sequence":"first","affiliation":[{"name":"Shu Chien-Gene Lay Department of Bioengineering, University of California San Diego, La Jolla, CA 92093"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuang","family":"Wu","sequence":"additional","affiliation":[{"name":"Shu Chien-Gene Lay Department of Bioengineering, University of California San Diego, La Jolla, CA 92093"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hyehyun","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, University of California - Davis, Davis, CA, 95616"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tong-You Wade","family":"Wei","sequence":"additional","affiliation":[{"name":"Division of Cardiology, Department of Medicine, University of California San Diego, La Jolla, CA 92093"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian","family":"Kang","sequence":"additional","affiliation":[{"name":"Institute of Engineering in Medicine, University of California San Diego, La Jolla, CA 92093"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anahita","family":"Ataran","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Division of Cardiovascular Research, Washington University School of Medicine, St. Louis, MO 63110"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fangyuan","family":"Gao","sequence":"additional","affiliation":[{"name":"School of Medicine, University of California, Irvine, CA 92697"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dorota","family":"Skowronska-Krawczyk","sequence":"additional","affiliation":[{"name":"School of Medicine, University of California, Irvine, CA 92697"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kenneth B.","family":"Margulies","sequence":"additional","affiliation":[{"name":"Cardiovascular Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ali","family":"Javaheri","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Division of Cardiovascular Research, Washington University School of Medicine, St. Louis, MO 63110"},{"name":"John Cochran Veterans Affairs Hospital, St. Louis, MO 63106"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Sun","sequence":"additional","affiliation":[{"name":"Division of Cardiology, Department of Medicine, University of California San Diego, La Jolla, CA 92093"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John Y-J","family":"Shyy","sequence":"additional","affiliation":[{"name":"Institute of Engineering in Medicine, University of California San Diego, La Jolla, CA 92093"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erkin","family":"Seker","sequence":"additional","affiliation":[{"name":"Department of Electrical and Computer Engineering, University of California - Davis, Davis, CA 95616"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1373-3206","authenticated-orcid":false,"given":"Lingyan","family":"Shi","sequence":"additional","affiliation":[{"name":"Shu Chien-Gene Lay Department of Bioengineering, University of California San Diego, La Jolla, CA 92093"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026082607151105000_2026.04.11.717872v2.1","doi-asserted-by":"crossref","unstructured":"J. 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