ABSTRACT
The contractile behavior of cardiomyocytes can be monitored by measuring their action potentials, and the analysis is essential for screening the safety of potential drugs. However, immobilizing cardiac cells on a specific electrode is considerably complicated. In this study, we demonstrate that scanning electrochemical microscopy (SECM) can be used to analyze rapid topographic changes in beating cardiomyocytes in a standard culture dish. Various cardiomyocyte contraction parameters and oxygen consumption based on cell respiration could be determined from SECM data. We also confirmed that cellular changes induced by adding the cardiotonic agent digoxin were conveniently monitored by this SECM system. These results show that SECM can be a potentially powerful tool for use in drug development for cardiovascular diseases.
Subject(s)
Heart/physiology , Microscopy, Electrochemical, Scanning/methods , Myocytes, Cardiac/metabolism , Oxygen Consumption , Animals , Cardiotonic Agents/pharmacology , Cell Respiration/drug effects , Heart/drug effects , Muscle Contraction/drug effects , Myocytes, Cardiac/cytology , Myocytes, Cardiac/drug effects , Oxygen Consumption/drug effects , Rats , TemperatureABSTRACT
Scanning electrochemical microscopy (SECM) is useful for analyzing various cellular responses. We have combined a micropipette (MP) with SECM to perform quantitative solution delivery to single cells. In this system, since the concentrations of electrochemical mediators are changed by the volume of solution delivered from the MP, we constructed a feedback control system to regulate MP delivery by SECM-detected signals. Cellular responses induced by MP delivery could be monitored by the SECM, and cell apoptosis was successfully detected by adding a kinase inhibitor of two orders of magnitude less than what is required in the conventional method. The SECM-based MP can activate a target cell, requiring a minimal amount of agent, and can continually examine target cell responses. This system improves the accuracy of delivery from the MP and is useful for single-cell analysis.
