ArticleNature protocols2011
Determining the number of specific proteins in cellular compartments by quantitative microscopy.
Article in Nature protocols, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed.
- Measuring the Degree of Labeling of Antibody-Dye Conjugates with a Single-Molecule-Sensitive Digital Flow Cytometer.Analytical chemistry · 2026Article
- Characterization of a Single-Molecule Sensitive Digital Flow Cytometer for Amplification-Free Digital Assays.ACS nano · 2025Article
- Direct imaging with multidimensional labelling and high-content analysis allows quantitative categorization and characterizations of individual small extracellular vesicles and nanoparticles (sEVPs).Journal of extracellular vesicles · 2024Article
- Comparison of EV characterization by commercial high-sensitivity flow cytometers and a custom single-molecule flow cytometer.Journal of extracellular vesicles · 2024Article
- Payload distribution and capacity of mRNA lipid nanoparticles.Nature communications · 2022Article
- Microphysiological Modeling of the Structure and Function of Neuromuscular Transmitter Release Sites.Frontiers in synaptic neuroscience · 2022Review
- High-Throughput Counting and Superresolution Mapping of Tetraspanins on Exosomes Using a Single-Molecule Sensitive Flow Technique and Transistor-like Semiconducting Polymer Dots.Angewandte Chemie (International ed. in English) · 2021Article
- Sizing Extracellular Vesicles Using Membrane Dyes and a Single Molecule-Sensitive Flow Analyzer.Analytical chemistry · 2021Article
- Special Delivery: Potential Mechanisms of Botulinum Neurotoxin Uptake and Trafficking within Motor Nerve Terminals.International journal of molecular sciences · 2020Review
- Cell-Compatible Nanoprobes for Imaging Intracellular Phosphatase Activities.Chembiochem : a European journal of chemical biology · 2019Article
- Single-Molecule Flow Platform for the Quantification of Biomolecules Attached to Single Nanoparticles.Analytical chemistry · 2018Article
- Quantitative microscopy based on single-molecule fluorescence.Current opinion in chemical biology · 2017Review
- Synaptic vesicles contain small ribonucleic acids (sRNAs) including transfer RNA fragments (trfRNA) and microRNAs (miRNA).Scientific reports · 2015Article
- Overlapping functions of stonin 2 and SV2 in sorting of the calcium sensor synaptotagmin 1 to synaptic vesicles.Proceedings of the National Academy of Sciences of the United States of America · 2015Article
- Direct interrogation of DNA content distribution in nanoparticles by a novel microfluidics-based single-particle analysis.Nano letters · 2014Article
- Live-cell imaging of receptors around postsynaptic membranes.Nature protocols · 2014Article
- Individual synaptic vesicles from the electroplaque of Torpedo californica, a classic cholinergic synapse, also contain transporters for glutamate and ATP.Physiological reports · 2014Article
- Single-axonal organelle analysis method reveals new protein-motor associations.ACS chemical neuroscience · 2013Article
- Robust polarity establishment occurs via an endocytosis-based cortical corralling mechanism.The Journal of cell biology · 2013Article
- Single-molecule fluorescence quantification with a photobleached internal standard.Analytical chemistry · 2012Article
Corrections and comments
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Authors and funding
7 authors.
Funding
Abstract
This protocol describes a method for determining both the average number and variance of proteins, in the few to tens of copies, in isolated cellular compartments such as organelles and protein complexes. Other currently available protein quantification techniques either provide an average number, but lack information on the variance, or they are not suitable for reliably counting proteins present in the few to tens of copies. This protocol entails labeling of the cellular compartment with fluorescent primary-secondary antibody complexes, total internal reflection fluorescence microscopic imaging of the cellular compartment, digital image analysis and deconvolution of the fluorescence intensity data. A minimum of 2.5 d is required to complete the labeling, imaging and analysis of a set of samples. As an illustrative example, we describe in detail the procedure used to determine the copy number of proteins in synaptic vesicles. The same procedure can be applied to other organelles or signaling complexes.
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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.