Photometry unlocks 3D information from 2D localization microscopy data.
basic_science · Level V
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- Record sourced from PubMed, PMID 27869814.
- Also identified by DOI 10.1038/nmeth.4073.
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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.
Medical subject headings
- Imaging, Three-Dimensional
- Microscopy
- Molecular Imaging
- Photometry
- Serum Albumin, Bovine
- rab3 GTP-Binding Proteins