The goal of the project is to develop a general optical imaging technology for studying vital small bio-molecules (e.g., amino acids, nucleic acids, fatty acids, glucose, neurotransmitters and drugs) inside live cells and tissues, an important but otherwise intractable goal. To do so, we propose to couple the emerging stimulated Raman scattering (SRS) microscopy, which is capable of producing concentration maps of chemical bonds in biological samples, with three distinct classes of small vibrational tags with characteristic Raman transitions, including alkyne moieties (i.e., C=C triple bond), deuterium isotope and 13C isotope. While alkyne tags are generally applicable to any small biomolecules, deuterium isotope and 13C isotope will be particularly useful for labeling amino acids. When spectrally targeting these vibrational tags labeled to small bio-molecules, SRS microscopy is ideally suited for probing functional metabolism of living systems at microscopic level, as proved in our recent publications. We have laid out systematic plans as to how to crystallize this concept into a mature and general technical platform. Accompanied by the technical development, several biomedical applications are being proposed (some with exciting preliminary data) including imaging neurogenesis in living brain tissues by monitoring the metabolic incorporation of alkyne-tagged deoxyribonucleoside into newly born neurons, multicolor chemical imaging by developing a color palette of metabolite library, monitoring protein synthesis in nervous systems with deuterium-labeled amino acids, and probing intracellular protein degradation during Huntingtin aggregation with 13C-labeled phenylalanine. If successfully implemented, we will establish a new imaging platform of bioorthogonal nonlinear Raman microscopy that could allow us to interrogate a broad spectrum of small bio-molecules with superb sensitivity, specificity, biocompatibility and multiplex ability. The resulting bioorthogonal nonlinear Raman microscopy might do for small bio-molecules what fluorescence imaging of organic dyes and fluorescent proteins has done for larger molecular species, bringing small bio-molecules under the illumination of modern light microscopy.

Public Health Relevance

The unprecedented ability to visualize protein synthesis and protein degradation inside health and disease brain tissues should be able to offer important insights regarding the misregulation of protein metabolism in neurodegenerative diseases such as Alzheimer's disease, Parkinson disease and Huntington disease. In addition, tumors are known to exhibit drastically aberrant metabolic activities compared to healthy tissues. Thus, our unique technique to image small metabolites could also be adopted to probe the microenvironment of abnormal tumor metabolism.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
1R01EB020892-01
Application #
8940638
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Conroy, Richard
Project Start
2015-06-01
Project End
2019-05-31
Budget Start
2015-06-01
Budget End
2016-05-31
Support Year
1
Fiscal Year
2015
Total Cost
Indirect Cost
Name
Columbia University (N.Y.)
Department
Chemistry
Type
Graduate Schools
DUNS #
049179401
City
New York
State
NY
Country
United States
Zip Code
10027
Zeng, Chen; Hu, Fanghao; Long, Rong et al. (2018) A ratiometric Raman probe for live-cell imaging of hydrogen sulfide in mitochondria by stimulated Raman scattering. Analyst 143:4844-4848
Shi, Lingyan; Zheng, Chaogu; Shen, Yihui et al. (2018) Optical imaging of metabolic dynamics in animals. Nat Commun 9:2995
Wei, Lu; Min, Wei (2018) Electronic Preresonance Stimulated Raman Scattering Microscopy. J Phys Chem Lett 9:4294-4301
Hu, Fanghao; Zeng, Chen; Long, Rong et al. (2018) Supermultiplexed optical imaging and barcoding with engineered polyynes. Nat Methods 15:194-200
Long, Rong; Zhang, Luyuan; Shi, Lingyan et al. (2018) Two-color vibrational imaging of glucose metabolism using stimulated Raman scattering. Chem Commun (Camb) 54:152-155
Gaschler, Michael M; Hu, Fanghao; Feng, Huizhong et al. (2018) Determination of the Subcellular Localization and Mechanism of Action of Ferrostatins in Suppressing Ferroptosis. ACS Chem Biol 13:1013-1020
Cheng, Qian; Wei, Lu; Liu, Zhe et al. (2018) Operando and three-dimensional visualization of anion depletion and lithium growth by stimulated Raman scattering microscopy. Nat Commun 9:2942
Shen, Yihui; Zhao, Zhilun; Zhang, Luyuan et al. (2017) Metabolic activity induces membrane phase separation in endoplasmic reticulum. Proc Natl Acad Sci U S A 114:13394-13399
Zhang, Luyuan; Min, Wei (2017) Bioorthogonal chemical imaging of metabolic changes during epithelial-mesenchymal transition of cancer cells by stimulated Raman scattering microscopy. J Biomed Opt 22:1-7
Wei, Lu; Chen, Zhixing; Shi, Lixue et al. (2017) Super-multiplex vibrational imaging. Nature 544:465-470

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