Real-time Surface Plasmon Resonance Imaging Research Articles

Surface plasmon resonance biosensor using hydrogel-AuNP supramolecular spheres for determination of prostate cancer-derived exosomes.

Based on the hydrogel-AuNP supramolecular sphere (H-Au), a label-free and real-time surface plasmon resonance imaging biosensor has been developed for highly sensitive and specific determination of prostate cancer cell-derived exosomes. After integrating the signal amplification effect of themass cumulative hydrogel and theLSPR effect of AuNPs with high specific aptamer, theSPRi biosensor for exosome detection exhibited a wide linear range from 1.00 × 105 to 1.00 × 107 particles/mL with a limit of detection of1.00 × 105 particles/mL. Most importantly, with a strong correlation between the SPRi signal and the t-PSA value measured by the clinical chemiluminescence immunoassay, this biosensor displayed excellent practicability for human serumanalysis, which exhibits great potential applications in disease diagnosis and bioanalysis. Prostate cancer has been one of the most threatening diseases in human life and health nowadays. In particular, as cancer metastasizes, it is more likely to cause fracture, paraplegia, and even fatal consequences. However, the predominant t-PSA test needs further improvement for the deficiencies of limited specificity and sensitivity, which is prone to false positive. As one of the noninvasive markers of liquid biopsies, exosome has the potential to be a substitute for t-PSA, which can provide specific and predictive information in disease diagnosis and prognosis. Herein, based on the hydrogel-AuNP supramolecular sphere (H-Au), a label-free and real-time surface plasmon resonance biosensor has been developed for highly sensitive and specific detection of prostate cancer cell-derived exosomes. After integrating the signal amplification effect of mass cumulative hydrogel and LSPR effect of AuNPs with high specific aptamer, this developed SPRi biosensor for exosome detection exhibited a wide linear range from 1.00 × 105 to 1.00 × 107 particles/mL with a limit of detection down to 1.00 × 105 particles/mL. Most importantly, with a strong correlation between the SPRi signal and the t-PSA value measured by the clinical chemiluminescence immunoassay, this biosensor displayed excellent practicability in human serum, which exhibited great potential applications in disease diagnosis and bioanalysis.

Real-time SPR imaging based on a large area beam from a wavelength-swept laser.

We demonstrate a real-time surface plasmon resonance imaging (SPRi) system based on a wavelength-swept laser. Compared to conventional spectral-modulation SPRi using white light source and spectral filtering, the proposed system has a higher scan rate to detect rapid changes in refractive index and a higher output power for large-area illumination. This SPRi system achieves scan rates faster than 12Hz, simultaneously obtaining SPR dip positions over full illumination fields of 12×12 mm. Using the wavelength-swept laser, two-dimensional biomolecular array imaging can be acquired with a high dynamic detection range (7.67×10-3 refractive index unit (RIU)) as well as high sensitivity (6501nm/RIU) and resolution (1.89×10-6 RIU).

Surface Plasmon Resonance Imaging Microscopy of Liposomes and Liposome-Encapsulated Gold Nanoparticles

Liposomes are small vesicles that can be used in various targeting applications as carrier vehicles. In this paper, we show that real-time surface plasmon resonance imaging microscopy (SPRI microscopy) can be used to detect diffraction patterns of these singular vesicles in water phase at room temperature and without any additives. The diffraction pattern intensities, related to the particle size, are shown to follow the log-normal distribution in a cumulative distribution function (CDF) that is very well in accordance with the normal size distribution of liposomes prepared with the extrusion method. In addition, this distribution is further analyzed to determine the number of gold nanoparticle (GNP) encapsulated liposomes in a set of liposomal adsorption events. Thus, we obtain the encapsulation efficiency and present a method to study the intrinsic properties of liposomes and other soft nanomaterials.

Single-Nanoparticle Near-Infrared Surface Plasmon Resonance Microscopy for Real-Time Measurements of DNA Hybridization Adsorption

A novel 814 nm near-infrared surface plasmon resonance (SPR) microscope is used for the real-time detection of the sequence-selective hybridization adsorption of single DNA-functionalized gold nanoparticles. The objective-coupled, high numerical aperture SPR microscope is capable of imaging in situ the adsorption of single polystyrene and gold particles with diameters ranging from 450 to 20 nm onto a 90 μm × 70 μm area of a gold thin film with a time resolution of approximately 1-3 s. Initial real-time SPR imaging (SPRI) measurements were performed to detect the accumulation of 40 nm gold nanoparticles for 10 min onto a gold thin film functionalized with a 100% complementary DNA surface at concentrations from 5 pM to 100 fM by counting individual particle binding events. A 100% noncomplementary DNA surface exhibited virtually no nanoparticle adsorption. In contrast, in a second set of SPRI measurements, two component complementary/noncomplementary mixed DNA monolayers that contained a very small percentage of complementary sequences ranging from 0.1 to 0.001%, showed both permanent and transient hybridization adsorption of the gold nanoparticles that could be tracked both temporally and spatially with the SPR microscope. These experiments demonstrate that SPR imaging measurements of single biofunctionalized nanoparticles can be incorporated into bioaffinity biosensing methods at subpicomolar concentrations.

Sensitivity Enhancement of Surface Plasmon Resonance Imaging Using Periodic Metallic Nanowires

A nanowire-mediated surface plasmon resonance (SPR) imaging is numerically investigated for enhanced sensitivity. The results calculated by rigorous coupled-wave analysis present that interplays between localized surface plasmons and surface plasmon polaritons contribute to sensitivity enhancement. Compared to conventional thin film-based SPR imaging measurement, an optimal nanowire structure can provide sensitivity enhancement by 3.44 times as well as highly linear detection property for quantification of surface reactions of interests. This paper demonstrates the potential and limitation for a highly sensitive, label-free, and real-time SPR imaging sensor based on periodic metallic nanowires.

Full-field and real-time surface plasmon resonance imaging thermometry

The feasibility of surface plasmon resonance (SPR) imaging thermometry is tested as a potential tool for full-field and real-time temperature field mapping for thermally transient liquid mediums. Using the well-known Kretschmann's analysis [Physik 241, 313 (1971)]. parametric examinations are performed to delineate the effects of important optical properties, including seven different prism materials with different refractive index values and seven different measured dielectric constants for thin gold (Au) films (approximately 47.5 nm in thickness), on the temperature dependence of SPR reflectance intensity variations. Furthermore, a laboratory-implemented real-time SPR thermometry system demonstrates the full-field mapping capabilities for transient temperature field developments in the near-wall region when a hot water droplet (80 degrees C) contacts the Au metal surface (20 degrees C) and spreads either in an air- or in a water-surrounded environment.

Determination of Ribonuclease H Surface Enzyme Kinetics by Surface Plasmon Resonance Imaging and Surface Plasmon Fluorescence Spectroscopy

The kinetics of the ribonuclease H (RNase H) surface hydrolysis of RNA-DNA heteroduplexes formed on DNA microarrays was studied using a combination of real-time surface plasmon resonance imaging (SPRI) and surface plasmon fluorescence spectroscopy (SPFS). Time-dependent SPRI and SPFS data at various enzyme concentrations were quantitatively analyzed using a simple model that couples diffusion, enzyme adsorption, and surface enzyme kinetics. This model is characterized by a set of three rate constants, enzyme adsorption (k(a)), enzyme desorption (k(d)), enzyme catalysis (k(cat)), and one dimensionless diffusion parameter (beta). Values of k(a) = 3.15 (+/-0.20) x 10(6) M(-1).s(-1), k(d) = 0.10 (+/-0.05) s(-1), and k(cat) = 0.95 (+/-0.10) s(-1) were determined from fitting all of the SPRI and SPFS data sets. One of the most interesting kinetic parameters is the surface RNase H hydrolysis reaction rate constant (k(cat)), which was found to be approximately 10 times slower than that observed in solution, but approximately 100 times faster than that recently observed for the exonuclease III surface hydrolysis of double-stranded DNA microarrays (k(cat) = 0.009 s(-1)). Moreover, the surface coverage of the intermediate enzyme-substrate complex (ES) was found to be extremely small during the course of the reaction because k(cat) is much larger than the product of k(a) and the bulk enzyme concentration.

Real-time surface plasmon resonance imaging measurements for the multiplexed determination of protein adsorption/desorption kinetics and surface enzymatic reactions on peptide microarrays.

The kinetics of protein adsorption/desorption onto peptide microarrays was studied using real-time surface plasmon resonance (SPR) imaging. S protein binding interactions were examined using an array composed of five different peptides: N terminal and C terminal immobilized wild-type S peptide (S1 and S2), an alternate binding sequence derived by phage display (LB2), an NVOC-protected S peptide, and a FLAG peptide control sequence (F). Kinetic measurements of the S protein-S1 peptide interaction were analyzed to determine a desorption rate constant (k(d)) of 1.1 (+/-0.08) x 10(-2) s(-1), an adsorption rate constant (k(a)) of 1.9 (+/-0.05) x 10(5) M(-1) s(-1), and an equilibrium adsorption constant (K(Ads)) of 1.7 (+/-0.08) x 10(7) M(-1). SPR imaging equilibrium measurements of S protein to S1 peptide were performed to independently confirm the kinetically determined value of K(Ads). Rate constants for the S2 and LB2 peptides on the array were measured as follows: 1.6 (+/-0.04) x 10(5) M(-1) s(-1) (k(a)) and 1.1 (+/-0.07) x 10(-2) s(-1) (k(d)) for S2, 1.2 (+/-0.05) x 10(5) M(-1) s(-1) (k(a)) and 1.1 (+/-0.03) x 10(-2) s(-1) (k(d)) for LB2. In addition to S protein adsorption/desorption, real-time SPR imaging of peptide arrays was applied to study the surface enzymatic activities of the protease factor Xa. Enzymatic cleavage of the substrate peptide (P1) was shown to follow first-order kinetics and proceed at a rate 10 times faster than that of the mutant peptide (P2), with cleavage velocities of 5.6 (+/-0.3) x 10(-4) s(-1) for P1 and 5.7 (+/-0.3) x 10(-5) s(-1) for P2.