Microfluidic organ-on-a-chip model of the outer blood-retinal barrier with clinically relevant read-outs for tissue permeability and vascular structure.

The outer blood-retinal barrier (oBRB) tightly controls the transport processes between the neural tissue of the retina and the underlying blood vessel network. The barrier is formed by the retinal pigment epithelium (RPE), its basal membrane and the underlying choroidal capillary bed. Realistic three-dimensional cell culture based models of the oBRB are needed to study mechanisms and potential treatments of visual disorders such as age-related macular degeneration that result from dysfunction of the barrier tissue. Ideally, such models should also include clinically relevant read-outs to enable translation of experimental findings in the context of pathophysiology. Here, we report a microfluidic organ-on-a-chip model of the oBRB that contains a monolayer of human immortalized RPE and a microvessel of human endothelial cells, separated by a semi-permeable membrane. Confluent monolayers of both cell types were confirmed by fluorescence microscopy. The three-dimensional vascular structures within the chip were imaged by optical coherence tomography: a medical imaging technique, which is routinely applied in ophthalmology. Differences in diameters and vessel density could be readily detected. Upon inducing oxidative stress by treating with hydrogen peroxide (H2O2), a dose dependent increase in barrier permeability was observed by using a dynamic assay for fluorescence tracing, analogous to the clinically used fluorescence angiography. This organ-on-a-chip of the oBRB will allow future studies of complex disease mechanisms and treatments for visual disorders using clinically relevant endpoints in vitro.

Assessment of a new prototype hydrogel CO( 2 ) sensor; comparison with air tonometry.

OBJECTIVE: Gastrointestinal ischemia is always accompanied by an increased luminal CO(2). Currently, air tonometry is used to measure luminal CO(2). To improve the response time a new sensor was developed, enabling continuous CO(2) measurement. It consists of a pH-sensitive hydrogel which swells and shrinks in response to luminal CO(2), which is measured by the pressure sensor. We evaluated the potential clinical value of the sensor during an in vitro and in vivo study. METHODS: The response time to immediate, and stepwise change in pCO(2) was determined between 5 and 15 kPa, as well as temperature sensitivity between 25 and 40 degrees C at two pCO(2) levels. Three sensors were compared to air tonometry (Tonocap) in healthy volunteers using a stepwise incremental exercise test, followed by a period of hyperventilation and an artificial CO(2)-peak. RESULTS: The in vitro response time to CO(2) increase and decrease was mean 5.9 and 6.6 min. The bias, precision and reproducibility were +5%, 3% and 2%, resp. Increase of 1 degrees C at constant pCO(2) decreased sensor signal by 8%. In vivo tests: The relation with the Tonocap was poor during the exercise test. The response time of the sensor was 3 min during hyperventilation and the CO(2) peak. CONCLUSION: The hydrogel carbon dioxide sensor enabled fast and accurate pCO(2) measurement in a controlled environment but is very temperature dependent. The current prototype hydrogel sensor is still too unstable for clinical use, and should therefore be improved.

pH sensitivity of Si--C linked organic monolayers on crystalline silicon surfaces.

The electrochemical behavior of Si--C linked organic monolayers is studied in electrolyte-insulator-Si devices, under conditions normally encountered in potentiometric biosensors, to gain fundamental knowledge on the behavior of such Si electrodes under practical conditions. This is done via titration experiments, Mott-Schottky data analysis, and data fitting using a site-binding model. The results are compared with those of native SiO(2) layers and native SiO(2) layers modified with hexamethyldisilazane. All samples display pH sensitivity. The number of Si--OH groups on the alkylated samples is calculated to be less than 0.7 % of that of a pure SiO(2) insulator, which still causes a pH sensitivity of approximately 25 mV per pH unit in the pH range: 4-7. The alkylated samples hardly suffer from response changes during up- and down-going titrations, which indicates that very little oxide is additionally formed during the measurements. The pK(a) values of all samples with monolayers (4.0-4.4) are lower than that of native SiO(2) (6.0). The long-term drift (of approximately 1 mV h(-1)) is moderate. The results indicate that biosensors composed of alkylated Si substrates are feasible if a cross-sensitivity towards pH in the sensor signal is taken into account.

Si-C linked organic monolayers on crystalline silicon surfaces as alternative gate insulators.

Herein, the influence of silicon surface modification via Si-C(n)H(2n+1) (n=10,12,16,22) monolayer-based devices on p-type 100 and n-type 100 silicon is studied by forming MIS (metal-insulator-semiconductor) diodes using a mercury probe. From current density-voltage (J-V) and capacitance-voltage (C-V) measurements, the relevant parameters describing the electrical behavior of these diodes are derived, such as the diode ideality factor, the effective barrier height, the flatband voltage, the barrier height, the monolayer dielectric constant, the tunneling attenuation factor, and the fixed charge density (Nf). It is shown that the J-V behavior of our MIS structures could be precisely tuned via the monolayer thickness. The use of n-type silicon resulted in lower diode ideality factors as compared to p-type silicon. A similar flatband voltage, independent of monolayer thickness, was found, indicating similar properties for all silicon-monolayer interfaces. An exception was the C10-based monolayer device on p-type silicon. Furthermore, low values of N(f) were found for monolayers on p-type silicon (approximately 6 x 10(11) cm(-2)). These results suggest that Si--C linked monolayers on flat silicon may be a viable material for future electronic devices.

An efficient method for the fabrication of temperature-sensitive hydrogel microactuators.

In this article a new method for the photolithographical deposition of temperature-sensitive hydrogels is presented. The method can be used in conjunction with standard 365 nm UV-photolithography to accurately dimension and position temperature-sensitive hydrogel microactuators in a highly parallel fashion. A number of characteristics of the hydrogels were investigated. These include: the photolithographical reproduction quality, the effect of the crosslinking density in the hydrogels on their swelling behavior, the swelling hysteresis behavior, the effect of dimensional constraints on the swelling of the hydrogels and the effect of copolymerization with an ionizable comonomer on the temperature behavior of the hydrogels. The method presents a considerable improvement in the microfabrication of temperature-sensitive hydrogel microactuators and has potential for the mass-fabrication of these interesting microactuators.

Stimulus-sensitive hydrogels and their applications in chemical (micro)analysis.

In this tutorial review the use of stimulus-sensitive hydrogels as sensors and actuators for (micro)analytical applications is discussed. The first part of the article is aimed at making the reader familiar with stimulus-sensitive hydrogels, their chemical composition and their chemo-physical behavior. The prior art in the field, that comprises a number of sensors ranging from metal ion-sensitive sensors to antigen-sensitive sensors and a few actuators, is also treated in this part. The second part of the article focusses on the use of stimulus-sensitive hydrogels for microsensors and microactuators as well as their application in micro total analysis systems. The benefits of stimulus-sensitive hydrogels, their miniaturisation and the use of 365 nm UV-photolithography as a fast economical manufacturing technique are discussed.

Fascicular selectivity in transverse stimulation with a nerve cuff electrode: a theoretical approach.

The performance of cathode-anode configurations in a cuff electrode to stimulate a single fascicle in a nerve trunk has been investigated theoretically. A three-dimensional volume conductor model of a nerve trunk with four fascicles in a cuff electrode and a model of myelinated nerve fiber stimulation were used to calculate the recruitment of 15 m fibers in each fascicle. The effect of a monopole, a transverse bipole (anode opposite the cathode), and a narrow transverse tripole (guarded cathode) in selectively stimulating 15 m fibers in each fascicle has been quantified and presented as recruitment curves. It is predicted that selective fascicle stimulation is advanced most by stimulation with a bipole in a plane perpendicular to the axis of the nerve trunk. Monopoles and conventional longitudinal tripoles perform less well, as does a longitudinal tripole with an additional "steering" anode. Apart from transverse bipolar stimulation an additional anode may be used to maximally fit the area of excitation to the topography of the fascicle to be recruited. As compared to monopolar and longitudinal tripolar stimulation, the slope of the recruitment curves in transverse bipolar stimulation is reduced considerably, thus allowing improved fine tuning of nerve (and thus force) recruitment. Another advantage of this method is a minimal number of cable connections to the cuff electrode. The cost of the improved selectivity is an increased stimulation current.

Optimization of an electrolyte conductivity detector for measuring low ion concentrations.

The optimization process of a planar interdigitated conductivity detector for measuring very low electrolyte concentrations for use in a lab-on-chip gas detection system is described. An electrical equivalent of the sensor is given, which includes the double layer capacitance dependency on the electrolyte concentration, resulting in a better description of the impedance of the sensor. The cell constant of the sensor is minimized to reduce the cell resistance in low specific conductivity solutions under the restriction of a small electrode area (> or = 0.1 cm(2)) for fast measurement, prescribed by the ammonia detection system. The small size makes it suitable for integration in micro channels. The developed sensor has a cell constant of 7.9 m(-1) resulting in a maximum resistance for deionized water of 177 k Omega at a frequency of 1 kHz.