Cell-based biosensors based on light-addressable potentiometric sensors for single cell monitoring

Biosensor National Special Laboratory, Key Laboratory of Biomedical Engineering of the Ministry of Education, Department of Biomedical Engineering, PO Box 1590, Zhejiang University, Hangzhou 310027, China.
Biosensors & Bioelectronics (Impact Factor: 6.41). 04/2005; 20(9):1757-63. DOI: 10.1016/j.bios.2004.06.037
Source: PubMed


Cell-based biosensors incorporate cells as sensing elements that convert changes in immediate environment to signals for processing. This paper reports an investigation on light-addressable potentiometric sensor (LAPS) to be used as a possible cell-base biosensor that will enable us to monitor extracellular action potential of single living cell under stimulant. In order to modify chip surface and immobilize cells, we coat a layer of poly-L-ornithine and laminin on surface of LAPS chip on which rat cortical cells are grown well. When 10 microg/ml acetylcholine solution is administrated, the light pointer is focused on a single neuronal cell and the extracellular action potential of the targeted cell is recorded with cell-based biosensor based on LAPS. The results demonstrate that this kind of biosensor has potential to monitor electrophysiology of living cell non-invasive for a long term, and to evaluate drugs primarily.

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    • "Such responses can then be converted to useful information by combining them with conventional detection for changes in cellular metabolism, cytotoxic responses, and bioavailability of medicinal substances. Therefore, the biosensing technology has been applied as an essential tool to various bio-analytical sectors including drug discovery, toxicology, pharmacology, bio-assay, pathogen and toxin screening, environmental monitoring, and bio-security2345. "
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    ABSTRACT: For monitoring of human cellular response to repetitive bacterial stimulations (e.g., Pseudomonas aeruginosa in a lysate form), we devised a chemiluminescent immuno-analytical system for toll-like receptor 1 (TLR1) as marker present on cell surfaces (e.g., A549). Upon stimulation, TLR1 recognizes pathogen-associated molecular patterns of the infectious agent and are then up-regulated via activation of the nuclear factor-κB (NF-κB) pathway. In this study, the receptor density was quantified by employing an antibody specific to the target receptor and by producing a chemiluminometric signal from an enzyme labeled to the binder. The activated status was then switched back to normal down-regulated stage, by changing the culture medium to one containing animal serum. The major factors affecting activation were the stimulation dose of the bacterial lysate, stimulation timing during starvation, and up- and down-regulation time intervals. Reiterative TLR regulation switching up to three times was not affected by either antibody remained after immunoassay or enzyme substrate (e.g., hydrogen peroxide) in solution. This immuno-analysis for TLRs could be unique to acquire accumulated response of the human cells to repeated stimulations and, therefore, can eventually apply to persistency testing of the cellular regulation in screening of anti-inflammatory substances.
    Scientific Reports 08/2014; 4(6011). DOI:10.1038/srep06011 · 5.58 Impact Factor
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    • "We chose n-type silicon (100) as the substrate to fabricate the LAPS chip (Xu et al., 2005; Liu et al., 2006). The silicon was covered with a 30-nm-thick SiO 2 layer on the positive side, and thermally oxidized at 1180 °C for 20 min. "
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    ABSTRACT: Neurochip based on light-addressable potentiometric sensor (LAPS), whose sensing elements are excitable cells, can monitor electrophysiological properties of cultured neuron networks with cellular signals well analyzed. Here we report a kind of neurochip with rat pheochromocytoma (PC12) cells hybrid with LAPS and a method of de-noising signals based on wavelet transform. Cells were cultured on LAPS for several days to form networks, and we then used LAPS system to detect the extracellular potentials with signals de-noised according to decomposition in the time-frequency space. The signal was decomposed into various scales, and coefficients were processed based on the properties of each layer. At last, signal was reconstructed based on the new coefficients. The results show that after de-noising, baseline drift is removed and signal-to-noise ratio is increased. It suggests that the neurochip of PC12 cells coupled to LAPS is stable and suitable for long-term and non-invasive measurement of cell electrophysiological properties with wavelet transform, taking advantage of its time-frequency localization analysis to reduce noise.
    Journal of Zhejiang University SCIENCE B 05/2010; 11(5):323-31. DOI:10.1631/jzus.B0900349 · 1.28 Impact Factor
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    • "The measurement setup is illustrated in Fig.4 and introduced elsewhere (Xu et al., 2005), with a Fig.2 "
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    ABSTRACT: Titanium dioxide (TiO(2)) thin film was deposited on the surface of the light addressable potentiometric sensor (LAPS) to modify the sensor surface for the non-labeled detection of DNA molecules. To evaluate the effect of ultraviolet (UV) treatment on the silanization level of TiO(2) thin film by 3-aminopropyltriethoxysilane (APTS), fluorescein isothiocyanate (FITC) was used to label the amine group on the end of APTS immobilized onto the TiO(2) thin film. We found that, with UV irradiation, the silanization level of the irradiated area of the TiO(2) film was improved compared with the non-irradiated area under well-controlled conditions. This result indicates that TiO(2) can act as a coating material on the biosensor surface to improve the effect and efficiency of the covalent immobilization of biomolecules on the sensor surface. The artificially synthesized probe DNA molecules were covalently linked onto the surface of TiO(2) film. The hybridization of probe DNA and target DNA was monitored by the recording of I-V curves that shift along the voltage axis during the process of reaction. A significant LAPS signal can be detected at 10 micromol/L of target DNA sample.
    Journal of Zhejiang University SCIENCE B 11/2009; 10(11):860-6. DOI:10.1631/jzus.B0920090 · 1.28 Impact Factor
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