Marian Baclayon, TUD

Title: Combined Fluorescence and Cryo-Transmission Electron Microscopy on SiN Nanochambers to Elucidate the End-On Interaction of Microtubules and Dynein
Session: 14:30


We developed nanochambers with SiN windows, which are transparent to both photons and electrons. These chambers are compatible for in-vitro assays that are normally performed on glass, are functionalizable using gold-thiol chemistry, and can be vitrified for high-resolution electron microscopy (EM). We apply this technology to elucidate the end-on interaction between microtubules and dynein motors, which plays important roles in cell division, including centrosome positioning and chromosome segregation. Using fluorescence microscopy, we aim to determine the dynamics of this process by fixing dynein to gold barriers or pillars and observe how microtubules interact end-on with dynein. Using high-resolution cryo-EM imaging on the same chamber allows us to quantify the number of motors that can interact with a single microtubule, understand how dynein stabilizes the microtubule end, and determine how it can exert pushing and pulling forces.


to be announced

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