Three-dimensional sub-100 nm resolution fluorescence microscopy of thick samples.
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.
Juette
Manuel F
MF
Institute for Molecular Biophysics, The Jackson Laboratory, 600 Main Street, Bar Harbor, Maine 04609, USA.
Gould
Travis J
TJ
Lessard
Mark D
MD
Mlodzianoski
Michael J
MJ
Nagpure
Bhupendra S
BS
Bennett
Brian T
BT
Hess
Samuel T
ST
Bewersdorf
Joerg
J
D001703:Biophysics / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D019793:Fluorescein / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000494:pharmacology; D007089:Image Enhancement; D007090:Image Interpretation, Computer-Assisted / Q000379:methods; D021621:Imaging, Three-Dimensional / Q000295:instrumentation / Q000379:methods*; D007834:Lasers; D008027:Light; D008853:Microscopy / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D012984:Software
D005456:Fluorescent Dyes; D019793:Fluorescein