Abstract:
Reproductive processes are dynamic and involve extensive morphological remodeling and cell-cell interactions. Live imaging of organs enhances our understanding of how biological processes occur in real time. Slice culture is a type of organ culture where thick slices are collected from an organ and cultured for several days. Slice culture is a useful and easy-to-implement technique for live imaging of reproductive events at cellular resolution. Here we describe a pipeline of live imaging on slice culture to visualize the process of urethra closure in mouse embryonic penis as a proof of principle. In combination with genetic reporter mice, nuclear stains, and exposure experiments, we demonstrate the feasibility of slice culture on a reproductive organ. We also provide a step-by-step protocol and troubleshooting guide to facilitate the adoption of slice culture with live imaging in other reproductive organs. Lastly, we discuss potential utilities and experiments that could be implemented with slice culture in reproductive sciences.
摘要:
生殖过程是动态的,涉及广泛的形态重塑和细胞间相互作用。器官的实时成像增强了我们对生物过程如何实时发生的理解。切片培养是一种器官培养,其中从器官中收集厚切片并培养几天。切片培养是一种有用且易于实施的技术,用于以细胞分辨率对生殖事件进行实时成像。在这里,我们描述了在切片培养上进行实时成像的管道,以可视化小鼠胚胎阴茎中尿道闭合的过程作为原理的证明。结合基因报告小鼠,核污渍,和暴露实验,我们证明了在生殖器官上进行切片培养的可行性。我们还提供了逐步的方案和故障排除指南,以促进在其他生殖器官中采用带有实时成像的切片培养。最后,我们讨论了在生殖科学中使用切片培养可以实现的潜在效用和实验。