Tunable lifetime multiplexing using luminescent nanocrystals

Journal name:
Nature Photonics
Volume:
8,
Pages:
32–36
Year published:
DOI:
doi:10.1038/nphoton.2013.322
Received
Accepted
Published online

Abstract

Optical multiplexing plays an important role in applications such as optical data storage1, document security2, molecular probes3, 4 and bead assays for personalized medicine5. Conventional fluorescent colour coding is limited by spectral overlap and background interference, restricting the number of distinguishable identities. Here, we show that tunable luminescent lifetimes τ in the microsecond region can be exploited to code individual upconversion nanocrystals. In a single colour band, one can generate more than ten nanocrystal populations with distinct lifetimes ranging from 25.6 µs to 662.4 µs and decode their well-separated lifetime identities, which are independent of both colour and intensity. Such ‘τ-dots’ potentially suit multichannel bioimaging, high-throughput cytometry quantification, high-density data storage, as well as security codes to combat counterfeiting. This demonstration extends the optical multiplexing capability by adding the temporal dimension of luminescent signals, opening new opportunities in the life sciences, medicine and data security.

At a glance

Figures

  1. Lifetime tuning scheme and time-resolved confocal images for NaYF4:Yb,Tm upconversion nanocrystals.
    Figure 1: Lifetime tuning scheme and time-resolved confocal images for NaYF4:Yb,Tm upconversion nanocrystals.

    Each pixel was excited for 200 µs, followed by a delayed detection window of up to 3.8 ms to record its time-gated luminescence decay (40 ms exposure time to allow 10 times integration). The colour tone (hue) for each pixel represents its lifetime value decoded from the decay curve. The nanocrystals in the images from left to right have Tm doping concentrations of 4, 2, 1, 0.5 and 0.2 mol%, respectively, as well as 20 mol% Yb dopants.

  2. Results for [tau]-dots-labelled Giardia cysts measured by the time-resolved scanning cytometry system.
    Figure 2: Results for τ-dots-labelled Giardia cysts measured by the time-resolved scanning cytometry system.

    a,b, Lifetime histograms obtained from cysts labelled with different lifetime-encoded τ-dots (Yb/Tm co-doping concentration (mol%:mol%) of 20:1 for a and 20:4 for b). The scanning cytometry allows retrieval of each individual target cyst for luminescence as well as bright-field imaging confirmation. c, Typical recorded luminescence images for the same cyst under 4 h continuous laser excitation. All images were captured with a 100 ms exposure time.

  3. Concept of [tau]-dots-encoded microspheres as the lifetime multiplexing suspension arrays.
    Figure 3: Concept of τ-dots-encoded microspheres as the lifetime multiplexing suspension arrays.

    a, The synthesized monodispersed Tm upconversion nanocrystals can be embedded into the shell of porous microspheres, which can be decoded by the time-resolved scanning cytometry system, for example. b,c, Typical TEM image of the nanocrystals (b) and SEM image of a microsphere incorporating the nanocrystals (c).

  4. Results for [tau]-dots-encoded populations of microspheres carrying unique lifetime identities.
    Figure 4: Results for τ-dots-encoded populations of microspheres carrying unique lifetime identities.

    a, The mechanism of upconversion energy transfer, by adjusting the co-dopant concentration of the sensitizer/emitter, can generate eight lifetime populations of microspheres in the Tm blue-emission band. Symbols α and β represent cubic and hexagonal crystal phases, respectively. The numeral besides each histogram is the mean lifetime ± lifetime CV from Gaussian distribution fitting. The blocks in the axis above represent the lifetime resources (±3σ) occupied by each population. The open spaces suggest more populations could be engineered. b, Two-dimensional (intensity versus lifetime) scattered plots showing that all lifetime populations are independent of the intensities of individual microcarriers.

  5. Demonstration of lifetime-encoded document security and photonic data storage.
    Figure 5: Demonstration of lifetime-encoded document security and photonic data storage.

    ac, Three overlapping patterns are printed with different Tm τ-dots: (CYb:CTm) 20:4 for the ‘Macquarie University’ logo, 20:1 for the Sydney Opera House image, and 20:0.5 for the Sydney Harbour Bridge image. Intensity-based luminescence imaging only gives a complex picture (a), but time-resolved scanning separates the patterns based on the lifetime components of every pixel (b), so that genuine multiplexing information contained in the same overlapping space of the document can be decoded (c; pseudocolour is used to indicate the luminescence lifetime for each pixel). Scale bars (all images), 5 mm.

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Author information

Affiliations

  1. Advanced Cytometry Laboratories, MQ Photonics Research Centre and MQ BioFocus Research Centre, Macquarie University, Sydney, New South Wales 2109, Australia

    • Yiqing Lu,
    • Jiangbo Zhao,
    • Run Zhang,
    • Yujia Liu,
    • Deming Liu,
    • Ewa M. Goldys,
    • Jie Lu,
    • Yu Shi,
    • James A. Piper &
    • Dayong Jin
  2. Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, New South Wales 2109, Australia

    • Run Zhang,
    • Anwar Sunna,
    • Jie Lu,
    • Yu Shi &
    • Dayong Jin
  3. Department of Biomedical Engineering, College of Engineering, Peking University, Beijing 100871, China

    • Yujia Liu,
    • Xusan Yang &
    • Peng Xi
  4. School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, China

    • Yujia Liu
  5. Newport Instruments, 3345 Hopi Place, San Diego, California 92117-3516, USA

    • Robert C. Leif
  6. Department of Electronic Engineering, Tsinghua University, Beijing 100084, China

    • Yujing Huo
  7. Olympus Australia, 82 Waterloo Road, North Ryde, New South Wales 2113, Australia

    • Jian Shen
  8. Purdue University Cytometry Laboratories, Bindley Bioscience Center, Purdue University, West Lafayette, Indiana 47907, USA

    • J. Paul Robinson &
    • Dayong Jin

Contributions

D.J., J.A.P., R.C.L. and J.P.R. conceived the project. D.J. designed the experiments and supervised the research. Y.Lu, J.Z. and D.J. were primarily responsible for data collection and analysis. Y.Lu, E.M.G. and D.J. prepared figures and wrote the main manuscript text. Y.Lu, J.Z., R.Z., D.L. and D.J. were primarily responsible for the Supplementary Information. All authors contributed to data analysis, discussions and manuscript preparation.

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The authors declare no competing financial interests.

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