ArticlePloS one2014
Quantitative measurement of brightness from living cells in the presence of photodepletion.
Article in PloS one, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
14 citing papers in PubMed.
- Continuous distribution of cancer cells in the cell cycle unveiled by AI-segmented imaging of 37,000 HeLa FUCCI cells.Heliyon · 2024Article
- Assembly mechanism of the inflammasome sensor AIM2 revealed by single molecule analysis.Nature communications · 2023Article
- Benchmarking of novel green fluorescent proteins for the quantification of protein oligomerization in living cells.PloS one · 2023Article
- Empirical Bayes method using surrounding pixel information for number and brightness analysis.Biophysical journal · 2021Article
- 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
- Protein oligomerization and mobility within the nuclear envelope evaluated by the time-shifted mean-segmented Q factor.Methods (San Diego, Calif.) · 2019Article
- Using enhanced number and brightness to measure protein oligomerization dynamics in live cells.Nature protocols · 2019Article
- Calibration-free In Vitro Quantification of Protein Homo-oligomerization Using Commercial Instrumentation and Free, Open Source Brightness Analysis Software.Journal of visualized experiments : JoVE · 2018Article
- Optimal fluorescent protein tags for quantifying protein oligomerization in living cells.Scientific reports · 2018Article
- Fluorescence fluctuation spectroscopy reveals differential SUN protein oligomerization in living cells.Molecular biology of the cell · 2018Article
- Quantitative Brightness Analysis of Protein Oligomerization in the Nuclear Envelope.Biophysical journal · 2017Article
- Association of Endophilin B1 with Cytoplasmic Vesicles.Biophysical journal · 2016Article
- Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
6 authors.
Funding
Abstract
The brightness of fluorescently labeled proteins provides an excellent marker for identifying protein interactions in living cells. Quantitative interpretation of brightness, however, hinges on a detailed understanding of the processes that affect the signal fluctuation of the fluorescent label. Here, we focus on the cumulative influence of photobleaching on brightness measurements in cells. Photobleaching within the finite volume of the cell leads to a depletion of the population of fluorescently labeled proteins with time. The process of photodepletion reduces the fluorescence signal which biases the analysis of brightness data. Our data show that even small reductions in the signal can introduce significant bias into the analysis of the data. We develop a model that quantifies the bias and introduce an analysis method that accurately determines brightness in the presence of photodepletion as verified by experiments with mammalian and yeast cells. In addition, photodepletion experiments with the fluorescent protein EGFP reveal the presence of a photoconversion process, which leads to a marked decrease in the brightness of the EGFP protein. We also identify conditions where the effect of EGFP's photoconversion on brightness experiments can be safely ignored.
Indexed as
Identifiers
What OpenQuestion holds
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.