ArticleBiophysical journal2007
Deconvolving single-molecule intensity distributions for quantitative microscopy measurements.
Article in Biophysical journal, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 55 papers.
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Who cites it
55 citing papers in PubMed.
- Imaging nanoscopic co-assemblies of α-synuclein and Tau in human samples.Nature communications · 2026Article
- Measuring the Degree of Labeling of Antibody-Dye Conjugates with a Single-Molecule-Sensitive Digital Flow Cytometer.Analytical chemistry · 2026Article
- Fluorescence-free single-molecule microscopy via electronic resonance stimulated Raman scattering.Nature communications · 2026Article
- Autophosphorylation of oncoprotein TEL-ABL in myeloid and lymphoid cells confers resistance to the allosteric ABL inhibitor asciminib.Science signaling · 2025Article
- Characterization of a Single-Molecule Sensitive Digital Flow Cytometer for Amplification-Free Digital Assays.ACS nano · 2025Article
- Single-molecule lipid biosensors mitigate inhibition of endogenous effector proteins.The Journal of cell biology · 2025Article
- Single molecule Lipid Biosensors Mitigate Inhibition of Endogenous Effector Proteins.bioRxiv : the preprint server for biology · 2025Article
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- Comparison of EV characterization by commercial high-sensitivity flow cytometers and a custom single-molecule flow cytometer.Journal of extracellular vesicles · 2024Article
- A statistical simulation model to guide the choices of analytical methods in arrayed CRISPR screen experiments.PloS one · 2024Article
- Physically informed Monte Carlo simulation of dual-wedge prism-based spectroscopic single-molecule localization microscopy.Journal of biomedical optics · 2024Article
- Monte Carlo simulations for the evaluation of oligomerization data in TOCCSL experiments.Biophysical journal · 2023Article
- Quantitative imaging of vesicle-protein interactions reveals close cooperation among proteins.Journal of extracellular vesicles · 2023Article
- Non-invasive chimeric HaloTag labeling to study clustering and diffusion of membrane proteins.STAR protocols · 2022Article
- Payload distribution and capacity of mRNA lipid nanoparticles.Nature communications · 2022Article
- Article
- Evaluating the Effect of Dye-Dye Interactions of Xanthene-Based Fluorophores in the Fluorosequencing of Peptides.Bioconjugate chemistry · 2022Article
- A particle size threshold governs diffusion and segregation of PAR-3 during cell polarization.Cell reports · 2022Article
- Rigor and reproducibility: status and challenges for single vesicle analysis.Extracellular vesicles and circulating nucleic acids · 2022Article
- Sizing Extracellular Vesicles Using Membrane Dyes and a Single Molecule-Sensitive Flow Analyzer.Analytical chemistry · 2021Article
Corrections and comments
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Authors and funding
6 authors.
Funding
Abstract
In fluorescence microscopy, images often contain puncta in which the fluorescent molecules are spatially clustered. This article describes a method that uses single-molecule intensity distributions to deconvolve the number of fluorophores present in fluorescent puncta as a way to "count" protein number. This method requires a determination of the correct statistical relationship between the single-molecule and single-puncta intensity distributions. Once the correct relationship has been determined, basis histograms can be generated from the single-molecule intensity distribution to fit the puncta distribution. Simulated data were used to demonstrate procedures to determine this relationship, and to test the methodology. This method has the advantages of single-molecule measurements, providing both the mean and variation in molecules per puncta. This methodology has been tested with the avidin-biocytin binding system for which the best-fit distribution of biocytins in the sample puncta was in good agreement with a bulk determination of the avidin-biocytin binding ratio.
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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.