Photometry unlocks 3D information from 2D localization microscopy data

Nat Methods. 2017 Jan;14(1):41-44. doi: 10.1038/nmeth.4073. Epub 2016 Nov 21.

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.

MeSH terms

  • Algorithms
  • Animals
  • Cattle
  • Computer Simulation
  • Humans
  • Imaging, Three-Dimensional / methods*
  • Microscopy / methods*
  • Molecular Imaging / methods*
  • Photometry / methods*
  • Photons
  • Serum Albumin, Bovine / metabolism*
  • Single-Cell Analysis / methods
  • rab3 GTP-Binding Proteins / metabolism*

Substances

  • Serum Albumin, Bovine
  • rab3 GTP-Binding Proteins