ArticlePloS one2015
Quantitative Brightness Analysis of Fluorescence Intensity Fluctuations in E. Coli.
Article in PloS one, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 20 citations in OpenAlex.
- Investigating the effects of temporal correlation on accurate molecular brightness determination.Biophysical journal · 2025Article
- Statistical analysis of the autocorrelation function in fluorescence correlation spectroscopy.Biophysical journal · 2024Article
- Autocorrelation function of finite-length data in fluorescence correlation spectroscopy.Biophysical journal · 2023Article
- Automated analysis of mitochondrial dimensions in mesenchymal stem cells: Current methods and future perspectives.Heliyon · 2023Review
- Number and Brightness Analysis: Visualization of Protein Oligomeric State in Living Cells.Advances in experimental medicine and biology · 2021Article
- Identifying Heteroprotein Complexes in the Nuclear Envelope.Biophysical journal · 2020Article
- Quantitative modeling of self-oligomerization of proteins in the nuclear envelope by fluorescence fluctuation analysis.Analytical biochemistry · 2019Article
- Protein oligomerization and mobility within the nuclear envelope evaluated by the time-shifted mean-segmented Q factor.Methods (San Diego, Calif.) · 2019Article
- Fluorescence fluctuation spectroscopy reveals differential SUN protein oligomerization in living cells.Molecular biology of the cell · 2018Article
- Scanning number and brightness yields absolute protein concentrations in live cells: a crucial parameter controlling functional bio-molecular interaction networks.Biophysical reviews · 2018Review
- Eph-ephrin signaling modulated by polymerization and condensation of receptors.Proceedings of the National Academy of Sciences of the United States of America · 2017Article
- Quantitative Brightness Analysis of Protein Oligomerization in the Nuclear Envelope.Biophysical journal · 2017Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
The brightness measured by fluorescence fluctuation spectroscopy specifies the average stoichiometry of a labeled protein in a sample. Here we extended brightness analysis, which has been mainly applied in eukaryotic cells, to prokaryotic cells with E. coli serving as a model system. The small size of the E. coli cell introduces unique challenges for applying brightness analysis that are addressed in this work. Photobleaching leads to a depletion of fluorophores and a reduction of the brightness of protein complexes. In addition, the E. coli cell and the point spread function of the instrument only partially overlap, which influences intensity fluctuations. To address these challenges we developed MSQ analysis, which is based on the mean Q-value of segmented photon count data, and combined it with the analysis of axial scans through the E. coli cell. The MSQ method recovers brightness, concentration, and diffusion time of soluble proteins in E. coli. We applied MSQ to measure the brightness of EGFP in E. coli and compared it to solution measurements. We further used MSQ analysis to determine the oligomeric state of nuclear transport factor 2 labeled with EGFP expressed in E. coli cells. The results obtained demonstrate the feasibility of quantifying the stoichiometry of proteins by brightness analysis in a prokaryotic cell.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.