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.
Subscribe to Journal
Get full journal access for 1 year
only $8.25 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
The data that support the plots within this paper and other findings of this study are available from the corresponding author upon reasonable request.
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.
Gage, S. H. Modern dark-field microscopy and history of its development. Trans. Am. Microsc. Soc. 39, 95–141 (1920).
Hecht, E. Optics 3rd edn (Addison-Wesley, 1998).
Murphy, D. B. & Davidson, M. W. Fundamentals of Light Microscopy and Electronic Imaging 2nd edn (Wiley-Blackwell, 2013).
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).
Ueno, H. et al. Simple dark-field microscopy with nanometer spatial precision and microsecond temporal resolution. Biophys. J. 98, 2014–2023 (2010).
Kudo, S., Magariyama, Y. & Aizawa, S. Abrupt changes in flagellar rotation observed by laser dark-field microscopy. Nature 346, 677–680 (1990).
Dunn, A. R. & Spudich, J. A. Dynamics of the unbound head during myosin V processive translocation. Nat. Struct. Mol. Biol. 14, 246–248 (2007).
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).
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).
Sönnichsen, C., Franzl, T., Wilk, T., von Plessen, G. & Feldmann, J. Plasmon resonances in large noble-metal clusters. New J. Phys. 4, 93 (2002).
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).
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).
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).
Braslavsky, I. et al. Objective-type dark-field illumination for scattering from microbeads. Appl. Opt. 40, 5650–5657 (2001).
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).
Taylor, M. A. & Bowen, W. P. Enhanced sensitivity in dark-field microscopy by optimizing the illumination angle. Appl. Opt. 52, 5718–5723 (2013).
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).
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).
Balci, S., Karademir, E., Kocabas, C. & Aydinli, A. Direct imaging of localized surface plasmon polaritons. Opt. Lett. 36, 3401–3403 (2011).
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).
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).
Vukusic, P., Sambles, J. R. & Lawrence, C. R. Structural colour: colour mixing in wing scales of a butterfly. Nature 404, 457–457 (2000).
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).
Kolle, M. et al. Mimicking the colourful wing scale structure of the Papilio blumei butterfly. Nat. Nanotechnol. 5, 511–515 (2010).
Heavens, O. S. Optical Properties of Thin Solid Films (Dover Publications, 1965).
Johnson, P. B. & Christy, R. W. Optical constants of the noble metals. Phys. Rev. B 6, 4370–4379 (1972).
Hopkins, H. H. On the diffraction theory of optical images. Proc. R. Soc. A. 217, 408–432 (1953).
Born, M. et al. Principles of Optics: Electromagnetic Theory of Propagation, Interference and Diffraction of Light (Cambridge Univ. Press, 1999).
Goodman, J. W. Statistical Optics (Wiley, 2015).
Shirasaki, Y., Supran, G., Bawendi, M. G. & Bulović, V. Emergence of colloidal quantum-dot light-emitting technologies. Nat. Photon. 7, 13–23 (2012).
Mashford, B. S. et al. High-efficiency quantum-dot light-emitting devices with enhanced charge injection. Nat. Photon. 7, 407–412 (2013).
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).
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.
The authors declare no competing interests.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
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
Nature Communications (2021)
Bandwidth limits of luminescent solar concentrators as detectors in free-space optical communication systems
Light: Science & Applications (2021)
Nature Communications (2021)
Nature Photonics (2020)