An Integrated Optical Biosensor (IOBS)

Abstract

We have demonstrated an integrated optical biosensor (IOBS) employed as differential refractometer with a resolution down to n_C.=5 × 10^?5. The minimum volume required for measurements is V=L_x?L_y?L_z=1?10^?4 microlitres where L_x?L_y=1mm?0.1mm is the illuminated grating area and L_z is the penetration depth of the evanescent field of the guided mode into the cover medium.

The sensor described was successfully used to monitor enzyme activities by measuring the change of refractive index of a substrate converted by an enzyme. Because the sensor responds not only to refractive index changes of a cover medium but also to thickness changes of absorbed layers it should be possible to measure biological interactions such as between antigen/antibody, inhibitor/enzyme or receptor/cell by coupling of either respective biomolecule onto the waveguiding film. Thus we have shown in preliminary experiments that on coupling covalently to the grating rabbit anti-human-IgG (both of similar molecular weight around 150 000) a pronounced attenuation of the intensity of the guided mode was observed due to the increased adlayer thickness (to be published).

It should be possible to miniaturize the unit using channel waveguides fabricated by a combination of photolithography and etching techniques with the aim to obtain implantable clinical integrated optical biosensors.

The sensitivity can be further improved electronically by measuring directly the intensity of the guided mode using a second grating as an output coupler. This will increase the signal to noise ratio substantially.     

Chemical sensor based on a long-period fibre grating modified by a functionalized polydimethylsiloxane coating

A chemical sensor based on a coated long-period grating has been prepared and characterized. Designer coatings based on polydimethylsiloxane were prepared by the incorporation of diphenylsiloxane and titanium cross-linker in order to provide enhanced sensitivity for a variety of key environmental pollutants and optimal refractive index of the coating. Upon microextraction of the analyte into the polymer matrix, an increase in the refractive index of the coating resulted in a change in the attenuation spectrum of the long-period grating. The grating was interrogated using ring-down detection as a means to amplify the optical loss and to gain stability against misalignment and power fluctuations. Chemical differentiation of cyclohexane and xylene was achieved and a detection limit of 300 ppm of xylene vapour was realized.     

Conducting polymer diffraction gratings on gold surfaces created by microcontact printing and electropolymerization at submicron length scales

Conducting polymer diffraction gratings on Au substrates have been created using microcontact printing of C18-alkanethiols, followed by electropolymerization of either poly(aniline) (PANI) or poly(3,4-ethylenedioxythiophene) (PEDOT). Soft-polymer replicas of simple diffraction grating masters (1200 lines/mm) were used to define the alkanethiol template for polymer growth. Growth of PANI and PEDOT diffraction gratings was followed in real time, through in situ tapping-mode atomic force microscopy, and by monitoring diffraction efficiency (DE) as a function of grating depth. DE increased as grating depth increased, up to a limiting efficiency (13-26%, with white light illumination), defined by the combined optical properties of the grating and the Au substrate, and ultimately limited by the loss of resolution due to coalescence of the polymer films. Grating efficiency is strongly dependent upon the grating depth and the refractive index contrast between the grating material and the surrounding solutions. Both PEDOT and PANI gratings show refractive index changes as a function of applied potential, consistent with changes in refractive index brought about by the doping/dedoping of the conducting polymer. The DE of PANI gratings are strongly dependent on the pH of the superstrate solution; the maximum sensitivity (DeltaDE/DeltapH) is achieved with PANI gratings held at +0.4 V versus Ag/AgCl, where the redox chemistry is dominated by the acid-base equilibrium between the protonated (emeraldine salt) and deprotonated (emeraldine base) forms of PANI. Simulations of DE were conducted for various combinations of conducting polymer refractive index and grating depth, to compute sensitivity parameters, which are maximized when the grating depth is ca. 50% of its maximum obtainable depth.     

固定光路可变焦宽调角表面等离子共振成像装置

设计了一种折反式棱镜耦合单元, 用于简化表面等离子共振系统的角度调节操作, 并基于此单元构建了固定光路可变焦宽调角表面等离子共振成像装置. 该装置通过旋转折反式棱镜耦合单元即可实现角度调节, 调节范围可达40°~80°, 结合1~2倍的变焦成像系统, 可对1~4 cm²传感芯片成像, 且可分辨至少3600 Point/cm². 以蔗糖溶液为研究对象, 考察了该装置的灵敏度, 结果表明其灵敏度可达7×10^-6 RIU(Refractive index unit), 共振角与折射率的线性相关系数为0.99953. 将该装置用于研究抗牛血清白蛋白抗体与其它蛋白的实时相互作用, 所得结果与理论相符. 该表面等离子共振成像装置将在实时并行分析方面具有广阔的应用前景.     

An integrated disposable dye clad leaky waveguide sensor for micro-TAS applications

An integrated, disposable, dye clad leaky waveguide (DCLW) device has been fabricated and tested for both refractive index and fluorescence detection in mu-TAS applications. The chip comprises the required flow geometry and optical coupling elements in a robust device that is relatively simple and inexpensive to fabricate. Disposable DCLW chips were fabricated at room temperature by spin-coating both the dye and silica sol-gel waveguiding layers on a polymer substrate which contained injection moulded grating coupler. These devices have been designed to increase the interaction of the evanescent field light at the channel wall and with the sample in the channel. The DCLW device has been used to detect changes in the refractive index of different percentages of glycerol solutions and to detect low concentrations down to 10(-12) M fluorescein using a grating coupler.     

Large-scale resonance amplification of optical sensing of volatile compounds with chemoresponsive visible-region diffraction gratings

Micropatterning of the vapochromic charge-transfer salt, [Pt(CNC6H4C10H21)4][Pd(CN)4], on transparent platforms yields transmissive chemoresponsive diffraction gratings. Exposure of the gratings to volatile organic compounds (VOCs) such as chloroform and methanol leads to VOC uptake by the porous material comprising the grating lattice or framework, and a change in the material's complex refractive index, ?. The index change is accompanied by a change in the degree of index contrast between the lattice and the surrounding medium (in this case, air), and a change in the diffraction efficiency of the grating. When a monochromatic light source that is not absorbed by the lattice material is employed as a probe beam, only changes in the real component of ? are sensed. Under these conditions, the grating behaves as a nonselective, but moderately sensitive, sensor for those VOCs capable of permeating the porous lattice material. When a probe color is shifted to a wavelength coincident with the vapochromic charge-transfer transition of the lattice material, the sensor response is selectively amplified by up to 3.5 orders of magnitude, resulting in greatly enhanced sensitivity and some degree of chemical specificity. On the basis of studies at four probe wavelengths, the amplification effect is dominated by resonant changes in the imaginary component of the refractive index. The observed wavelength- and analyte-dependent amplification effects are quantitatively well described by a model that combines a Kramers-Kronig analysis with an effective-medium treatment of dielectric effects.     

A novel C-shaped, gold nanoparticle coated, embedded polymer waveguide for localized surface plasmon resonance based detection

In this study, a novel embedded optical waveguide based sensor which utilizes localized surface plasmon resonance of gold nanoparticles coated on a C-shaped polymer waveguide is being reported. The sensor, as designed, can be used as an analysis chip for detection of minor variations in the refractive index of its microenvironment, which makes it suitable for wide scale use as an affinity biosensor. The C-shaped waveguide coupled with microfluidic channel was fabricated by single step patterning of SU8 on an oxidized silicon wafer. The absorbance due to the localized surface plasmon resonance (LSPR) of SU8 waveguide bound gold nano particle (GNP) was found to be linear with refractive index changes between 1.33 and 1.37. A GNP coated C-bent waveguide of 200 μ width with a bend radius of 1 mm gave rise to a sensitivity of ~5 ΔA/RIU at 530 nm as compared to the ~2.5 ΔA/RIU (refractive index units) of the same dimension bare C-bend SU8 waveguide. The resolution of the sensor probe was ~2 × 10(-4) RIU.     

Optimisation of an integrated optical evanescent wave absorbance sensor for the determination of chlorinated hydrocarbons in water

The suitability of an integrated optical chemical sensor for the determination of highly volatile chlorinated hydrocarbons in aqueous solutions has been proven. The analytes are detected by NIR absorption spectrometry in the evanescent field of an integrated optical strip waveguide generated in a BGG31 (Schott, Germany) glass substrate, which is coated with a hydrophobic polymer superstrate as sensing layer. It has been shown that the sensitivity increases when the refractive index of the superstrate is increased from 1.333 up to 1.46. Different UV-cured polysiloxanes with low cross sensitivity to water have been prepared. Due to the good light transmission properties of the IO-sensors prepared by this method, quantitative measurements have been performed with the model system trichloroethene (TCE) in water. A detection limit of 22 ppm has been found and the sensor response times (t(90)-value) are between five and fourteen minutes for a coating thickness of around 30 microm. The sensor response is totally reversible. The analyte desorbes in air within 2 min. The enrichment of trichloroethene in the polysiloxane coating can be described by film diffusion through the aqueous boundary layer as rate determining step.     

Fiber-optic surface plasmon resonance for vapor phase analyses

Fiber-optic sensors based on surface plasmon resonance (SPR) for direct refractive index (RI) measurements of samples with the RI between 1.00 and 1.30 are described. Most applications of SPR sensors are designed to function near the refractive index of water (1.3330 RI). The RI changes of aqueous solution (RI, ca. 1.34) can easily be monitored by silica-fiber (RI, 1.4601 at 550 nm) based SPR sensor. With regard to gas species detection, the fiber-optic SPR sensor must be modified for sensitivity to changes in refractive index near 1.0008 (i.e., RI of air). However, the silica waveguide has a prohibitively high RI for unmodified monitoring of the RI changes of gas. The silica-fiber based SPR probe design presented here is based upon the modification of the probe geometry to the ability to tune the SPR coupling wavelength/angle pair. In this study, the tapered silica-based fiber SPR sensors are shown to directly determine the RI changes of gas species and the density change of dry air.     

Analysis of the kinetics of diffraction efficiency during the holographic grating recording in azobenzene functionalized polymers

The laser-assisted holographic grating recording process in films of azobenzene functionalized polymers is usually studied by observation of the efficiency of light scattering on a developing in time diffraction grating. Various possible mechanisms contributing to grating formation as well as the bulk or surface origin (bulk refractive index and/or relief grating) of light scattering make the analysis of kinetics of grating recording, from the light scattering data only, difficult and ambiguous. To fully explain experimentally observed various and complex (frequently nonexponential) kinetics of the first-order light diffraction intensity, we considered a simple single-exponential growth of the two phase gratings in the same polymer film. In modeling we assumed that the bulk refractive index grating Deltan(t) and the surface relief grating Deltad(t) differ considerably in their growth rates and we allowed for a nonstationary phase shift Deltaphi(t) between them which was experimentally observed during the recording process. The origin of the nonstationary phase shift is a result of a slow shift of interference pattern due to delicate symmetry breaking in illumination conditions (e.g., difference in beam intensities and deviation of exact symmetrical beam incidence angles on the sample). Changing only such parameters as stationary amplitudes of refractive index and relief gratings for a span of phase shifts (0-pi) between them, we obtained a series of kinetic responses which we discuss and interpret. The various examples of temporal evolution of diffraction efficiency for the same grating formation kinetics, modeled in our work, supply evidence that great care must be taken to properly interpret the experimental results.     

Fibre optic coupler as a detector for microfluidic applications

A new construction of a fibre optic coupler is presented in the paper. Two polymer optical fibres were used to build a coupler in which coupling efficiency of optical power depends on the refractive index of liquid delivered to a microchannel formed by the fibres. The coupler was tested as a detector in saccharose concentration measurements, and was used in absorbance measurements. A red light emitting diode and a spectrometer were used as a light source and a photodetector, respectively. Experiments confirmed that the coupler can be used for the real time monitoring of the changes in the refractive index of a saccharose solution exhibiting repeatable changes in the signal, with no hysteresis. Absorbance tests were performed with a solution of bromothymol blue at different pH.     

Measurement uncertainty in analytical studies based on surface plasmon resonance

The sensitivity of surface plasmon resonance (SPR) transducers depends on the thickness and spatial organization of interfacial structures at their surfaces. This is because the response of the SPR sensor is determined by integrating the distance-dependent refractive index (spatial interfacial architectures), weighted by the square of the electromagnetic field, from zero to infinite distance. The effect of SPR transducer sensitivity variation on the accuracy of SPR analysis is considered. Our quantitative estimation (based on the results of refractometric studies) gave a value for sensitivity variation of about 3% for the formation of a self-assembled thiocyanate layer or a trypsin-soybean trypsin inhibitor surface complex. The estimated accuracy in measured variation (i.e., by 0.01) for the refractive index of the external medium was 3 × 10⁻⁴. This restriction, which follows immediately from the physical mechanism of the SPR phenomenon, should be taken into account when analyzing data obtained with the above technique.     

Observation of spatial distribution of laser-induced refractive index change in dye-doped liquid crystals

Spatial distributions of photothermal refractive index changes in dye-doped liquid crystals were determined by an optical interferometric method. The refractive index change of the order of 10^-1, including spatial distribution, was estimated by the described experimental technique. The absolute value of the refractive index change was proportional to the pump beam power, and the diameter of the index distribution was slightly larger than that of the pump laser beam due to heat conduction.     

适用于高空间频率透射式全息聚合物分散液晶光栅的材料研究

提出了适用于空间频率为3000 lp/mm的透射式全息聚合物分散液晶(HF-THPDLC)光栅的材料体系,并在反应动力学方面进行了深入的分析.为了能够得到高衍射效率并具有良好表面形貌的HF-THPDLC 光栅,首先确定了体系的平均官能度,使得预聚单体和液晶的扩散时间、液晶的成核时间以及于聚合物的凝胶时间达到最佳匹配状态...     

Quantitative characterization of the optical properties of absorbing polymer films: Comparative investigation of the internal reflection intensity analysis method

Nondestructive, three‐dimensional refractive‐index measurements are used for the determination of both the crystallinity and orientation in thin polymer films. The prism wave‐guide coupler is particularly suited for three‐dimensional isotropic and anisotropic thin‐film studies because of the quantitative character of the information obtained and the ease of data acquisition. It has been limited, however, to determining only the refractive index of transparent or weakly absorbing thin‐film samples. On the basis of thin‐film optics, this study develops a new internal reflection intensity analysis (IRIA) method, which uses the intensity information rather than the conventional mode angle values to acquire both the refractive index and the extinction coefficient over a range of transparent to highly absorbing polymer films. Therefore, the IRIA method overcomes the limitations of this prism wave‐guide coupler technique, which can only measure the refractive index of a weakly absorbing sample. With a Metricon PC‐2010 as the skeletal framework, a prototype instrument has been developed to apply and test the IRIA method. A study comparing both the refractive index and extinction coefficient obtained with ellipsometry, ultraviolet–visible/near‐infrared reflectometry, and IRIA for solvent blue 59 dyed polystyrene films confirms that the IRIA method is effective for obtaining the three‐dimensional refractive indices and extinction coefficients of polymer films. In addition, the refractive index and extinction coefficient spectrum (400–800 nm) of solvent blue 59 have been determined with the effective media theory. Furthermore, the three‐dimensional complex refractive indices of highly absorbing black electrical tape, inaccessible to other optical measurement because of its surface character, has been determined by the IRIA method. © 2003 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 41: 842–855, 2003     

Active control of interfacial properties

Recent studies aimed at establishing principles for active control of the physicochemical properties of interfaces have made substantial progress towards demonstrating spatial and temporal control of interfacial properties of both liquids and solids. Light-active and redox-active surfactants have been shown to permit large (>20 mN/m) and reversible changes in the surface tensions of liquids on time-scales of seconds. These changes can be directed to localized regions of liquids with sub-millimeter spatial resolution, thus providing new means to create controlled gradients in surfactant-based properties of liquids (e.g. gradients in surface tension). Progress has also been reported in the electrowetting of liquids on the surfaces of solids, although it is still not possible to use an external electric field to cause an aqueous solution to wet a hydrophobic surface.     

Optimisation of an integrated optical evanescent wave absorbance sensor for the determination of chlorinated hydrocarbons in water

The suitability of an integrated optical chemical sensor for the determination of highly volatile chlorinated hydrocarbons in aqueous solutions has been proven. The analytes are detected by NIR absorption spectrometry in the evanescent field of an integrated optical strip waveguide generated in a BGG31 (Schott, Germany) glass substrate, which is coated with a hydrophobic polymer superstrate as sensing layer. It has been shown that the sensitivity increases when the refractive index of the superstrate is increased from 1.333 up to 1.46. Different UV-cured polysiloxanes with low cross sensitivity to water have been prepared. Due to the good light transmission properties of the IO-sensors prepared by this method, quantitative measurements have been performed with the model system trichloroethene (TCE) in water. A detection limit of 22 ppm has been found and the sensor response times (t₉₀-value) are between five and fourteen minutes for a coating thickness of around 30 m. The sensor response is totally reversible. The analyte desorbes in air within 2 min. The enrichment of trichloroethene in the polysiloxane coating can be described by film diffusion through the aqueous boundary layer as rate determining step.     

A surface plasmon resonance sensor on a compact disk-type microfluidic device

A surface plasmon resonance (SPR) sensor on a compact disk (CD)-type microfluidic device was developed to miniaturize the elements of a complete analytical system, pump and valves. The CD-type microfluidic device was fabricated by attaching a polydimethylsiloxane disk plate that contained microchannels and reservoirs to a flat polycarbonate disk plate that contained grating films with a thin layer of Au. The optical system of the SPR sensor and the theory for its operation are based on the principle of a grating coupled-type SPR. The sample and reagent solutions in the reservoirs on the CD-type microfluidic device were sequentially introduced into the detection chamber by centrifugal force generated by the rotation of the microfluidic device. The variation of resonance wavelength was dependent on the refractive index of the sample solution. This CD-type SPR sensor was successfully used in an immunoassay of immunoglobulin A (IgA). The anti-IgA, blocking reagent, sample and washing solution in the reservoirs were sequentially introduced into the detection chamber by changing the frequency of rotation of the microfluidic device. IgA in the sample solution was adsorbed to the anti-IgA immobilized on the Au thin layer in the detection chamber and was then detected by the SPR sensor.     

Allylated cyclodextrins as effective affinity materials in chemical sensing of volatile aromatic hydrocarbons using an optical planar Bragg grating sensor

We report on the application of perallyl-substituted α-, β- and γ-cyclodextrins to an optical planar Bragg grating refractive index sensor for the effective sensitization of the sensor for airborne volatile aromatic hydrocarbons. Thereby, the emphasis of this work lies on the comparison of the different cyclodextrin types regarding their suitability as affinity material assessed by the sensors sensitivity and response behavior. The opto-chemical sensor device showed an immediate and quick response to the application of the investigated analytes benzene, toluene and m-xylene as well as a linear dependence on the concentration of those analytes. Studies on the sensors sensitivity depending on the applied cyclodextrin types revealed a generally higher sensitivity for the sensor sensitized with perallyl-substituted β-cyclodextrins. Here, the sensor systems detection limit was found to 60 ± 4 ppm for benzene, 18 ± 3 ppm for toluene and 3.8 ± 0.5 ppm for m-xylene. The response time and recovery time were found to approximately 30 s and 40 s, respectively, depending on the applied cyclodextrin and the chosen analyte.     

Lab-in-a-tube: on-chip integration of glass optofluidic ring resonators for label-free sensing applications

The fabrication of tubular rolled-up optofluidic ring resonators (RU-OFRRs) based on glass (SiO(2)) material with high quality factors is reported. A novel methodology combining lab-on-a-chip fabrication methods and rolled-up nanotech is presented for the fabrication of fully integrated tubular optofluidic sensors. The microfluidic integration of several RU-OFRRs on one chip is solved by enclosing the microtubes with a patterned robust SU-8 polymeric matrix. A viewport on each microtube enables exact excitation and monitoring of whispering gallery modes with a photoluminescence spectroscopy system under constant ambient conditions, while exchanging the content of the RU-OFRR with liquids of different refractive indices. The refractrometric sensor capabilities are investigated regarding signal stability, sensitivity and reliability. The sensitivity of the integrated RU-OFRR, which is the response of the modes to the change in refractive index of the liquid, is up to 880 nm/refractive index units (RIU).