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J Struct Biol ; 213(4): 107791, 2021 12.
Artículo en Inglés | MEDLINE | ID: mdl-34520869

RESUMEN

Cryo-electron tomography is the highest resolution tool available for structural analysis of macromolecular complexes within their native cellular environments. At present, data acquisition suffers from low throughput, in part due to the low probability of positioning a cell such that the subcellular structure of interest is on a region of the electron microscopy (EM) grid that is suitable for imaging. Here, we photo-micropatterned EM grids to optimally position endothelial cells so as to enable high-throughput imaging of cell-cell contacts. Lattice micropatterned grids increased the average distance between intercellular contacts and thicker cell nuclei such that the regions of interest were sufficiently thin for direct imaging. We observed a diverse array of membranous and cytoskeletal structures at intercellular contacts, demonstrating the utility of this technique in enhancing the rate of data acquisition for cellular cryo-electron tomography studies.


Asunto(s)
Comunicación Celular , Microscopía por Crioelectrón/métodos , Tomografía con Microscopio Electrónico/métodos , Imagenología Tridimensional/métodos , Uniones Intercelulares/ultraestructura , Microscopía Electrónica de Transmisión/métodos , Cadherinas/metabolismo , Núcleo Celular/metabolismo , Núcleo Celular/ultraestructura , Células Cultivadas , Células Endoteliales/citología , Células Endoteliales/metabolismo , Células Endoteliales/ultraestructura , Matriz Extracelular/metabolismo , Matriz Extracelular/ultraestructura , Humanos , Uniones Intercelulares/metabolismo , Microscopía de Fuerza Atómica/métodos , Microscopía Confocal/métodos , Reproducibilidad de los Resultados
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