Microfluidic biosensing systems. Part II. Monitoring the dynamic production of glucose and ethanol from microchip-immobilised yeast cells using enzymatic chemiluminescent micro-biosensors

A microfluidic flow injection (microFIA) system was employed for handling and monitoring of cell-released products from living cells immobilised on silicon microchips. The dynamic release of glucose and ethanol produced from sucrose by immobilised Saccharomyces cerevisiae cells was determined using microchip biosensors (micro-biosensors) with either co-immobilised glucose oxidase-horseradish peroxidase (GOX-HRP), or alcohol oxidase-horseradish peroxidase (AOX-HRP), catalysing a series of reactions ending up with chemiluminescence (CL) generated from HRP-catalysed oxidation of luminol in presence of p-iodophenol (PIP). The yeast cells were attached by first treating them with polyethylenimine (PEI) followed by adsorption to the microchip surface. The cell loss during assaying was evaluated qualitatively using scanning electron microscopy (SEM), showing that no cells were lost after 35 min liquid handling of the cell chip at 10 microl min(-1). The enzymes were immobilised on microchips via PEI-treatment followed by glutaraldehyde (GA) activation. The GOX-HRP micro-biosensors could be used during five days without any noticeable decrease in response, while the AOX-HRP micro-biosensors showed continuously decreasing activity, but could still be used employing calibration correction. The glucose and ethanol released from the immobilised yeast chips were quantitatively monitored, by varying the incubation time with sucrose, showing the possibilities and advantages of using a microfluidic system set-up for cell-based assays.     

Peculiarities of the regulation of fermentation and respiration in the crabtree-negative,xylose-fermenting yeastPichia stipitis

Applied Biochemistry and Biotechnology - The respiration ofPichia stipitis was not repressed by either high concentrations of fermentable sugars or oxygen limitation. Fermentation was not induced...     

Langmuir Probe Diagnostics of a Low Temperature Non-Isothermal Plasma in a Weak Magnetic Field

This article presents measurements by a cylindrical Langmuir probe in the plasma of a DC cylindrical magnetron discharge át the pressure 1.5 Pa that aim at the experimental assessment of the influence of a weak magnetic field to the estimation of the electron density when using conventional methods of probe data interpretation. The probe data was obtained under the presence of a weak magnetic field in the range 1.10^?2?5.10^?2 T. The influence of the magnetic field on the electron probe current is experimentally assessed for two cylindrical probes with different radii, 50 μm and 21 μm. This assessment is based on comparison of the values of the electron density estimated from the electron current part with the values of the positive ion density estimated from the positive ion current part of the probe characteristic respectively by assuming that at the magnetic field strengths used in the present study the probe positive ion currents are possible to be assumed as uninfluenced by the magnetic field. For interpretation of the probe positive ion current two theories are used and compared to each other: the radial motion model by Allen, Boyd and Reynolds [10] and Chen [11] and the model that accounts for the collisions of positive ions with neutrals in the probe space charge sheath that we call Chen-Talbot model [8]. At lower magnetic field 3 · 10^?2 T the positive ion density values interpreted by using the Chen-Talbot model [8] are in better agreement with the values of electron density compared to those obtained by using the theory [10,11]; therefore the model [8] is used for calculation of the positive ion density from the probe data at higher magnetic fields. The comparison of the positive ion and electron density values calculated from the same probe data at higher magnetic fields shows that up to the magnetic field strength 4 . 10^?2 T with the probe 100 μm and up to 5 . 10^?2 T with the probe 42 μm in diameter respectively the decrease of the magnitude of the electron current at the space potential due to the magnetic field does not exceed the error limits that are usual for Langmuir probe measurements (absolute error ±20%).