ArticleChemphyschem : a European journal of chemical physics and physical chemistry2016
Addressing the Requirements of High-Sensitivity Single-Molecule Imaging of Low-Copy-Number Proteins in Bacteria.
Article in Chemphyschem : a European journal of chemical physics and physical chemistry, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed, 14 citations in OpenAlex.
- The translesion polymerase Pol Y1 is a constitutive component of the B. subtilis replication machinery.Nucleic acids research · 2024Article
- Single-molecule tracking of PprI inFrontiers in microbiology · 2023Article
- Computerized fluorescence microscopy of microbial cells.World journal of microbiology & biotechnology · 2021Review
- Identification of Multiple Kinetic Populations of DNA-Binding Proteins in Live Cells.Biophysical journal · 2019Article
- Guidelines for DNA recombination and repair studies: Mechanistic assays of DNA repair processes.Microbial cell (Graz, Austria) · 2019Review
- Method to identify and minimize artifacts induced by fluorescent impurities in single-molecule localization microscopy.Journal of biomedical optics · 2018Article
- Bacterial cell wall nanoimaging by autoblinking microscopy.Scientific reports · 2018Article
- Single-Molecule DNA Polymerase Dynamics at a Bacterial Replisome in Live Cells.Biophysical journal · 2016Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
Single-molecule fluorescence super-resolution imaging and tracking provide nanometer-scale information about subcellular protein positions and dynamics. These single-molecule imaging experiments can be very powerful, but they are best suited to high-copy number proteins where many measurements can be made sequentially in each cell. We describe artifacts associated with the challenge of imaging a protein expressed in only a few copies per cell. We image live Bacillus subtilis in a fluorescence microscope, and demonstrate that under standard single-molecule imaging conditions, unlabeled B. subtilis cells display punctate red fluorescent spots indistinguishable from the few PAmCherry fluorescent protein single molecules under investigation. All Bacillus species investigated were strongly affected by this artifact, whereas we did not find a significant number of these background sources in two other species we investigated, Enterococcus faecalis and Escherichia coli. With single-molecule resolution, we characterize the number, spatial distribution, and intensities of these impurity spots.
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Registered trials
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