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Dual-color 3D superresolution microscopy by combined spectral-demixing and biplane imaging.

  • Winterflood CM Randall Division of Cell and Molecular Biophysics, King's College London, London, United Kingdom.
  • Platonova E Randall Division of Cell and Molecular Biophysics, King's College London, London, United Kingdom; Institute of Biochemistry, ETH Zurich, Zurich, Switzerland.
  • Albrecht D Randall Division of Cell and Molecular Biophysics, King's College London, London, United Kingdom; Institute of Biochemistry, ETH Zurich, Zurich, Switzerland.
  • Ewers H Randall Division of Cell and Molecular Biophysics, King's College London, London, United Kingdom; Institute of Chemistry and Biochemistry, Free University of Berlin, Berlin, Germany. Electronic address: helge.ewers@fu-berlin.de.
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  • 2015-07-09
Published in:
  • Biophysical journal. - 2015
Animals
Ankyrins
Carbocyanines
Cell Line, Tumor
Cells, Cultured
Color
Fluorescent Dyes
Hippocampus
Humans
Image Processing, Computer-Assisted
Imaging, Three-Dimensional
Microscopy
Neurons
Optical Imaging
Rats, Sprague-Dawley
Spectrin
English Multicolor three-dimensional (3D) superresolution techniques allow important insight into the relative organization of cellular structures. While a number of innovative solutions have emerged, multicolor 3D techniques still face significant technical challenges. In this Letter we provide a straightforward approach to single-molecule localization microscopy imaging in three dimensions and two colors. We combine biplane imaging and spectral-demixing, which eliminates a number of problems, including color cross-talk, chromatic aberration effects, and problems with color registration. We present 3D dual-color images of nanoscopic structures in hippocampal neurons with a 3D compound resolution routinely achieved only in a single color.
Language
  • English
Open access status
bronze
Identifiers
Persistent URL
https://roar.hep-bejune.ch/global/documents/253901
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