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Fast multicolor 3D imaging using aberration-corrected multifocus microscopy

Sara Abrahamsson, Jiji Chen, Bassam Hajj, Sjoerd Stallinga, Alexander Y Katsov, Jan Wisniewski, Gaku Mizuguchi, Pierre Soule, Florian Mueller, Claire Dugast Darzacq, Xavier Darzacq, Carl Wu, Cornelia I Bargmann, David A Agard, Maxime Dahan & Mats G L Gustafsson

Conventional acquisition of three-dimensional (3D) microscopy data requires sequential z scanning and is often too slow to capture biological events. We report an aberration-corrected multifocus microscopy method capable of producing an instant focal stack of nine 2D images. Appended to an epifluorescence microscope, the multifocus system enables high-resolution 3D imaging in multiple colors with single-molecule sensitivity, at speeds limited by the camera readout time of a single image.

Nature Methods (2012) doi:10.1038/nmeth.2277