Current-to-voltage convertor for measurement of oxygen

A highly sensitive, miniature, inexpensive circuit for the measurement of PO2 in vivo has been described. The circuit is constructed from a current-to-voltage convertor, clamping circuit, differential amplifier, and reverse voltage and overvoltage protector. The design of the circuit allows us to apply voltage bias to the measuring electrode while grounding the preparation. The clamping circuit holds the selected bias voltage constant while the differential amplifier subtracts this bias potential from the PO2 signal yielding an output voltage that is proportional to the current sensed by the oxygen electrode. The circuit is protected from reverse voltage and overvoltage.     

Self-referencing optrodes for measuring spatially resolved, real-time metabolic oxygen flux in plant systems

The ability to non-invasively measure metabolic oxygen flux is a very important tool for physiologists interested in a variety of questions ranging from basic metabolism, growth/development, and stress adaptation. Technologies for measuring oxygen concentration near the surface of cells/tissues include electrochemical and optical techniques. A wealth of knowledge was gained using these tools for quantifying real-time physiology. Fiber-optic microprobes (optrodes) have recently been developed for measuring oxygen in a variety of biomedical and environmental applications. We have adopted the use of these optical microsensors for plant physiology applications, and used the microsensors in an advanced sensing modality known as self-referencing. Self-referencing is a non-invasive microsensor technique used for measuring real-time flux of analytes. This paper demonstrates the use of optical microsensors for non-invasively measuring rhizosphere oxygen flux associated with respiration in plant roots, as well as boundary layer oxygen flux in phytoplankton mats. Highly sensitive/selective optrodes had little to no hysteresis/calibration drift during experimentation, and an extremely high signal-to-noise ratio. We have used this new tool to compare various aspects of rhizosphere oxygen flux for roots of Glycine max, Zea mays, and Phaseolus vulgaris, and also mapped developmentally relevant profiles and distinct temporal patterns. We also characterized real-time respiratory patterns during inhibition of cytochrome and alternative oxidase pathways via pharmacology. Boundary layer oxygen flux was also measured for a phytoplankton mat during dark:light cycling and exposure to pharamacological inhibitors. This highly sensitive technology enables non-invasive study of oxygen transport in plant systems under physiologically relevant conditions.     

Design, operation, and modeling of a membrane photobioreactor to study the growth of the Cyanobacterium Arthrospira platensis in space conditions

A membrane photobioreactor was designed, implemented and used to grow the cyanobacterium Arthrospira platensis PCC 8005 in batch mode. Growth was followed directly by monitoring optical density and indirectly by measuring pressure increase due to the oxygen produced and separated from the liquid phase by diffusion through a hydrophobic membrane, and pH increase due to carbon consumption. When the pressure attained an upper limit, valves opened automatically, and the oxygen in the gas chamber was flushed out with nitrogen. As expected, two growth phases were observed, a short exponential phase followed by a linear phase, indicating limitation by light transfer. Growth rate during the second phase was measured easily and accurately, and consistency of optical density, pressure and pH data values was checked using a model of the system. Pressure measurement was found best suited to monitoring and measuring growth rate in space in terms of accuracy, precision and reliability.     

A versatile chamber for simultaneous measurements of oxygen exchange and fluorescence in filamentous and thallous algae as well as higher plants

A new chamber was developed for a simultaneous measurement of fluorescence kinetics and oxygen exchange in filamentous and thallous algae as well as in small leaves of water plants. Algal filaments or thalli are kept by a stainless grid close to the bottom window of the chamber in the sample compartment. The grid separates the object from the electrode compartment with the oxygen electrode at the top. This compartment accommodates, in addition, a magnetic stirrer that provides efficient circulation of the medium between the sample and the electrode. This magnetic bar spins on a fixed axis and is driven by an electronically commutated magnetic field produced by four coils which are arranged around the chamber. This design yields a very favourable signal to noise ratio in the oxygen electrode records. Consequently, measurements can be performed even of algae with very low photosynthetic rates such as marine low-light red algae or algae under severe stress. For irradiation of the samples and for fluorescence measurements a fibre optic light guide is used facing the window of the chamber. The four branches of a commercially available light guide serve the following purposes: collection of sample fluorescence and supply of measuring, actinic, and saturating light, respectively.     

A versatile chamber for simultaneous measurements of oxygen exchange and fluorescence in filamentous and thallous algae as well as higher plants

A new chamber was developed for a simultaneous measurement of fluorescence kinetics and oxygen exchange in filamentous and thallous algae as well as in small leaves of water plants. Algal filaments or thalli are kept by a stainless grid close to the bottom window of the chamber in the sample compartment. The grid separates the object from the electrode compartment with the oxygen electrode at the top. This compartment accommodates, in addition, a magnetic stirrer that provides efficient circulation of the medium between the sample and the electrode. This magnetic bar spins on a fixed axis and is driven by an electronically commutated magnetic field produced by four coils which are arranged around the chamber. This design yields a very favourable signal to noise ratio in the oxygen electrode records. Consequently, measurements can be performed even of algae with very low photosynthetic rates such as marine low-light red algae or algae under severe stress. For irradiation of the samples and for fluorescence measurements a fibre optic light guide is used facing the window of the chamber. The four branches of a commercially available light guide serve the following purposes: collection of sample fluorescence and supply of measuring, actinic, and saturating light, respectively.This revised version was published online in March 2005 with corrections to the page numbers.     

Red blood cell velocity and oxygen tension measurement in cerebral microvessels by double-wavelength photoexcitation.

Because the regulation of microcirculation in the cerebral cortex cannot be analyzed without measuring the blood flow dynamics and oxygen concentration in cerebral microvessels, we developed a fluorescence and phosphorescence system for estimating red blood cell velocity and oxygen tension in cerebral microcirculation noninvasively and continuously with high spatial resolution. Using red blood cells labeled with fluorescent isothiocyanate to visualize red cell distribution and using the oxygen quenching of Pd-meso-tetra-(4-carboxyphenyl)-porphyrin phosphorescence to measure oxygen tension enabled simultaneous measurement of blood velocity and oxygen tension. We examined how the measurement accuracy was affected by the spatial resolution and by the excitation laser light passing through the targeted microvessel and exciting the oxygen probe dye in the tissue beneath it. Focusing the excitation light into the microvessel stabilized the phosphorescence lifetime at each spatial resolution; moreover, it greatly reduced phosphorescence from the brain tissue. Animal experiments involving acute hemorrhagic shock demonstrated the feasibility of our system by showing that the changes in venular velocity and oxygen tension are synchronized to the change in mean arterial pressure. Our system measures the red cell velocity and oxygen concentration in the cerebral microcirculation by using the differences in luminescence and wavelength between fluorescence and phosphorescence, making it possible to easily acquire information about cerebral microcirculatory distribution and oxygen tension simultaneously.     

THE TOTAL LUMINOUS EFFICIENCY OF LUMINOUS BACTERIA

Methods are described for measuring the light emitted by an emulsion of luminous bacteria of given thickness, and calculating the light emitted by a single bacterium, measuring 1.1 x 2.2 micra, provided there is no absorption of light in the emulsion. At the same time, the oxygen consumed by a single bacterium was measured by recording the time for the bacteria to use up .9 of the oxygen dissolved in sea water from air (20 per cent oxygen). The luminescence intensity does not diminish until the oxygen concentration falls below 2 per cent, when the luminescence diminishes rapidly. Above 2 per cent oxygen (when the oxygen dissolving in sea water from pure oxygen at 760 mm. Hg pressure = 100 per cent) the bacteria use equal amounts of oxygen in equal times, while below 2 per cent oxygen it seems very likely that rate of oxygen absorption is proportional to oxygen concentration. By measuring the time for a tube of luminous bacteria of known concentration saturated with air (20 per cent oxygen) to begin to darken (2 per cent oxygen) we can calculate the oxygen absorbed by one bacterium per second. The bacteria per cc. are counted on a blood counting slide or by a centrifugal method, after measuring the volume of a single bacterium (1.695 x 10^–12 cc.). Both methods gave results in good agreement with each other. The maximum value for the light from a single bacterium was 24 x 10^–14 lumens or 1.9 x 10^–14 candles. The maximum value for lumen-seconds per mg. of oxygen absorbed was 14. The average value for lumen-seconds per mg. O₂ was 9.25. The maximum values were selected in calculating the efficiency of light production, since some of the bacteria counted may not be producing light, although they may still be using oxygen. The "diet" of the bacteria was 60 per cent glycerol and 40 per cent peptone. To oxidize this mixture each mg. of oxygen would yield 3.38 gm. calories or 14.1 watts per second. 1 lumen per watt is therefore produced by a normal bacterium which emits 14 lumen-seconds per mg. O₂ absorbed. Since the maximum lumens per watt are 640, representing 100 per cent efficiency, the total luminous efficiency if .00156. As some of the oxygen is used in respiratory oxidation which may have nothing to do with luminescence, the luminescence efficiency must be higher than 1 lumen per watt. Experiments with KCN show that this substance may reduce the oxygen consumption to 1/20 of its former value while reducing the luminescence intensity only ¼. A partial separation of respiratory from luminescence oxidations is therefore effected by KCN, and our efficiency becomes 5 lumens per watt, or .0078. This is an over-all efficiency, based on the energy value of the "fuel" of the bacteria, regarded as a power plant for producing light. It compares very favorably with the 1.6 lumens per watt of a tungsten vacuum lamp or the 3.9 lumens per watt of a tungsten nitrogen lamp, if we correct the usual values for these illuminants, based on watts at the lamp terminals, for a 20 per cent efficiency of the power plant converting the energy of coal fuel into electric current. The specific luminous emission of the bacteria is 3.14 x 10^–6 lumens per cm². One bacterium absorbs 215,000 molecules of oxygen per second and emits 1,280 quanta of light at λ_max = 510µµ. If we suppose that a molecule of oxygen uniting with luminous material gives rise to the emission of 1 quantum of light energy, only 1/168 of the oxygen absorbed is used in luminescence. On this basis the efficiency becomes 168 lumens per watt or 26.2 per cent.     

A method for continuous in vivo measurement of hemolymph conductivity in crabs

A miniaturized conductivity measuring device has been developed to monitor in vivo hemolymph conductivity in crabs. The device measures the conductivity by means of implanted platinum electrodes, and transmits the data via radiotelemetry to a receiving station through 6 ft of water. Crabs used in these investigations survived the insertion of the conductivity probe and appeared to behave and feed normally for up to one week with the probe in position. Blue crabs, Callinectes sapidus Rathbun, which were acclimated to either 5 or 35 ‰ were abruptly transferred to 35 and 5 ‰, respectively and the rate of change of hemolymph conductivity sucessfully monitored. The time required for the conductivity (ionic composition) change in the hemolymph of the crabs after transfer varied from 12 to 16 h for crabs transferred from 35 to 5 ‰ to less than an hour when the change was from 5 to 35 ‰.     

Continuous measurement of glucose utilization in heart myoblasts

Understanding quantitative aspects of cell energy metabolism and how it is influenced by environment is central to biology, medicine, and biotechnology. Most methods used for measuring metabolic fluxes associated with energy metabolism require considerable personnel effort or high maintenance instrumentation. The microphysiometer is a commercially available instrument that measures acid extrusion rates, which are commonly used for drug screening. With the addition of oxygen sensors, the instrument can also be used to measure cell oxygen consumption rates and thereby calculate glycolytic fluxes. In the work described here, oxygen consumption and acid extrusion rates were used to measure glucose utilization by the H9c2 rat heart myoblast cell line and these results are compared with fluxes measured with a radiometric assay. Both assays were used to investigate changes in H9c2 energy metabolism due to cell stimulation with carbachol and insulin. The results demonstrate the utility of the microphysiometer method for measuring both transient and sustained changes in partitioning of glucose utilization between glycolysis and oxidation in live cells.     

Photobioreactor for cultivation and real‐time,in‐situ measurement of O2 and CO2 exchange rates,growth dynamics,and of chlorophyll fluorescence emission of photoautotrophic microorganisms

A detailed knowledge about the dynamics of phytoplanktonic photosynthesis and respiration is crucial for the determination of primary productivity in open oceans as well as for biotechnological applications. The dynamics are best studied in photobioreactors that are able to simulate natural conditions in such, that light can be modulated not only diurnally but also mimicking effects of solar elevation angle from sunrise to sunset, variable cloudiness, light modulation in refractory sun flecks due to water waves, or light intermittence due to turbulent flow in dense suspensions. In addition, high performance photobioreactors ought to be able to monitor in real time photosynthetic and respiratory activities as well as culture growth. Here, we demonstrate performance of a newly designed bench‐top laboratory photobioreactor that meets these needs, with a study of green alga Scenedesmus quadricauda. The algal suspension was exposed to simulated daily variations of total photosynthetic active irradiance and spectral profile, with a larger proportion of red photons in the morning and evening hours. The instrument monitored automatically the culture growth by measuring the optical densities at 735 nm and 680 nm and by measuring steady state and maximal chlorophyll fluorescence emission yields. The photochemical yields were estimated from chlorophyll fluorescence data. These widely used but rather indirect yield estimates were confronted with direct measurements of oxygen evolution and consumption quantum yields. The CO₂ fluxes in and out of the culture media as well as the dissolved CO₂ in algal suspension were also recorded. The experiments demonstrated potential of the new photobioreactor to reveal minute modulations in gas exchange rates as well as to yield data for calculation of photon requirement of oxygen evolution in the suspension volume that is key technological parameter for planning of large scale photobioreactors as well as key optimization parameter for strain selection.     

Photosynthesis, Respiration and Post-illumination Fixation of CO2 by Corn Leaves as Influenced by Light and Oxygen Concentration

The effect of oxygen concentration on the rate of CO₂-uptake in continuous and intermittent light was studied as well as the CO₂-fixation during a short dark period after light was turned off. In addition the dark respiration and the CO₂-compensation point of attached and detached corn leaves were determined. Leaves of 4 to 22-day old plants were used as experimental material. A closed circuit system of an infrared carbon dioxide analyzer was employed to measure the rate of CO₂-exchange. It was found that in an atmosphere consisting of 100 % oxygen, there was about 50 per cent inhibition of the rate of CO₂-uptake in continuous and intermittent light compared to that in an atmosphere consisting of 21% oxygen. The same was true of the rate of CO₂-fixation in darkness during a short period after the light was turned off. Since the response to oxygen concentration of the CO₂-uptake in light and of the CO₂-fixation in darkness after the light was turned off were similar, it is concluded that the fixation of CO₂ in the short dark period represents an over- shoot of photosynthesis. The rate of dark respiration was little affected by the oxygen concentration in the ranges used in the experiments. The carbon dioxide compensation point which has been observed in leaves of 4 to 14-day old plants was not influenced by either oxygen concentration or light intensity. Since the changes in the rate of CO₂-uptake due to changes in the concentration of oxygen and light intensity had no effect on the CO₂-compensation point, it is concluded that a reabsorption of respiratory CO₂ by photosynthesis could not account for the low value of this point. These results are interpreted as a further corroboration of the statement that the leaves of corn lack the process of photorespiration and that dark respiration is inhibited in light. It was observed that the rate of the CO₂-uptake gradually increased in plants which were from 4 to 22-days old. The inhibitory effect of high concentration of oxygen on the rate of CO₂-uptake was relatively higher in old plants than in young ones.     

Photosynthesis, Photorespiration and Respiration of Detached Spruce Twigs as Influenced by Oxygen Concentration and Light Intensity

The effects of oxygen concentration and light intensity on the rates of apparent photosynthesis, true photosynthesis, photorespiration and dark respiration of detached spruce twigs were determined by means of an infra-red carbon dioxide analyzer (IRCA). A closed circuit system IRCA was filled with either 1 per cent of oxygen in nitrogen, air (21 % O₂) or pure oxygen (100 % O₂). Two light intensities 30 × 10³ erg · cm ^?2· s^?1 and 120 × 10³ erg · cm^?2· s^?1 were applied. It has been found that the inhibitory effect of high concentration of oxygen on the apparent photosynthesis was mainly a result of a stimulation of the rate of CO₂ production in light (photorespiration). In the atmosphere of 100 % O₂, photorespiration accounts for 66–80 per cent of total CO₂ uptake (true photosynthesis). Owing to a strong acceleration of photorespiration by high oxygen concentrations, the rate of true photosynthesis calculated as the sum of apparent photosynthesis and photorespiration was by several times less inhibited by oxygen than the rate of apparent photosynthesis. The rates of dark respiration were essentially unaffected by the oxygen concentrations used in the experiments. An increase in the intensity of light from 30 × 10³ erg · cm^?3· s^?1 to 120 · 10³ erg · cm^?2· s^?1 enhanced the rate of photorespiration in the atmospheres of 21 and 100 % oxygen but not in 1 % O₂. The rate of apparent photosynthesis, however, was little affected by light intensity in an atmosphere of 1 % oxygen.     

Capsule Design for Blue Light Therapy against Helicobacter pylori

A photo-medical capsule that emits blue light for Helicobacter pylori treatment was described in this paper. The system consists of modules for pH sensing and measuring, light-emitting diode driver circuit, radio communication and microcontroller, and power management. The system can differentiate locations by monitoring the pH values of the gastrointestinal tract, and turn on and off the blue light according to the preset range of pH values. Our experimental tests show that the capsule can operate in the effective light therapy mode for more than 32 minutes and the wireless communication module can reliably transmit the measured pH value to a receiver located outside the body.     

Light‐Driven BiVO4–C Fuel Cell with Simultaneous Production of H2O2

Photoelectrochemical (PEC) systems have been researched for decades due to their great promise to convert sunlight to fuels. The majority of the research on PEC has been using light to split water to hydrogen and oxygen, and its performance is limited by the need of additional bias. Another research direction on PEC using light, is to decompose organic materials while producing electricity. In this work, the authors report a new type of unassisted PEC system that uses light, water and oxygen to simultaneously produce electricity and hydrogen peroxide (H₂O₂) on both the photoanode and cathode, which is essentially a light‐driven fuel cell with H₂O₂ as the main product at the two electrodes, meanwhile achieving a maximum power density of 0.194 mW cm^‐2, an open circuit voltage of 0.61 V, and a short circuit current density of 1.09 mA cm^‐2. The electricity output can be further used as a sign for cell function when accompanied by a detector such as a light‐emitting diode (LED) light or a multimeter. This is the first work that shows H₂O₂ two‐side generation with a strict key factors study of the system, with a clear demonstration of electricity output ability using low‐cost earth abundant materials on both sides, which represents an exciting new direction for PEC systems.     

Implementing conventional logic unconventionally: photochromic molecular populations as registers and logic gates

In this paper we detail experimental methods to implement registers, logic gates and logic circuits using populations of photochromic molecules exposed to sequences of light pulses. Photochromic molecules are molecules with two or more stable states that can be switched reversibly between states by illuminating with appropriate wavelengths of radiation. Registers are implemented by using the concentration of molecules in each state in a given sample to represent an integer value. The register's value can then be read using the intensity of a fluorescence signal from the sample. Logic gates have been implemented using a register with inputs in the form of light pulses to implement 1-input/1-output and 2-input/1-output logic gates. A proof of concept logic circuit is also demonstrated; coupled with the software workflow describe the transition from a circuit design to the corresponding sequence of light pulses.     

Electromedical patient protection circuit]

As protection against too high an input voltage, anti-parallel diodes have long been incorporated in the input circuit of electromedical measuring instruments. This protective principle also functions in the other direction and can be used in an appropriate circuit design to protect the patient against earth fault currents, originating from the appliance. After a short review of present electromedical protection systems, the author describes the new protective circuit and initiates a discussion about it and still open questions.     

基于ADS8332的生理信号采集系统电路设计

本文介绍了医学生理信号的的特点,根据其特点设计出了针对医学生理信号进行采集的电路。该电路设计实现了基于16位的高精度A/D 转换芯片ADS8332 的医学生理信号采集系统,以基于Cortex-M3 内核的32 位微控制器STM32F103RD作为采集控制芯片,该系统电路可以实现对最多16 个通道的模拟医学生理信号进行采集。此电路设计充分利用了ADS8332 的高精度、低功耗、高采样速率的特点以及片上集成功能,并配合了信号调理电路和相应的抗干扰措施,因此保证了医学生理信号的采集精度和系统的稳定性。     

System for measuring oxygen consumption rates of mammalian cells in static culture under hypoxic conditions

Estimating the oxygen consumption rates (OCRs) of mammalian cells in hypoxic environments is essential for designing and developing a three‐dimensional (3‐D) cell culture system. However, OCR measurements under hypoxic conditions are infrequently reported in the literature. Here, we developed a system for measuring OCRs at low oxygen levels. The system injects nitrogen gas into the environment and measures the oxygen concentration by an optical oxygen microsensor that consumes no oxygen. The developed system was applied to HepG2 cells in static culture. Specifically, we measured the spatial profiles of the local dissolved oxygen concentration in the medium, then estimated the OCRs of the cells. The OCRs, and also the pericellular oxygen concentrations, decreased nonlinearly as the oxygen partial pressure in the environment decreased from 19% to 1%. The OCRs also depended on the culture period and the matrix used for coating the dish surface. Using this system, we can precisely estimate the OCRs of various cell types under environments that mimic 3‐D culture conditions, contributing crucial data for an efficient 3‐D culture system design. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 32:189–197, 2016     

Phase fluorometric sterilizable optical oxygen sensor

We report here on a low-cost, optical oxygen sensor as an attractive alternative to the widely used amperometric Clark-type oxygen electrode for measuring dissolved oxygen tensions in cell cultures and bioreactor. Our sensor is based on the defferential quenching of the fluorescence lifetime of chromophore in response to the partial pressure of oxygen. This is measured as a phase shift in fluorescence emission from the chromophore due to oxygen quenching when excited by an intensity modulated beam of light. In this article we demonstrate the advantages of lifetime-based optical methods over both intensity based optical methods and amperometric electrodes. Our sensor is particularly suitable for measuring dissolved oxygen in bioreactors. It is autoclavable, is free of maintenance requirements, and solvents the problems of long-term stability, calibration drifts, and reliable measurement of low oxygen tensions in dense microbial cultures that limit the utility of Clark-type elcectordes. (c) 1994 John Wiley & Sons, Inc.     

Correlation of stable oxygen isotope temperature record with light attenuation profiles in reef-dwellingTridacna shells

Molluscs are known to record environmental changes in their carbonate shells in detail. This paper reports the findings of a high-resolution analysis of stable oxygen isotopic compositions and light transmission properties of a shell of the reef-dwelling Pacific giant clamTridacna gigas. Our findings reveal that the annual growth rates and the longevity ofTridacna specimens can be readily determined by measuring the annual light attenuation pattern within the shell. Annual seasonal changes in water temperature are reflected with high resolution in the stable oxygen isotope ratios and in the light attenuation values of the aragonite shell. The inner shell ofT. gigas deposited below the pallial line revealing undisturbed shell accretion with high growth rates shows the maximum seasonal oxygen isotope range and the highest resolution in light attenuation changes. We suggest that this is the best part of the shell to reconstruct former seasonal surface water temperatures in tropical environments. Scanning electron microscopy (SEM) studies suggest that the annual growth patterns observed in transmitted light are generated by a complex pattern of daily growth increments with varying sizes of skeletal crystallites and varying amounts of organic carbon.