{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T21:18:29Z","timestamp":1792790309575,"version":"4.5.0"},"posted":{"date-parts":[[2026,9,15]]},"group-title":"Physiology","reference-count":70,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T00:00:00Z","timestamp":1789430400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"award":["R21AG086974"],"award-info":[{"award-number":["R21AG086974"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"name":"NIH Common Fund","award":["R01AG081270"],"award-info":[{"award-number":["R01AG081270"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Hevolution\/AFAR","award":["AGR00030264"],"award-info":[{"award-number":["AGR00030264"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,15]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Mitochondrial dysfunction is a hallmark of aging, yet how mitochondrial states are remodeled across tissues and subcellular compartments\n                  <jats:italic>in vivo<\/jats:italic>\n                  remains elusive. Progress has been limited, in part, because mitochondrial physiology is highly sensitive to experimental perturbations, underscoring the need for minimally disruptive measurement strategies. Here, we establish a tissue-resolved,\n                  <jats:italic>in vivo<\/jats:italic>\n                  framework for the quantitative analysis of mitochondrial states in live, intact\n                  <jats:italic>Caenorhabditis elegans<\/jats:italic>\n                  without confounding effects from mounting-induced hypoxia. This platform couples two-photon fluorescence lifetime imaging microscopy (2p-FLIM) with a custom segmentation pipeline, MitoSLIT, to track functional and structural features across multiple tissues and single neurons. By integrating membrane potential-associated TMRM intensity, lifetime-based microenvironmental metrics, and morphological descriptors, we uncover localized metabolic heterogeneity masked by conventional intensity analysis. Leveraging this framework, we mapped physiological aging against mitochondrial shifts induced by acute stress and fission-fusion mutations. Our analyses reveal that mitochondrial aging is highly tissue-specific, executing distinct trajectories across cell types. Extending the framework to genetically identified neurons revealed age-dependent divergence between somatic and axonal mitochondrial states, accompanied by structural remodeling and a late shift in optical redox ratio. Together, our findings demonstrate that mitochondrial populations do not converge on a uniform bioenergetic endpoint during aging, but rather follow highly compartmentalized, tissue-specific spatiotemporal trajectories\n                  <jats:italic>in vivo<\/jats:italic>\n                  .\n                <\/jats:p>","DOI":"10.64898\/2026.09.09.750228","type":"posted-content","created":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T05:15:16Z","timestamp":1789535716000},"source":"Crossref","is-referenced-by-count":0,"title":["Multiparametric\n                  <i>in vivo<\/i>\n                  mapping reveals tissue-specific mitochondrial aging trajectories"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9521-3476","authenticated-orcid":false,"given":"Wenyu","family":"Tang","sequence":"first","affiliation":[{"name":"Department of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0954-0834","authenticated-orcid":false,"given":"Haoyu","family":"Xu","sequence":"additional","affiliation":[{"name":"Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven","family":"Beck","sequence":"additional","affiliation":[{"name":"Department of Physiology and Aging, University of Florida, Gainesville, FL, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2718-6533","authenticated-orcid":false,"given":"Marcus T","family":"Cicerone","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA, USA"},{"name":"Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7442-754X","authenticated-orcid":false,"given":"Sung Min","family":"Han","sequence":"additional","affiliation":[{"name":"Department of Physiology and Aging, University of Florida, Gainesville, FL, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2234-5686","authenticated-orcid":false,"given":"Wei-Wen","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026091809400707000_2026.09.09.750228v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2024.04.037"},{"key":"2026091809400707000_2026.09.09.750228v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2022.11.001"},{"key":"2026091809400707000_2026.09.09.750228v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/s41574-021-00626-7"},{"key":"2026091809400707000_2026.09.09.750228v1.4","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00058.2021"},{"key":"2026091809400707000_2026.09.09.750228v1.5","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-019-03024-5"},{"key":"2026091809400707000_2026.09.09.750228v1.6","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2017.10.038"},{"key":"2026091809400707000_2026.09.09.750228v1.7","doi-asserted-by":"publisher","DOI":"10.1016\/j.semcdb.2023.02.010"},{"issue":"2","key":"2026091809400707000_2026.09.09.750228v1.8","first-page":"734","article-title":"Tissue-specific implications of mitochondrial alterations in aging","volume":"5","year":"2013","journal-title":"Front Biosci (Elite Ed"},{"key":"2026091809400707000_2026.09.09.750228v1.9","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2016.106"},{"key":"2026091809400707000_2026.09.09.750228v1.10","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymeth.2010.01.033"},{"key":"2026091809400707000_2026.09.09.750228v1.11","doi-asserted-by":"publisher","DOI":"10.1002\/cptx.10"},{"key":"2026091809400707000_2026.09.09.750228v1.12","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejcb.2023.151371"},{"key":"2026091809400707000_2026.09.09.750228v1.13","doi-asserted-by":"publisher","DOI":"10.1038\/s44318-025-00632-9"},{"key":"2026091809400707000_2026.09.09.750228v1.14","doi-asserted-by":"publisher","DOI":"10.1039\/D3CS00456B"},{"key":"2026091809400707000_2026.09.09.750228v1.15","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccr.2024.215893"},{"key":"2026091809400707000_2026.09.09.750228v1.16","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccr.2025.216456"},{"key":"2026091809400707000_2026.09.09.750228v1.17","doi-asserted-by":"publisher","DOI":"10.1038\/s43586-024-00358-8"},{"key":"2026091809400707000_2026.09.09.750228v1.18","doi-asserted-by":"publisher","DOI":"10.1039\/d5cs00280j"},{"key":"2026091809400707000_2026.09.09.750228v1.19","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-021-01108-4"},{"key":"2026091809400707000_2026.09.09.750228v1.20","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-bioeng-071516-044730"},{"key":"2026091809400707000_2026.09.09.750228v1.21","doi-asserted-by":"publisher","DOI":"10.1016\/j.addr.2023.115081"},{"key":"2026091809400707000_2026.09.09.750228v1.22","doi-asserted-by":"publisher","DOI":"10.1038\/s42003-024-06057-0"},{"key":"2026091809400707000_2026.09.09.750228v1.23","doi-asserted-by":"publisher","DOI":"10.1016\/j.semcdb.2023.04.006"},{"key":"2026091809400707000_2026.09.09.750228v1.24","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-3495(99)77214-0"},{"key":"2026091809400707000_2026.09.09.750228v1.25","doi-asserted-by":"publisher","DOI":"10.1038\/s41514-021-00061-y"},{"issue":"12","key":"2026091809400707000_2026.09.09.750228v1.26","first-page":"060613","article-title":"Spectral scanning and fluorescence lifetime imaging microscopy (FLIM) enable separation and characterization of c. elegans autofluorescence in the cuticle and gut","volume":"13","year":"2024","journal-title":"Biol. 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