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    脂双层记录: Orbit产品系列

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    CardioExcyte 96 SOL:用光遗传的手段起搏心肌细胞

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2020 - Cell Monitoring using Multi-Frequency Impedance Recordings for Label-free and Time-resolved Cell Response Analysis

 Icon CE   CardioExcyte 96 poster, Safety Pharmacology Society Virtual Meeting 2020   logo pdf   (1.6 MB)

Abstract:

Reading the impedance of planar gold-film electrodes that are used as growth substrate for adherent cells reveals changes in electrode coverage or cell morphology. Thus, real time impedance data provide insights in various cell characteristics, such as cell morphology, proliferation, lateral migration or cytotoxicity even over prolonged periods of time. A crucial advantage over standard assays is the continuous cell monitoring. Advanced information content is obtained by using multi-frequency impedance readouts as they allow zooming in on changes in membrane topography, cell-cell or cell- matrix junctions deconvoluting the complex whole cell response, for instance, to G-proteincoupled receptor (GPCRs) activation. Multi-frequency impedance measurements were used to monitor cell adhesion, cell specific structural changes, proliferation and cytotoxicity. The multi-frequency character of the data allows selecting the most sensitive frequency for a particular application, cell type or detail of the cell response after data acquisition is complete.
Impedance spectra were calculated for HeLa, HEK293 and CHO cells to demonstrate cell-type specific differences under stationary conditions. Data acquisition was performed using the CardioExcyte96 impedance recording system. Cells covering the electrode impede the applied current due to insulating properties of their membranes and the individual tightness of their cell junctions. Cell responses to experimental challenges will induce corresponding impedance changes. Multi- frequency impedance analysis provides a stronger basis for datainterpretation as it highlights the changes in specific cell structures. HeLa, HEK293 and CHO cells show different proliferation curves. Frequency dependent impedance data reveal individual drifts of the most sensitive frequency, i.e. of the maximal responses indicating cell type specific changes in cell junctions and proliferation dynamics. The expected concentration dependent toxicity was observed for DMSO, Escin and Triton-X by multi-frequency impedance recordings showing decreasing impedance values with exposure time and drifting of the maximal responses to lower frequencies. GPCR-mediated signal transduction was investigated using cells recombinantly expressing the dopamine D2 receptor. Agonist mode measurements using the endogeneous agonist dopamine and antagonist mode measurements using the specific antagonist haloperidole returned potencies that are in line with the literature. Multi-frequency impedance data allow for non-invasive continuous monitoring of cells with the option to zoom in on certain cell aspects or to tailor the readout to individual cell types for improved data interpretation.

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