{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T20:00:35Z","timestamp":1772136035708,"version":"3.50.1"},"reference-count":43,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,3,23]],"date-time":"2021-03-23T00:00:00Z","timestamp":1616457600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2021,3,23]],"date-time":"2021-03-23T00:00:00Z","timestamp":1616457600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"NIH","award":["R01CA241618"],"award-info":[{"award-number":["R01CA241618"]}]},{"name":"NIH","award":["R01EB023232"],"award-info":[{"award-number":["R01EB023232"]}]},{"name":"NSF","award":["CBET 18-41539"],"award-info":[{"award-number":["CBET 18-41539"]}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The mitochondrion is one of the key organelles for maintaining cellular homeostasis. External environmental stimuli and internal regulatory processes may alter the metabolism and functions of mitochondria. To understand these activities of mitochondria, it is critical to probe the key metabolic molecules inside these organelles. In this study, we used label-free chemical imaging modalities including coherent anti-Stokes Raman scattering and multiphoton-excited fluorescence to investigate the mitochondrial activities in living cancer cells. We found that hypothermia exposure tends to induce fatty-acid (FA) accumulation in some mitochondria of MIAPaCa-2 cells. Autofluorescence images show that the FA-accumulated mitochondria also have abnormal metabolism of nicotinamide adenine dinucleotide hydrogen, likely induced by the dysfunction of the electron transport chain. We also found that when the cells were re-warmed to physiological temperature after a period of hypothermia, the FA-accumulated mitochondria changed their structural features. To the best of our knowledge, this is the first time that FA accumulation in mitochondria was observed in live cells. Our research also demonstrates that multimodal label-free chemical imaging is an attractive tool to discover abnormal functions of mitochondria at the single-organelle level and can be used to quantify the dynamic changes of these organelles under perturbative conditions.<\/jats:p>","DOI":"10.1038\/s41598-021-85795-1","type":"journal-article","created":{"date-parts":[[2021,3,23]],"date-time":"2021-03-23T07:04:44Z","timestamp":1616483084000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Tracking the formation and degradation of fatty-acid-accumulated mitochondria using label-free chemical imaging"],"prefix":"10.1038","volume":"11","author":[{"given":"Chi","family":"Zhang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stephen A.","family":"Boppart","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2021,3,23]]},"reference":[{"key":"85795_CR1","doi-asserted-by":"publisher","first-page":"2","DOI":"10.1002\/jrs.2425","volume":"41","author":"L-D Chiu","year":"2010","unstructured":"Chiu, L.-D., Ando, M. & Hamaguchi, H.-O. Study of the \u2018Raman spectroscopic signature of life\u2019in mitochondria isolated from budding yeast. J. Raman Spectrosc. 41, 2\u20133 (2010).","journal-title":"J. Raman Spectrosc."},{"key":"85795_CR2","doi-asserted-by":"publisher","first-page":"28","DOI":"10.1073\/pnas.1107524108","volume":"109","author":"M Okada","year":"2012","unstructured":"Okada, M. et al. Label-free Raman observation of cytochrome c dynamics during apoptosis. Proc. Natl. Acad. Sci. U. S. A. 109, 28\u201332 (2012).","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"85795_CR3","doi-asserted-by":"publisher","first-page":"e41990","DOI":"10.1371\/journal.pone.0041990","volume":"7","author":"NA Brazhe","year":"2012","unstructured":"Brazhe, N. A., Treiman, M., Brazhe, A. R., Maksimov, G. V. & Sosnovtseva, O. V. Mapping of redox state of mitochondrial cytochromes in live cardiomyocytes using Raman microspectroscopy. PLoS ONE 7, e41990 (2012).","journal-title":"PLoS ONE"},{"key":"85795_CR4","doi-asserted-by":"publisher","first-page":"827","DOI":"10.1021\/jp035693v","volume":"108","author":"J-X Cheng","year":"2004","unstructured":"Cheng, J.-X. & Xie, X. S. Coherent anti-Stokes Raman scattering microscopy: Instrumentation, theory, and applications. J. Phys. Chem. B 108, 827\u2013840 (2004).","journal-title":"J. Phys. Chem. B"},{"key":"85795_CR5","doi-asserted-by":"publisher","first-page":"883","DOI":"10.1146\/annurev.anchem.1.031207.112754","volume":"1","author":"CL Evans","year":"2008","unstructured":"Evans, C. L. & Xie, X. S. Coherent anti-Stokes Raman scattering microscopy: Chemical imaging for biology and medicine. Annu. Rev. Anal. Chem. 1, 883\u2013909 (2008).","journal-title":"Annu. Rev. Anal. Chem."},{"key":"85795_CR6","doi-asserted-by":"publisher","first-page":"1857","DOI":"10.1126\/science.1165758","volume":"322","author":"CW Freudiger","year":"2008","unstructured":"Freudiger, C. W. et al. Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy. Science 322, 1857\u20131861 (2008).","journal-title":"Science"},{"key":"85795_CR7","doi-asserted-by":"publisher","first-page":"2282","DOI":"10.1021\/ar400331q","volume":"47","author":"D Zhang","year":"2014","unstructured":"Zhang, D., Wang, P., Slipchenko, M. N. & Cheng, J.-X. Fast vibrational imaging of single cells and tissues by stimulated Raman scattering microscopy. Acc. Chem. Res. 47, 2282\u20132290 (2014).","journal-title":"Acc. Chem. Res."},{"key":"85795_CR8","doi-asserted-by":"publisher","first-page":"3623","DOI":"10.1021\/ja210081h","volume":"134","author":"D Fu","year":"2012","unstructured":"Fu, D. et al. Quantitative chemical imaging with multiplex stimulated Raman scattering microscopy. J. Am. Chem. Soc. 134, 3623\u20133626 (2012).","journal-title":"J. Am. Chem. Soc."},{"key":"85795_CR9","doi-asserted-by":"publisher","first-page":"241","DOI":"10.1364\/OL.31.000241","volume":"31","author":"EO Potma","year":"2006","unstructured":"Potma, E. O., Evans, C. L. & Xie, X. S. Heterodyne coherent anti-Stokes Raman scattering (CARS) imaging. Opt. Lett. 31, 241\u2013243 (2006).","journal-title":"Opt. Lett."},{"key":"85795_CR10","doi-asserted-by":"publisher","first-page":"3812","DOI":"10.1021\/ac100222c","volume":"82","author":"PD Chowdary","year":"2010","unstructured":"Chowdary, P. D. et al. High speed nonlinear interferometric vibrational analysis of lipids by spectral decomposition. Anal. Chem. 82, 3812\u20133818 (2010).","journal-title":"Anal. Chem."},{"key":"85795_CR11","doi-asserted-by":"publisher","first-page":"123905","DOI":"10.1103\/PhysRevLett.92.123905","volume":"92","author":"DL Marks","year":"2004","unstructured":"Marks, D. L. & Boppart, S. A. Nonlinear interferometric vibrational imaging. Phys. Rev. Lett. 92, 123905 (2004).","journal-title":"Phys. Rev. Lett."},{"key":"85795_CR12","doi-asserted-by":"publisher","first-page":"1543","DOI":"10.1364\/OL.31.001543","volume":"31","author":"GW Jones","year":"2006","unstructured":"Jones, G. W., Marks, D. L., Vinegoni, C. & Boppart, S. A. High-spectral-resolution coherent anti-Stokes Raman scattering with interferometrically detected broadband chirped pulses. Opt. Lett. 31, 1543\u20131545 (2006).","journal-title":"Opt. Lett."},{"key":"85795_CR13","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41467-018-05401-3","volume":"9","author":"L Shi","year":"2018","unstructured":"Shi, L. et al. Optical imaging of metabolic dynamics in animals. Nat. Commun. 9, 1\u201317 (2018).","journal-title":"Nat. Commun."},{"key":"85795_CR14","doi-asserted-by":"publisher","first-page":"2202","DOI":"10.1194\/jlr.D300022-JLR200","volume":"44","author":"X Nan","year":"2003","unstructured":"Nan, X., Cheng, J.-X. & Xie, X. S. Vibrational imaging of lipid droplets in live fibroblast cells with coherent anti-Stokes Raman scattering microscopy. J. Lipid Res. 44, 2202\u20132208 (2003).","journal-title":"J. Lipid Res."},{"key":"85795_CR15","doi-asserted-by":"publisher","first-page":"4502","DOI":"10.1021\/acs.analchem.6b04699","volume":"89","author":"C Zhang","year":"2017","unstructured":"Zhang, C., Li, J., Lan, L. & Cheng, J.-X. Quantification of lipid metabolism in living cells through the dynamics of lipid droplets measured by stimulated Raman scattering imaging. Anal. Chem. 89, 4502\u20134507 (2017).","journal-title":"Anal. Chem."},{"key":"85795_CR16","doi-asserted-by":"publisher","first-page":"1666","DOI":"10.1016\/j.bpj.2012.01.057","volume":"102","author":"W Dou","year":"2012","unstructured":"Dou, W., Zhang, D., Jung, Y., Cheng, J.-X. & Umulis, D. M. Label-free imaging of lipid-droplet intracellular motion in early Drosophila embryos using femtosecond-stimulated Raman loss microscopy. Biophys. J. 102, 1666\u20131675 (2012).","journal-title":"Biophys. J."},{"key":"85795_CR17","doi-asserted-by":"publisher","first-page":"5787","DOI":"10.1063\/1.1829162","volume":"85","author":"DL Marks","year":"2004","unstructured":"Marks, D. L., Vinegoni, C., Bredfeldt, J. S. & Boppart, S. A. Interferometric differentiation between resonant coherent anti-Stokes Raman scattering and nonresonant four-wave-mixing processes. Appl. Phys. Lett. 85, 5787\u20135789 (2004).","journal-title":"Appl. Phys. Lett."},{"key":"85795_CR18","doi-asserted-by":"publisher","first-page":"14658","DOI":"10.1073\/pnas.0703594104","volume":"104","author":"T Hellerer","year":"2007","unstructured":"Hellerer, T. et al. Monitoring of lipid storage in Caenorhabditis elegans using coherent anti-Stokes Raman scattering (CARS) microscopy. Proc. Natl. Acad. Sci. U.S.A. 104, 14658\u201314663 (2007).","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"85795_CR19","doi-asserted-by":"publisher","first-page":"3677","DOI":"10.1021\/acs.analchem.5b04468","volume":"88","author":"C Di Napoli","year":"2016","unstructured":"Di Napoli, C. et al. Quantitative spatiotemporal chemical profiling of individual lipid droplets by hyperspectral CARS microscopy in living human adipose-derived stem cells. Anal. Chem. 88, 3677\u20133685 (2016).","journal-title":"Anal. Chem."},{"key":"85795_CR20","doi-asserted-by":"publisher","first-page":"11981","DOI":"10.1002\/ange.201406029","volume":"126","author":"P Wang","year":"2014","unstructured":"Wang, P. et al. Imaging lipid metabolism in live Caenorhabditis elegans using fingerprint vibrations. Angew. Chem. Int. Ed 126, 11981\u201311986 (2014).","journal-title":"Angew. Chem. Int. Ed"},{"key":"85795_CR21","doi-asserted-by":"publisher","first-page":"1120","DOI":"10.1016\/j.bbalip.2014.02.003","volume":"1841","author":"Y Yu","year":"2014","unstructured":"Yu, Y., Ramachandran, P. V. & Wang, M. C. Shedding new light on lipid functions with CARS and SRS microscopy. Biochim. Biophys. Acta 1841, 1120\u20131129 (2014).","journal-title":"Biochim. Biophys. Acta"},{"key":"85795_CR22","doi-asserted-by":"publisher","first-page":"393","DOI":"10.1016\/j.cmet.2014.01.019","volume":"19","author":"S Yue","year":"2014","unstructured":"Yue, S. et al. Cholesteryl ester accumulation induced by PTEN loss and PI3K\/AKT activation underlies human prostate cancer aggressiveness. Cell Metab. 19, 393\u2013406 (2014).","journal-title":"Cell Metab."},{"key":"85795_CR23","doi-asserted-by":"publisher","first-page":"303","DOI":"10.1016\/j.stem.2016.11.004","volume":"20","author":"J Li","year":"2017","unstructured":"Li, J. et al. Lipid desaturation is a metabolic marker and therapeutic target of ovarian cancer stem cells. Cell Stem Cell 20, 303\u2013314 (2017).","journal-title":"Cell Stem Cell"},{"key":"85795_CR24","doi-asserted-by":"publisher","first-page":"2817","DOI":"10.1007\/s00216-011-4953-z","volume":"400","author":"WA Benalcazar","year":"2011","unstructured":"Benalcazar, W. A. & Boppart, S. A. Nonlinear interferometric vibrational imaging for fast label-free visualization of molecular domains in skin. Anal. Bioanal. Chem. 400, 2817\u20132825 (2011).","journal-title":"Anal. Bioanal. Chem."},{"key":"85795_CR25","doi-asserted-by":"publisher","first-page":"201103","DOI":"10.1063\/1.3028346","volume":"93","author":"I Rocha-Mendoza","year":"2008","unstructured":"Rocha-Mendoza, I., Langbein, W. & Borri, P. Coherent anti-Stokes Raman microspectroscopy using spectral focusing with glass dispersion. Appl. Phys. Lett. 93, 201103 (2008).","journal-title":"Appl. Phys. Lett."},{"key":"85795_CR26","doi-asserted-by":"publisher","first-page":"800","DOI":"10.1002\/jrs.2264","volume":"40","author":"W Langbein","year":"2009","unstructured":"Langbein, W., Rocha-Mendoza, I. & Borri, P. Coherent anti-Stokes Raman micro-spectroscopy using spectral focusing: Theory and experiment. J. Raman Spectrosc. 40, 800\u2013808 (2009).","journal-title":"J. Raman Spectrosc."},{"key":"85795_CR27","doi-asserted-by":"publisher","first-page":"4634","DOI":"10.1021\/jp308938t","volume":"117","author":"D Fu","year":"2013","unstructured":"Fu, D., Holtom, G., Freudiger, C., Zhang, X. & Xie, X. S. Hyperspectral imaging with stimulated Raman scattering by chirped femtosecond lasers. J. Phys. Chem. B 117, 4634\u20134640 (2013).","journal-title":"J. Phys. Chem. B"},{"key":"85795_CR28","doi-asserted-by":"publisher","first-page":"849","DOI":"10.1016\/j.media.2012.01.002","volume":"16","author":"P Coup\u00e9","year":"2012","unstructured":"Coup\u00e9, P., Munz, M., Manj\u00f3n, J. V., Ruthazer, E. S. & Collins, D. L. A CANDLE for a deeper in vivo insight. Med. Image Anal. 16, 849\u2013864 (2012).","journal-title":"Med. Image Anal."},{"key":"85795_CR29","doi-asserted-by":"publisher","first-page":"534","DOI":"10.1038\/nphoton.2016.94","volume":"10","author":"H Tu","year":"2016","unstructured":"Tu, H. et al. Stain-free histopathology by programmable supercontinuum pulses. Nat. Photonics 10, 534\u2013540 (2016).","journal-title":"Nat. Photonics"},{"key":"85795_CR30","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41467-018-04470-8","volume":"9","author":"S You","year":"2018","unstructured":"You, S. et al. Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy. Nat. Commun. 9, 1\u20139 (2018).","journal-title":"Nat. Commun."},{"key":"85795_CR31","doi-asserted-by":"publisher","first-page":"15943","DOI":"10.1021\/acs.analchem.0c03366","volume":"92","author":"C Zhang","year":"2020","unstructured":"Zhang, C. & Boppart, S. A. Dynamic signatures of lipid droplets as new markers to quantify cellular metabolic changes. Anal. Chem. 92, 15943\u201315952 (2020).","journal-title":"Anal. Chem."},{"key":"85795_CR32","doi-asserted-by":"publisher","first-page":"1062","DOI":"10.1126\/science.1219855","volume":"337","author":"RJ Youle","year":"2012","unstructured":"Youle, R. J. & Van Der Bliek, A. M. Mitochondrial fission, fusion, and stress. Science 337, 1062\u20131065 (2012).","journal-title":"Science"},{"key":"85795_CR33","doi-asserted-by":"publisher","first-page":"755","DOI":"10.1038\/s41556-018-0133-0","volume":"20","author":"V Eisner","year":"2018","unstructured":"Eisner, V., Picard, M. & Hajn\u00f3czky, G. Mitochondrial dynamics in adaptive and maladaptive cellular stress responses. Nat. Cell Biol. 20, 755\u2013765 (2018).","journal-title":"Nat. Cell Biol."},{"key":"85795_CR34","doi-asserted-by":"publisher","first-page":"1208","DOI":"10.1161\/CIRCRESAHA.112.265819","volume":"111","author":"DA Kubli","year":"2012","unstructured":"Kubli, D. A. & Gustafsson, \u00c5. B. Mitochondria and mitophagy: the yin and yang of cell death control. Circ. Res. 111, 1208\u20131221 (2012).","journal-title":"Circ. Res."},{"key":"85795_CR35","doi-asserted-by":"publisher","first-page":"9","DOI":"10.1038\/nrm3028","volume":"12","author":"RJ Youle","year":"2011","unstructured":"Youle, R. J. & Narendra, D. P. Mechanisms of mitophagy. Nat. Rev. Mol. Cell Biol. 12, 9\u201314 (2011).","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"85795_CR36","doi-asserted-by":"publisher","first-page":"245","DOI":"10.1016\/j.abb.2007.03.034","volume":"462","author":"I Kim","year":"2007","unstructured":"Kim, I., Rodriguez-Enriquez, S. & Lemasters, J. J. Selective degradation of mitochondria by mitophagy. Arch. Biochem. Biophys 462, 245\u2013253 (2007).","journal-title":"Arch. Biochem. Biophys"},{"key":"85795_CR37","doi-asserted-by":"publisher","first-page":"R170","DOI":"10.1016\/j.cub.2018.01.004","volume":"28","author":"S Pickles","year":"2018","unstructured":"Pickles, S., Vigi\u00e9, P. & Youle, R. J. Mitophagy and quality control mechanisms in mitochondrial maintenance. Curr. Biol. 28, R170\u2013R185 (2018).","journal-title":"Curr. Biol."},{"key":"85795_CR38","doi-asserted-by":"publisher","first-page":"7541","DOI":"10.1016\/S0021-9258(19)81272-3","volume":"249","author":"MP Lee","year":"1974","unstructured":"Lee, M. P. & Gear, A. R. The effect of temperature on mitochondrial membrane-linked reactions. J. Biol. Chem. 249, 7541\u20137549 (1974).","journal-title":"J. Biol. Chem."},{"key":"85795_CR39","doi-asserted-by":"publisher","first-page":"e0208453","DOI":"10.1371\/journal.pone.0208453","volume":"13","author":"ME Pamenter","year":"2018","unstructured":"Pamenter, M. E., Lau, G. Y. & Richards, J. G. Effects of cold on murine brain mitochondrial function. PLoS ONE 13, e0208453 (2018).","journal-title":"PLoS ONE"},{"key":"85795_CR40","doi-asserted-by":"publisher","first-page":"177","DOI":"10.1172\/JCI114409","volume":"85","author":"N Watmough","year":"1990","unstructured":"Watmough, N. et al. Impaired mitochondrial beta-oxidation in a patient with an abnormality of the respiratory chain. Studies in skeletal muscle mitochondria. J. Clin. Invest. 85, 177\u2013184 (1990).","journal-title":"J. Clin. Invest."},{"key":"85795_CR41","doi-asserted-by":"publisher","first-page":"12380","DOI":"10.1074\/jbc.RA119.008680","volume":"294","author":"Y Wang","year":"2019","unstructured":"Wang, Y. et al. Mitochondrial fatty acid oxidation and the electron transport chain comprise a multifunctional mitochondrial protein complex. J. Biol. Chem. 294, 12380\u201312391 (2019).","journal-title":"J. Biol. Chem."},{"key":"85795_CR42","doi-asserted-by":"publisher","first-page":"e0189456","DOI":"10.1371\/journal.pone.0189456","volume":"12","author":"JA Zoladz","year":"2017","unstructured":"Zoladz, J. A. et al. Effect of temperature on fatty acid metabolism in skeletal muscle mitochondria of untrained and endurance-trained rats. PLoS ONE 12, e0189456 (2017).","journal-title":"PLoS ONE"},{"key":"85795_CR43","doi-asserted-by":"publisher","first-page":"7681","DOI":"10.1021\/jp902231y","volume":"113","author":"MN Slipchenko","year":"2009","unstructured":"Slipchenko, M. N., Le, T. T., Chen, H. & Cheng, J.-X. High-speed vibrational imaging and spectral analysis of lipid bodies by compound Raman microscopy. J. Phys. Chem. B 113, 7681\u20137686 (2009).","journal-title":"J. Phys. Chem. B"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-85795-1.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-85795-1","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-85795-1.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,1,29]],"date-time":"2023-01-29T22:06:05Z","timestamp":1675029965000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-85795-1"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,3,23]]},"references-count":43,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["85795"],"URL":"https:\/\/doi.org\/10.1038\/s41598-021-85795-1","relation":{"has-preprint":[{"id-type":"doi","id":"10.21203\/rs.3.rs-119989\/v1","asserted-by":"object"}]},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,3,23]]},"assertion":[{"value":"2 December 2020","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"4 March 2021","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"23 March 2021","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"Stephen Boppart is a co-founder and consultant of LiveBx, Champaign, IL, which is licensing intellectual property from the University of Illinois at Urbana-Champaign to develop novel optical sources and label-free multimodal multiphoton imaging platforms for biological and medical applications. Chi Zhang declares no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"6671"}}