| 1. |
- Dahlin, Andreas, et al.
(författare)
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Synchronized Quartz Crystal Microbalance and Nanoplasmonic Sensing of Biomolecular Recognition Reactions
- 2008
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Ingår i: ACS Nano. - : American Chemical Society (ACS). - 1936-086X .- 1936-0851. ; 2:10, s. 2174-2182
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Tidskriftsartikel (refereegranskat)abstract
- We present a method providing synchronized measurements using the two techniques: quartz crystal microbalance with dissipation (QCM-D) monitoring and localized surface plasmon resonance (LSPR). This was achieved by letting a thin gold film perforated with short-ranged ordered plasmon-active nanoholes act as one of the electrodes of a QCM-D crystal. This enabled transmission-mode optical spectroscopy to be used to temporally resolve colorimetric changes of the LSPR active substrate induced upon bionnolecular binding events. The LSPR response could thus be compared with simultaneously obtained changes in resonance frequency, Delta f, and energy dissipation, AD, of the QCM-D device. Since the LSPR technique is preferentially sensitive to changes within the voids of the nanoholes, while the QCM-D technique is preferentially sensitive to reactions on the planar region between the holes, a surface chemistry providing the same binding kinetics on both gold and silica was used. This was achieved by coating the substrate with poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG), which was shown to bind in the same manner on silica and gold modified with a carboxyl-terminated thiol. In this way, the combined setup provided new information about structural changes upon PLL-g-PEG adsorption. We also demonstrate subsequent binding of NeutrAvidin and an immunoreaction utilizing biotin-modified IgG. The combined information from the synchronized measurements was also used in a new way to estimate the sensing volume of the LSPR sensor.
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| 2. |
- Höök, Fredrik, 1966, et al.
(författare)
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Supported lipid bilayers, tethered lipid vesicles, and vesicle fusion investigated using gravimetric, plasmonic, and microscopy techniques
- 2007
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Ingår i: Biointerphases. - : American Vacuum Society. - 1559-4106 .- 1934-8630. ; 3:2
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Tidskriftsartikel (refereegranskat)abstract
- This article summarizes our most recent contributions to the rapidly growing field of supported lipid assemblies with emphasis on current studies addressing both fundamental and applied aspects of supported lipid bilayer (SLB) and tethered lipid vesicles (TLVs) to be utilized in sensing applications. The new insights obtained from combining the quartz crystal microbalance with dissipation monitoring technique with surface plasmon resonance are described, and we also present recent studies in which nanoplasmonic sensing has been used in studies of SLBs and TLVs. To gain full control over the spatial arrangement of TLVs in both two and three dimensions, we have developed a method for site-selective and sequence-specific sorting of DNA-tagged vesicles to surfaces modified with complementary DNA. The combination of this method with nanoplasmonic sensing formats is covered as well as the possibility of using DNA-modified vesicles for the detection of unlabeled DNA targets on the single-molecule level. Finally, a new method for membrane fusion induced by hybridization of vesicle-anchored DNA is demonstrated, including new results on content mixing obtained with vesicle populations encapsulating short, complementary DNA strands.
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| 3. |
- Jonsson, Magnus, 1981, et al.
(författare)
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Nanoplasmonic biosensing with focus on short-range ordered nanoholes in thin metal films
- 2008
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Ingår i: Biointerphases. - : American Vacuum Society. - 1559-4106 .- 1934-8630. ; 3:3, s. FD30-FD40
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Tidskriftsartikel (refereegranskat)abstract
- The resonance conditions for excitation of propagating surface plasmons at planar metal/dielectric interfaces and localized surface plasmons associated with metal nanostructures are both sensitive to changes in the interfacial refractive index. This has made these phenomena increasingly popular as transducer principles in label-free sensing of biomolecular recognition reactions. In this article, the authors review the recent progress in the field of nanoplasmonic bioanalytical sensing in general, but set particular focus on certain unique possibilities provided by short-range ordered nanoholes in thin metal films. Although the latter structures are formed in continuous metal films, while nanoparticles are discrete entities, these two systems display striking similarities with respect to sensing capabilities, including bulk sensitivities, and the localization of the electromagnetic fields. In contrast, periodic arrays of nanoholes formed in metal films, most known for their ability to provide wavelength-tuned enhanced transmission, show more similarities with conventional propagating surface plasmon resonance. However, common for both short-range ordered and periodic nanoholes formed in metal films is that the substrate is electrically conductive. Some of the possibilities that emerge from sensor templates that are both electrically conductive and plasmon active are discussed and illustrated using recent results on synchronized nanoplasmonic and quartz crystal microbalance with dissipation monitoring of supported lipid bilayer formation and subsequent biomolecular recognition reactions. Besides the fact that this combination of techniques provides an independent measure of biomolecular structural changes, it is also shown to contribute with a general means to quantify the response from nanoplasmonic sensors in terms of bound molecular mass. c 2008 American Vacuum Society.
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| 4. |
- Jonsson, Magnus P., et al.
(författare)
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Simultaneous Nanoplasmonic and Quartz Crystal Microbalance Sensing: Analysis of Biomolecular Conformational Changes and Quantification of the Bound Molecular Mass
- 2008
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Ingår i: Analytical Chemistry. - : American Chemical Society. - 0003-2700 .- 1520-6882. ; 80:21, s. 7988-7995
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Tidskriftsartikel (refereegranskat)abstract
- This paper presents a study of supported lipid bilayer (SIB) formation and subsequent protein binding using a sensor that combines localized surface plasmon resonance (LSPR) and quartz crystal microbalance with dissipation (QCM-D) monitoring. The LSPR activity arises from silicon oxide (SiOx) coated nanometric apertures in a thin gold film, which also serves as the active electrode of a QCM-D crystal. Both transducer principles provide signatures for the formation of a SLB upon adsorption and subsequent rupture of adsorbed lipid vesicles. However, the two techniques are sensitive over different regions of the sample: LSPR primarily inside and on the rim of the holes and QCM-D primarily on the planar areas between the holes. Although the dimension of the lipid vesicles is on the same order as the dimension of the nanoholes, it is concluded from the response of the combined system that vesicle rupture in the nanoholes and on the planar region between the holes is synchronized. Furthermore, by determining the thickness of the SLB from the QCM-D response, the characteristic decay length of the LSPR field intensity could be determined. This made it possible not only to determine the mass and refractive index of the homogeneous SLB but also to postulate a generic means to quantify the LSPR response in terms of mass-uptake also for nonhomogeneous films. This is exemplified by measuring the adsorbed lipid mass during vesicle adsorption, yielding the critical lipid vesicle coverage at which spontaneous rupture into a planar bilayer occurs. The generic applicability and versatility of the method is demonstrated from specific protein binding to a functionalized SLB. From the absolute refractive index of the protein, provided from the LSPR data alone, it was possible to determine both the effective thickness of the protein film and the molecular mass (or number) of bound protein.
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| 5. |
- Jonsson, Magnus, et al.
(författare)
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Supported Lipid Bilayer Formation and Lipid-Membrane-Mediated Biorecognition Reactions Studied with a New Nanoplasmonic Sensor Template
- 2007
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Ingår i: Nano Letters. - : American Chemical Society (ACS). - 1530-6992 .- 1530-6984. ; 7:11, s. 3462-3468
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Tidskriftsartikel (refereegranskat)abstract
- This paper presents the use of the localized surface plasmon resonance (LSPR) sensor concept to probe the formation of macroscopic and laterally mobile supported lipid bilayers (SLBs) on SiOx-encapsulated nanohole-containing Au and Ag films. A comparison between Au- and Ag-based sensor templates demonstrates a higher sensitivity for Au-based templates with respect to both bulk and interfacial refractive index (RI) changes in aqueous solution. The lateral mobility of SLBs formed on the SiOx-encapsulated nanohole templates was analyzed using fluorescence recovery after photobleaching (FRAP), demonstrating essentially complete (>96%) recovery, but a reduction in diffusivity of about 35% compared with SLBs formed on flat SiOx substrates. Furthermore, upon SLB formation, the temporal variation in extinction peak position of the LSPR active templates display a characteristic shape, illustrating what, to the best of our knowledge, is the first example where the nanoplasmonic concept is shown capable of probing biomacromolecular structural changes without the introduction of labels. With a signal-to-noise ratio better than 500 upon protein binding to the cell-membrane mimics, the sensor concept is also proven competitive with state-of-the-art label-free sensors.
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| 6. |
- Jönsson, Peter, et al.
(författare)
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A Method Improving the Accuracy of Fluorescence Recovery after Photobleaching Analysis
- 2008
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Ingår i: Biophysical Journal. - : Biophysical Society. - 0006-3495 .- 1542-0086. ; 95:11, s. 5334-5348
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Tidskriftsartikel (refereegranskat)abstract
- Fluorescence recovery after photobleaching has been an established technique of quantifying the mobility of molecular species in cells and cell membranes for more than 30 years. However, under nonideal experimental conditions, the current methods of analysis still suffer from occasional problems; for example, when the signal/noise ratio is low, when there are temporal fluctuations in the illumination, or when there is bleaching during the recovery process. We here present a method of analysis that overcomes these problems, yielding accurate results even under nonideal experimental conditions. The method is based on circular averaging of each image, followed by spatial frequency analysis of the averaged radial data, and requires no prior knowledge of the shape of the bleached area. The method was validated using both simulated and experimental fluorescence recovery after photobleaching data, illustrating that the diffusion coefficient of a single diffusing component can be determined to within similar to 1%, even for small signal levels (100 photon counts), and that at typical signal levels (5000 photon counts) a system with two diffusion coefficients can be analyzed with less than 10% error.
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| 7. |
- Zhou, Ye, et al.
(författare)
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Quantitative interpretation of gold nanoparticle-based bioassays designed for detection of immunocomplex formation
- 2007
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Ingår i: BIOINTERPHASES. - : American Vacuum Society. - 1559-4106 .- 1934-8630. ; 2:1
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Tidskriftsartikel (refereegranskat)abstract
- The authors present in this paper how the extended Mie theory can be used to translate not only end-point data but also temporal variations of extinction peak-position changes, Delta lambda(peak)(t), into absolute mass uptake, Gamma(t), upon biomacromolecule binding to localized surface plasmon resonance (SPR) active nanoparticles (NPs). The theoretical analysis is applied on a novel sensor template composed of a three-layer surface architecture based on (i) a self-assembled monolayer of HS(CH2)(15)COOH, (ii) a 1:1 mixture of biotinylated and pure poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG), and (iii) NeutrAvidin. Assisted by independent estimations of the thickness of the three-layer architecture using quartz crystal microbalance with dissipation (QCM-D) monitoring, excellent agreement with parallel mass-uptake estimations using planar SPR is obtained. Furthermore, unspecific binding of serum to PLL-g-PEG was shown to be below the detection limit, making the surface architecture ideally suited for label-free detection of immunoreactions. To ensure that the immunocomplex formation occurred within the limited sensing depth (similar to 10 nm) of the NPs, a compact model system composed of a biotinylated human recombinant single-chain antibody fragment (empty set similar to 2 nm) directed against cholera toxin was selected. By tracking changes in the centroid (center of mass) of the extinction peak, rather than the actual peak position, signal-to-noise levels and long-term stability upon cholera toxin detection are demonstrated to be competitive with results obtained using conventional SPR and state-of-the-art QCM-D data. (C) 2007 American Vacuum Society.
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| 8. |
- Åsberg, Peter, et al.
(författare)
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Hydrogels from a water-soluble Zwitterionic polythiophene : dynamics under pH change and biomolecular interactions observed using quartz crystal microbalance with dissipation monitoring
- 2005
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Ingår i: Langmuir. - : American Chemical Society (ACS). - 0743-7463 .- 1520-5827. ; 21:16, s. 7292-7298
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Tidskriftsartikel (refereegranskat)abstract
- The water-soluble zwitterionic polythiophene, poly(3-((S)-5-amino-5- carboxyl-3-oxapentyl)-2,5-thiophene) hydrochloride (POWT), is a conjugated polyelectrolyte (CPE) with properties well suited for biochip applications. CPEs readily form hydrogels when exposed to water-based buffer solutions or biomolecule solutions. In this work, we used in situ quartz crystal microbalance with dissipation (QCM-D) monitoring to collect information on the interaction between POWT films exposed to buffers with different pH and POWT/DNA chains. Our data show that POWT swells significantly when exposed to low-pH buffers, such as pH 4 acetate, this is seen as an increase in thickness and decrease in viscosity obtained via a Voight-based modeling of combined f and D QCM-D measurements. The magnitude of thickness and viscosity change upon changing from a pH 10 carbonate buffer to pH 4 acetate is 100% increase in thickness and 50% decrease in viscosity. The response of the hydrogel under pH change is well correlated with fluorescence data from POWT films on glass. The state of the hydrogel is important during interaction with biomolecules; illustrated by the observation that a swollen CPE hydrogel adsorbs a higher amount of DNA than a compacted one. In agreement with previous results, the QCM-D data confirmed that the POWT/DNA hydrogel sense complementary DNA specifically and with negligible binding of noncomplementary DNA. These results are important for efficient constructions of biochips in water environments using this class of materials.
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