Article | Published:

Supermultiplexed optical imaging and barcoding with engineered polyynes

Nature Methods volume 15, pages 194200 (2018) | Download Citation

Abstract

Optical multiplexing has a large impact in photonics, the life sciences and biomedicine. However, current technology is limited by a 'multiplexing ceiling' from existing optical materials. Here we engineered a class of polyyne-based materials for optical supermultiplexing. We achieved 20 distinct Raman frequencies, as 'Carbon rainbow', through rational engineering of conjugation length, bond-selective isotope doping and end-capping substitution of polyynes. With further probe functionalization, we demonstrated ten-color organelle imaging in individual living cells with high specificity, sensitivity and photostability. Moreover, we realized optical data storage and identification by combinatorial barcoding, yielding to our knowledge the largest number of distinct spectral barcodes to date. Therefore, these polyynes hold great promise in live-cell imaging and sorting as well as in high-throughput diagnostics and screening.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    & Advances in fluorescence labeling strategies for dynamic cellular imaging. Nat. Chem. Biol. 10, 512–523 (2014).

  2. 2.

    et al. Applying systems-level spectral imaging and analysis to reveal the organelle interactome. Nature 546, 162–167 (2017).

  3. 3.

    et al. Multi-target spectrally resolved fluorescence lifetime imaging microscopy. Nat. Methods 13, 257–262 (2016).

  4. 4.

    & Fluorescent cell barcoding in flow cytometry allows high-throughput drug screening and signaling profiling. Nat. Methods 3, 361–368 (2006).

  5. 5.

    et al. MicroRNA expression profiles classify human cancers. Nature 435, 834–838 (2005).

  6. 6.

    , & Multiplexed detection of pathogen DNA with DNA-based fluorescence nanobarcodes. Nat. Biotechnol. 23, 885–889 (2005).

  7. 7.

    , , & Suspension arrays based on nanoparticle-encoded microspheres for high-throughput multiplexed detection. Chem. Soc. Rev. 44, 5552–5595 (2015).

  8. 8.

    , & Five-dimensional optical recording mediated by surface plasmons in gold nanorods. Nature 459, 410–413 (2009).

  9. 9.

    et al. Tunable lifetime multiplexing using luminescent nanocrystals. Nat. Photonics 8, 32–36 (2014).

  10. 10.

    et al. Programmable microfluidic synthesis of over one thousand uniquely identifiable spectral codes. Adv. Opt. Mater. 5, 1600548 (2017).

  11. 11.

    et al. Facile and rapid one-step mass preparation of quantum-dot barcodes. Angew. Chem. Int. Ed. Engl. 47, 5577–5581 (2008).

  12. 12.

    , , & Quantum-dot-tagged microbeads for multiplexed optical coding of biomolecules. Nat. Biotechnol. 19, 631–635 (2001).

  13. 13.

    , & Nanoparticles with Raman spectroscopic fingerprints for DNA and RNA detection. Science 297, 1536–1540 (2002).

  14. 14.

    , , & Glass-bead-based parallel detection of DNA using composite Raman labels. Small 2, 375–380 (2006).

  15. 15.

    , , & Carbon-atom wires: 1-D systems with tunable properties. Nanoscale 8, 4414–4435 (2016).

  16. 16.

    The era of carbon allotropes. Nat. Mater. 9, 868–871 (2010).

  17. 17.

    , , , & Carbyne from first principles: chain of C atoms, a nanorod or a nanorope. ACS Nano 7, 10075–10082 (2013).

  18. 18.

    & Synthesis of polyynes to model the sp-carbon allotrope carbyne. Nat. Chem. 2, 967–971 (2010).

  19. 19.

    et al. Synthesis, structure, and nonlinear optical properties of diarylpolyynes. Org. Lett. 7, 51–54 (2005).

  20. 20.

    , , , & Carbon nanowires: phonon and pi-electron confinement. Phys. Rev. B 74, 153418 (2006).

  21. 21.

    et al. Absolute Raman intensity measurements and determination of the vibrational second hyperpolarizability of adamantyl endcapped polyynes. J. Raman Spectrosc. 43, 1293–1298 (2012).

  22. 22.

    et al. Alkyne-tag Raman imaging for visualization of mobile small molecules in live cells. J. Am. Chem. Soc. 134, 20681–20689 (2012).

  23. 23.

    et al. Multicolor live-cell chemical imaging by isotopically edited alkyne vibrational palette. J. Am. Chem. Soc. 136, 8027–8033 (2014).

  24. 24.

    et al. Multiplexed multicolor Raman imaging of live cells with isotopically modified single walled carbon nanotubes. J. Am. Chem. Soc. 130, 13540–13541 (2008).

  25. 25.

    et al. Evidence for solution-state nonlinearity of sp-carbon chains based on IR and Raman spectroscopy: violation of mutual exclusion. J. Am. Chem. Soc. 131, 4239–4244 (2009).

  26. 26.

    et al. Super-multiplex vibrational imaging. Nature 544, 465–470 (2017).

  27. 27.

    et al. Label-free biomedical imaging with high sensitivity by stimulated Raman scattering microscopy. Science 322, 1857–1861 (2008).

  28. 28.

    et al. Live-cell imaging of alkyne-tagged small biomolecules by stimulated Raman scattering. Nat. Methods 11, 410–412 (2014).

  29. 29.

    et al. A sensitive and specific Raman probe based on bisarylbutadiyne for live cell imaging of mitochondria. Bioorg. Med. Chem. Lett. 25, 664–667 (2015).

  30. 30.

    , & Encoded microcarriers for high-throughput multiplexed detection. Angew. Chem. Int. Ed. Engl. 45, 6104–6117 (2006).

  31. 31.

    , , & Dye-labeled polystyrene latex microspheres prepared via a combined swelling-diffusion technique. J. Colloid Interface Sci. 363, 137–144 (2011).

  32. 32.

    & Intracellular microlasers. Nat. Photonics 9, 572–576 (2015).

  33. 33.

    et al. Structure and chain polarization of long polyynes investigated with infrared and Raman spectroscopy. J. Raman Spectrosc. 44, 1398–1410 (2013).

  34. 34.

    et al. High-speed molecular spectral imaging of tissue with stimulated Raman scattering. Nat. Photonics 6, 845–851 (2012).

  35. 35.

    et al. Microsecond scale vibrational spectroscopic imaging by multiplex stimulated Raman scattering microscopy. Light Sci. Appl. 4, e265 (2015).

  36. 36.

    et al. Stimulated Raman scattering flow cytometry for label-free single-particle analysis. Optica 4, 103–109 (2017).

Download references

Acknowledgements

We are grateful for the discussion with L. Brus, Y. Shen and Z. Chen. W.M. acknowledges support from NIH Director's New Innovator Award (1DP2EB016573), R01 (EB020892), the US Army Research Office (W911NF-12-1-0594), and the Camille and Henry Dreyfus Foundation.

Author information

Author notes

    • Fanghao Hu
    •  & Chen Zeng

    These authors contributed equally to this work.

Affiliations

  1. Department of Chemistry, Columbia University, New York, New York, USA.

    • Fanghao Hu
    • , Chen Zeng
    • , Rong Long
    • , Yupeng Miao
    • , Lu Wei
    • , Qizhi Xu
    •  & Wei Min
  2. Kavli Institute for Brain Science, Columbia University, New York, New York, USA.

    • Wei Min

Authors

  1. Search for Fanghao Hu in:

  2. Search for Chen Zeng in:

  3. Search for Rong Long in:

  4. Search for Yupeng Miao in:

  5. Search for Lu Wei in:

  6. Search for Qizhi Xu in:

  7. Search for Wei Min in:

Contributions

F.H. performed the spectroscopy, microscopy and biological studies and analyzed the data with the help of Y.M., L.W. and Q.X.; C.Z. performed the chemical synthesis together with R.L.; F.H. and W.M. conceived the concept; F.H., C.Z. and W.M. designed the experiments and wrote the manuscript with input from all authors.

Competing interests

Columbia University has filed a patent application (US 62/540,953) based on this study.

Corresponding author

Correspondence to Wei Min.

Integrated supplementary information

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–13, Supplementary Tables 1–3 and Supplementary Note 1

  2. 2.

    Life Sciences Reporting Summary

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nmeth.4578