Kanagawa Institute for Technology, Atsugi, Japan.
Exploratory Research Center on Life and Living Systems, National Institute of Natural Sciences, Okazaki, Japan.
Methods Mol Biol. 2025;2872:37-50. doi: 10.1007/978-1-0716-4224-5_3.
Plant cells have a unique organization of microtubules during mitosis and cytokinesis. However, it is difficult to observe cell division in plant cells because of the distortion of images caused by thick samples. We addressed this issue by two-photon excitation coupled with spinning-disk confocal microscopy. Herein, we describe the method for observing microtubules and nuclei during cell division in tobacco BY-2 cells by two-photon spinning-disk confocal microscopy. The protocol includes the transformation of BY-2 cells with a plasmid for the expression of tubulin and histone markers, preparation of cells for microscopy, and operation of the two-photon spinning-disk microscope.
植物细胞在有丝分裂和胞质分裂过程中有独特的微管组织。然而，由于厚样品导致图像失真，观察植物细胞的细胞分裂变得很困难。我们通过双光子激发与旋转盘共聚焦显微镜相结合来解决这个问题。在此，我们描述了通过双光子旋转盘共聚焦显微镜观察烟草 BY-2 细胞中微管和核分裂的方法。该方案包括用表达微管蛋白和组蛋白标记物的质粒转化 BY-2 细胞、为显微镜制备细胞以及操作双光子旋转盘显微镜。
