Nature Methods
- 5, 527 - 529 (2008)
Published online: 11 May 2008; | doi:10.1038/nmeth.1211
Three-dimensional sub–100 nm resolution fluorescence microscopy of thick samplesManuel F Juette1, 2, Travis J Gould3, 4, Mark D Lessard1, Michael J Mlodzianoski1, Bhupendra S Nagpure1, 3, Brian T Bennett5, Samuel T Hess3, 4 & Joerg Bewersdorf11
Institute for Molecular Biophysics, The Jackson Laboratory, 600 Main Street, Bar Harbor, Maine 04609, USA. 2
Department of Biophysical Chemistry, University of Heidelberg, and Department of New Materials and Biosystems, Max Planck Institute for Metals Research, Heisenbergstr. 3, 70569 Stuttgart, Germany. 3
Department of Physics and Astronomy, University of Maine, 120 Bennett Hall, Orono, Maine 04469, USA. 4
Institute for Molecular Biophysics, University of Maine, 5737 Jenness Hall, Orono, Maine 04469, USA. 5
Active Motif, Inc., 1914 Palomar Oaks Way, Carlsbad, California 92008, USA.
Correspondence should be addressed to Joerg Bewersdorf joerg.bewersdorf@jax.org 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  30 75nm 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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