6533b829fe1ef96bd128af32
RESEARCH PRODUCT
Three-dimensional multiple-particle tracking with nanometric precision over tunable axial ranges
Andrea BarberisTiziana RavasengaLorenzo ScipioniGiuseppe SancataldoMartí DuocastellaLuca LanzanoAlberto Diasprosubject
0301 basic medicineOptical devicesMaterials scienceComplex system02 engineering and technologyTracking (particle physics)Deformable mirrorlaw.invention03 medical and health sciencesOpticsPosition (vector)lawAtomic and Molecular PhysicsElectronicImaging systemsDepth of fieldOptical and Magnetic MaterialsFluorescence microscopy; Imaging systems; Microscopy; Optical devices; Three-dimensional image processing; Electronic; Optical and Magnetic Materials; Atomic and Molecular Physics; and OpticsFluorescence microscopyMicroscopybusiness.industryThree-dimensional image processingFluorescence microscopy; Imaging systems; Microscopy; Optical devices; Three-dimensional image processing; Electronic Optical and Magnetic Materials; Atomic and Molecular Physics and Optics021001 nanoscience & nanotechnologyAtomic and Molecular Physics and OpticsElectronic Optical and Magnetic MaterialsNumerical apertureLens (optics)030104 developmental biologyTemporal resolutionand Optics0210 nano-technologybusinessFluorescence microscopy Imaging systems Microscopy Optical devices Three-dimensional image processing Electronic Optical and Magnetic Materials Atomic and Molecular Physics and Opticsdescription
The precise localization of nanometric objects in three dimensions is essential to identify functional diffusion mechanisms in complex systems at the cellular or molecular level. However, most optical methods can achieve high temporal resolution and high localization precision only in two dimensions or over a limited axial (z) range. Here we develop a novel wide-field detection system based on an electrically tunable lens that can track multiple individual nanoscale emitters in three dimensions over a tunable axial range with nanometric localization precision. The optical principle of the technique is based on the simultaneous acquisition of two images with an extended depth of field while encoding the z position of the emitters via a lateral shift between images. We provide a theoretical framework for this approach and demonstrate tracking of free diffusing beads and GABAA receptors in live neurons. This approach allows getting nanometric localization precision up to an axial range above 10 µm with a high numerical aperture lens-quadruple that of a typical 3D tracking system. Synchronization or complex fitting procedures are not requested here, which leads to a suitable architecture for localizing single molecules in four dimensions, namely, three dimensions in real-time.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2017-03-13 |