A microdevice to locally electroporate embryos with high efficiency and reduced cell damage

• DOI: 10.1242/dev.106633
• Publication Date: 2014-05-13T08:27:30Z
• Authors: Elsa Mazari; Xuan Zhao; Isabelle Migeotte; Jérôme Collignon; Charlie Gosse; Aitana Perea-Gomez
• Keywords: Live cell imaging

The ability to follow and modify cell behaviour with accurate spatiotemporal resolution is a prerequisite study morphogenesis in developing organisms. Electroporation, the delivery of exogenous molecules into targeted populations through electric permeation plasma membrane, has been used this aim different model systems. However, current localised electroporation strategies suffer from insufficient reproducibility mediocre survival when applied small delicate organisms such as early post-implantation mouse embryos. We introduce here microdevice achieve high efficiency reduced damage. In silico simulations using simple electrical embryos indicated that dielectric guide-based design would improve on existing alternatives. Such device was microfabricated its capacities tested by targeting distal visceral endoderm (DVE), migrating population essential for anterior-posterior axis establishment. Transfection efficiently reproducibly restricted fewer than four cells without compromising embryo survival. Combining mosaic expression fluorescent markers live imaging transgenic revealed that, like leading DVE cells, non-leading ones send long basal projections intercalate during their migration. Finally, we show use our microsystem can be extended variety embryological contexts, preimplantation stages organ explants. Hence, have experimentally validated an approach delivering tailor-made tool embryo. Furthermore, delineated comprehensive strategy development ad hoc devices.