ArticleBiophysical journal2000
Imaging and tracking of single GFP molecules in solution.
Article in Biophysical journal, 2000. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 79 papers.
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Who cites it
79 citing papers in PubMed.
- SlimVar for rapid in vivo single-molecule tracking of chromatin regulators in plants.Nature communications · 2025Article
- Photobleaching Analysis of Fluorescent Proteins in Two-Photon Microscopy at High Repetition Rates.Optics communications · 2025Article
- Nascent mitochondrial proteins initiate the localized condensation of cytosolic protein aggregates on the mitochondrial surface.Proceedings of the National Academy of Sciences of the United States of America · 2023Article
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- Answering some questions about structured illumination microscopy.Philosophical transactions. Series A, Mathematical, physical, and engineering sciences · 2022Article
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- Ultrarapid cryo-arrest of living cells on a microscope enables multiscale imaging of out-of-equilibrium molecular patterns.Science advances · 2021Article
- Split-HaloTag imaging assay for sophisticated microscopy of protein-protein interactionsPlant communications · 2021Article
- Semiconductor Nanocrystals for Biological Imaging and Fluorescence Spectroscopy.Advances in experimental medicine and biology · 2021Article
- Optogenetic Imaging of Protein Activity Using Two-Photon Fluorescence Lifetime Imaging Microscopy.Advances in experimental medicine and biology · 2021Review
- Rhenium (I) Complexes as Probes for Prokaryotic and Fungal Cells by Fluorescence Microscopy: Do Ligands Matter?Frontiers in chemistry · 2019Review
- Single-Nanoparticle Tracking with Angstrom Localization Precision and Microsecond Time Resolution.Biophysical journal · 2018Article
- FRET-enhanced photostability allows improved single-molecule tracking of proteins and protein complexes in live mammalian cells.Nature communications · 2018Article
- PhotoGate microscopy to track single molecules in crowded environments.Nature communications · 2017Article
- High-resolution real-time dual-view imaging with multiple point of view microscopy.Biomedical optics express · 2016Article
- Investigating the usage of point spread functions in point source and microsphere localization.Proceedings of SPIE--the International Society for Optical Engineering · 2016Article
- Article
- Investigation of the numerics of point spread function integration in single molecule localization.Optics express · 2015Article
- Rapid isolation and single-molecule analysis of ribonucleoproteins from cell lysate by SNAP-SiMPull.RNA (New York, N.Y.) · 2015Article
- Imaging live cells at the nanometer-scale with single-molecule microscopy: obstacles and achievements in experiment optimization for microbiology.Molecules (Basel, Switzerland) · 2014Review
19 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Visualization and tracking of single fluorescent molecules is a recent development in optical microscopy holding great promise for the study of cell biological processes. However, all experimental strategies realized so far confined the observation to extremely thin interfacial layers. The detection and characterization of single molecules in three-dimensionally extended systems such as living cells has yet to be accomplished. We show, here, for the first time that single protein molecules can be visualized and tracked in three-dimensional (3D) samples at room temperature. Using a wide-field fluorescence microscope equipped with an Ar(+)-laser and a low-light-level CCD camera, single molecules of the green fluorescent protein (GFP) were detected in gels and viscous solutions at depths of up to approximately 10 microm from the interface. A time resolution of 5 ms was achieved by a high-speed framing mode. The two-dimensional localization accuracy was determined to be approximately 30 nm. The number of photons emitted by single GFP molecules before photodestruction was found to be < or = 4 * 10(5). Freely diffusing GFP molecules could be tracked over up to nine images acquired at a frame rate of approximately 80 Hz. From the trajectories, the diffusion coefficients of single GFP molecules were derived and found to agree well with expectation and microphotolysis measurements. Our results imply that the visualization and tracking of single molecules in living cells is possible.
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