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Nanophotonic approaches to investigate the spatiotemporal organization of biological membranes

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A hot topic in cell biology is to understand the specific nanometer-scale organization and distribution of the surface machinery of living cells and its role regulating the spatiotemporal control of different cellular processes. Cell adhesion, pathogen recognition or lipid-mediated signaling, all fundamentally important processes in immunology, are governed by molecular interactions occurring at the nanoscale. From the technical point of view, the quest for optical imaging of biological processes at the nanoscale has driven in recent years a swift development of a large number of microscopy techniques based on far-field optics. These super-resolution methods are providing new capabilities for probing biology at the nanoscale by fluorescence. While these techniques conveniently use lens-based microscopy, the attainable resolution and/or localization precision severely depend on the sample fluorescence properties. True nanoscale optical resolution free from these constrains can alternatively be obtained by interacting with fluorophores in the near-field. Indeed, near-field scanning optical microscopy (NSOM) using subwavelength aperture probes is one of the earliest approaches sought to achieve nanometric optical resolution [1]. More recently, photonic antennas have emerged as excellent alternative candidates to further improve the resolution of NSOM by amplifying electromagnetic fields into regions of space much smaller than the wavelength of light [2]. In this talk I will describe our efforts towards the fabrication of different nanoantenna probe configurations as well as 2D antenna arrays for applications in nano-imaging and spectroscopy of living cells [3-5]. For nanoscale imaging, we have recently pushed the limits of spatial resolution by demonstrating dual colour imaging of individual fluorescent molecules with true 20nm spatial resolution and sub-nanometre localization accuracy using antenna probes [4]. In parallel, we have recently demonstrated that photonic antennas allow the recording of individual lipid diffusion on living cell membranes in regions as small as 20nm in size [5]. Finally, I will summarize our efforts towards dual–colour fluorescent cross correlation spectroscopy in living cells with a spatial confinement of 50nm.


[1] P. Hinterdorfer, M.F. Garcia-Parajo, Y. Dufrene, “Single-molecule imaging of cell surfaces using near-field nanoscopy”, Acc. Chem. Res. 45, 327-336, 2012. [2] M.F. Garcia-Parajo, “Photonic antennas focus in on biology”. Nature Photonics, 2, 201-204, 2008. [3] D. Punj, M. Mivelle, S. B. Moparthi, T. S. van Zanten, H. Rigneault, N. F. van Hulst, M. F. García-Parajó, J. Wenger, “A plasmonic ‘antenna-in-box’ platform for enhanced single-molecule analysis at micromolar concentrations”, Nature Nanotechnol. 8, 512-516, 2013. [4] M. Mivelle, T. S. van Zanten, M. F. Garcia-Parajo, “Hybrid photonic antennas for subnanometer multicolor localization and nanoimaging of single molecules”, Nano Lett. 4, 4895-4900, 2014. [5] V. Flauraud, T. S. van Zanten, M. Mivelle, C. Manzo, M. F. Garcia Parajo, J. Brugger, “Large-scale arrays of bowtie nanoaperture antennas for nanoscale dynamics in living cell membranes”, Nano Lett. 15, 4176-4182 2015.

This talk is part of the BSS Formal Seminars series.

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