Using lattice light sheet microscopy to determine how the Spindle Assembly Checkpoint turns off

独立的动点蛋白通过产生一种由MAD2、Bub3和BUBR1纺锤体组装检查点蛋白组成的抑制蛋白来防止后期。稳定附着在微管上的动点不再产生抑制物，但我们不知道微管附着是如何做到这一点的。一个附着的微管就足够了，还是需要几个？微管“剥离”抑制蛋白的能力足够快吗？为了回答这些问题，我们需要实时成像微管附着在动心率上，并量化动心率时MAD2和BUBR1的变化。我们现在可以使用晶格光片显微镜和内源标记蛋白质来完成这项工作。

Unattached kinetochores prevent anaphase by generating an inhibitor composed of the MAD2, BUB3 and BUBR1 Spindle Assembly Checkpoint proteins. Kinetochores that stably attach to microtubules no longer generate the inhibitor but we do not understand how microtubule attachment does this. Is one attached microtubule sufficient, or are several required? Is the ability of microtubules to 'strip' kinetochores of the inhibitor proteins fast enough? To answer these questions we need to image microtubule attachment to kinetochores in real-time and quantify the changes in MAD2 and BUBR1 at kinetochores. We can now do this using lattice-lightsheet microscopy and endogenously tagged proteins.