A Caltech Library Service

Live-Cell Bioorthogonal Chemical Imaging: Stimulated Raman Scattering Microscopy of Vibrational Probes

Wei, Lu and Hu, Fanghao and Chen, Zhixing and Shen, Yihui and Zhang, Luyuan and Min, Wei (2016) Live-Cell Bioorthogonal Chemical Imaging: Stimulated Raman Scattering Microscopy of Vibrational Probes. Accounts of Chemical Research, 49 (8). pp. 1494-1502. ISSN 0001-4842. PMCID PMC5704954.

[img] PDF - Accepted Version
See Usage Policy.


Use this Persistent URL to link to this item:


Innovations in light microscopy have tremendously revolutionized the way researchers study biological systems with subcellular resolution. In particular, fluorescence microscopy with the expanding choices of fluorescent probes has provided a comprehensive toolkit to tag and visualize various molecules of interest with exquisite specificity and high sensitivity. Although fluorescence microscopy is currently the method of choice for cellular imaging, it faces fundamental limitations for studying the vast number of small biomolecules. This is because common fluorescent labels, which are relatively bulky, could introduce considerable perturbation to or even completely alter the native functions of vital small biomolecules. Hence, despite their immense functional importance, these small biomolecules remain largely undetectable by fluorescence microscopy. To address this challenge, a bioorthogonal chemical imaging platform has recently been introduced. By coupling stimulated Raman scattering (SRS) microscopy, an emerging nonlinear Raman microscopy technique, with tiny and Raman-active vibrational probes (e.g., alkynes and stable isotopes), bioorthogonal chemical imaging exhibits superb sensitivity, specificity, and biocompatibility for imaging small biomolecules in live systems. In this Account, we review recent technical achievements for visualizing a broad spectrum of small biomolecules, including ribonucleosides and deoxyribonucleosides, amino acids, fatty acids, choline, glucose, cholesterol, and small-molecule drugs in live biological systems ranging from individual cells to animal tissues and model organisms. Importantly, this platform is compatible with live-cell biology, thus allowing real-time imaging of small-molecule dynamics. Moreover, we discuss further chemical and spectroscopic strategies for multicolor bioorthogonal chemical imaging, a valuable technique in the era of “omics”. As a unique tool for biological discovery, this platform has been applied to studying various metabolic processes under both physiological and pathological states, including protein synthesis activity of neuronal systems, protein aggregations in Huntington disease models, glucose uptake in tumor xenografts, and drug penetration through skin tissues. We envision that the coupling of SRS microscopy with vibrational probes would do for small biomolecules what fluorescence microscopy of fluorophores has done for larger molecular species.

Item Type:Article
Related URLs:
URLURL TypeDescription CentralArticle
Wei, Lu0000-0001-9170-2283
Hu, Fanghao0000-0002-8659-4027
Min, Wei0000-0003-2570-3557
Additional Information:© 2016 American Chemical Society. Received: May 2, 2016. Published: August 3, 2016. We appreciate helpful discussions with Meng Wang, Colin Nuckolls, Louis Brus, Ann McDermott, Ronald Breslow, Virginia Cornish, Rafael Yuste, Sunney Xie, and Steven Boxer. This work is supported by NIH Director’s New Innovator Award (Grant 1DP2EB016573), 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. Y. Shen acknowledges support from HHMI International Student Research Fellowship. The authors declare the following competing financial interest(s): Columbia University has filed a patent application based on this work.
Funding AgencyGrant Number
NIHR01 EB020892
Army Research Office (ARO)W911NF-12-1-0594
Alfred P. Sloan FoundationUNSPECIFIED
Camille and Henry Dreyfus FoundationUNSPECIFIED
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
Issue or Number:8
PubMed Central ID:PMC5704954
Record Number:CaltechAUTHORS:20180608-113519551
Persistent URL:
Official Citation:Live-Cell Bioorthogonal Chemical Imaging: Stimulated Raman Scattering Microscopy of Vibrational Probes Lu Wei, Fanghao Hu, Zhixing Chen, Yihui Shen, Luyuan Zhang, and Wei Min Accounts of Chemical Research 2016 49 (8), 1494-1502 DOI: 10.1021/acs.accounts.6b00210
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:86920
Deposited By: George Porter
Deposited On:11 Jun 2018 19:46
Last Modified:09 Mar 2020 13:19

Repository Staff Only: item control page