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Optical trapping of individual human immunodeficiency viruses in culture fluid reveals heterogeneity with single-molecule resolution

Nature Nanotechnology volume 9, pages 624630 (2014) | Download Citation

Abstract

Optical tweezers use the momentum of photons to trap and manipulate microscopic objects, contact-free, in three dimensions. Although this technique has been widely used in biology and nanotechnology to study molecular motors, biopolymers and nanostructures, its application to study viruses has been very limited, largely due to their small size. Here, using optical tweezers that can simultaneously resolve two-photon fluorescence at the single-molecule level, we show that individual HIV-1 viruses can be optically trapped and manipulated, allowing multi-parameter analysis of single virions in culture fluid under native conditions. We show that individual HIV-1 differs in the numbers of envelope glycoproteins by more than one order of magnitude, which implies substantial heterogeneity of these virions in transmission and infection at the single-particle level. Analogous to flow cytometry for cells, this fluid-based technique may allow ultrasensitive detection, multi-parameter analysis and sorting of viruses and other nanoparticles in biological fluid with single-molecule resolution.

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Acknowledgements

This work was supported by a National Institutes of Health (NIH) Director's New Innovator Award (1DP2OD008693-01, to W.C.), a National Science Foundation CAREER Award (CHE1149670, to W.C.) and also in part by a research grant from the March of Dimes Foundation (5-FY10-490, to W.C.). The authors thank A. Ono and A. Telesnitsky for discussions and Cheng Lab members, especially M. DeSantis, for critical reading of the manuscript. The MATLAB code for analysis of transmission electron microscopy images of polystyrene beads was provided by M. DeSantis. The following reagents were obtained through the AIDS Research and Reference Reagent Program, Division of AIDS, National Institute of Allergy and Infectious Diseases (NIAID), NIH: pNL4-3 from M. Martin; pNL4-3.Luc.R-E- from N. Landau; pEGFP–Vpr from W. C. Greene; TZM-bl cells from J. C. Kappes, X. Wu and Tranzyme Inc; b12 antibody from D. Burton and C. Barbas.

Author information

Author notes

    • Ximiao Hou

    Present address: College of Life Sciences, Northwest A&F University, Shaanxi, China

Affiliations

  1. Department of Pharmaceutical Sciences, University of Michigan, 428 Church Street, Ann Arbor, Michigan 48109, USA

    • Yuanjie Pang
    • , Hanna Song
    • , Jin H. Kim
    • , Ximiao Hou
    •  & Wei Cheng

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Contributions

W.C. conceived and directed the project. H.S. and J.H.K. prepared experimental materials. Y.P., H.S., J.H.K., X.H. and W.C. conducted the experiments. Y.P., H.S., J.H.K., X.H. and W.C. performed the analysis. Y.P. and W.C. wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Wei Cheng.

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DOI

https://doi.org/10.1038/nnano.2014.140

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