Three-dimensional sub-100 nm resolution fluorescence microscopy of thick samples.

Juette, Manuel F; Gould, Travis J; Lessard, Mark D; Mlodzianoski, Michael J; Nagpure, Bhupendra S; Bennett, Brian T; Hess, Samuel T; Bewersdorf, Joerg · Nat Methods · 2008

basic_science · Level V

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Abstract

Imaging volumes as thick as whole cells at three-dimensional (3D) super-resolution is required to reveal unknown features of cellular organization. We report a light microscope that generates images with translationally invariant 30 x 30 x 75 nm resolution over a depth of several micrometers. This method, named biplane (BP) FPALM, combines a double-plane detection scheme with fluorescence photoactivation localization microscopy (FPALM) enabling 3D sub-diffraction resolution without compromising speed or sensitivity.

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