Barazia Andrew, Monks Colin
Intelligent Imaging Innovations Inc., Denver, CO, USA.
Methods Mol Biol. 2022;2415:245-252. doi: 10.1007/978-1-0716-1904-9_18.
Mitosis is one of the most fundamental processes of life, allowing organisms to grow, develop, and evolve. Acquiring microscopic images and understanding the detailed mechanism of this process is critical in the fields of cell and developmental biology. Modern fluorescence microscopy is the standard for imaging specific molecules and proteins as they interact during this complicated process. However, researchers must take care to ensure that they are maintaining the basal cell processes during mitosis without disruption by placing the sample on a microscope. In addition, mitosis in itself is an incredibly dynamic process that requires both high-speed and high-resolution imaging (McIntosh and Hays. Biology. 5:55, 2016). The Lattice LightSheet is an advanced system, developed in the lab of Eric Betzig (Chen et al. Science. 346:1257998), that offers imaging speeds in the volumes/second while still resolving fine, intracellular structures. Here we describe how to prepare cell culture samples for ideal mitotic imaging on this cutting-edge light sheet fluorescence microscope.
有丝分裂是生命最基本的过程之一，使生物体得以生长、发育和进化。获取微观图像并了解这一过程的详细机制在细胞生物学和发育生物学领域至关重要。现代荧光显微镜是对特定分子和蛋白质在这一复杂过程中相互作用进行成像的标准工具。然而，研究人员必须注意，在将样本放置在显微镜上时，要确保在有丝分裂过程中维持基础细胞过程不受干扰。此外，有丝分裂本身是一个极其动态的过程，需要高速和高分辨率成像（麦金托什和海斯。生物学。5:55，2016）。晶格光片是埃里克·贝齐格实验室开发的一种先进系统（陈等人。科学。346:1257998），它能在每秒成像多个体积的同时，仍能分辨精细的细胞内结构。在这里，我们描述如何为在这种前沿的光片荧光显微镜上进行理想的有丝分裂成像制备细胞培养样本。
