title: Dual-color 3D superresolution microscopy by combined spectral-demixing and biplane imaging.
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.
authors.affiliation: Randall Division of Cell and Molecular Biophysics, King's College London, London, United Kingdom.
authors.forename: Christian M
authors.initials: CM
authors.lastname: Winterflood
authors.affiliation: Randall Division of Cell and Molecular Biophysics, King's College London, London, United Kingdom; Institute of Biochemistry, ETH Zurich, Zurich, Switzerland.
authors.forename: Evgenia
authors.initials: E
authors.lastname: Platonova
authors.affiliation: Randall Division of Cell and Molecular Biophysics, King's College London, London, United Kingdom; Institute of Biochemistry, ETH Zurich, Zurich, Switzerland.
authors.forename: David
authors.initials: D
authors.lastname: Albrecht
authors.affiliation: 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.
authors.forename: Helge
authors.initials: H
authors.lastname: Ewers
mesh_terms: D000818:Animals; D017487:Ankyrins / Q000378:metabolism; D002232:Carbocyanines; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D003116:Color; D005456:Fluorescent Dyes; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008853:Microscopy / Q000379:methods*; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D061848:Optical Imaging / Q000379:methods; D017207:Rats, Sprague-Dawley; D013049:Spectrin / Q000378:metabolism
chemical_list: C490613:ANK3 protein, human; C569686:Alexa Fluor 647; C490614:Ank3 protein, rat; D017487:Ankyrins; D002232:Carbocyanines; D005456:Fluorescent Dyes; D013049:Spectrin