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J Neurosci Methods ; 257: 194-203, 2016 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-26432934

RESUMEN

BACKGROUND: Multi-electrode arrays (MEAs) allow non-invasive multi-unit recording in-vitro from cultured neuronal networks. For sufficient neuronal growth and adhesion on such MEAs, substrate preparation is required. Plating of dissociated neurons on a uniformly prepared MEA's surface results in the formation of spatially extended random networks with substantial inter-sample variability. Such cultures are not optimally suited to study the relationship between defined structure and dynamics in neuronal networks. To overcome these shortcomings, neurons can be cultured with pre-defined topology by spatially structured surface modification. Spatially structuring a MEA surface accurately and reproducibly with the equipment of a typical cell-culture laboratory is challenging. NEW METHOD: In this paper, we present a novel approach utilizing micro-contact printing (µCP) combined with a custom-made device to accurately position patterns on MEAs with high precision. We call this technique AP-µCP (accurate positioning micro-contact printing). COMPARISON WITH EXISTING METHODS: Other approaches presented in the literature using µCP for patterning either relied on facilities or techniques not readily available in a standard cell culture laboratory, or they did not specify means of precise pattern positioning. CONCLUSION: Here we present a relatively simple device for reproducible and precise patterning in a standard cell-culture laboratory setting. The patterned neuronal islands on MEAs provide a basis for high throughput electrophysiology to study the dynamics of single neurons and neuronal networks.


Asunto(s)
Técnicas de Cultivo de Célula/instrumentación , Microelectrodos , Microtecnología/instrumentación , Neuronas/fisiología , Impresión/instrumentación , Potenciales de Acción , Animales , Astrocitos/fisiología , Adhesión Celular , Recuento de Células , Técnicas de Cultivo de Célula/métodos , Diseño de Equipo , Hipocampo/citología , Hipocampo/fisiología , Inmunohistoquímica , Microscopía Electrónica de Rastreo , Microscopía de Contraste de Fase , Microtecnología/métodos , Neuronas/citología , Impresión/métodos , Ratas , Reproducibilidad de los Resultados , Propiedades de Superficie
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