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1.
Nanotechnology ; 27(46): 465201, 2016 Nov 18.
Artículo en Inglés | MEDLINE | ID: mdl-27734808

RESUMEN

The scanning tunnelling microscope (STM) may be used as a low-energy, electrical nanosource of surface plasmon polaritons and light. In this article, we demonstrate that the optimum mode of operation of the STM for maximum photon emission is completely different in air than in vacuum. To this end, we investigate the emission of photons, the variation in the relative tip-sample distance and the measured current as a function of time for an STM operating in air. Contrary to the case of an STM operating in vacuum, the measured current between the tip and sample for an STM in air is very unstable (rapidly fluctuating in time) when the applied voltage between the tip and sample is in the ∼1.5-3 V range (i.e., in the energy range of visible photons). The photon emission occurs in short (50 µs) bursts when the STM tip is closest to the sample. The current instabilities are shown to be a key ingredient for producing intense light emission from an STM operating in air (photon emission rate several orders of magnitude higher than for stable current). These results are explained in terms of the interplay between the tunnel current and the electrochemical current in the ubiquitous thin water layer that exists when working in air.

2.
Biophys J ; 92(6): 2150-61, 2007 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-17172306

RESUMEN

Fluorescence labeling is the prevailing imaging technique in cell biology research. When they involve statistical investigations on a large number of cells, experimental studies require both low magnification to get a reliable statistical population and high contrast to achieve accurate diagnosis on the nature of the cells' perturbation. Because microscope objectives of low magnification generally yield low collection efficiency, such studies are limited by the fluorescence signal weakness. To overcome this technological bottleneck, we proposed a new method based on metal-coated substrates that enhance the fluorescence process and improve collection efficiency in epifluorescence observation and that can be directly used with a common microscope setup. We developed a model based on the dipole approximation with the aim of simulating the optical behavior of a fluorophore on such a substrate and revealing the different mechanisms responsible for fluorescence enhancement. The presence of a reflective surface modifies both excitation and emission processes and additionally reshapes fluorescence emission lobes. From both theoretical and experimental results, we found the fluorescence signal emitted by a molecular cyanine 3 dye layer to be amplified by a factor approximately 30 when fluorophores are separated by a proper distance from the substrate. We then adapted our model to the case of homogeneously stained micrometer-sized objects and demonstrated mean signal amplification by a factor approximately 4. Finally, we applied our method to fluorescence imaging of dog kidney cells and verified experimentally the simulated results.


Asunto(s)
Técnicas de Cultivo de Célula/métodos , Materiales Biocompatibles Revestidos/química , Aumento de la Imagen/métodos , Metales/química , Microscopía Fluorescente/métodos , Fotometría/métodos , Espectrometría de Fluorescencia/métodos , Reproducibilidad de los Resultados , Sensibilidad y Especificidad
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