Design and Application of Multi Functional Light Addressable Cell-based Biosensors
|Keywords||Cytosensor The cells physiological parameters detection Light addressable potentiometric sensor (LAPS) Cellular microenvironment Myocardial cell chip for the differentiation of stem cells The light addressable cell impedance detection|
Living cells on the response to the stimulus signal is reflected in the changes in the parameters of the variety of cells, for example cell metabolism, and action potential and cell impedance changes. The cell sensor (Cell-based Biosensors, CBB) use of living cells as a sensing unit, through the two transducer cells in response to the parameter is converted to electrical signals detected. The light addressable cell sensor the (Light addressable cell-based biosensors, LACBB) as a special cell sensor light addressable potentiometric sensor (Light addressable potentiometric sensor, LAPS) as the two transducers using laser cell addressing cultured cells, can effectively solve the sensors of other types of cells need to locate and difficult. The start key technologies from the cell sensor design and instrument making, based on LAPS multifunctional optical addressing cell sensor to detect cell impedance extracellular potential application in the extracellular microenvironment study: combined the impedance detection technology and LAPS technology, a light-addressable cell impedance sensor (Light addressable cell impedance sensor LACIS) for the detection of cell attachment impedance characteristics; the combined is LAPS extracellular potential detection technology and stem cell technology designed myocardial cell chip for the differentiation of stem cells and for drug screening; extracellular potential and ion detection combined cellular electrophysiological parameters and metabolic parameters simultaneously detect the LAPS cell sensor; Finally, the combination of the LAPS ion detection technology and instrument development technology, design and complete can Real-time detection of a variety of cell metabolism parameters of the cellular microenvironment detector. Innovation and contribution of this article is to: 1. Proposed a method to detect cell impedance LAPS-based light-addressable. Cell impedance can reflect cell migration, proliferation, and the decline of physiological processes, is an important cell physiological parameters. The traditional cell impedance detection using metal electrode, and the reaction is the average of all the cells in contact with the electrode impedance. In this paper, the electrolyte - insulator - semiconductor (EIS) structure of the device, combined with LAPS principle of light-addressable and electrochemical impedance detection technology, first proposed a light-addressable detection cell impedance method, cells found by laser address, the local cell impedance variation detecting light region rather than the average of all cells change in impedance. This cell impedance detection method is more suitable for the detection of a small number of cells or even single cells, can also be applied to the cell impedance tomography. 2. Proposed based on LAPS stem cell differentiation myocardial cell chip design methods and applied to drug screening using mouse embryonic stem cells as a new source of cells and optimize the structure of LAPS sensor design and the LAPS stem cell differentiation of cardiomyocytes chips for cell electric Detection of Physiology. Stable extracellular action potential signal stem cells induced to differentiate into cardiomyocytes by LAPS detection and analysis of the of four drugs role of cardiac myocyte electrophysiology. Myocardial cells based on the differentiation of the stem cells of the LAPS chip research can be an effective solution to traditional extracellular potential detection sensors, such as micro-electrode array (Micro-electrode array, MEA), field-effect transistor (Field effect transistor, FET), in cellular localization cultured on difficulties, and to verify the new cell source of the stem cells can be used as a cell sensor. 3 based LAPS is a multifunctional cell sensor design method to achieve the multifunction detection of myocardial cell potential and extracellular ion. In order to be able to more accurately describe the physiological state of the cells under the action of external stimulation response, it is necessary to evaluate a variety of physiological activities, so developers cell sensor that can detect a variety of cell physiological parameter of great significance. This article is designed multifunctional cell sensors based on LAPS myocardial cells, for example, to achieve the simultaneous detection of the extracellular potential and extracellular acidification rate for the same time characterization the electrical physiological state of the cells and metabolic levels. Together to complete the system design and manufacture of the cellular microenvironment detection instrument, the initial realization of the functions of the automatic detection of the cellular microenvironment. The traditional the cellular microenvironment physiological dollars shortcomings in its detection parameters single instrument expensive, and difficult to achieve high throughput detection. The technology, based on the sensitive film sensor packaging technology, the fluid path design, the detection circuit design and prototype machining technology, is designed and completed LAPS the cellular microenvironment detection instrument used to measure the components of the extracellular microenvironment caused by cell metabolism changes. The instrument has eight channel measurement capability, and can measure a variety of parameters, compared to the cost micro physiological count was significantly reduced, the initial cell test that the apparatus can be effectively used for the automatic detection of the cellular microenvironment.