利用共聚焦延时显微镜对胚胎发生进行整体成像。

In toto imaging of embryogenesis with confocal time-lapse microscopy.

作者信息

Megason Sean G

机构信息

Department of Systems Biology, Harvard Medical School, 200 Longwood Ave, WAB536, Boston, MA, 02115, USA.

出版信息

Methods Mol Biol. 2009;546:317-32. doi: 10.1007/978-1-60327-977-2_19.

Abstract

Microscopy has been one of the most direct and powerful tools since the beginning of biological research. Continued advances such as confocal and two-photon fluorescence microscopy and fluorescent proteins now make imaging useful at a variety of spatial scales (molecules, circuits, cells, tissues, and even whole embryos) and temporal scales (<seconds to days). Zebrafish is uniquely poised to benefit from these continued technological improvements because of its inherent suitability for both imaging and genetics. This chapter presents an approach called "in toto imaging". The goal of in toto imaging is to image and track every single cell movement and division that forms a tissue or organ. This approach is powerful for understanding how cell lineage, shape changes, and movements control the morphogenesis of a tissue. When used with transgenic lines, in toto imaging can be used to "digitize" data at single cell level over time from a living organism. This quantitative, digitized data can then serve as the basis for forming models of how biological circuits orchestrate developmental processes.

摘要

自生物学研究伊始,显微镜检查一直是最直接且强大的工具之一。诸如共聚焦和双光子荧光显微镜以及荧光蛋白等技术的持续进步,使得成像在各种空间尺度(分子、回路、细胞、组织乃至整个胚胎)和时间尺度(<秒至天)上都变得有用。由于斑马鱼在成像和遗传学方面具有内在的适用性,它特别适合从这些持续的技术改进中受益。本章介绍一种称为“整体成像”的方法。整体成像的目标是对形成组织或器官的每一个细胞的运动和分裂进行成像和追踪。这种方法对于理解细胞谱系、形状变化和运动如何控制组织的形态发生非常有力。当与转基因品系一起使用时,整体成像可用于随时间在单细胞水平上对来自活生物体的数据进行“数字化”。然后,这些定量的、数字化的数据可作为构建生物回路如何协调发育过程模型的基础。

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