ArticleScientific reports2015
3D tracking of single nanoparticles and quantum dots in living cells by out-of-focus imaging with diffraction pattern recognition.
Article in Scientific reports, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed.
- Highly inclined light sheet allows volumetric super-resolution imaging of efflux pumps distribution in bacterial biofilms.Scientific reports · 2024Article
- Particle Localization Using Local Gradients and Its Application to Nanometer Stabilization of a Microscope.ACS nano · 2022Article
- Myosin V fluorescence imaging dataset for single-molecule localization and tracking.Data in brief · 2019Article
- Axial electrokinetic trapping of anisotropic particles.Scientific reports · 2019Article
- Article
- Dissecting myosin-5B mechanosensitivity and calcium regulation at the single molecule level.Nature communications · 2018Article
- Sliding of a single lac repressor protein along DNA is tuned by DNA sequence and molecular switching.Nucleic acids research · 2018Article
- Enhancing optical microscopy illumination to enable quantitative imaging.Scientific reports · 2018Article
- Holographic Traction Force Microscopy.Scientific reports · 2018Article
- A Novel Methodology for Characterizing Cell Subpopulations in Automated Time-lapse Microscopy.Frontiers in bioengineering and biotechnology · 2018Article
- Heterotypic endosomal fusion as an initial trigger for insulin-induced glucose transporter 4 (GLUT4) translocation in skeletal muscle.The Journal of physiology · 2017Article
- Optical microscope illumination analysis using through-focus scanning optical microscopy.Optics letters · 2017Article
- Live-cell single-molecule labeling and analysis of myosin motors with quantum dots.Molecular biology of the cell · 2017Article
Corrections and comments
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Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Live cells are three-dimensional environments where biological molecules move to find their targets and accomplish their functions. However, up to now, most single molecule investigations have been limited to bi-dimensional studies owing to the complexity of 3d-tracking techniques. Here, we present a novel method for three-dimensional localization of single nano-emitters based on automatic recognition of out-of-focus diffraction patterns. Our technique can be applied to track the movements of single molecules in living cells using a conventional epifluorescence microscope. We first demonstrate three-dimensional localization of fluorescent nanobeads over 4 microns depth with accuracy below 2 nm in vitro. Remarkably, we also establish three-dimensional tracking of Quantum Dots, overcoming their anisotropic emission, by adopting a ligation strategy that allows rotational freedom of the emitter combined with proper pattern recognition. We localize commercially available Quantum Dots in living cells with accuracy better than 7 nm over 2 microns depth. We validate our technique by tracking the three-dimensional movements of single protein-conjugated Quantum Dots in living cell. Moreover, we find that important localization errors can occur in off-focus imaging when improperly calibrated and we give indications to avoid them. Finally, we share a Matlab script that allows readily application of our technique by other laboratories.
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