Cells and tissues often display pronounced spatial and dynamical metabolic heterogeneity. Common glucose-imaging techniques report glucose uptake or catabolism activity, yet do not trace the functional utilization of glucose-derived anabolic products. Here we report a microscopy technique for the optical imaging, via the spectral tracing of deuterium (STRIDE), of diverse macromolecules derived from glucose. Based on stimulated Raman-scattering imaging, STRIDE visualizes the metabolic dynamics of newly synthesized macromolecules, such as DNA, protein, lipids and glycogen, via the enrichment and distinct spectra of carbon–deuterium bonds transferred from the deuterated glucose precursor. STRIDE can also use spectral differences derived from different glucose isotopologues to visualize temporally separated glucose populations using a pulse–chase protocol. We also show that STRIDE can be used to image glucose metabolism in many mouse tissues, including tumours, brain, intestine and liver, at a detection limit of 10 mM of carbon–deuterium bonds. STRIDE provides a high-resolution and chemically informative assessment of glucose anabolic utilization.
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The authors declare that all data supporting the results of this study are available within the paper and its Supplementary Information. All raw and processed images generated in this work are available from the corresponding author on reasonable request.
Saltiel, A. R. & Kahn, C. R. Insulin signalling and the regulation of glucose and lipid metabolism. Nature 414, 799–806 (2001).
Hensley, C. T. et al. Metabolic heterogeneity in human lung tumors. Cell 164, 681–694 (2016).
Vander Heiden, M. G. & DeBerardinis, R. J. Understanding the intersections between metabolism and cancer biology. Cell 168, 657–669 (2017).
Pipeleers, D. G. Heterogeneity in pancreatic β-cell population. Diabetes 41, 777–781 (1992).
Halpern, K. B. et al. Single-cell spatial reconstruction reveals global division of labour in the mammalian liver. Nature 542, 1–5 (2017).
Gambhir, S. S. Molecular imaging of cancer with positron emission tomography. Nat. Rev. Cancer 2, 683–693 (2002).
Walker-Samuel, S. et al. In vivo imaging of glucose uptake and metabolism in tumors. Nat. Med. 19, 1067–1072 (2013).
Rodrigues, T. B. et al. Magnetic resonance imaging of tumor glycolysis using hyperpolarized 13C-labeled glucose. Nat. Med. 20, 93–97 (2013).
Sugiura, Y. et al. Visualization of in vivo metabolic flows reveals accelerated utilization of glucose and lactate in penumbra of ischemic heart. Sci. Rep. 6, 32361 (2016).
Guillermier, C., Poczatek, J. C., Taylor, W. R. & Steinhauser, M. L. Quantitative imaging of deuterated metabolic tracers in biological tissues with nanoscale secondary ion mass spectrometry. Int. J. Mass Spectrom. 422, 42–50 (2017).
Zou, C., Wang, Y. & Shen, Z. 2-NBDG as a fluorescent indicator for direct glucose uptake measurement. J. Biochem. Biophys. Methods 64, 207–215 (2005).
Hu, F. et al. Vibrational imaging of glucose uptake activity in live cells and tissues by stimulated Raman scattering. Angew. Chem. Int. Ed. 54, 9821–9825 (2015).
Nelson, D. & Cox, M. Lehninger Principles of Biochemistry 4th edn (W. H. Freeman, 2005).
Chen, Y. J. et al. Differential incorporation of glucose into biomass during Warburg metabolism. Biochemistry 53, 4755–4757 (2014).
Lunt, S. Y. & Vander Heiden, M. G. Aerobic glycolysis: meeting the metabolic requirements of cell proliferation. Annu. Rev. Cell Dev. Biol. 27, 441–464 (2011).
Otero, Y. F., Stafford, J. M. & McGuinness, O. P. Pathway-selective insulin resistance and metabolic disease: the importance of nutrient flux. J. Biol. Chem. 289, 20462–20469 (2014).
Uyeda, K. & Repa, J. J. Carbohydrate response element binding protein, ChREBP, a transcription factor coupling hepatic glucose utilization and lipid synthesis. Cell Metab. 4, 107–110 (2006).
Freudiger, C. W. et al. Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy. Science 322, 1857–1861 (2008).
Min, W., Freudiger, C. W., Lu, S. & Xie, X. S. Coherent nonlinear optical imaging: beyond fluorescence microscopy. Annu. Rev. Phys. Chem. 62, 507–530 (2011).
Chung, C.-Y. & Potma, E. O. Biomolecular imaging with coherent nonlinear vibrational microscopy. Annu. Rev. Phys. Chem. 64, 77–99 (2013).
Cheng, J.-X. & Xie, X. S. Vibrational spectroscopic imaging of living systems: an emerging platform for biology and medicine. Science 350, aaa8870 (2015).
Zhao, Z., Shen, Y., Hu, F. & Min, W. Applications of vibrational tags in biological imaging by Raman microscopy. Analyst 142, 4018–4029 (2017).
Wei, L. et al. Live-cell bioorthogonal chemical imaging: stimulated Raman scattering microscopy of vibrational probes. Acc. Chem. Res. 49, 1494–1502 (2016).
Li, M., Huang, W. E., Gibson, C. M., Fowler, P. W. & Jousset, A. Stable isotope probing and Raman spectroscopy for monitoring carbon flow in a food chain and revealing metabolic pathway. Anal. Chem. 85, 1642–1649 (2013).
Li, J. & Cheng, J.-X. Direct visualization of de novo lipogenesis in single living cells. Sci. Rep. 4, 6807 (2015).
Long, R. et al. Two-color vibrational imaging of glucose metabolism by stimulated Raman scattering. Chem Commun. 54, 152–155 (2017).
Sun, R. C. et al. Noninvasive liquid diet delivery of stable isotopes into mouse models for deep metabolic network tracing. Nat. Commun. 8, 1646 (2017).
Longhi, G., Zerbi, G., Paterlini, G., Ricard, L. & Abbate, S. Conformational dependence of CH(CD)-strechings in d-glucose and some deuterated derivatives as revealed by infrared and Raman spectroscopy. Carbohydr. Res. 161, 1–22 (1987).
Orringer, D. A. et al. Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy. Nat. Biomed. Eng. 1, 0027 (2017).
Jung, Y., Tam, J., Jalian, H. R., Anderson, R. R. & Evans, C. L. Longitudinal, 3D in vivo imaging of sebaceous glands by coherent anti-stokes Raman scattering microscopy: normal function and response to cryotherapy. J. Invest. Dermatol. 135, 39–44 (2015).
Yoshikawa, F. et al. Opalin, a transmembrane sialylglycoprotein located in the central nervous system myelin paranodal loop membrane. J. Biol. Chem. 283, 20830–20840 (2008).
Bercury, K. K. & Macklin, W. B. Dynamics and mechanisms of CNS myelination. Dev. Cell 32, 447–458 (2015).
Jurevics, H. & Morell, P. Cholesterol for synthesis of myelin is made locally, not imported into brain. J. Neurochem. 64, 895–901 (1995).
Cannon, B. & Nedergaard, J. Brown adipose tissue: function and physiological significance. Physiol. Rev. 84, 277–359 (2004).
Brehar, F. M. et al. The development of xenograft glioblastoma implants in nude mice brain. J. Med. Life 1, 275–286 (2008).
Flores, C. A., Hing, S. A., Wells, M. A. & Koldovsky, O. Rates of triolein absorption in suckling and adult rats. Am. J. Physiol. Liver Physiol. 257, G823–G829 (1989).
Lindquist, S. & Hernell, O. Lipid digestion and absorption in early life: an update. Curr. Opin. Clin. Nutr. Metab. Care 13, 314–320 (2010).
Pácha, J. Development of intestinal transport function in mammals. Physiol. Rev. 80, 1633–1667 (2000).
Alberts, B. et al. Molecular Biology of the Cell 4th edn (Garland Science, 2002).
Gaietta, G. et al. Multicolor and electron microscopic imaging of connexin trafficking. Science 296, 503–507 (2002).
Dieterich, D. C. et al. In situ visualization and dynamics of newly synthesized proteins in rat hippocampal neurons. Nat. Neurosci. 13, 897–905 (2010).
Wei, L. et al. Imaging complex protein metabolism in live organisms by stimulated Raman scattering microscopy with isotope labeling. ACS Chem. Biol. 10, 901–908 (2015).
Lewis, C. A. et al. Tracing compartmentalized nadph metabolism in the cytosol and mitochondria of mammalian cells. Mol. Cell 55, 253–263 (2014).
Liu, L. et al. Malic enzyme tracers reveal hypoxia-induced switch in adipocyte NADPH pathway usage. Nat. Chem. Biol. 12, 345–352 (2016).
Kudryavtseva, M. V., Sakuta, G. A., Stein, G. I. & Kudryavtsev, BN. The metabolic zonation of glycogen synthesis in rat liver after fasting and refeeding. Tissue Cell 24, 31–35 (1992).
Jungermann, K. & Katz, N. Functional specialization of different hepatocyte populations. Physiol. Rev. 69, 708–764 (1989).
Fu, D. et al. In vivo metabolic fingerprinting of neutral lipids with hyperspectral stimulated Raman scattering microscopy. J. Am. Chem. Soc. 136, 8820–8828 (2014).
Zhang, L. & Min, W. Bioorthogonal chemical imaging of metabolic changes during epithelial-mesenchymal transition of cancer cells by stimulated Raman scattering microscopy. J. Biomed. Opt. 22, 1–7 (2017).
Hou, J., Williams, J., Botvinick, E., Potma, E. & Tromberg, B. J. Visualization of breast cancer metabolism using multimodal non-linear optical microscopy of cellular lipids and redox state. Cancer Res. 78, 2503–2512 (2018).
Hong, W. et al. Antibiotic susceptibility determination within one cell cycle at single-bacterium level by stimulated Raman metabolic imaging. Anal. Chem. 90, 3737–3743 (2018).
Saar, B. G. et al. Video-rate molecular imaging in vivo with stimulated Raman scattering. Science 330, 1368–1370 (2010).
Saar, B. G., Johnston, R. S., Freudiger, C. W., Xie, X. S. & Seibel, E. J. Coherent Raman scanning fiber endoscopy. Opt. Lett. 36, 2396 (2011).
Shen, Y., Xu, F., Wei, L., Hu, F. & Min, W. Live-cell quantitative imaging of proteome degradation by stimulated Raman scattering. Angew. Chem. Int. Ed. 53, 5596–5599 (2014).
Wei, L. et al. Live-cell imaging of alkyne-tagged small biomolecules by stimulated Raman scattering. Nat. Methods 11, 410–412 (2014).
Lu, F.-K. et al. Label-free DNA imaging in vivo with stimulated Raman scattering microscopy. Proc. Natl Acad. Sci. USA 112, 11624–11629 (2015).
We thank F. Hu and C. Zheng for suggestions on this manuscript. W.M. acknowledges support from a National Institutes of Health Director’s New Innovator Award (1DP2EB016573), NIH R01 (grant EB020892), the Alfred P. Sloan Foundation, the Camille and Henry Dreyfus Foundation and a Pilot and Feasibility grant from the New York Obesity Nutrition Research Center.
The authors declare no competing interests.
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Zhang, L., Shi, L., Shen, Y. et al. Spectral tracing of deuterium for imaging glucose metabolism. Nat Biomed Eng 3, 402–413 (2019). https://doi.org/10.1038/s41551-019-0393-4
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