Metal‐Enhanced Fluorescence: Application to High‐Throughput Screening and Drug Discovery

Aslan, Kadir; Gryczyński, Ignacy; Malicka, Joanna; Lakowicz, Joseph R.; Geddes, Chris D.

doi:10.1002/0471728780.ch14

Cited by 7 publications

(12 citation statements)

References 39 publications

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“…SIFs are routinely used in MEF-based biosensing applications [15, 19, 20]. In this regard, the homogeneous deposition of SIFs onto planar substrates is crucial for quantitative determination of target biomolecules.…”

Section: Resultsmentioning

confidence: 99%

“…Since the fluorescence emission from SIFs is significantly larger than those from blank glass slides, the contribution of fluorophore / protein coverage to the increased fluorescence emission has to be determined for accurate evaluation of metal nanoparticle-deposited surfaces for MEF applications. Despite the presence of numerous publications on MEF phenomenon [16-18] and MEF-based applications [15, 19, 20], the quantitative comparison of fluorophore or protein surface coverage on SIFs and blank glass slides was never investigated.…”

Section: Introductionmentioning

confidence: 99%

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Quantitative Comparison of Protein Surface Coverage on Glass Slides and Silver Island Films in Metal-Enhanced Fluorescence-based Biosensing Applications

Grell¹,

Paredes²,

Das³

et al. 2010

Nano BioMed ENG

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The use of Metal-Enhanced Fluorescence (MEF) phenomenon in fluorescence-based bioassays affords for increased sensitivity to be realized by incorporating metal nanoparticles onto planar surfaces. The close-range interactions of metal-fluorophores result in increased fluorescence emission from the bioassays, which in turn affords for the detection of target biomolecules at lower concentrations. Moreover, the use of silver nanoparticles increases the photostability of fluorophores improving the detectability of fluorescence emission under prolonged use of excitation light. Although numerous reports on MEF-based biosensing applications exist, the contribution of protein coverage on Silver Island Films (SIFs) on the increased fluorescence emission was never investigated. This work presents our findings on the quantitative comparison of protein surface coverage on SIFs and blank glass slides. In this regard, identical protein bioassay for a model protein (biotinylated bovine serum albumin, b-BSA) on these surfaces is constructed and the relative extent of protein surface coverage on SIFs and blank glass slides was determined using radio-labeled biomolecules. It was found that the total scintillation counts on SIFs and blank glass slides were similar for BSA concentrations ranging from 1 μM to 1 pM, which implies that increased fluorescence in MEF-based biosensing applications is only due to metal-fluorophore interactions.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Quantitative Comparison of Protein Surface Coverage on Glass Slides and Silver Island Films in Metal-Enhanced Fluorescence-based Biosensing Applications

Grell¹,

Paredes²,

Das³

et al. 2010

Nano BioMed ENG

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“…Biotin-avidin interactions are one of the strongest biological interactions in nature [32], and are typically employed in proof-of-principle demonstration of new assay techniques [5, 37, 38]. The biotin-avidin interactions typically reach >95% completion in 10–20 min for all concentrations [39] and subsequently we did not seek longer incubation times.…”

Section: Resultsmentioning

confidence: 99%

Rapid and Sensitive Colorimetric ELISA Using Silver Nanoparticles, Microwaves and Split Ring Resonator Structures

Addae¹,

Pinard²,

Çağlayan³

et al. 2010

Nano BioMed ENG

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We report a new approach to colorimetric Enzyme-Linked Immunosorbent Assay (ELISA) that reduces the total assay time to < 2 min and the lower-detection-limit by 100-fold based on absorbance readout. The new approach combines the use of silver nanoparticles, microwaves and split ring resonators (SRR). The SRR structure is comprised of a square frame of copper thin film (30 μm thick, 1 mm wide, overall length of ~9.4 mm on each side) with a single split on one side, which was deposited onto a circuit board (2x2 cm 2 ). A single micro-cuvette (10 μl volume capacity) was placed in the split of the SRR structures. Theoretical simulations predict that electric fields are focused in and above the micro-cuvette without the accumulation of electrical charge that breaks down the copper film. Subsequently, the walls and the bottom of the micro-cuvette were coated with silver nanoparticles using a modified Tollen's reaction scheme. The silver nanoparticles served as a mediator for the creation of thermal gradient between the bioassay medium and the silver surface, where the bioassay is constructed. Upon exposure to low power microwave heating, the bioassay medium in the micro-cuvette was rapidly and uniformly heated by the focused electric fields. In addition, the creation of thermal gradient resulted in the rapid assembly of the proteins on the surface of silver nanoparticles without denaturing the proteins. The proof-of-principle of the new approach to ELISA was demonstrated for the detection of a model protein (biotinylated-bovine serum albumin, b-BSA). In this regard, the detection of b-BSA with bulk concentrations (1 µM to 1 pM) was carried out on commercially available 96-well high throughput screening (HTS) plates and silver nanoparticle-deposited SRR structures at room temperature and with microwave heating, respectively. While the room temperature bioassay (without microwave heating) took 70 min to complete, the identical bioassay took < 2 min to complete using the SRR structures (with microwave heating). A lower detection limit of 0.01 nM for b-BSA (100-fold lower than room temperature ELISA) was observed using the SRR structures. Keywords

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“…Metal-enhanced fluorescence (MEF), a phenomenon where the quantum yield and photostability of weakly fluorescing species are dramatically increased, is becoming a powerful tool for the fluorescence-based applications of drug discovery, 1,2 high-throughput screening, 3,4 immunoassays 5 and proteinprotein detection. 1,4,6 In all of these examples of MEF, [1][2][3][4][5][6] primarily quartz or glass substrates with deposited silver nanostructures have been used. This has been because the chemistries of the surface of glass (and quartz) are well established and therefore the covalent immobilization of silver nanostructures onto glass less arduous and is indeed reproducibly reliable.…”

Section: Introductionmentioning

confidence: 99%

“…This has been because the chemistries of the surface of glass (and quartz) are well established and therefore the covalent immobilization of silver nanostructures onto glass less arduous and is indeed reproducibly reliable. [1][2][3][4][5][6] Polymeric substrates may, however, provide a suitable, cheap and versatile alternative to the use of chemically benign glass substrates. 7 Although often unsuitable for shortwavelength fluorescence-based detection strategies due to intrinsic polymer fluorescence, several polymers, such as poly(methyl methacrylate) and polycarbonate (PC), typically do not suffer from these detractions and are suitable for use in longer-wavelength (l > 520 nm) fluorescence-based detection schemes.…”

Section: Introductionmentioning

confidence: 99%

Metal-enhanced fluorescence from silver nanoparticle-deposited polycarbonate substrates

2006

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Metal‐Enhanced Fluorescence: Application to High‐Throughput Screening and Drug Discovery

Cited by 7 publications

References 39 publications

Quantitative Comparison of Protein Surface Coverage on Glass Slides and Silver Island Films in Metal-Enhanced Fluorescence-based Biosensing Applications

Quantitative Comparison of Protein Surface Coverage on Glass Slides and Silver Island Films in Metal-Enhanced Fluorescence-based Biosensing Applications

Rapid and Sensitive Colorimetric ELISA Using Silver Nanoparticles, Microwaves and Split Ring Resonator Structures

Metal-enhanced fluorescence from silver nanoparticle-deposited polycarbonate substrates

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