Division of Molecular Biology, Ruđer Bošković Institute, Zagreb, Croatia.
Methods Mol Biol. 2025;2872:3-19. doi: 10.1007/978-1-0716-4224-5_1.
Stimulated emission depletion (STED) microscopy is a powerful super-resolution imaging technique that only recently entered the field of mitosis, where it proved to be invaluable for studying various microtubule classes, kinetochore-microtubule attachments and chromosome segregation errors. Here, we describe immunofluorescence combined with STED microscopy as a method for analyzing microtubules and kinetochore-microtubule attachments in human mitotic spindles. We also describe live-cell STED microscopy as a method for single-plane short-term imaging of transient processes in crowded spindle areas. Finally, we outline image analysis approaches for the quantitative assessment of microtubule bundles within the spindle.
受激发射耗散（STED）显微镜是一种强大的超分辨率成像技术，最近才进入有丝分裂领域，在该领域，它在研究各种微管类、动粒-微管连接和染色体分离错误方面非常有价值。在这里，我们描述了免疫荧光结合 STED 显微镜作为一种分析人有丝分裂纺锤体中微管和动粒-微管连接的方法。我们还描述了活细胞 STED 显微镜作为一种在拥挤的纺锤体区域中单平面短期成像瞬时过程的方法。最后，我们概述了用于定量评估纺锤体中微管束的图像分析方法。
