The mechanics of filament networks depend on both the individual filament properties and the network architecture. Despite the importance of filament networks in biology, there exists no technique which can precisely localize filaments and cross-links in three dimensions and simultaneously resolve their dynamics. Thermal noise imaging is a three-dimensional scanning probe technique that utilizes the confined thermal motion of an optically trapped nanoparticle to noninvasively probe the sample. Here, we apply the technique to a stably crosslinked network of microtubules. The filament axes are localized with a precision of 4nm. Analysis also reveals fluctuations of individual filaments and properties of cross-links.
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