ArticleCytometry. Part A : the journal of the International Society for Analytical Cytology2013
Calcium-phosphate microprecipitates mimic microparticles when examined with flow cytometry.
Article in Cytometry. Part A : the journal of the International Society for Analytical Cytology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 42 citations in OpenAlex.
- Conformational plasticity of human acid-sensing ion channel 1a.Nature structural & molecular biology · 2026Article
- Direct detection of circulating donor-derived extracellular vesicles in kidney transplant recipients.Scientific reports · 2022Article
- An imaging flow cytometry-based methodology for the analysis of single extracellular vesicles in unprocessed human plasma.Communications biology · 2022Article
- A Role for Extracellular Vesicles in SARS-CoV-2 Therapeutics and Prevention.Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology · 2021Review
- Centrifugation Removes a Population of Large Vesicles, or "Macroparticles," Intermediate in Size to RBCs and Microvesicles.International journal of molecular sciences · 2021Article
- Therapeutic Potentials of Extracellular Vesicles for the Treatment of Diabetes and Diabetic Complications.International journal of molecular sciences · 2020Review
- Article
- Optimisation of imaging flow cytometry for the analysis of single extracellular vesicles by using fluorescence-tagged vesicles as biological reference material.Journal of extracellular vesicles · 2019Article
- Systematic Methodological Evaluation of a Multiplex Bead-Based Flow Cytometry Assay for Detection of Extracellular Vesicle Surface Signatures.Frontiers in immunology · 2018Article
- A novel fluorescent probe-based flow cytometric assay for mineral-containing nanoparticles in serum.Scientific reports · 2017Article
- Saline is a more appropriate solution for microvesicles for flow cytometric analyses.Oncotarget · 2017Article
- Detection of platelet vesicles by flow cytometry.Platelets · 2017Review
- Phosphatidylethanolamine is progressively exposed in RBCs during storage.Transfusion medicine (Oxford, England) · 2017Article
- Renal extracellular vesicles: from physiology to clinical application.The Journal of physiology · 2016Review
- Applying extracellular vesicles based therapeutics in clinical trials - an ISEV position paper.Journal of extracellular vesicles · 2015Article
- Circulating membrane-derived microvesicles in redox biology.Free radical biology & medicine · 2014Review
- Standardization of sample collection, isolation and analysis methods in extracellular vesicle research.Journal of extracellular vesicles · 2013Article
Corrections and comments
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Authors and funding
4 authors at 3 institutions in 1 country.
Funding
Abstract
There are increased levels of circulating microparticles (MPs) in several disease states. Flow cytometry is a common method to examine MPs, but their small size necessitates the use of markers to distinguish specifically MPs from artifact. Annexin V, which binds phosphatidylserine, is a commonly used marker for MP detection. Annexin V requires millimolar calcium ion for optimum binding. Ca(++) can precipitate with phosphate in phosphate-buffered saline (PBS). Calcium-phosphate microprecipitates were formed by titrating Ca(++) into PBS and examined using flow cytometry. Calcium-phosphate microprecipitates were compared with MPs derived from aged donor blood units. Microprecipitates were ∼0.7-0.9 μm in diameter compared with standard beads of known size. The microprecipitates disappeared with the addition of Ca(++) chelator. When we added fluorescently labeled antibodies to microprecipitates, the median fluorescent signal increased with increasing Ca(++) concentration regardless of specificity of the antibody. When repeated with a biological sample, there was an apparent increase in the fluorescent signal that returned to baseline after Ca(++) chelation. The flow cytometry signal of calcium-phosphate microprecipitates overlaps with the MP signal. Since Ca(++) is essential for annexin V binding, it is essential to avoid artifacts from calcium-phosphate microprecipitates when using any buffer or biological fluid containing phosphate. This also highlights the potential utility of flow cytometry for the analysis of crystals in biological fluids.
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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.