Evanescent-field detection

For almost 20 years, the optical waveguide sensor has been used for the label-free, evanescent-field detection of chemical or biological reactions taking place in the close vicinity of the waveguide surface. This includes numerous applications within the fields of chemical and biological sensing, where in... 

The reduction of dimensions also reduces volumes which are accessible to the detector. Thus, detection principles related to geometric dimensions of the detector cell ai e not ideally suited for coupling to microsystems, whereas surface sensitive principles, such as electrochemical methods or optical methods utilizing the evanescent field of a waveguide, or methods which can be focussed on a small amount of liquid, such as electrochemiluminescence (ECE), ai e better suited. This is why electrochemiluminescence detectors ai e combined to microsystems. Moreover ECE has found wide applications in biochemistry because of its high sensitivity, relatively simplicity and feasibility under mild conditions. 

In addition, typical methods of sensing are total internal reflection fluorescence or monitoring of fluorescence resonance energy transfer6,7. The second class is a direct optical detection principle which relies either on reflectometry or refractometry. The latter is connected to evanescent field... 

Interferometric sensors frequently have also been applied to biosensor measurements. Thereby, the evanescent field technique (Mach-Zehnder interferometer) has been compared with other optical detection principles regarding information on layer structure and in case of biosensing30. The... 

The background problem can be further overcome when using a surface-confined fluorescence excitation and detection scheme at a certain angle of incident light, total internal reflection (TIR) occurs at the interface of a dense (e.g. quartz) and less dense (e.g. water) medium. An evanescent wave is generated which penetrates into the less dense medium and decays exponentially. Optical detection of the binding event is restricted to the penetration depth of the evanescent field and thus to the surface-bound molecules. Fluorescence from unbound molecules in the bulk solution is not detected. In contrast to standard fluorescence scanners, which detect the fluorescence after hybridization, evanescent wave technology allows the measurement of real-time kinetics (www.zeptosens.com, www.affinity-sensors.com). 

The enhanced evanescent field, as confirmed by the coupling enhancement results noted above, has been shown to significantly enhance detection sensitivity in preliminary experiments using dye in water solution at 800 nm. In addition, a... 

The type of matrix used for immobilization of the recognition element for bacterial cell detection is crucial to achieve high sensitivity. Two important conditions should be considered specifically for bacterial detection (1) the accessibility of the recognition elements in the immobilization matrix for bacteria binding on the sensor surface and (2) to obtain the binding of the analytes within the most sensitive region of the evanescent field, immediately adjacent to the sensor surface. 

The circles in Fig. 16.6a show the calculated sensitivity, AX/Am for these different cases. As can be seen, the innermost holes are the most sensitive to any refractive index changes in the local environment as opposed to the holes that are further away from the cavity. These results can be explained by noting that the evanescent field is largest inside the innermost holes and decreases inside holes that are situated further away from the cavity. This is important to note because targeting only the inner most holes for functionalization allows for the lowest possible limit of mass detection for this device. In the case where only the inner two holes are functionalized we find that the resonance shifts by 3.5 nm when 1 fg of DNA binds to the resonator. Therefore, a mass change of 10 ag would result in a mass surface density of 0.84 ng cm 2 and an approximate shift of 0.01 nm, which can be experimentally detected. We therefore take this as the potential LOD of the device. 

A disposable, patterned, planar waveguide with a number of individual wells has been reported for a one-step homogeneous immunoassay of IgG.<133) The device is fabricated by an ion-exchange process, etching, and covalent reagent immobilization. The sample fills the waveguide by capillary action. The sample well, as well as fluorescent and nonfluorescent control wells are excited by an evanescent field, and individually scanned. The IgG detection limit is in the 10range. 

The fact that ATR-IR spectroscopy uses an evanescent field and therefore probes only the volume very close to the IRE has important consequences for its application in heterogeneous catalysis, in investigations of films of powder catalysts. The catalyst particle size and packing affect the size of the detectable signals from the catalyst and bulk phase. Furthermore, if the catalyst layer is much thicker than the penetration depth of the evanescent field, diffusion of reactants and products may influence the observed signals. In fast reactions, gradients may exist within the catalyst layer, and ATR probes only the slice closest to the IRE. 

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