title: Photometry unlocks 3D information from 2D localization microscopy data.
abstract: We developed a straightforward photometric method, temporal, radial-aperture-based intensity estimation (TRABI), that allows users to extract 3D information from existing 2D localization microscopy data. TRABI uses the accurate determination of photon numbers in different regions of the emission pattern of single emitters to generate a z-dependent photometric parameter. This method can determine fluorophore positions up to 600 nm from the focal plane and can be combined with biplane detection to further improve axial localization.
authors.affiliation: Department of Biotechnology and Biophysics, University of Würzburg, Würzburg, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Franke
authors.affiliation: Department of Biotechnology and Biophysics, University of Würzburg, Würzburg, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Sauer
authors.affiliation: Department of Biotechnology and Biophysics, University of Würzburg, Würzburg, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: van de Linde
mesh_terms: D000465:Algorithms; D000818:Animals; D002417:Cattle; D003198:Computer Simulation; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008853:Microscopy / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods*; D010783:Photometry / Q000379:methods*; D017785:Photons; D012710:Serum Albumin, Bovine / Q000378:metabolism*; D059010:Single-Cell Analysis / Q000379:methods; D020692:rab3 GTP-Binding Proteins / Q000378:metabolism*
chemical_list: D012710:Serum Albumin, Bovine; D020692:rab3 GTP-Binding Proteins