Character and rapidity of protein diffusion in intracellular fluids are key determinants of the dynamics and steady state of a plethora of biochemical reactions. So far, an anomalous diffusion in cytoplasmic fluids with viscoelastic and even glassy characteristics has been reported in a variety of organisms on several length scales and timescales. Here, we show that the contiguous fluid of former cytoplasm and nucleoplasm features an anisotropically varying diffusion of macromolecules during eukaryotic cell division. In metaphase, diffusion in the contiguous nucleocytoplasmic fluid appears less anomalous along the spindle axis as compared to perpendicular directions. As a consequence, the long-time diffusion of macromolecules preferentially points along the spindle axis, leading to prolonged residence of macromolecules in the spindle region. Based on our experimental data, we suggest that anisotropic diffusion facilitates the encounter and interaction of spindle-associated proteins, e.g., during the formation of a dynamic spindle matrix.
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