Ultra-High Resolution 3D Imaging of Whole Cells.
Huang F., Sirinakis G., Allgeyer ES., Schroeder LK., Duim WC., Kromann EB., Phan T., Rivera-Molina FE., Myers JR., Irnov I., Lessard M., Zhang Y., Handel MA., Jacobs-Wagner C., Lusk CP., Rothman JE., Toomre D., Booth MJ., Bewersdorf J.
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.