Live-cell stimulated Raman scattering imaging of alkyne-tagged biomolecules.

Alkynes can be metabolically incorporated into biomolecules including nucleic acids, proteins, lipids, and glycans. In addition to the clickable chemical reactivity, alkynes possess a unique Raman scattering within the Raman-silent region of a cell. Coupling this spectroscopic signature with Raman microscopy yields a new imaging modality beyond fluorescence and label-free microscopies. The bioorthogonal Raman imaging of various biomolecules tagged with an alkyne by a state-of-the-art Raman imaging technique, stimulated Raman scattering (SRS) microscopy, is reported. This imaging method affords non-invasiveness, high sensitivity, and molecular specificity and therefore should find broad applications in live-cell imaging.

[1]  Christian Matthäus,et al.  Noninvasive imaging of intracellular lipid metabolism in macrophages by Raman microscopy in combination with stable isotopic labeling. , 2012, Analytical chemistry.

[2]  G. Charron,et al.  Palmitoylome profiling reveals S-palmitoylation-dependent antiviral activity of IFITM3. , 2010, Nature chemical biology.

[3]  Eric C Le Ru,et al.  Single-molecule surface-enhanced Raman spectroscopy. , 2012, Annual review of physical chemistry.

[4]  Srinjan Basu,et al.  Label-free live-cell imaging of nucleic acids using stimulated Raman scattering microscopy. , 2012, Chemphyschem : a European journal of chemical physics and physical chemistry.

[5]  Anne B. Neef,et al.  Dynamic metabolic labeling of DNA in vivo with arabinosyl nucleosides , 2011, Proceedings of the National Academy of Sciences.

[6]  Satoshi Kawata,et al.  Imaging of EdU, an alkyne-tagged cell proliferation probe, by Raman microscopy. , 2011, Journal of the American Chemical Society.

[7]  Wei Min,et al.  Vibrational imaging of newly synthesized proteins in live cells by stimulated Raman scattering microscopy , 2013, Proceedings of the National Academy of Sciences.

[8]  X. Xie,et al.  Video-Rate Molecular Imaging in Vivo with Stimulated Raman Scattering , 2010, Science.

[9]  Timothy J. Mitchison,et al.  A chemical method for fast and sensitive detection of DNA synthesis in vivo , 2008, Proceedings of the National Academy of Sciences.

[10]  Bo Zhang,et al.  Label-free chemical imaging in vivo: three-dimensional non-invasive microscopic observation of amphioxus notochord through stimulated Raman scattering (SRS) , 2012 .

[11]  Mortazavi,et al.  Supporting Online Material Materials and Methods Figs. S1 to S13 Tables S1 to S3 References Label-free Biomedical Imaging with High Sensitivity by Stimulated Raman Scattering Microscopy , 2022 .

[12]  G. Charron,et al.  Bioorthogonal chemical reporters for analyzing protein lipidation and lipid trafficking. , 2011, Accounts of chemical research.

[13]  W. Zinth,et al.  Femtosecond stimulated Raman microscopy , 2007 .

[14]  R. Dasari,et al.  Surface-enhanced Raman scattering and biophysics , 2001 .

[15]  Dan Fu,et al.  Quantitative chemical imaging with multiplex stimulated Raman scattering microscopy. , 2012, Journal of the American Chemical Society.

[16]  John Paul Pezacki,et al.  Chemical contrast for imaging living systems: molecular vibrations drive CARS microscopy , 2011, Nature chemical biology.

[17]  X. Xie,et al.  Living Cells as Test Tubes , 2006, Science.

[18]  X. Xie,et al.  Rapid, Label-Free Detection of Brain Tumors with Stimulated Raman Scattering Microscopy , 2013, Science Translational Medicine.

[19]  Mingzi M. Zhang,et al.  Robust fluorescent detection of protein fatty-acylation with chemical reporters. , 2009, Journal of the American Chemical Society.

[20]  A. Salic,et al.  Metabolic labeling and direct imaging of choline phospholipids in vivo , 2009, Proceedings of the National Academy of Sciences.

[21]  Wei Min,et al.  Coherent nonlinear optical imaging: beyond fluorescence microscopy. , 2011, Annual review of physical chemistry.

[22]  Wei Min,et al.  RNAi Screening for Fat Regulatory Genes with SRS Microscopy , 2010, Nature Methods.

[23]  Wei Min,et al.  Highly specific label-free molecular imaging with spectrally tailored excitation stimulated Raman scattering (STE-SRS) microscopy. , 2011, Nature photonics.

[24]  Zhong-Qun Tian,et al.  A bioorthogonal Raman reporter strategy for SERS detection of glycans on live cells. , 2013, Angewandte Chemie.

[25]  Markus Grammel,et al.  Chemical reporters for biological discovery. , 2013, Nature chemical biology.

[26]  M. Resh,et al.  Trafficking and signaling by fatty-acylated and prenylated proteins , 2006, Nature chemical biology.

[27]  Ping Wang,et al.  Label-free quantitative imaging of cholesterol in intact tissues by hyperspectral stimulated Raman scattering microscopy. , 2013, Angewandte Chemie.

[28]  T. Huser,et al.  Lipid-cell interactions in human monocytes investigated by doubly-resonant coherent anti-Stokes Raman scattering microscopy. , 2011, Journal of biomedical optics.

[29]  Satoshi Kawata,et al.  Alkyne-tag Raman imaging for visualization of mobile small molecules in live cells. , 2012, Journal of the American Chemical Society.

[30]  Jürgen Popp,et al.  Quantitative detection of C-deuterated drugs by CARS microscopy and Raman microspectroscopy. , 2011, The Analyst.

[31]  R. Deschenes,et al.  Palmitoylation: policing protein stability and traffic , 2007, Nature Reviews Molecular Cell Biology.

[32]  E. Schuman,et al.  Fluorescence visualization of newly synthesized proteins in mammalian cells. , 2006, Angewandte Chemie.

[33]  F. Gasparri,et al.  A novel method based on click chemistry, which overcomes limitations of cell cycle analysis by classical determination of BrdU incorporation, allowing multiplex antibody staining , 2008, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[34]  Scott T. Clarke,et al.  Cell type specific applicability of 5‐ethynyl‐2′‐deoxyuridine (EdU) for dynamic proliferation assessment in flow cytometry , 2009, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[35]  Benjamin Bird,et al.  Label-free imaging of human cells: algorithms for image reconstruction of Raman hyperspectral datasets. , 2010, The Analyst.

[36]  N. Shah,et al.  Surface-enhanced Raman spectroscopy. , 2008, Annual review of analytical chemistry.