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Luminescent surfaces with tailored angular emission for compact dark-field imaging devices

Abstract

Dark-field microscopy is a standard imaging technique widely employed in biology that provides high image contrast for a broad range of unstained specimens1. Unlike bright-field microscopy, it accentuates high spatial frequencies and can therefore be used to emphasize and resolve small features. However, the use of dark-field microscopy for reliable analysis of blood cells, bacteria, algae and other marine organisms often requires specialized, bulky microscope systems, as well as expensive additional components, such as dark-field-compatible objectives or condensers2,3. Here, we propose to simplify and downsize dark-field microscopy equipment by generating the high-angle illumination cone required for dark-field microscopy directly within the sample substrate. We introduce a luminescent photonic substrate with a controlled angular emission profile and demonstrate its ability to generate high-contrast dark-field images of micrometre-sized living organisms using standard optical microscopy equipment. This new type of substrate forms the basis for miniaturized lab-on-chip dark-field imaging devices that are compatible with simple and compact light microscopes.

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Fig. 1: The concept of SLED.
Fig. 2: The fabricated SLED surface.
Fig. 3: Optical characteristics of the light-emitting surfaces.
Fig. 4: Application of the SLED surfaces to image colloids and marine microorganisms.
Fig. 5: Modelling to compare image formation with SLED illumination and bright-field illumination.

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Data availability

The data that support the plots within this paper and other findings of this study are available from the corresponding author upon reasonable request.

Code availability

The MATLAB codes used to model the surfaces’ emission properties and partially coherent imaging are available for download from https://github.com/mathiaskolle/substrate-luminescence-enabled-darkfield-imaging.

References

  1. Gage, S. H. Modern dark-field microscopy and history of its development. Trans. Am. Microsc. Soc. 39, 95–141 (1920).

    Article  Google Scholar 

  2. Hecht, E. Optics 3rd edn (Addison-Wesley, 1998).

  3. Murphy, D. B. & Davidson, M. W. Fundamentals of Light Microscopy and Electronic Imaging 2nd edn (Wiley-Blackwell, 2013).

  4. Noda, N. & Kamimura, S. A new microscope optics for laser dark-field illumination applied to high precision two-dimensional measurement of specimen displacement. Rev. Sci. Instrum. 79, 023704 (2008).

    Article  ADS  Google Scholar 

  5. Ueno, H. et al. Simple dark-field microscopy with nanometer spatial precision and microsecond temporal resolution. Biophys. J. 98, 2014–2023 (2010).

    Article  ADS  Google Scholar 

  6. Kudo, S., Magariyama, Y. & Aizawa, S. Abrupt changes in flagellar rotation observed by laser dark-field microscopy. Nature 346, 677–680 (1990).

    Article  ADS  Google Scholar 

  7. Dunn, A. R. & Spudich, J. A. Dynamics of the unbound head during myosin V processive translocation. Nat. Struct. Mol. Biol. 14, 246–248 (2007).

    Article  Google Scholar 

  8. Nishiyama, M., Muto, E., Inoue, Y., Yanagida, T. & Higuchi, H. Substeps within the 8 nm step of ATPase cycle of single kinesin molecules. Nat. Cell Biol. 3, 425–428 (2001).

    Article  Google Scholar 

  9. Yasuda, R., Noji, H., Yoshida, M., Kinosita, K. Jr & Itoh, H. Resolution of distinct rotational substeps by submillisecond kinetic analysis of F1-ATPase. Nature 410, 898–904 (2001).

    Article  ADS  Google Scholar 

  10. Sönnichsen, C., Franzl, T., Wilk, T., von Plessen, G. & Feldmann, J. Plasmon resonances in large noble-metal clusters. New J. Phys. 4, 93 (2002).

    Article  ADS  Google Scholar 

  11. Rosman, C. et al. A new approach to assess gold nanoparticle uptake by mammalian cells: combining optical dark-field and transmission electron microscopy. Small 23, 3683–3690 (2012).

    Article  Google Scholar 

  12. Ma, J., Liu, Y., Gao, P. F., Zou, H. Y. & Huang, C. Z. Precision improvement in dark-field microscopy imaging by using gold nanoparticles as an internal reference: a combined theoretical and experimental study. Nanoscale 8, 8729–8736 (2016).

    Article  ADS  Google Scholar 

  13. Von Olshausen, P. & Rohrbach, A. Coherent total internal reflection dark-field microscopy: label-free imaging beyond the diffraction limit. Opt. Lett. 38, 4066–4069 (2013).

    Article  ADS  Google Scholar 

  14. Braslavsky, I. et al. Objective-type dark-field illumination for scattering from microbeads. Appl. Opt. 40, 5650–5657 (2001).

    Article  ADS  Google Scholar 

  15. Kim, S., Blainey, P. C., Schroeder, C. M. & Xie, X. S. Multiplexed single-molecule assay for enzymatic activity on flow-stretched DNA. Nat. Methods 4, 397–399 (2007).

    Article  Google Scholar 

  16. Taylor, M. A. & Bowen, W. P. Enhanced sensitivity in dark-field microscopy by optimizing the illumination angle. Appl. Opt. 52, 5718–5723 (2013).

    Article  ADS  Google Scholar 

  17. Zheng, G., Cui, X. & Yang, C. Surface-wave-enabled darkfield aperture for background suppression during weak signal detection. Proc. Natl Acad. Sci. USA 107, 9043–9048 (2010).

    Article  ADS  Google Scholar 

  18. Zhang, J., Pitter, M. C., Liu, S., See, C. & Somekh, M. G. Surface-plasmon microscopy with a two-piece solid immersion lens: bright and dark fields. Appl. Opt. 45, 7977–7986 (2006).

    Article  ADS  Google Scholar 

  19. Balci, S., Karademir, E., Kocabas, C. & Aydinli, A. Direct imaging of localized surface plasmon polaritons. Opt. Lett. 36, 3401–3403 (2011).

    Article  ADS  Google Scholar 

  20. Wei, F., O, Y. W., Li, G., Cheah, K. W. & Liu, Z. Organic light-emitting-diode-based plasmonic dark-field microscopy. Opt. Lett. 37, 4359–4361 (2012).

    Article  ADS  Google Scholar 

  21. Coropceanu, I. & Bawendi, M. G. Core/shell quantum dot based luminescent solar concentrators with reduced reabsorption and enhanced efficiency. Nano Lett. 14, 4097–4101 (2014).

    Article  ADS  Google Scholar 

  22. Vukusic, P., Sambles, J. R. & Lawrence, C. R. Structural colour: colour mixing in wing scales of a butterfly. Nature 404, 457–457 (2000).

    Article  ADS  Google Scholar 

  23. Vukusic, P., Sambles, J. R., Lawrence, C. R. & Wakely, G. Sculpted-multilayer optical effects in two species of Papilio butterfly. Appl. Opt. 40, 1116–1125 (2001).

    Article  ADS  Google Scholar 

  24. Kolle, M. et al. Mimicking the colourful wing scale structure of the Papilio blumei butterfly. Nat. Nanotechnol. 5, 511–515 (2010).

    Article  ADS  Google Scholar 

  25. Heavens, O. S. Optical Properties of Thin Solid Films (Dover Publications, 1965).

  26. Johnson, P. B. & Christy, R. W. Optical constants of the noble metals. Phys. Rev. B 6, 4370–4379 (1972).

    Article  ADS  Google Scholar 

  27. Hopkins, H. H. On the diffraction theory of optical images. Proc. R. Soc. A. 217, 408–432 (1953).

    MathSciNet  MATH  ADS  Google Scholar 

  28. Born, M. et al. Principles of Optics: Electromagnetic Theory of Propagation, Interference and Diffraction of Light (Cambridge Univ. Press, 1999).

  29. Goodman, J. W. Statistical Optics (Wiley, 2015).

  30. Shirasaki, Y., Supran, G., Bawendi, M. G. & Bulović, V. Emergence of colloidal quantum-dot light-emitting technologies. Nat. Photon. 7, 13–23 (2012).

    Article  ADS  Google Scholar 

  31. Mashford, B. S. et al. High-efficiency quantum-dot light-emitting devices with enhanced charge injection. Nat. Photon. 7, 407–412 (2013).

    Article  ADS  Google Scholar 

  32. Anikeeva, P. O., Halpert, J. E., Bawendi, M. G. & Bulović, V. Electroluminescence from a mixed red-green-blue colloidal quantum dot monolayer. Nano Lett. 7, 2196–2200 (2007).

    Article  ADS  Google Scholar 

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Acknowledgements

We thank E. Shirman and T. Shirman for their guidance in designing the multiple-step moulding process used for fabricating the micropatterned bottom reflectors. C.A.C.C. and M.K. acknowledge support from the National Science Foundation through the ‘Designing Materials to Revolutionize and Engineer our Future’ programme (DMREF-1922321) and from the US Army Research Office through the Institute for Soldier Nanotechnologies at MIT under contract no. W911NF-13-D-0001. P.T.C.S. and C.J.R. acknowledge support from NIH 9P41EB015871.

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Authors

Contributions

M.K. and C.A.C.C. conceived the research. C.A.C.C. designed and built the dark-field devices from the master that M.R.J.S. created. K.B. and Y.K. provided advice to optimize the microfabrication process. M.K. and C.A.C.C. wrote the MATLAB code for optical modelling. C.J.R. and P.T.C.S. provided advice to build the optical characterization set-up that C.A.C.C., C.J.R. and S.N. implemented onto a microscope for emission characterization of the SLED devices. I.C. and M.G.B. synthesized the QDs that were used in the SLED devices. C.A.C.C. and M.K. wrote the manuscript. All authors commented on the manuscript.

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Correspondence to Mathias Kolle.

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Supplementary Figs. 1–6, Supplementary Discussion and details on modelling.

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Chazot, C.A.C., Nagelberg, S., Rowlands, C.J. et al. Luminescent surfaces with tailored angular emission for compact dark-field imaging devices. Nat. Photonics 14, 310–315 (2020). https://doi.org/10.1038/s41566-020-0593-1

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