Surface Plasmon Resonance Chip Research Articles

Azide-Functionalized Naphthoxyloside as a Tool for Glycosaminoglycan Investigations.

We present a xylosylated naphthoxyloside carrying a terminal azide functionality that can be used for conjugation using click chemistry. We show that this naphthoxyloside serves as a substrate for β4GalT7 and induces the formation of soluble glycosaminoglycan (GAG) chains with physiologically relevant lengths and sulfation patterns. Finally, we demonstrate its usefulness by conjugation to the Alexa Fluor 647 and TAMRA fluorophores and coupling to a surface plasmon resonance chip for interaction studies with the hepatocyte growth factor known to interact with the GAG heparan sulfate.

Interfacing Live Cells with Surfaces: A Concurrent Control Technique for Quantifying Surface Ligand Activity.

Surface ligand activity is a key design parameter for successfully interfacing surfaces with cells─whether in the context of in vitro investigations for understanding cellular signaling pathways or more applied applications in drug delivery and medical implants. Unlike other crucial surface parameters, such as stiffness and roughness, surface ligand activity is typically based on a set of assumptions rather than directly measured, giving rise to interpretations of cell adhesion that can vary with the assumptions made. To fill this void, we have developed a concurrent control technique for directly characterizing in vitro ligand surface activity. Pairs of gold-coated glass chips were biofunctionalized with RGD ligand in a parallel workflow: one chip for in vitro applications and the other for surface plasmon resonance (SPR)-based RGD activity characterization. Recombinant αVβ3 integrins were injected over the SPR chip surface as mimics of the cellular-membrane-bound receptors and the resulting binding kinetics parameterized to quantify surface ligand activity. These activity measurements were correlated with cell morphological features, measured by interfacing MDA-MB-231 cells with the in vitro chip surfaces on the live cell microscope. We demonstrate how the interpretation of a cell phenotype based on direct activity measurements can vary markedly from interpretations based on assumed activity. The SPR concurrent control approach has multiple advantages due to the fact that SPR is a standardized technique and has the sensitivity to measure ligand activity across the most relevant range of extracellular surface densities, while the in vitro chip design can be used with all commonly used light microscopy modalities (e.g., phase contrast, DIC, and fluorescence) so that a wide range of phenotypic and molecular markers can be correlated to the ligand surface activity.

Molecular mechanism and thermodynamic study of Rosuvastatin interaction with human serum albumin using a surface plasmon resonance method combined with a multi-spectroscopic, and molecular modeling approach

Rosuvastatin (ROS) is an anti-cholesterol drug belonging to statin drugs. A multi-spectroscopic approach combined with a molecular modeling technique was used to assess ROS association with human serum albumin (HSA). Besides, an HSA immobilized surface plasmon resonance (SPR) chip was used to obtain kinetic parameters (ka, kd, and KD). Fluorescence quenching titrations revealed that ROS interacts with HSA via a dynamic, exothermic, enthalpy-driven mechanism. Hydrogen bonds and van der Waals interactions as the most prevalent bonding forces contribute to ROS-HSA complex formation. ROS binding to HSA alters HSA conformation. The SPR results indicated that ROS and HSA have a strong interaction possessing an equilibrium constant (KD) of 1.55 × 10−8 M at 298 K. A competitive analysis of site markers showed that ROS has a higher tendency to bind to the warfarin binding site (site IIA), which may explain why warfarin has a higher anticoagulant effect in ROS users. FRET analysis indicated that non-radiation energy transfer occurred between ROS and HSA. According to molecular docking studies, ROS prefers binding sites IB and IIA while the ROS-HSA complex stabilizes due to the hydrogen bond and π-π interaction. The presence of hydrogen-bond donors and acceptors, as well as aromatic ROS moieties, facilitates such interactions.

Surface Plasmon Resonance for Protease Detection by Integration of Homogeneous Reaction.

The heterogeneous assays of proteases usually require the immobilization of peptide substrates on the solid surface for enzymatic hydrolysis reactions. However, immobilization of peptides on the solid surface may cause a steric hindrance to prevent the interaction between the substrate and the active center of protease, thus limiting the enzymatic cleavage of the peptide. In this work, we reported a heterogeneous surface plasmon resonance (SPR) method for protease detection by integration of homogeneous reaction. The sensitivity was enhanced by the signal amplification of streptavidin (SA)-conjugated immunoglobulin G (SA-IgG). Caspase-3 (Cas-3) was determined as the model. A peptide labeled with two biotin tags at the N- and C-terminals (bio-GDEVDGK-bio) was used as the substrate. In the absence of Cas-3, the substrate peptide was captured by neutravidin (NA)-covered SPR chip to facilitate the attachment of SA-IgG by the avidin-biotin interaction. However, once the peptide substrate was digested by Cas-3 in the aqueous phase, the products of bio-GDEVD and GK-bio would compete with the substrate to bond NA on the chip surface, thus limiting the attachment of SA-IgG. The method integrated the advantages of both heterogeneous and homogeneous assays and has been used to determine Cas-3 inhibitor and evaluate cell apoptosis with satisfactory results.

Continuous polymerase chain reaction microfluidics integrated with a gold-capped nanoslit sensing chip for Epstein-Barr virus detection

We present the first combination of a microfluidic polymerase chain reaction (PCR) with a gold nanoslit-based surface plasmon resonance (SPR) sensor for detecting the DNA sequence of latent membrane protein 1 (LMP1). The PCR microchannel was produced through a laser scribing technique, and the SPR nanoslit chip was manufactured via hot-embossing nanoimprinting lithography. Afterward, the LMP1 DNA probe was adsorbed onto the SPR chip of the integrated device through electrostatic interactions for further detection. The device can complete the analytical procedure in around 36 min, while the traditional machine requires 105 min to achieve similar signals under the same PCR thermal cycles. The calibration curve with serially diluted LMP1 DNA exhibited the accuracy (R2 > 0.99) and sensitivity (limit of detection: ∼10−11 g/mL) of the device. Moreover, extracted DNA from Epstein-Barr virus (EBV)-positive cells were directly detected through the integrated chip. In brief, this all-in-one chip can amplify gene fragments at the front-end and detect them at the back-end, decreasing the time required for the analysis without compromising accuracy or sensitivity. We believe this label-free, real-time, low-cost device has enormous potential for rapid detection of various viruses, such as EBV and COVID-19.

Real‐Time Detection of Fibrinogen via Imprinted Recognition Sites

AbstractIn this study, the fibrinogen imprinted surface plasmon resonance (SPR) biosensor was developed for the monitoring of fibrinogen levels in patient serum samples using a microcontact imprinting method. The fibrinogen imprinted and non‐imprinted polymeric films on the SPR chip surface were prepared by the microcontact imprinting approach and characterized by ellipsometer, contact angle measurements, and Fourier transform infrared spectroscopy. The limit of detection and quantification values of the fibrinogen imprinted SPR biosensors in the linear range of 0.05–9.0 mg/mL were 0.00066 and 0.00219 mg/mL, respectively. In all kinetic analyzes, the response time was 11 min for equilibration, adsorption, and desorption cycles. The selectivity studies of SPR biosensors were performed in the presence of fibrinogen, myoglobin, hemoglobin, and immunoglobulin G in a competitive manner. For the validation studies of the fibrinogen imprinted SPR biosensor, the fibrinogen level of diluted patient serum samples was determined by enzyme‐linked immunosorbent assay (ELISA). The experimental results show that the serum samples of patients in high and low‐risk groups were identified with the recovery of about % 97–99 according to both SPR biosensor and ELISA analysis results.

Patulin Imprinted Nanoparticles Decorated Surface Plasmon Resonance Chips for Patulin Detection

In this study, the patulin imprinted and the non-imprinted nanoparticles are synthesized by the two-phase mini emulsion polymerization method and characterized by zeta-size analysis, Fourier transform infrared spectroscopy, and scanning electron microscopy. Afterwards, the patulin imprinted and the non-imprinted nanoparticles are attached on the surface of surface plasmon resonance (SPR) chips. The patulin imprinted and the non-imprinted SPR nanosensors are characterized by using atomic force microscope, ellipsometer, and contact angle measurements. Kinetic studies for patulin detection are carried out in the concentration range of 0.5 nmolar–750 nmolar. The limit of detection and the limit of quantification values are obtained as 0.011 nmolar and 0.036 nmolar, respectively. In all kinetic analysis, the response time is 13 min for equilibration, adsorption, and desorption cycles. The selectivity studies of the patulin imprinted and the non-imprinted SPR nanosensors are determined in the presence of ochratoxin A and aflatoxin B1. In order to demonstrate the applicability, validation studies of the patulin imprinted SPR nanosensor are performed by liquid chromatography-tandem mass spectrometry (LC-MS).

Surface plasmon resonance aptasensor based on niobium carbide MXene quantum dots for nucleocapsid of SARS-CoV-2 detection.

A novel label-free surface plasmon resonance (SPR) aptasensor has been constructed for the detection of N-gene of SARS-CoV-2 by using thiol-modified niobium carbide MXene quantum dots (Nb2C-SH QDs) as the bioplatform for anchoring N-gene-targeted aptamer. In the presence of SARS-CoV-2 N-gene, the immobilized aptamer strands changed their conformation to specifically bind with N-gene. It thus increased the contact area or enlarged the distance between aptamer and the SPR chip, resulting in a change of the SPR signal irradiated by the laser (He-Ne) with the wavelength (λ) of 633 nm. Nb2C QDs were derived from Nb2C MXene nanosheets via a solvothermal method, followed by functionalization with octadecanethiol through a self-assembling method. Subsequently, the gold chip for SPR measurements was modified with Nb2C-SH QDs via covalent binding of the Au-S bond also by self-assembling interaction. Nb2C-SH QDs not only resulted in high bioaffinity toward aptamer but also enhanced the SPR response. Thus, the Nb2C-SH QD-based SPR aptasensor had low limit of detection (LOD) of 4.9 pg mL−1 toward N-gene within the concentration range 0.05 to 100 ng mL−1. The sensor also showed excellent selectivity in the presence of various respiratory viruses and proteins in human serum and high stability. Moreover, the Nb2C-SH QD-based SPR aptasensor displayed a vast practical application for the qualitative analysis of N-gene from different samples, including seawater, seafood, and human serum. Thus, this work can provide a deep insight into the construction of the aptasensor for detecting SARS-CoV-2 in complex environments.Graphical abstract A novel label-free surface plasmon resonance aptasensor has been constructed to detect sensitively and selectively the N-gene of SARS-CoV-2 by using thiol-modified niobium carbide MXene quantum dots as the scaffold to anchor the N-gene-targeted aptamer.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00604-021-04974-z.

Bovine Serum Albumin Protein Detection by a Removable SPR Chip Combined with a Specific MIP Receptor

Nowadays, the development of simple, fast, and low-cost selective sensors to detect substances of interest is of great importance in several application fields. Among this kind of sensors, those based on surface plasmon resonance (SPR) represent a promising category, since they are highly sensitive, versatile, and label-free. In this work, an SPR probe, based on a poly(methyl methacrylate) (PMMA) slab waveguide covered by a gold nanofilm, combined with a specific molecularly imprinted polymer (MIP) receptor for bovine serum albumin (BSA) protein, has been realized and experimentally characterized. The obtained experimental results have shown a limit of detection (LOD) equal to about 8.5 × 10−9 M. This value is smaller than the one achieved by another SPR probe, based on a D-shaped plastic optical fiber (POF), functionalized with the same MIP receptor; more specifically, the obtained LOD was reduced by about three orders of magnitude with respect to the POF configuration. Moreover, concerning the D-shaped POF configuration, no manufacturing process is present in the proposed sensor configuration. In addition, fibers are used only to connect the simple sensor chip with a light source and a detector, promoting a bio-chemical sensing approach based on disposable, low-cost, and removable chips.

Sensitive and selective detection of amitrole based on molecularly imprinted nanosensor.

SPR sensor used for amitrole detection was prepared without using any modification. Molecularly imprinted SPR sensor enabled high selectivity for amitrole pesticide. Amino acid-based functional monomer MATrp was integrated as a recognition element. Tailor-made SPR sensor enables real-time monitoring of amitrole pesticide. Synthetic recognition sites provided by MATrp were prepared without labeling.

Theoretical analysis of sensitivity enhancement of surface plasmon resonance biosensor with zinc oxide and blue phosphorus/MoS2 heterostructure

In this paper, an ultrasensitive surface plasmon resonance (SPR) biosensor structure (CaF2 Prism /ZnO/Au/BlueP-MoS2/Sensing medium) based on angular interrogation technique has been studied to see the effect of Blue Phosphorus/MoS2 heterostructure with an adhesive layer of zinc oxide (ZnO). The adhesive layer is ZnO used between calcium fluoride (CaF2) prism and gold (Au) layer. Transfer matrix method is used for numerical analysis at operating wavelength 633 nm. The layers ZnO and Blue Phosphorus/MoS2 heterostructure are optimized to achieve the maximum sensitivity with minimum reflectance. The performance parameters, such as sensitivity (2280RIU−1), quality factor (56.211RIU−1), detection accuracy (0.2810), full width at half maximum (4.0561), and limit of detection (4.3859 × 10−6) are calculated for proposed SPR biosensor at room temperature. For two layers of BlueP/MoS2 heterostructure, the maximum sensitivity is 2350RIU−1. The result shows that the sensitivity of proposed structure is 17.5% greater as compared to the conventional structure. Transverse magnetic (TM) field is plotted, and the value of penetration depth is calculated as 145.25 nm for proposed SPR sensor. The proposed SPR sensor could be very useful in the SPR chip to detect biomolecules or analyte in visible range.

Surface Plasmon Resonance for In-Plane Birefringence Measurement of Anisotropic Thin Organic Film

The measurement of in-plane birefringence (\(\Delta {n}\)) of ultrathin film is challenging due to a significant deviation of physical properties of materials in ultrathin regime as compared to that in bulk state. Surface plasmon resonance (SPR) phenomenon can be employed to measure change in refractive index of ultrathin film at a very high resolution. This article discusses simulation of SPR phenomenon in Kretschmann configuration for the measurement of \(\Delta {n}\) in organic thin film exhibiting nematic-like ordering on the two-dimensional gold surface. The distribution of plasmonic field on the gold surface was found to be anisotropic. This suggested that the coupling plasmonic field with that of organic thin film exhibiting nematic-like ordering on the gold surface will be non-isotropic. Therefore, a nonzero difference in resonance angle (RA) was obtained from SPR measurement performed along the optic-axis (OA) and orthogonal to OA of the in-plane nematic ordering (\(\Delta \theta\)). A calibration surface showing the variation of (\(\Delta \theta\)) as a function of \(\Delta {n}\) and thickness of thin organic film consisting of shape anisotropic tilted molecules exhibiting nematic-like ordering on gold surface was obtained. This calibration surface was employed for the measurement of \(\Delta {n}\) of single layer of Langmuir–Blodgett films of cadmium stearate (CdSA) and 4’-octyl-4-biphenylcarbonitrile (8CB) deposited on SPR chips. The thickness of the LB films was estimated using X-ray reflectivity measurement, and \(\Delta \theta\) was measured using a home built SPR instrument. The \(\Delta {n}\) values were found to be 0.012 and 0.022 for ultrathin films of CdSA and 8CB molecules, respectively.

Preparation of surface plasmon resonance-based nanosensor for curcumin detection.

In this study, the curcumin imprinted and the non-imprinted poly(2-hydroxyethyl methacrylate-N-methacryloyl-L-tryptophan) (poly(HEMA-MATrp)) nanoparticle based surface plasmon resonance (SPR) nanosensors were prepared for the detection of curcumin and characterized by zeta-size analysis, Fourier transform infrared spectroscopy, and scanning electron microscopy. After, the curcumin imprinted and the non-imprinted nanoparticles are attached on the surface of SPR chips. The curcumin imprinted and the non-imprinted SPR nanosensors are characterized by using atomic force microscope, ellipsometer, and contact angle measurements. Kinetic studies were carried out with curcumin aqueous solution at a concentration range of 0.01-150 mg/L using the curcumin imprinted and the non-imprinted SPR nanosensors. In all kinetic analysis, the response time is 14 min for equilibration, adsorption, and desorption cycles. The limit of detection and limit of quantification for the curcumin imprinted SPR nanosensors was 0.0012 mg/L and 0.0040 mg/L, respectively. The validity of the curcumin imprinted SPR nanosensors in real samples was carried out using liquid chromatography-tandem mass spectrometry (LC-MS).

Highly sensitive antimonene-coated black phosphorous-based surface plasmon-resonance biosensor for DNA hybridization: design and numerical analysis

The design of an ultrasensitive surface plasmon resonance (SPR) sensor for the detection of deoxyribose nucleic acid (DNA) hybridization is highly desirable for biomedical applications. We have proposed an SPR sensor architecture (Prism-SF10/Al/Si/BP/Antimonene/phosphate buffer saline solution) and performed its numerical analysis for DNA hybridization using angular interrogation at 633 nm. Antimonene is used as a biorecognition element layer for better attachment of single stranded (ss), i.e., ss probe DNA on its surface, due to its higher binding energy for ssDNA (even greater than graphene). Silicon and black phosphorous are utilized to enhance the electromagnetic field at the sensing layer interface. Aluminum is used as an surface plasmon active metal and it also improves the quality factor. Remarkable sensitivity (202.37 deg / refractive index unit) is attained for this work, which is greater than any other SF10 prism-based SPR sensor proposed to date. The performance of the SPR biosensor is meticulously evaluated as limit-of-detection (0.0028 nM) and figure-of-merit (0.037 nM − 1) with respect to the concentration of complementary target ssDNA in the sensing medium. This versatile biosensor shows great application to detect deadly diseases via DNA hybridization in the biomedical field if developed as an SPR chip for experimental verification.

Detection of amoxicillin residues in egg extract with a molecularly imprinted polymer on gold microchip using surface plasmon resonance and quartz crystal microbalance methods.

In this study, surface plasmon resonance (SPR) and quartz crystal microbalance (QCM) sensors were prepared for the detection of amoxicillin from the commercial and local chicken eggs by using molecular imprinting technique. Amoxicillin imprinted poly(hydroxyethyl methacrylate-methacrylic acid) polymeric film was synthesized onto the surface of the SPR and QCM chips by ultra violet polymerization to determine lower concentrations of amoxicillin. Ellipsometry, contact angle analysis, and atomic force microscopy measurements were used for the surface morphology of the polymeric film layer. The ellipsometric thickness of AMOX imprinted and nonimprinted SPR and QCM chip surfaces were measured as 35 ± 0.9nm,32.89 ± 1.9nm,30 ± 0.6nm, and 28 ± 0.22nm,respectively. Contact angles of bare gold surfaces, AMOX imprinted SPR and QCM chip surfaces were measured to be as 82.3° ± 0.15, 79.2° ± 0.14, 75.01° ± 1.07, and 69.11° ± 0.89, respectively. The range of linearity was measured as 0.1 to 10ng/mL for amoxicillin imprinted SPR and QCM sensors. The maximum residue limit of AMOX in eggs is at 10µg/kg in accordance with the "Positive List System for Agricultural Chemical Residues in Foods." The response time for the test, including adsorption, desorption, and regeneration, was approximately 45min. The limit of detections for SPR and QCM sensors were found to be 0.0005 and 0.0023ng/mL, respectively. The reusabilities of amoxicillin imprinted SPR and QCM sensors were observed by the equilibration-binding-regeneration. Validation studies of the AMOX imprinted SPR and QCM sensors were performed by liquid chromatography-tandem mass spectrometry.

Microfluidic Sensing System with a Multichannel Surface Plasmon Resonance Chip: Damage-Free Characterization of Cells by Pattern Recognition.

The development of a versatile sensing strategy for the damage-free characterization of cultured cells is of great importance for both fundamental biological research and industrial applications. Here, we present a pattern-recognition-based cell-sensing approach using a multichannel surface plasmon resonance (SPR) chip. The chip, in which five cysteine derivatives with different structures are immobilized on Au films, is capable of generating five unique SPR sensorgrams for the cell-secreted molecules that are contained in cell culture media. An automatic statistical program was built to acquire kinetic parameters from the SPR sensorgrams and to select optimal parameters as "pattern information" for subsequent multivariate analysis. Our system rapidly (∼10 min) provides the complex information by merely depositing a small amount of cell culture media (∼25 μL) onto the chip, and the amount of information obtained is comparable to that furnished by a combination of conventional laborious biochemical assays. This noninvasive pattern-recognition-based cell-sensing approach could potentially be employed as a versatile tool for characterizing cells.

Reusable surface plasmon resonance biosensor chip for the detection of H1N1 influenza virus

We developed a reusable magnetic surface plasmon resonance (SPR) sensor chip for detecting various target molecules repeatedly in a conventional SPR system. Here, ferromagnetic patterns on a SPR sensor chip were utilized to trap a layer of magnetic particles, and they were utilized as a solid substrate for SPR sensing in a conventional SPR system. After a sensing experiment, the used magnetic particles were removed by external magnetic fields, and a new layer of magnetic particles was immobilized to the SPR sensor chip for additional sensing measurements. Since magnetic particles were trapped on the ferromagnetic patterns, we could use our reusable SPR chip for SPR sensing measurements in a traditional SPR system without any applied magnetic fields. Significantly, ferromagnetic patterns on the sensor chip surface deflected the strong external fields, so that the large aggregation of magnetic particles on the sensor surface was reduced. We demonstrated using a single reusable SPR sensor chip to measure the nucleoprotein (NP) of H1N1 influenza virus solution ranging repeatedly for more than 7 times without significant signal degradation. Also, different target molecules could be repeatedly measured in a single SPR chip. Since our reusable SPR sensor chip can be repeatedly used in a conventional SPR system without any chemical processes for refreshment, the cost for SPR sensing should be significantly reduced. In this case, our reusable SPR sensor chip can be a major breakthrough and can be used for versatile practical applications of SPR sensors.

Antibody Epitope and Affinity Determination of the Myocardial Infarction Marker Myoglobin by SPR-Biosensor Mass Spectrometry.

Myoglobin (MG) is a biomarker for heart muscle injury, making it a potential target protein for early detection of myocardial infarction. Elevated myoglobin levels alone have low specificity for acute myocardial infarction (AMI) but in combination with cardiac troponin T have been considered highly efficient diagnostic biomarkers. Myoglobin is a monomeric heme protein with a molecular weight of 17 kDa that is found in skeletal and cardiac tissue as an intracellular storage unit of oxygen. MG consists of eight α-helices connected by loops and a heme group responsible for oxygen-binding. Monoclonal antibodies are widely used analytical tools in biomedical research and have been employed for immunoanalytical detection of MG. However, the epitope(s) recognized by MG antibodies have been hitherto unknown. Precise molecular identification of the epitope(s) recognized by antibodies is of key importance for the development of MG as a diagnostic biomarker. The epitope of a monoclonal MG antibody was identified by proteolytic epitope extraction mass spectrometry in combination with surface plasmon resonance (SPR) biosensor analysis. The MG antibody was immobilized both on an affinity microcolumn and a gold SPR chip. The SPR kinetic analysis provided an affinity-binding constant KD of 270 nM for MG. Binding of a tryptic peptide mixture followed by elution of the epitope from the SPR-MS affinity interface by mild acidification provided a single-epitope peptide located at the C-terminus [146-153] [YKELGFQG] of MG. The specificity and affinity of the epitope were ascertained by synthesis and affinity-mass spectrometric characterization of the epitope peptide.

Sensitivity Improvement of Surface Plasmon Resonance Sensor on Using BlueP/MoS2 Heterostructure and Antimonene

Here, an ultrasensitive surface plasmon resonance (SPR) sensor based on angular interrogation using antimonene and BlueP/MoS2 heterostructure is proposed. Antimonene is used as an interacting layer for analytes due to its higher binding energy for analyte adsorption. BlueP/MoS2 enhances the sensitivity by manipulating charge transfer to metal/dielectric interface. Sensitivity for the proposed SPR is analyzed by tuning the number of heterostructure, as well as antimonene layers for wide range of refractive index. Highest sensitivity of 198.4°/RIU is achieved for three layers of BlueP/MoS2. Results show significant sensitivity for proposed design in comparison to conventional and graphene based SPR sensor. Proposed SPR sensor can be successfully developed as SPR chip for biomolecule/analyte detection in visible range.

Platinum substrate for surface plasmon microscopy at small angles.

Platinum is reported as the main component of the substrate in surface plasmon microscopy of the metal-dielectric interface for small-angle measurements. In the absence of a narrow dip in the angular spectrum of platinum, the refractive index of the dielectric medium or the thickness of a deposited layer is proven deducible from the observed sharp peak, close to the critical angle. The sensitivities of refractive index and thickness measurements using platinum are compared with that of a gold surface plasmon resonance chip. Furthermore, the thickness of a structured layer of (3-Aminopropyl)triethoxysilane on the platinum substrate is measured to be 0.7 nm, demonstrating the high sensitivity of the technique.