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Fluorescence nanoscopy, or super-resolution microscopy, has become an important tool in cell biological research. However, because of its usually inferior resolution in the depth direction (50-80 nm) and rapidly deteriorating resolution in thick samples, its practical biological application has been effectively limited to two dimensions and thin samples. Here, we present the development of whole-cell 4Pi single-molecule switching nanoscopy (W-4PiSMSN), an optical nanoscope that allows imaging of three-dimensional (3D) structures at 10- to 20-nm resolution throughout entire mammalian cells. We demonstrate the wide applicability of W-4PiSMSN across diverse research fields by imaging complex molecular architectures ranging from bacteriophages to nuclear pores, cilia, and synaptonemal complexes in large 3D cellular volumes.

Nuclear pore complexes are massive protein gateways that control molecular exchange between the nucleus and cytoplasm. In this issue of Cell , Singh et al. provide the first high-resolution views of the elusive nuclear basket, which extends deep into the nucleus to coordinate functions from genome organization to mRNP export. [ABSTRACT FROM AUTHOR]

Results show that during nuclear transport of macromolecules in mitosis, the nuclear pore complex is capable of inhibit the Kap121p-mediated import pathway. Data indicate that the inhibitory action is triggered by structural rearrangements within the nuclear pore complex exposing Kap121p binding site on Nup539 nucleoporin.