Fluorescence Enhancements on Silver Colloid Coated Surfaces

Łukomska, Joanna; Malicka, Joanna; Gryczyński, Ignacy; Lakowicz, Joseph R.

doi:10.1023/b:jofl.0000031823.04926.75

Cited by 57 publications

(42 citation statements)

References 26 publications

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“…As expected, no difference in the emission spectra was detected. This is consistent with earlier results using Texas red-labeled bovine serum albumin [35]. The intensity of fluorescence at 575 nm was enhanced 4 times.…”

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confidence: 93%

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Decreasing photobleaching by silver island films: application to muscle

Muthu

Gryczyński

et al. 2007

Analytical Biochemistry

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Recently it has become possible to study interactions between proteins at the level of single molecules. This requires collecting data from an extremely small volume, small enough to contain one molecule-typically of the order of attoliters (10 −18 L). Collection of data from such a small volume with sufficiently high signal-to-noise ratio requires that the rate of photon detection per molecule be high. This calls for a large illuminating light flux, which in turn leads to rapid photobleaching of the fluorophores that are labeling the proteins. To decrease photobleaching, we measured fluorescence from a sample placed on coverslips coated with silver island films (SIF). SIF reduce photobleaching because they enhance fluorescence brightness and significantly decrease fluorescence lifetime. Increase in the brightness effectively decreases photobleaching because illumination can be attenuated to obtain the same fluorescence intensity. Decrease of lifetime decreases photobleaching because short lifetime minimizes the probability of oxygen attack while the fluorophore is in the excited state. The decrease of photobleaching was demonstrated in skeletal muscle. Myofibrils were labeled lightly with rhodamine-phalloidin, placed on coverslips coated with SIF, illuminated by total internal reflection, and observed through a confocal aperture. We show that SIF causes the intensity of phalloidin fluorescence to increase 4-to 5-fold and its fluorescence lifetime to decrease on average 23-fold. As a consequence, the rate of photobleaching of four or five molecules of actin of a myofibril on Olympus coverslips coated with SIF decreased at least 30-fold in comparison with photobleaching on an uncoated coverslip. Significant decrease of photobleaching makes the measurement of signal from a single cross-bridge of contracting muscle feasible. Keywords Photobleaching; Silver island films; MicroscopyRecently it has become possible to study single protein molecules in a cell. This avoids problems associated with averaging responses from an assembly of molecules with different kinetics and problems associated with diluting of proteins in vitro. The difficulties of studying large assemblies of molecules are well illustrated by the problem of muscle contraction. Contraction results from the interactions of myosin cross-bridges with actin. Myosin crossbridges act asynchronously, i.e., at any time during muscle contraction each one is in a different part of the mechanochemical cycle. Therefore measurement taken from an assembly of crossbridges at any time during contraction is an average value, likely to obscure details of crossbridge kinetics. Furthermore, it is desirable to detect signal from a working muscle, because contractile proteins may behave differently in solution and in whole muscle, where protein ⋆ Dedicated to Professor Avraham Oplatka on the occasion of his birthday. To be able to obtain information from the individual molecules in muscle, it is necessary to collect data from an extremely small volume, small enough to...

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confidence: 93%

“…The observed increase of brightness can be caused by the (i) increase of rhodamine quantum yield, (ii) the decrease in self-quenching [38], or (iii) the enhancement of the local electromagnetic field [35].…”

Section: Discussionmentioning

confidence: 99%

Decreasing photobleaching by silver island films: application to muscle

Muthu

Gryczyński

et al. 2007

Analytical Biochemistry

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“…19). The signal on SCCS is almost twice as strong as that on SIFs [67]. Next, we measured life-times and again found that the intensity decays on SCCS are more affected than on SIFs (Fig.…”

Section: Comparison Of Fluorescence Enhancements On Sifs and Sccsmentioning

confidence: 76%

Advances in Surface-Enhanced Fluorescence

Lakowicz

Geddes

Gryczyński

et al. 2004

Journal of Fluorescence

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306

227

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We report recent achievements in metal-enhanced fluorescence from our laboratory. Several fluorophore systems have been studied on metal particle-coated surfaces and in colloid suspensions. In particular, we describe a distance dependent enhancement on silver island films (SIFs), release of self-quenching of fluorescence near silver particles, and the applications of fluorescence enhancement near metalized surfaces to bioassays. We discuss a number of methods for various shaped silver particle deposition on surfaces.

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“…3-aminopropyltriethoxysilane (APS) was obtained from Fluka. The silver colloids used in these experiments were prepared by chemical reduction of silver nitrate with trisodium citrate using a modified Lee and Meisel method [21][22][23]. According to this method the sodium citrate first acts as a reducing agent, and later the negative citrate ions are adsorbed on silver nanoparticles.…”

Section: Methodsmentioning

confidence: 99%

Enhancement of FITC Fluorescence by Silver Colloids and Silver Island Films

et al. 2008

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Silver colloids and silver island films as fluorescence enhancement systems are the subject of this paper. Enhancement of fluorescence of molecules placed near metallic nanoparticles is a result of resonance interaction of exciting and emitted lights with surface plasmons. Two models explaining this phenomenon are presented. Processes of silver colloids and silver island films forming are described. Results of extinction, absorption and scattering studies of fluorescence enhancement systems are presented. The influence of these systems, i.e. silver colloids and silver island films on the fluorescence of fluorescein-5-isothiocyanate was studied. 7-fold increase in fluorescence intensity of fluorescein-5-isothiocyanate molecules placed near silver nanoparticles was observed.

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Fluorescence Enhancements on Silver Colloid Coated Surfaces

Cited by 57 publications

References 26 publications

Decreasing photobleaching by silver island films: application to muscle

Decreasing photobleaching by silver island films: application to muscle

Advances in Surface-Enhanced Fluorescence

Enhancement of FITC Fluorescence by Silver Colloids and Silver Island Films

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