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Bioorthogonal chemical imaging of metabolic activities in live mammalian hippocampal tissues with stimulated Raman scattering

Hu, Fanghao and Lamprecht, Michael R. and Wei, Lu and Morrison, Barclay and Min, Wei (2016) Bioorthogonal chemical imaging of metabolic activities in live mammalian hippocampal tissues with stimulated Raman scattering. Scientific Reports, 6 . Art. No. 39660 . ISSN 2045-2322. PMCID PMC5175176. doi:10.1038/srep39660.

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Brain is an immensely complex system displaying dynamic and heterogeneous metabolic activities. Visualizing cellular metabolism of nucleic acids, proteins, and lipids in brain with chemical specificity has been a long-standing challenge. Recent development in metabolic labeling of small biomolecules allows the study of these metabolisms at the global level. However, these techniques generally require nonphysiological sample preparation for either destructive mass spectrometry imaging or secondary labeling with relatively bulky fluorescent labels. In this study, we have demonstrated bioorthogonal chemical imaging of DNA, RNA, protein and lipid metabolism in live rat brain hippocampal tissues by coupling stimulated Raman scattering microscopy with integrated deuterium and alkyne labeling. Heterogeneous metabolic incorporations for different molecular species and neurogenesis with newly-incorporated DNA were observed in the dentate gyrus of hippocampus at the single cell level. We further applied this platform to study metabolic responses to traumatic brain injury in hippocampal slice cultures, and observed marked upregulation of protein and lipid metabolism particularly in the hilus region of the hippocampus within days of mechanical injury. Thus, our method paves the way for the study of complex metabolic profiles in live brain tissue under both physiological and pathological conditions with single-cell resolution and minimal perturbation.

Item Type:Article
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URLURL TypeDescription CentralArticle
Hu, Fanghao0000-0002-8659-4027
Wei, Lu0000-0001-9170-2283
Additional Information:© 2016 The Author(s). This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit received: 30 September 2016, accepted: 25 November 2016. Published: 21 December 2016. We thank Y. Shen and L. Shi for helpful discussions. W. M. acknowledges support from NIH Director’s New Innovator Award (Grant 1DP2EB016573), NIH R01 (Grant EB020892), the US Army Research Office (Grant W911NF-12-1-0594), the Alfred P. Sloan Foundation, and the Camille and Henry Dreyfus Foundation. B.M. acknowledges support from the Army Research Laboratory (Grant W911NF-10-1-0526). M.R.L acknowledges support from a National Science Foundation Graduate Research Fellowship. Author Contributions: F.H., B.M. and W.M. conceived the concept and designed the experiments. F.H., M.R.L. and L.W. performed the experiments. F.H. analyzed the data and wrote the manusript. B.M. and W.M. oversaw the study and edited the manuscript. All authors reviewed the manusript. The authors declare no competing financial interests.
Funding AgencyGrant Number
NIHR01 EB020892
Army Research Office (ARO)W911NF-12-1-0594
Alfred P. Sloan FoundationUNSPECIFIED
Camille and Henry Dreyfus FoundationUNSPECIFIED
Army Research LaboratoryW911NF-10-1-0526
NSF Graduate Research FellowshipUNSPECIFIED
PubMed Central ID:PMC5175176
Record Number:CaltechAUTHORS:20180608-112524422
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Official Citation:Hu, F. et al. Bioorthogonal chemical imaging of metabolic activities in live mammalian hippocampal tissues with stimulated Raman scattering. Sci. Rep. 6, 39660; doi: 10.1038/srep39660 (2016).
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:86919
Deposited By: George Porter
Deposited On:08 Jun 2018 23:28
Last Modified:15 Nov 2021 20:43

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