302 research outputs found
Direct optical sensing of single unlabeled small proteins and super-resolution microscopy of their binding sites
More than twenty years ago, scientists succeeded in pushing the limits of
optical detection to single molecules using fluorescence. This breakthrough has
revolutionized biophysical measurements, but restrictions in photophysics and
labeling protocols have motivated many efforts to achieve fluorescence-free
single-molecule sensitivity in biological studies. Although several interesting
mechanisms using vibrational spectroscopy, photothermal detection, plasmonics
or microcavities have been proposed for biosensing at the single-protein level,
no method has succeeded in direct label-free detection of single proteins.
Here, we present the first results using interferometric detection of
scattering (iSCAT) from single proteins without the need for any label, optical
nanostructure or microcavity. Furthermore, we demonstrate super-resolution
imaging of protein binding with nanometer localization precision. The ease of
iSCAT instrumentation promises a breakthrough for industrial usage as well as
fundamental laboratory experiments
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Novel concept of multi-channel fiber optic surface plasmon resonance sensor
A novel multi-channel fiber optic surface plasmon resonance (SPR) sensor is reported. The sensing structure consists of a single-mode optical fiber, covered with a thin gold layer, which supports a surface plasmon (SP), and a Bragg grating. The Bragg grating induces coupling between the forward-propagating fundamental core mode and the back-propagating SP-cladding mode. As the SP-cladding modes are highly sensitive to changes in the refractive index of the surrounding medium, the changes can be accurately measured by spectroscopy of these hybrid modes. Multichannel capability is achieved by employing a sequence of Bragg gratings of different periods and their reading via the wavelength division multiplexing. Theoretical analysis and optimization based on the coupled-mode theory (CMT) is carried out and performance characteristics of the sensor are determined
Generation of infrared surface plasmon resonances with high refractive index sensitivity utilizing tilted fiber Bragg gratings
We demonstrate the use of tilted fiber gratings to assist the generation of localized infrared surface Plasmons with short propagation lengths and a sensitivity of dλ/dn = 3365 nm in the aqueous index regime. It was also found that the resonances could be spectrally tuned over 1000 nm at the same spatial region with high coupling efficiency (in excess of 25 dB) by altering the polarization of the light illuminating the device. © 2007 Optical Society of America
Portable Surface Plasmon Resonance Biosensor for Detection of Nucleic Acids
AbstractWe present a portable surface plasmon resonance (SPR) sensor based on spectroscopy of surface plasmons on a special diffractive structure. The sensor combines a microfluidic cartridge incorporating the special diffractive structure functionalized with DNA probes and a compact SPR reader. We apply the sensor to detection of nucleic acids employing two most common immobilization methods: (a) biotinylated probes immobilized using the biotin-streptavidin chemistry and (b) thiolated probes attached directly to the surface of the SPR sensor. It is demonstrated that both these immobilization methods allow detecting short nucleic acids at levels below 100 pM
Low Cost Sensors Based on SPR in a Plastic Optical Fiber for Biosensor Implementation
This paper reports the fabrication and testing of two configurations of optical sensor systems based on Surface Plasmon Resonance (SPR) at the interface of a liquid sample and sandwiched structures realized starting from the exposed core of a Plastic Optical Fiber (POF). The proposed geometries have proven to be suitable for measuring the refractive indexes of liquids whose refractive index falls around 1.35. Furthermore, the proposed sensing head, being low cost and relatively easy to realize, may be very attractive for biosensor implementation
Physical characteristics of localized surface plasmons resulting from nano-scale structured multi-layer thin films deposited on D-shaped optical fiber
Novel surface plasmonic optical fiber sensors have been fabricated using multiple coatings deposited on a lapped section of a single mode fiber. UV laser irradiation processing with a phase mask produces a nano-scaled surface relief grating structure resembling nano-wires. The resulting individual corrugations produced by material compaction are approximately 20 μm long with an average width at half maximum of 100 nm and generate localized surface plasmons. Experimental data are presented that show changes in the spectral characteristics after UV processing, coupled with an overall increase in the sensitivity of the devices to surrounding refractive index. Evidence is presented that there is an optimum UV dosage (48 joules) over which no significant additional optical change is observed. The devices are characterized with regards to change in refractive index, where significantly high spectral sensitivities in the aqueous index regime are found, ranging up to 4000 nm/RIU for wavelength and 800 dB/RIU for intensity
Subtractive Inhibition Assay for the Detection of E. coli O157:H7 Using Surface Plasmon Resonance
A surface plasmon resonance (SPR) immunosensor was developed for the detection of E. coli O157:H7 by means of a new subtractive inhibition assay. In the subtractive inhibition assay, E. coli O157:H7 cells and goat polyclonal antibodies for E. coli O157:H7 were incubated for a short of time, and then the E. coli O157:H7 cells which bound antibodies were removed by a stepwise centrifugation process. The remaining free unbound antibodies were detected through interaction with rabbit anti-goat IgG polyclonal antibodies immobilized on the sensor chip using a BIAcore 3000 biosensor. The results showed that the signal was inversely correlated with the concentration of E. coli O157:H7 cells in a range from 3.0 × 104 to 3.0 × 108 cfu/mL with a detection limit of 3.0 × 104 cfu/mL. Compared with direct SPR by immobilizing antibodies on the chip surface to capture the bacterial cells and ELISA for E. coli O157:H7 (detection limit: both 3.0 × 105 cfu/mL in this paper), the detection limit of subtractive inhibition assay method was reduced by one order of magnitude. The method simplifies bacterial cell detection to protein-protein interaction, which has the potential for providing a practical alternative for the monitoring of E. coli O157:H7 and other pathogens
Analysis of noise in differential and ratiometric biosensing systems
This paper presents formulations to evaluate noise in differential and ratiometric measurements that are often performed in biosensing. These measurements are performed to improve signal to noise ratio of the sensing systems for sensitive detection of dynamic biological processes. The use of these formulations is discussed in the context of the differential intensity surface plasmon resonance (SPR) system that is widely used to characterise molecular interactions on a confined axial scale. Previous studies provide qualitative descriptions of the noise performance of such systems but lack rigorous characterisation. Here we present analytical expressions for quantitative evaluation of the noise in differential and ratiometric measurements by applying the rules of arithmetic operations on random variables. Such formulations provide the means for evaluating the signal to noise ratio of such systems. We present how correlated noise can be removed by performing differential or ratiometric processing. Applying these formulations, we also show how the sensitivity of the differential intensity SPR system changes during the experiment
Highly sensitive multipoint real-time kinetic detection of Surface Plasmon bioanalytes with custom CMOS cameras
Phase sensitive Surface Plasmon Resonance (SPR) techniques are a popular means of characterizing biomolecular interactions. However, limitations due to the narrow dynamic range and difficulty in adapting the method for multi-point sensing have restricted its range of applications. This paper presents a compact phase sensitive SPR technology using a custom CMOS camera. The system is exceptionally versatile enabling one to trade dynamic range for sensitivity without altering the optical system. We present results showing sensitivity over the array of better than 10−6 Refractive Index Units (RIU) over a refractive index range of 2×10−2 RIU, with peak sensitivity of 3×10−7 RIU at the center of this range. We also explain how simply altering the settings of polarization components can give sensitivity on the order of 10−8 RIU albeit at the cost of lower dynamic range. The consistent response of the custom CMOS camera in the system also allowed us to demonstrate precise quantitative detection of two Fibrinogen antibody–protein binding sites. Moreover, we use the system to determine reaction kinetics and argue how the multipoint detection gives useful insight into the molecular binding mechanisms
An integrated portable system for single chip simultaneous measurement of multiple disease associated metabolites
Metabolites, the small molecules that underpin life, can act as indicators of the physiological state of the body when their abundance varies, offering routes to diagnosis of many diseases. The ability to assay for multiple metabolites simultaneously will underpin a new generation of precision diagnostic tools. Here, we report the development of a handheld device based on complementary metal oxide semiconductor (CMOS) technology with multiple isolated micro-well reaction zones and integrated optical sensing allowing simultaneous enzyme-based assays of multiple metabolites (choline, xanthine, sarcosine and cholesterol) associated with multiple diseases. These metabolites were measured in clinically relevant concentration range with minimum concentrations measured: 25 μM for choline, 100 μM for xanthine, 1.25 μM for sarcosine and 50 μM for cholesterol. Linking the device to an Android-based user interface allows for quantification of metabolites in serum and urine within 2 min of applying samples to the device. The quantitative performance of the device was validated by comparison to accredited tests for cholesterol and glucose
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