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Cell culture assays for therapeutic drug screening today are fully automated. Vitality of the cells is monitored by different sensors. For such a system, we propose a new reader unit, which is capable of reading two different fluorescent sensors and electrical impedance in 24-well-plates. Main goals are to reduce cost, complexity and size while achieving a similar performance as the existing reader unit. To achieve this, measurement electronics and signal paths for frequency domain fluorescence and bio-impedance measurement are combined. Central component is an integrated circuit for impedance spectroscopy. A new compact and economic optical setup is developed to read two different sensor spots on the bottom of the well. Measurement errors introduced by different components like DFT leakage, and frequency dependent signal delays are evaluated and compensated. A set of commercially available fluorescence sensor spots is used to verify the read out performance. The results are usable, with noise slightly higher than commercial readers. To verify the impedance measurement accuracy, measurements of known resistances are conducted. In the relevant impedance and frequency range for biological applications a suitable accuracy is achieved. Due to the higher sampling rate of the new reader, the higher noise can be reduced through averaging. The new system is significantly smaller and cheaper to manufacture than commercially available devices., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Cellular assays have become a fundamental technique in scientific research, pharmaceutical drug screening or toxicity testing. Therefore, the requirements of technical developments for automated assays raised in the same rate. A novel measuring platform was developed, which combines automated assay processing with label-free high-content measuring and real-time monitoring of multiple metabolic and morphologic parameters of living cells or tissues. Core of the system is a test plate with 24 cell culture wells, each equipped with opto-chemical sensor-spots for the determination of cellular oxygen consumption and extracellular acidification, next to electrode-structures for electrical impedance sensing. An automated microscope provides the optical sensor read-out and allows continuous cell imaging. Media and drugs are supplied by a pipetting robot system. Therefore, assay can run over several days without personnel interaction. To demonstrate the performance of the platform in physiologic assays, we continuously recorded the kinetics of metabolic and morphologic parameters of MCF-7 breast cancer cells under the influence of the cytotoxin chloroacetaldehyde. The data point out the time resolved effect kinetics over the complete treatment period. Thereby, the measuring platform overcomes problems of endpoint tests, which cannot monitor the kinetics of different parameters of the same cell population over longer time periods., (Copyright © 2013 Elsevier B.V. All rights reserved.)

In vivo, the pH value and oxygen partial pressure are the most important physico-chemical parameters in the microenvironment of human tissues. In vitro, the extracellular acidification rate of cell cultures is an indicator of global cellular metabolism, while the rate of oxygen consumption is a measure of mitochondrial activity. Earlier approaches had the disadvantage that these two values had to be measured with two separate sensors at different loci within the tissue or cell culture. Furthermore, conventional Clark-type oxygen sensors are not very compatible for miniaturisation, making it impossible to measure at small cell volumes or even at the single cell level. We have, therefore, developed an ISFET based sensor structure which is able to measure both pH and oxygen partial pressure. This sensor structure was tested in vitro for simultaneous records of cellular acidification and respiration rates at the same site within the cell culture. This sensor is manufactured by a CMOS-process.

The pH in the cellular microenvironment (pH(M)) is an important regulator of cell-to-cell and cell-to-host interactions. Additionally the extracellular acidification rate of a cell culture is an important indicator of global cellular metabolism. In a new approach a biocompatible ion-sensitive field effect transistor (ISFET)-array was developed to measure the pH(M) close to a surface and the global extracellular acidification rate at the same time. This ISFET-array is part of a new multiparametric microsensor chip. The paper highlights some basic applications of this method for in-vitro measurements. Using a fluid perfusion system for cell culture media, it is possible to measure the pH(M) of few (five to ten) adherent tumor cells in a distance of 10-100 nm from the cell plasma membrane. Experiments showed a pH(M)-value of 6.68 +/- 0.06 pH. Further experiments suggest that both the low pH, and the extracellular acidification rate of the examined tumor cell line are mainly built up by glycolysis.

Microsensors provide instruments particularly suited for the noninvasive analysis of cell and tissue cultures. The outstanding benefit lies in the passive behaviour of continuously working transducers, which in turn allows the dynamic recording of function-specific cellular processes. The microsensor system presented in this paper is a modular arrangement of various planar and non-planar sensor elements surrounding small cell culture chambers. An optic access to the cultures (e.g. for high resolution light microscopy and spectro-photometric techniques) enables a parallel and comparative data acquisition. The system was originally designed for biomedical research in chemotherapy and pharmacology but it proved to be an effective device both for toxicological and environmental research.

A new method for on-line and real-time monitoring of concentration, growth and physiological state of cells in culture is described. This biosensor is based on impedance measurements of adherently growing cells on interdigitated electrode structures (IDES). The measurements can be performed for several days as there is no detectable electrical influence on the cells. The versatility of this new sensor is shown with some exemplary experiments. Cell density, growth and long-term behaviour of cells on the electrodes clearly change the impedance of the IDES. Both, the global influence of serum components (deprivation of foetal bovine serum) and the toxic effects of heavy metal ions (cadmium) result in changes of the sensor signal and can be visualized.