New methods for quantifying rapidity of action potential onset differentiate neuron types.

Two new methods for quantifying the rapidity of action potential onset have lower relative standard deviations and better distinguish neuron cell types than current methods. Action potentials (APs) in most central mammalian neurons exhibit sharp onset dynamics. The main views explaining such an abrupt onset differ. Some studies suggest sharp onsets reflect cooperative sodium channels activation, while others suggest they reflect AP backpropagation from the axon initial segment. However, AP onset rapidity is defined subjectively in these studies, often using the slope at an arbitrary value on the phase plot. Thus, we proposed more systematic methods using the membrane potential's second-time derivative ([Formula: see text]) peak width. Here, the AP rapidity was measured for four different cortical and hippocampal neuron types using four quantification methods: the inverse of full-width at the half maximum of the [Formula: see text] peak (IFWd2), the inverse of half-width at the half maximum of the [Formula: see text] peak (IHWd2), the phase plot slope, and the error ratio method. The IFWd2 and IHWd2 methods show the smallest variation among neurons of the same type. Furthermore, the AP rapidity, using the [Formula: see text] peak width methods, significantly differentiates between different types of neurons, indicating that AP rapidity can be used to classify neuron types. The AP rapidity measured using the IFWd2 method was able to differentiate between all four neuron types analyzed. Therefore, the [Formula: see text] peak width methods provide another sensitive tool to investigate the mechanisms impacting the AP onset dynamics.

Characterization of plasma-enhanced teflon AF for sensing benzene, toluene, and xylenes in water with near-IR surface plasmon resonance.

Near-IR surface plasmon resonance is used to characterize Teflon AF films for refractive index-based detection of the aromatic hydrocarbon contaminants benzene, toluene, and xylenes in water. The technique requires no sample preparation, and film sensitivity is found to be enhanced by oxygen plasma etching. A diffusion equation model is used to extract the diffusion and partition coefficients, which indicate film enrichment factors exceeding two orders of magnitude, permitting a limit of detection of 183, 105 and 55 ppb for benzene, toluene, and xylenes, respectively. The effect of other potential interfering contaminants is quantified.

A plastic total internal reflection photoluminescence device for enzymatic biosensing.

A total internal reflection photoluminescence (TIRPh) device employing an easily fabricated PMMA/PDMS waveguide system provides a detection limit comparable to the best reported results but without using an excitation filter. The optical mechanism is similar to total-internal-reflection-fluorescence (TIRF) but uses a ruthenium-based phosphorescent dye (Ru(dpp)3) deposited on the PMMA core, motivating the generalized term of photoluminescence to include both fluorescence and phosphorescence. An enzymatic hydrogen peroxide (H2O2) biosensor incorporating catalase was fabricated on the TIRPh platform without photolithography or etching. The O2-sensitive phosphorescence of Ru(dpp)3 was used as a transduction mechanism and catalase was used as a biocomponent for sensing. The H2O2 sensor exhibits a phosphorescence to scattered excitation light ratio of 76 ± 10 without filtering. The unfiltered device demonstrates a detection limit of (2.2 ± 0.6) µM with a linear range of 0.1 mM to 20 mM. The device is the first total internal reflection photoluminescence based enzymatic biosensor platform, and is promising for cost-effective, low excitation interference, field-portable sensing.

Fiber optic monooxygenase biosensor for toluene concentration measurement in aqueous samples.

Measurements of pollutants such as toluene are critical for the characterization of contaminated sites and for the monitoring of remediation processes and wastewater treatment effluents. Fiber optic enzymatic biosensors have the potential to provide cost-effective, real time, continuous, in situ measurements. In this study, a fiber optic enzymatic biosensor was constructed and characterized for the measurement of toluene concentrations in aqueous solutions. The biological recognition element was toluene ortho-monooxygenase (TOM), expressed by Escherichia coli TG1 carrying pBS(Kan)TOM, while an optical fiber coated with an oxygen-sensitive ruthenium-based phosphorescent dye served as the transducer. Toluene was detected based on the enzymatic reaction catalyzed by TOM, which resulted in the consumption of oxygen and changes in the phosphorescence intensity. The biosensor was found to have a limit of detection of 3 µM, a linear signal range up to 100 µM, and a response time of 1 h. The performance was reproducible with different biosensors (RSD=7.4%, n=8). The biosensor activity declined with each measurement and with storage time, particularly at elevated temperatures. This activity loss could be partially reversed by exposure to formate, suggesting that NADH consumption was the primary factor limiting lifetime. This is the first report of an enzymatic toluene sensor and of an oxygenase-based biosensor. Since many oxygenases have been reported, the design concept of this oxygenase-based biosensor has the potential to broaden biosensor applications in environmental monitoring.

Optofluidic intracavity spectroscopy of canine hemangiosarcoma.

The label-free technique of optofluidic intracavity spectroscopy (OFIS) uses light transmitted through a cellular body in a microfluidic optical resonator to distinguish different types of cells by their optical properties. The OFIS technique has differentiated canine hemangiosarcoma (HSA) cells from monocytes in peripheral blood mononuclear cells based on their distinctive transmission spectra. A single characteristic parameter indicative of strong multi-transverse-mode resonances was determined for each cell by forming a linear combination of the mean and standard deviation of the transmission spectra over one free spectral range, excluding the peaks of passive Fabry-Pérot cavities without cells. The difference in the characteristic parameters of HSA and monocyte samples was statistically highly significant with a p-value as low as 10(-6). The same method shows that the characteristic parameters of canine lymphoma and lymphocytes are distinct with p < 0.005. A receiver operating-characteristic curve constructed from t-distributions fit to the HSA and monocyte data indicates that 95% sensitivity and 98% specificity can be simultaneously achieved.

Optical and physical characterization of a local evanescent array coupled biosensor: Use of evanescent field perturbations for multianalyte sensing.

The evanescent field surrounding the core of an optical waveguide is very sensitive to refractive index changes near the core. This sensitivity can be exploited to form the basis for a quantitative sensor with high specificity and sensitivity. Selective probe molecules may be attached to the surface of a waveguide core and the evanescent field locally monitored as target analytes are introduced to the system. In this study, probe/analyte regions were simulated using lithographically patterned organic films with thicknesses of 60 nm and 130 nm. The evanescent field strength was measured quantitatively using near field scanning optical microscopy (NSOM). The presence of the organic material on the waveguide caused up to a 70% change in the intensity of the evanescent field over the patterned region and the excitation of a weakly bound higher order mode. The waveguide core and surrounding cladding were numerically simulated using the beam propagation method and these predictions are in quantitative agreement with the experimental results obtained using NSOM.

An optical waveguide array biosensor for serology - biomed 2010.

A label-free optical waveguide immunosensor was designed, fabricated and tested. Different from other popular resonance-based biosensors, such as surface-plasmon-resonance (SPR) or ring/disk resonance biosensors, the local evanescent array coupled (LEAC) biosensor relies on a local evanescent field shift mechanism and can be readily manufactured using trailing-edge integrated-circuit technology with chip-scale microfluidics technology to provide very low cost. The anticipated final form of the sensor technology will require no external equipment enabling disposable use for point-of-care disease detection in non-traditional health-care settings. The local detection ability enables the LEAC biosensor the capability to detect and identify multiple analytes on the same waveguide. On-chip detection is accomplished by integrating buried polysilicon detector arrays under a silicon nitride waveguide. Observations of antibody-antigen interactions using the LEAC biosensor are reported.

Label-free silicon photonic biosensor system with integrated detector array.

An integrated, inexpensive, label-free photonic waveguide biosensor system with multi-analyte capability has been implemented on a silicon photonics integrated circuit from a commercial CMOS line and tested with nanofilms. The local evanescent array coupled (LEAC) biosensor is based on a new physical phenomenon that is fundamentally different from the mechanisms of other evanescent field sensors. Increased local refractive index at the waveguide's upper surface due to the formation of a biological nanofilm causes local modulation of the evanescent field coupled into an array of photodetectors buried under the waveguide. The planar optical waveguide biosensor system exhibits sensitivity of 20%/nm photocurrent modulation in response to adsorbed bovine serum albumin (BSA) layers less than 3 nm thick. In addition to response to BSA, an experiment with patterned photoresist as well as beam propagation method simulations support the evanescent field shift principle. The sensing mechanism enables the integration of all optical and electronic components for a multi-analyte biosensor system on a chip.

Environmental applications of photoluminescence-based biosensors.

For monitoring and treatment of soil and water, environmental scientists and engineers require measurements of the concentration of chemical contaminants. Although laboratory-based methods relying on gas or liquid chromatography can yield very accurate measurements, they are also complex, time consuming, expensive, and require sample pretreatment. Furthermore, they are not readily adapted for in situ measurements.Sensors are devices that can provide continuous, in situ measurements, ideally without the addition of reagents. A biosensor incorporates a biological component coupled to a transducer, which translates the interaction between the analyte and the biocomponent into a signal that can be processed and reported. A wide range of transducers have been employed in biosensors, the most common of which are electrochemical and optical. In this contribution, we focus on photoluminescence-based biosensors of potential use in the applications described above.Following a review of photoluminescence and a discussion of the optoelectronic hardware part of these biosensor systems, we provide explanations and examples of optical biosensors for specific chemical groups: hydrocarbons and alcohols, halogenated organics, nitro-, phospho-, sulfo-, and other substituted organics, and metals and other inorganics. We also describe approaches that have been taken to describe chemical mixtures as a whole (biological oxygen demand and toxicity) since most environmental samples contain mixtures of unknown (and changing) composition. Finally, we end with some thoughts on future research directions that are necessary to achieve the full potential of environmental biosensors.

Evanescent field response to immunoassay layer thickness on planar waveguides.

The response of a compact photonic immunoassay biosensor based on a planar waveguide to variation in antigen (C-reactive protein) concentration as well as waveguide ridge height has been investigated. Near-field scanning optical microscope measurements indicate 1.7%nm and 3.3%nm top surface optical intensity modulation due to changes in effective adlayer thickness on waveguides with 16.5 and 10 nm ridge heights, respectively. Beam propagation method simulations are in good agreement with the experimental sensitivities as well as the observation of leaky mode interference both within and after the adlayer region.