Dual-Color 3D Superresolution Microscopy by Combined Spectral-Demixing and Biplane Imaging |
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| Authors: | Christian?M Winterflood Evgenia Platonova David Albrecht Helge Ewers |
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| Institution: | 1.Randall Division of Cell and Molecular Biophysics, King’s College London, London, United Kingdom;2.Institute of Biochemistry, ETH Zurich, Zurich, Switzerland;3.Institute of Chemistry and Biochemistry, Free University of Berlin, Berlin, Germany |
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| Abstract: | 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. |
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