2015
DOI: 10.1117/1.jbo.20.9.097001
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Reflected light microspectroscopy for single-nanoparticle biosensing

Abstract: Conventional and dark-field microscopy in the transmission mode is extensively used for single plasmonic nanoparticle (NP) imaging and spectral analysis. However, application of the transmission mode for realtime biosensing to single NP poses strict limitations on the size and material properties of the microfluidic system. This article proposes a simple optical technique based on reflected light microscopy to perform microspectroscopy of a single NP placed in a conventional, nontransparent liquid delivery sys… Show more

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Cited by 5 publications
(5 citation statements)
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“…The average number of NPs per cell was estimated by 3D scanning of the cells in random areas of the Petri dish. The 3D scanning was performed using back‐reflection setup and high‐NA‐×60‐oil objective . The z‐focus was controlled with PRIOR Pro Scan III accessory.…”
Section: Materials and Methods/experimentalmentioning
confidence: 57%
“…The average number of NPs per cell was estimated by 3D scanning of the cells in random areas of the Petri dish. The 3D scanning was performed using back‐reflection setup and high‐NA‐×60‐oil objective . The z‐focus was controlled with PRIOR Pro Scan III accessory.…”
Section: Materials and Methods/experimentalmentioning
confidence: 57%
“…The microscopy system was equipped with an oil immersion objective with a magnification of 100× and a variable numerical aperture (NA = 0.5-1.3) used for the reflected light microscopy (RLM) method. [29][30][31] The microscopy illumination was provided by a standard white light source (Nikon) with a 50 W halogen lamp and a 50/50 beam splitter providing a smooth illumination across the entire operating range (Fig. 1a).…”
Section: Microscopy Visualization Of Bacteria With Plasmonic Npsmentioning
confidence: 99%
“…As shown in the literature, high contrast imaging of plasmonic NPs in a highly diffusive medium can be obtained by using reflected light microscopy (RLM) with a high numerical aperture oil-immersion objective. [29][30][31] In Fig. 1c we present an image of the prepared bacteria-NP sample, wherein the highintensity bright spots correspond to the scattering from plasmonic NPs detected by RLM.…”
Section: Microscopy Visualization Of Bacteria With Plasmonic Npsmentioning
confidence: 99%
“…The speed and throughput of light scattering measurements should in principle be limited only by the available light power or maximum allowable local heating of the particle due to absorption. This has motivated the use of gold nanoparticle-functionalized probes in place of fluorescent probes for microarray labeling, which are either detected individually directly based on their light scattering or indirectly via silver deposition. Similar to the fluorescence-based techniques mentioned earlier, these nanoparticle-based techniques have also successfully enhanced the dynamic range of microarrays by several orders of magnitude and typically have a limit of detection of roughly 1 femtomolar. However, direct detection of nanoparticle labels has only been done with a high-NA lens, resulting in the same throughput limitations as single-molecule fluorescence.…”
mentioning
confidence: 99%
“…Interferometric detection is more resilient than dark-field to very weak signals since the scattered light amplitude is measured rather than intensity. Interferometric reflectance imaging sensing (IRIS) is one of a family of similar optical techniques for interferometric detection of nanoparticles immobilized on a substrate. ,, IRIS uses a substrate of polished silicon with a transparent thin film, typically thermally grown silicon dioxide. The substrate is imaged with a reflectance microscope with Köhler illumination from an LED source.…”
mentioning
confidence: 99%