Real-time physiological measurements of oxygen using a non-invasive self-referencing optical fiber microsensor.

Reactive molecular oxygen (O2) plays important roles in bioenergetics and metabolism and is implicated in biochemical pathways underlying angiogenesis, fertilization, wound healing and regeneration. Here we describe how to use the scanning micro-optrode technique (SMOT) to measure extracellular fluxes of dissolved O2. The self-referencing O2-specific micro-optrode (also termed micro-optode and optical fiber microsensor) is a tapered optical fiber with an O2-sensitive fluorophore coated onto the tip. The O2 concentration is quantified by fluorescence quenching of the fluorophore emission upon excitation with blue-green light. The micro-optrode presents high spatial and temporal resolutions with improved signal-to-noise ratio (in the picomole range). In this protocol, we provide step-by-step instructions for micro-optrode calibration, validation, example applications and data analysis. We describe how to use the technique for cells (Xenopus oocyte), tissues (Xenopus epithelium and rat cornea), organs (Xenopus gills and mouse skin) and appendages (Xenopus tail), and provide recommendations on how to adapt the approach to different model systems. The basic, user-friendly system presented here can be readily installed to reliably and accurately measure physiological O2 fluxes in a wide spectrum of biological models and physiological responses. The full protocol can be performed in ~4 h.

Technological developments for small-scale downstream processing of cell therapies.

Despite considerable regulatory and clinical hurdles, the development and use of cell-based therapies are gaining momentum. As more of these therapies move toward commercial approval and larger-scale distribution, associated manufacturing and processing technologies are being advanced. Modern technologies directed at downstream processing seek to distribute such therapies from the manufacturing site to the patient more efficiently and reliably. Novel small-scale downstream solutions boost the transformation of cell therapies from abstraction to reality.

SNAP: fabrication of long coupled microresonator chains with sub-angstrom precision.

Based on the recently-introduced Surface Nanoscale Axial Photonics (SNAP) platform, we demonstrate a chain of 30 coupled SNAP microresonators spaced by 50 micron along an optical fiber, which is fabricated with the precision of 0.7 angstrom and a standard deviation of 0.12 angstrom in effective microresonator radius. To the best of our knowledge, this result surpasses those achieved in other super-low-loss photonic technologies developed to date by two orders of magnitude. The chain exhibits bandgaps in both the discrete and continuous spectrum in excellent agreement with theory. The developed method enables robust fabrication of SNAP devices with sub-angstrom precision.

Rapid assessment of endpoint antioxidant capacity of red wines through microchemical methods using a kinetic matching approach.

Antioxidant capacity of food samples is usually assessed by different analytical methods, however the results attained even for the same method are strongly dependent on the selected reaction time and also on the standard compound used. To tackle this problem, we propose here a kinetic matching approach, associated to the conversion of results into equivalents of a common standard compound, as a universal way for expression of results. The methodology proposed was applied to methods based on different chemistries (Folin-Ciocalteu (F-C), CUPRAC, DPPH(•) and ABTS(•+) assays) and red wines (n=40) were chosen as a model of complex food sample. For implementation of the kinetic matching approach, the standard phenolic mixture (caffeic acid, (+)-catechin, hesperetin, morin and (-)-epigallocatechin gallate) was chosen for calibration in F-C, CUPRAC and DPPH(•) assays, while tannic acid was suitable for ABTS(•+) assay. Results showed that, for all methods, there was no statistical difference between results attained by the kinetic matching approach (after <10 min of reaction) and that at endpoint conditions (after 60 to 300 min). The repeatability and the reproducibility of the kinetic matching approach was <4.5%, for all antioxidant assays. The sample throughput increases from <18 (endpoint measurements) to >108 h(-1) using the proposed kinetic approach. Moreover, we have established here a way of converting results to equivalents of a common standard, providing values independent of its kinetic profile, by using the ratio between calibration sensitivities performed at endpoint conditions.

Fabrication of a flexible micro CO sensor for micro reformer applications.

Integration of a reformer and a proton exchange membrane fuel cell (PEMFC) is problematic due to the presence in the gas from the reforming process of a slight amount of carbon monoxide. Carbon monoxide poisons the catalyst of the proton exchange membrane fuel cell subsequently degrading the fuel cell performance, and necessitating the sublimation of the reaction gas before supplying to fuel cells. Based on the use of micro-electro-mechanical systems (MEMS) technology to manufacture flexible micro CO sensors, this study elucidates the relation between a micro CO sensor and different SnO2 thin film thicknesses. Experimental results indicate that the sensitivity increases at temperatures ranging from 100-300 °C. Additionally, the best sensitivity is obtained at a specific temperature. For instance, the best sensitivity of SnO2 thin film thickness of 100 nm at 300 °C is 59.3%. Moreover, a flexible micro CO sensor is embedded into a micro reformer to determine the CO concentration in each part of a micro reformer in the future, demonstrating the inner reaction of a micro reformer in depth and immediate detection.

Performance characteristics of a new generation pressure microsensor for physiologic applications.

A next generation fiber-optic microsensor based on the extrinsic Fabry-Perot interferometric (EFPI) technique has been developed for pressure measurements. The basic physics governing the operation of these sensors makes them relatively tolerant or immune to the effects of high-temperature, high-EMI, and highly-corrosive environments. This pressure microsensor represents a significant improvement in size and performance over previous generation sensors. To achieve the desired overall size and sensitivity, numerical modeling of diaphragm deflection was incorporated in the design, with the desired dimensions and calculated material properties. With an outer diameter of approximately 250 microm, a dynamic operating range of over 250 mmHg, and a sampling frequency of 960 Hz, this sensor is ideal for the minimally invasive measurement of physiologic pressures and incorporation in catheter-based instrumentation. Nine individual sensors were calibrated and characterized by comparing the output to a U.S. National Institute of Standards and Technology (NIST) Traceable reference pressure over the range of 0-250 mmHg. The microsensor performance demonstrated accuracy of better than 2% full-scale output, and repeatability, and hysteresis of better than 1% full-scale output. Additionally, fatigue effects on five additional sensors were 0.25% full-scale output after over 10,000 pressure cycles.