Minimization of detection volume by surface plasmon-coupled emission

Gryczyński, Zygmunt; Borejdo, Julian; Matveeva, Evgenia G.; Calander, Nils; Grygorczyk, Ryszard; Harper, J. S.; Gryczyński, Ignacy

doi:10.1117/12.673898

Cited by 5 publications

(8 citation statements)

References 35 publications

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“…This article reports application of SPCE to the study of muscle tissue. We have reported earlier that fluorescent microspheres were seen in a microscope under SPCE illumination (19), but this is the first account of an application to a live sample. The results make it clear that the long-term objective, to observe a single molecule of a contractile protein during contraction of muscle, is feasible.…”

Section: Discussionmentioning

confidence: 92%

“…SPCE has been described in free-standing configurations (16,17) and has been used to detect single molecules (18). Recently, it has been applied to a microscope (19)(20)(21). In the current application of this technique, the observational volume is made shallow by placing a sample on a thin metal film and illuminating it with the laser beam at the surface plasmon resonance (SPR) angle.…”

Section: Introductionmentioning

confidence: 99%

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Application of Surface Plasmon Coupled Emission to Study of Muscle

Borejdo

Gryczyński

Calander

et al. 2006

Biophysical Journal

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Muscle contraction results from interactions between actin and myosin cross-bridges. Dynamics of this interaction may be quite different in contracting muscle than in vitro because of the molecular crowding. In addition, each cross-bridge of contracting muscle is in a different stage of its mechanochemical cycle, and so temporal measurements are time averages. To avoid complications related to crowding and averaging, it is necessary to follow time behavior of a single cross-bridge in muscle. To be able to do so, it is necessary to collect data from an extremely small volume (an attoliter, 10(-18) liter). We report here on a novel microscopic application of surface plasmon-coupled emission (SPCE), which provides such a volume in a live sample. Muscle is fluorescently labeled and placed on a coverslip coated with a thin layer of noble metal. The laser beam is incident at a surface plasmon resonance (SPR) angle, at which it penetrates the metal layer and illuminates muscle by evanescent wave. The volume from which fluorescence emanates is a product of two near-field factors: the depth of evanescent wave excitation and a distance-dependent coupling of excited fluorophores to the surface plasmons. The fluorescence is quenched at the metal interface (up to approximately 10 nm), which further limits the thickness of the fluorescent volume to approximately 50 nm. The fluorescence is detected through a confocal aperture, which limits the lateral dimensions of the detection volume to approximately 200 nm. The resulting volume is approximately 2 x 10(-18) liter. The method is particularly sensitive to rotational motions because of the strong dependence of the plasmon coupling on the orientation of excited transition dipole. We show that by using a high-numerical-aperture objective (1.65) and high-refractive-index coverslips coated with gold, it is possible to follow rotational motion of 12 actin molecules in muscle with millisecond time resolution.

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Section: Discussionmentioning

confidence: 92%

Section: Introductionmentioning

confidence: 99%

Application of Surface Plasmon Coupled Emission to Study of Muscle

Borejdo

Gryczyński

Calander

et al. 2006

Biophysical Journal

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“…In terms of antenna concepts, a single circular nanoaperture such as a ZMW is simple to implement and robust, yet it appears far from being optimum. Another simple design to reduce the detection volume in single molecule fluorescence studies exploits the subwavelength confinement of light supported by surface plasmons at a metal–water interface . In this method that appears as the plasmonic form of TIRF, the sample is a thin metal film deposited on a glass substrate that is illuminated at the surface plasmon resonance angle.…”

Section: Plasmonic Optical Antennasmentioning

confidence: 99%

Plasmonic antennas and zero‐mode waveguides to enhance single molecule fluorescence detection and fluorescence correlation spectroscopy toward physiological concentrations

Punj

Ghenuche

Moparthi

et al. 2014

WIREs Nanomed Nanobiotechnol

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Single-molecule approaches to biology offer a powerful new vision to elucidate the mechanisms that underpin the functioning of living cells. However, conventional optical single molecule spectroscopy techniques such as Förster fluorescence resonance energy transfer (FRET) or fluorescence correlation spectroscopy (FCS) are limited by diffraction to the nanomolar concentration range, far below the physiological micromolar concentration range where most biological reaction occur. To breach the diffraction limit, zero-mode waveguides (ZMW) and plasmonic antennas exploit the surface plasmon resonances to confine and enhance light down to the nanometer scale. The ability of plasmonics to achieve extreme light concentration unlocks an enormous potential to enhance fluorescence detection, FRET, and FCS. Single molecule spectroscopy techniques greatly benefit from ZMW and plasmonic antennas to enter a new dimension of molecular concentration reaching physiological conditions. The application of nano-optics to biological problems with FRET and FCS is an emerging and exciting field, and is promising to reveal new insights on biological functions and dynamics.

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“…Rabbit psoas muscle was first prewashed with cold EDTArigor solution ͑50 mM KCl, 2 mM EDTA, 10 mM DTT, and 10 mM TRIS-HCl pH 7.6͒ for 1 2 h, followed by Ca-rigor so-lution ͑50 mM KCl, 2 mM MgCl 2 , 0.1 mM CaCl 2 , 10 mM DTT, and 10 mM TRIS-HCl pH 7.6͒. Myofibrils were made from muscle as previously described.…”

Section: Preparation Of Myofibrilsmentioning

confidence: 99%

“…PSPs are propagating charge density oscillations induced by light impinging on the metal surface at the surface plasmon resonance ͑SPR͒ angle. The resulting surfaceplasmon-assisted microscope 1,2 ͑SPAM͒ reduced the detection volume to a few attoliters ͑10 −18 L͒, making it possible to detect single cross-bridges in a skeletal muscle myofibril. 2 The photobleaching arises because signal from a single molecule must be measured with a high signal-to-noise ratio ͑SNR͒.…”

Section: Introductionmentioning

confidence: 99%

Decreasing photobleaching by silver nanoparticles on metal surfaces: application to muscle myofibrils

Muthu

Gryczyński

et al. 2008

J. Biomed. Opt.

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Recently it has become possible to study single protein molecules in a cell. However, such experiments are plagued by rapid photobleaching. We recently showed that the interaction of fluorophores with localized surface plasmon polaritons (LSPs) induced in the metallic nanoparticles led to a substantial reduction of photobleaching. We now investigate whether the photobleaching could be further reduced when the excited fluorophore interacts with the LSP excited in the metallic nanoparticles resident on mirrored surface. As an example we use myofibrils, subcellular structures within skeletal muscle. We compare nanoparticle-enhanced fluorescence of myofibrils in the presence and in the absence of a mirrored surface. The proximity of the mirrored surface led to enhancement of fluorescence and to a decrease in fluorescent lifetime, much greater than that observed in the presence of nanoparticles alone. We think that the effect is caused by the near-field interactions between fluorophores and LSP, and between fluorophores and propagating surface plasmons (PSPs) produced in the metallic surface by the nanoparticles. Photobleaching is decreased because fluorescence enhancement enables illumination with a weaker laser beam and because the decrease in fluorescence lifetime minimizes the probability of oxygen attack during the time a molecule is in the exited state.

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Minimization of detection volume by surface plasmon-coupled emission

Cited by 5 publications

References 35 publications

Application of Surface Plasmon Coupled Emission to Study of Muscle

Application of Surface Plasmon Coupled Emission to Study of Muscle

Plasmonic antennas and zero‐mode waveguides to enhance single molecule fluorescence detection and fluorescence correlation spectroscopy toward physiological concentrations

Decreasing photobleaching by silver nanoparticles on metal surfaces: application to muscle myofibrils

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