A Protocol for Improved Precision and Increased Confidence in Nanoparticle Tracking Analysis Concentration Measurements between 50 and 120 nm in Biological Fluids
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|Title:||A Protocol for Improved Precision and Increased Confidence in Nanoparticle Tracking Analysis Concentration Measurements between 50 and 120 nm in Biological Fluids||Authors:||Parsons, Martin E.M.
Szklanna, Paulina B
Guang, Matthew Ho Zhi
O’Connor, Hugh D.
Ní Áinle, Fionnuala
Maguire, Patricia B.
|Permanent link:||http://hdl.handle.net/10197/9955||Date:||3-Nov-2017||Online since:||2019-04-15T11:40:47Z||Abstract:||Nanoparticle tracking analysis (NTA) can be used to quantitate extracellular vesicles (EVs) in biological samples and is widely considered a useful diagnostic tool to detect disease. However, accurately profiling EVs can be challenging due to their small size and heterogeneity. Here, we aimed to provide a protocol to facilitate high-precision particle quantitation by NTA in plasma, the supernatant of activated purified platelets [the platelet releasate (PR)] and in serum, to increase confidence in NTA particle enumeration. The overall variance and the precision of NTA measurements were quantified by root mean square error and relative standard error. Using a bootstrapping approach, we found that increasing video replicates from 5 s × 60 s to 25 s × 60 s captures led to a reduction in overall variance and a reproducible increase in the precision of NTA particle-concentration quantitation for all three biofluids. We then validated our approach in an extended cohort of 32 healthy donors. Our results indicate that for vesicles sized between 50 and 120 nm, the precision of routine NTA measurements in serum, plasma, and PR can be significantly improved by increasing the number of video replicates captured. Our protocol provides a common platform to statistical compare particle size distribution profiles in the exosomal-vesicle size range across a variety of biofluids and in both healthy donor and patient groups.||Funding Details:||Health Research Board
Irish Research Council
Science Foundation Ireland
|Type of material:||Journal Article||Publisher:||Frontiers Media||Journal:||Frontiers in cardiovascular medicine||Volume:||4||Issue:||Article 68||Start page:||1||End page:||7||Copyright (published version):||2017 the Authors||Keywords:||Extracellular vesicles; Nanoparticle tracking analysis; Plasma; Serum; Platelets; Platelet releasate; Particle enumeration||DOI:||10.3389/fcvm.2017.00068||Language:||en||Status of Item:||Peer reviewed|
|Appears in Collections:||Conway Institute Research Collection|
Mathematics and Statistics Research Collection
Biomolecular and Biomedical Science Research Collection
Medicine Research Collection
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