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

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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.