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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 624–630 (2014)Cite this article

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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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Figure 1: Optical trapping of HIV virions in culture fluid.
Figure 2: Back focal plane interferometry to distinguish a single HIV-1 particle from aggregates in complete media.
Figure 3: Measurement of diameter for single HIV-1.
Figure 4: Aggregation of HIV-1 observed at high virion concentrations.
Figure 5: Optical trapping virometry.
Figure 6: Two-colour correlation analysis for EGFP–Vpr and envelope glycoproteins in single HIV-1 virions.

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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.

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  1. Ximiao Hou

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

Authors and Affiliations

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

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

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  1. Yuanjie Pang
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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.

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Correspondence to Wei Cheng.

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Pang, Y., Song, H., Kim, J. et al. Optical trapping of individual human immunodeficiency viruses in culture fluid reveals heterogeneity with single-molecule resolution. Nature Nanotech 9, 624–630 (2014). https://doi.org/10.1038/nnano.2014.140

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