Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Monovalent, reduced-size quantum dots for imaging receptors on living cells

Abstract

We describe a method to generate monovalent quantum dots (QDs) using agarose gel electrophoresis. We passivated QDs with a carboxy-terminated polyethylene-glycol ligand, yielding particles with half the diameter of commercial QDs, which we conjugated to a single copy of a high-affinity targeting moiety (monovalent streptavidin or antibody to carcinoembryonic antigen) to label cell-surface proteins. The small size improved access of QD-labeled glutamate receptors to neuronal synapses, and monovalency prevented EphA3 tyrosine kinase activation.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Overview of small monovalent QDs, and accessibility of sQDs to synapses.
Figure 2: Generation and characterization of monovalent sQDs.
Figure 3: Monovalent sQDs reduce EphA3 clustering and internalization.

Similar content being viewed by others

References

  1. Saxton, M.J. & Jacobson, K. Annu. Rev. Biophys. Biomol. Struct. 26, 373–399 (1997).

    Article  CAS  Google Scholar 

  2. Michalet, X. et al. Science 307, 538–544 (2005).

    Article  CAS  Google Scholar 

  3. Howarth, M., Takao, K., Hayashi, Y. & Ting, A.Y. Proc. Natl. Acad. Sci. USA 102, 7583–7588 (2005).

    Article  CAS  Google Scholar 

  4. Groc, L. et al. J. Neurosci. 27, 12433–12437 (2007).

    Article  CAS  Google Scholar 

  5. Pinaud, F., King, D., Moore, H.P. & Weiss, S. J. Am. Chem. Soc. 126, 6115–6123 (2004).

    Article  CAS  Google Scholar 

  6. Liu, W. et al. J. Am. Chem. Soc. 130, 1274–1284 (2008).

    Article  CAS  Google Scholar 

  7. Susumu, K. et al. J. Am. Chem. Soc. 129, 13987–13996 (2007).

    Article  CAS  Google Scholar 

  8. Worden, J.G., Shaffer, A.W. & Huo, Q. Chem. Commun. (Camb.) 7, 518–519 (2004).

    Article  Google Scholar 

  9. Sung, K.M., Mosley, D.W., Peelle, B.R., Zhang, S. & Jacobson, J.M. J. Am. Chem. Soc. 126, 5064–5065 (2004).

    Article  CAS  Google Scholar 

  10. Levy, R. et al. ChemBioChem 7, 592–594 (2006).

    Article  CAS  Google Scholar 

  11. Fu, A. et al. J. Am. Chem. Soc. 126, 10832–10833 (2004).

    Article  CAS  Google Scholar 

  12. Howarth, M. et al. Nat. Methods 3, 267–273 (2006).

    Article  CAS  Google Scholar 

  13. Pons, T., Uyeda, H.T., Medintz, I.L. & Mattoussi, H. J. Phys. Chem. B. 110, 20308–20316 (2006).

    Article  CAS  Google Scholar 

  14. Wimmer-Kleikamp, S.H., Janes, P.W., Squire, A., Bastiaens, P.I. & Lackmann, M. J. Cell Biol. 164, 661–666 (2004).

    Article  CAS  Google Scholar 

  15. Michaely, P., Li, W.P., Anderson, R.G., Cohen, J.C. & Hobbs, H.H. J. Biol. Chem. 279, 34023–34031 (2004).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

Funding was provided by the US National Institutes of Health (NIH) (P20GM072029-01), and the McKnight, the Dreyfus and the Sloan Foundations. M.H. was supported by an MIT-Merck fellowship, W.L. by a US National Science Foundation fellowship, M.G.B. by the Army Research Office DAAD 19-03-D0004, and K.D.W. and M.M.S. by the NIH (CA101830) and the MIT Biotechnology Training Program. We thank M. Lackmann, P. Janes, S. Manalis and A. Sparks for advice, J. Chan for technical assistance, and Tanabe for providing biotin.

Author information

Authors and Affiliations

Authors

Contributions

M.H., W.L., M.G.B. and A.Y.T. designed the experiments; M.H. and A.Y.T. wrote the paper; M.H., W.L., S.P., L.F.M. and Y.Z. performed the experiments; M.M.S. and K.D.W. generated the PEG-scFv.

Corresponding author

Correspondence to Alice Y Ting.

Ethics declarations

Competing interests

M.H., W.L., M.G.B. and A.Y.T. are authors on a submitted US patent application describing the quantum dots generated in this work.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–6, Supplementary Methods (PDF 890 kb)

Supplementary Video 1

Single-molecule imaging of wild-type LDL receptor with small monovalent QDs. COS7 cells transfected with wild-type AP-LDL receptor and BirA-ER were incubated with sQD-mSA1 and single QDs were imaged. The video is speeded up 3-fold. (MOV 963 kb)

Supplementary Video 2

Single-molecule imaging of mutant LDL receptor with small monovalent QDs. COS7 cells transfected with mutant AP-LDL receptor and BirA-ER were incubated with sQD-mSA1 and single QDs were imaged. The video is speeded up 3-fold. (MOV 847 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Howarth, M., Liu, W., Puthenveetil, S. et al. Monovalent, reduced-size quantum dots for imaging receptors on living cells. Nat Methods 5, 397–399 (2008). https://doi.org/10.1038/nmeth.1206

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth.1206

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing