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

未附着的着丝点通过产生由MAD2， BUB3和BUBR1纺锤体组装检查点蛋白组成的抑制剂来阻止后期。稳定附着在微管上的着丝点不再产生抑制剂，但我们不明白微管附着是如何做到这一点的。一个附着的微管是足够的，还是需要几个？微管“剥离”抑制蛋白的着丝点的能力是否足够快？为了回答这些问题，我们需要实时成像微管附着在着丝点上，并量化MAD2和BUBR1在着丝点上的变化。我们现在可以用点阵光片显微镜和内源性标记蛋白来做到这一点。

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.