Photometry unlocks 3D information from 2D localization microscopy data
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Updated: Mar 11, 2026

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy
Published on: December 9, 2013
Christian Franke1, Markus Sauer1, Sebastian van de Linde1
1Department of Biotechnology and Biophysics, University of Würzburg, Würzburg, Germany.
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.
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