High throughput transepithelial electrical resistance (TEER) measurements on perfused membrane-free epithelia.

Assessment of epithelial barrier function is critically important for studying healthy and diseased biological models. Here we introduce an instrument that measures transepithelial electrical resistance (TEER) of perfused epithelial tubes in the microfluidic OrganoPlate platform. The tubules are grown in microfluidic channels directly against an extracellular matrix, obviating the need for artificial filter membranes. We present TEER measurements on Caco-2 intestinal and renal proximal tubule epithelium. Forty tubules on one single plate were interrogated in less than a minute. We show that TEER measurement is significantly more sensitive than a fluorescent reporter leakage assay in response to staurosporine. We demonstrate a 40-channel time-lapse data acquisition over a 25 hour time period under flow conditions. We furthermore observed a 50% reduction in Caco-2 TEER values following exposure to a cocktail of inflammatory cytokines. To our best knowledge, this is the first instrument of its kind that allows routine TEER studies in perfused organ-on-a-chip systems without interference by artificial filter membranes. We believe the apparatus will contribute to accelerating routine adoption of perfused organ-on-a-chip systems in academic research and in industrial drug development.

Perfused microreactors for liver tissue engineering.

We developed scalable microreactors that foster the development of 3D microscopic pieces of tissue. By integrating microreactors, reservoirs, and pumps in the multiwell cell culture plate format, we created a high throughput cell culture system. However, in contrast to commonly used 2D static cell culture in multiwell plates, our new system allows 3D perfused cell culture. The system provides a means to conduct assays for toxicology and metabolism and can be used as a model for human diseases such as hepatic diseases, exposure-related pathologies, and cancer.

Development and calibration of field-effect transistor-based sensor array for measurement of hydrogen and ammonia gas mixtures in humid air.

A sensor array for analyzing hydrogen and ammonia gas mixtures in humid air has been developed, built into a rugged system, and calibrated for laboratory testing. The sensor array is comprised of four chemically sensitive field-effect transistors (CHEMFETs). Chemically sensitive layers for the sensors were developed and tested using a Kelvin probe. A combination of catalytic and noncatalytic thin layers (palladium and polyaniline) was selected for the four-sensor array. The work function responses of the CHEMFET sensor array to mixtures of hydrogen, ammonia, and humid air were measured. Chemometric multivariate methods, linear and nonlinear partial least squares, were used for the calibration of the sensor array using gas mixtures in the concentration range from 0 to 10 000 ppm hydrogen and ammonia in humid air. The sensor array for ammonia showed good sensitivity, selectivity, response time, and stability and is recommended for field deployment. In contrast, the sensor array for hydrogen, though highly sensitive to hydrogen, demonstrated inadequate stability, requiring further development before deployment is recommended.