Application Notes
Fibox 3 LCD-trace: "Dissolved Oxygen Determination: Dynamic measurements in wine with a micro-oxygenation system"
María del Alamo and Ignacio Nevares, UVaMOX, E. T. S. Ingenierías Agrarias, Universidad de Valladolid, 34071 Palencia, Spain
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- Micro-oxygenation processes happening while storing wine in barrels give quality red wines the necessary final trait. Dissolved oxygen (DO) levels reached during the usual storage months are so low that measurement systems requirements become very high. There are measurement systems with detection limits at ppb (µg/L) based on electrochemical probes but they consume oxygen during the measuring process which is a problem as carrying out a dynamic measurement, i. e. in a fluid in motion, becomes necessary. Measurement of DO in wine flow has several disadvantages that can be solved but at the expenses of time and money. The optical measurement system Fibox 3 LCD-trace allows to deal with the need of measurement in systems with low volume, at different atmospheric pressures, and the advantage of carrying it out in an inerted way.
SDR: "Co-Cultures of Caco-2 cells and E.coli - Optosensoric on-line monitoring of O2 uptake and extracellular acidification"
Martin Brischwein and Daniel Grundl, Heinz-Nixdorf-Lehrstuhl für Medizinische Elektronik, TU München
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- Oligogalacturonic acids (OGAs), obtained by hydrolysis of pectin, are known to inhibit the adhesion of pathogenic bacteria on intestinal cells and thus to prevent infectious diseases. To test the effects of OGAs, an in-vitro model consisting of Caco-2 cells (human small intestinum) and the E.coli C25 strain was established. The co-cultures were set up in a novel, label-free cell assay platform working with a 24-well plate. The SensorDish® Reader SDR allows to monitor oxygen and pH kinetically in each well with chemical-optical sensors to analyse the rates of cellular oxygen uptake and extracellular acidification.
CO2: "Non-invasive On-line CO2 Measurement with Chemical-optical CO2 Sensors - Chemical-optical CO2 sensors provide a versatile tool for on-line monitoring of dissolved CO2 in various scale and designs"
Athanas Apostolidis, PreSens Precision Sensing GmbH, 93053 Regensburg, Germany
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- Accurate CO2 measurement in the liquid phase has been a challenge for decades. Although off-gas measurement by means of IR-spectroscopy is a proven technology, the correlation of CO2-levels in gas and liquid pahse in biological systems is difficult.
SDR: "On-line Oxygen Monitoring in Cell Culture. Effects of mitochondrial modulators on O2dynamics of mammalian cells."
Lynn S. G. and LaPres J. J., Dept. of Biochemistry, Michigan State University, East Lansing, MI, USA
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- Real-time measurements of oxygen and pH in mammalian cell cultures are important to understand the metabolic dynamics in cell cultures. The SDR SensorDish® Reader allows the time course of cellular oxygen consumption to be followed in response to environmental conditions or toxicological insults. This type of monitoring could also be valuable for other applications such as tissue engineering and stem cell research.
SFR: "Characterization of Microbial Growth Profiles. On-line detection of oxygen and pH during culture in shake flasks."
Konstantin Schneider1, Verena Schütz1, Gernot Thomas John2 and Elmar Heinzle1
1) Biochemical Engineering Institute, Saarland University, 66123 Saarbrücken, Germany; 2) PreSens Precision Sensing GmbH, 93053 Regensburg, Germany
- Shake flasks are the most commonly used cultivation systems for process and media optimization, screening for new products and strains as well as for the supply of inoculum for bioreactor cultivation. Oxygen limitation is one of the major though often neglected problems in aerobic shake flask cultivation. The new technology of the Shake Flask Reader SFR is an ideal tool for non-invasive monitoring of the two culture parameters oxygen and pH, and in addition reveals a detailed picture of growth performance and profiles.
SFR: "Insect cell cultivation in orbitally shaken flasks with sensors. Oxygen mass transfer in insect cell-based high cell density cultivations."
N. Riesen, C. Ries, R. Eibl, D. Eibl; Zurich University of Applied sciences, School of Life Sciences and Facility Managment, Institute of Biotechnology, Biochemical Engineering and Cell Cultivation Techniques, Wädenswil, Switzerland
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- Insect cell-based processes have become increasingly influential for tool protein and Virus-like Particle (VLP) vaccine productions over the last years. At small scale insect cells are normally grown in non-instrumented orbitally shaken disposable flasks, in most cases Erlenmeyers, in order to perform seed train productions and early stage process developments. The Shake Flask Reader (SFR) with non-invasive optical sensors for pH and DO is an important contribution to easy quantification of oxygen mass transfer in Erlenmeyers recently made. In addition, this allows controlled processing at mL-scale as exemplified for described Sf-9 and Sf-21 cell mass propagation procedures.
SFR: "Accurate Insight into Oxygen Content of Shake Cultures"
Michael Findeis and Gernot T. John, PreSens GmbH, 93053 Regensburg, Germany
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- Numerous factors affect growth in cell culture in shake flasks. The free oxygen content plays a decisive role. Insufficient oxygen input from the environment limits growth. With the new SFR measurement system the dissolved oxygen content of the medium can be determined non-invasively and extremely accurately. Continous oxygen monitoring offers advantages for growing new sensitive cell lines, for media optimisation and for QA-related process control.
SDR: "Non-invasive On-line Oxygen Measurement. Bacterial toxicity tests for use in pharmacological agent"
Anna Vasilevskaya, Institute for Analytical Chemistry, University of Regensburg, Germany
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- Measurement of oxygen kinetics in respiration inhibition tests is of central importance for pharmacological agent research and toxicology testing of chemicals (REACH). The new SDR SensorDish® Reader makes non-invasive, simultaneous, on-line recording in 24-well plates possible for the first time ever. The study describes the use of the system in a bacterial-respiratory toxicity test.
FTC pH: "pH Regulation for a Liver Cell Bioreactor. Prototype development with a chemical-optical pH flow-through cell."
Marco Decker, Fachhochschule für Technik und Wirtschaft Berlin (FHTW), Berlin
- An important aspect of efficient liver cell bioreactors is the automated regulation of physio-chemical culture parameters. The present study describes the development of the first non-invasive pH regulation device for a perfusion bioreactor. The high performance of the system is based on a chemical-optical flow-through cell for pH detection and its combination with precision mass-flow controllers for gas. The new controller allows long time stable and contamination free on-line pH regulation in complex bioreactor systems - an important technical contribution for future clinical applications.
SDR: "Process Monitoring in Suspension-Adapted CHO Cell Cultures. Non-invasive on-line detection of pH and oxygen"
Harry Abts and Sarina Arain, Celonic GmbH, Karl-Heinz-Beckurts-Straße 13, Jülich, Germany, PreSens GmbH, Josef-Engert-Str. 11, 93053 Regensburg, Germany
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- On-line detection of oxygen and pH by using SDR at a small-scale can be used as a basis for latter cultivation in large-scale bioreactors to detect optimal growth conditions. Contamination-free on-line monitoring of culture parameters is not only important for scaling up production in process development, but it also could be valuable for other applications such as tissue engineering, stem cell research, and toxicological tests.
SDR: "On-line Oxygen Monitoring System - Non-conventional steroid fermentations in microtiter plates"
Marques M.P.C.1, Cabral J.M.S. 1, Fernandes P., Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Instituto Superior Técnico, Lisboa, Portugal
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- A key factor when optimising and developing bioprocesses is the monitoring of the fermentation, in order to quantify in real-time basis parameters like oxygen, pH and carbon dioxide, among others. This monitoring by using SDR allows the time course of the fermentation to be followed and process parameters to be adjusted to desired levels. On-line monitoring in stirred vessels, namely fermenters, is a widespread approach but it has seldom been applied in shaken systems due to technical limitations.
SDR: "A Novel On-line Monitoring System - For Non-Invasive Oxygen Detection in Chondrocyte Cultures"
Bettina Giere, Tierärztliche Hochschule Hannover, Germany
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- For optimisation of cell culture systems it is essential to monitor important metabolic parameters like oxygen and pH quantitatively and to readjust them to physiological values if necessary. Deviations from the physiological limits cause substantial changes for cell growth and differentiation. The new SDR SensorDish® Reader enables non-invasive on-line monitoring of oxygen and pH in cell culture media. This is exemplified by a novel 3D chondrocyte culture model.
SDR: "3D Cutures of Disc-Chondrocytes - Optosensoric On-line Monitoring of Oxygen and pH Kinetics"
Karin Benz and Jürgen Mollenhauer, Regenerative Medizin II, NMI Reutlingen, Markwiesenstraße 55, 72770 Reutlingen, Germany
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- Hydrogel-based 3D cell culture systems offer particular advantages for tissue-engineering in cell-based therapy for disc-degeneration. However, light microscopical control of both culture quality and morphological differentiation is not possible within this dense matrix. For a judgement of the quality, alternative parameters have to be considered. In the present study, availability of oxygen within culture medium and kinetics of the pH value are analyzed in this regard by using the SDR. Both parameters are key factors for a successful and reliable in vitro cultivation and differentiation.