A system for optical high resolution screening of electrical excitable cells
| Autor: | Lars Kaestner, Oliver Müller, Qinghai Tian, Roman Zantl, Valentin Kahl, Peter Lipp |
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| Rok vydání: | 2010 |
| Předmět: |
Male
Materials science Physiology Cell Culture Techniques Action Potentials Nanotechnology Calcium imaging Microscopy Animals Myocytes Cardiac Calcium Signaling Rats Wistar Electrodes Molecular Biology Cells Cultured FOIL method Fluorescent Dyes Total internal reflection fluorescence microscope Substrate (chemistry) Cell Biology Elasticity Electric Stimulation Voltage-Sensitive Dye Imaging Electronics Medical Extracellular Matrix Rats Electrophysiology Microscopy Fluorescence High-content screening Oil immersion Layer (electronics) Software Muscle Contraction Biomedical engineering |
| Zdroj: | Cell Calcium. 47:224-233 |
| ISSN: | 0143-4160 |
| DOI: | 10.1016/j.ceca.2009.11.012 |
| Popis: | The application of primary excitable cells for high content screening (HCS) requires a multitude of novel developments including cell culture and multi-well plates. Here we introduce a novel system combining optimised culture conditions of primary adult cardiomyocytes with the particular needs of excitable cells for arbitrary field stimulation of individual wells. The major advancements of our design were tested in calcium imaging experiments and comprise (i) each well of the plate can be subjected to individual pulse protocols, (ii) the software driving electrical stimulation can run as a stand-alone application but also as a plug-in in HCS software packages, (iii) the optical properties of the plastic substrate (foil) resemble those of glass coverslips fostering high resolution immersion-based microscopy, (iv) the bottom of the foil is coated with an oleophobic layer that prevents immersion oil from sticking, (v) the top of the foil is coated with an elastic film. The latter enables cardiomyocytes to display loaded contractions by mimicking the physiologically occurring local elastic network (e.g., extracellular matrix) and results in significantly increased contractions (with identical calcium transients) when compared to non-elastic substrates. Thus, our novel design and culture conditions represent an essential further step towards the application of primary cultured adult cardiomyocytes for HCS applications. |
| Databáze: | OpenAIRE |
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