期刊
PLOS ONE
卷 16, 期 4, 页码 -出版社
PUBLIC LIBRARY SCIENCE
DOI: 10.1371/journal.pone.0249603
关键词
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资金
- National Science Foundation Graduate Research International Experience [1829436]
- Fulbright Open Research Award - Netherlands-American Foundation
- Janos Bolyai Research Fellowship
- Netherlands Organisation for Scientific Research -Domain Applied and Engineering Sciences [VENI 15924]
- Office Of The Director
- Office Of Internatl Science &Engineering [1829436] Funding Source: National Science Foundation
In this study, the particle size distribution in biofluids was measured using microfluidic resistive pulse sensing (MRPS), revealing findings on optimal diluent, lower limit of detection, reproducibility, and agreement in concentration measurement. Operating guidelines were developed for reproducibly measuring the particle size distribution of extracellular vesicles (EVs) and other particles in biofluids with MRPS.
The particle size distribution (PSD) of extracellular vesicles (EVs) and other submicron particles in biofluids is commonly measured by nanoparticle tracking analysis (NTA) and tunable resistive pulse sensing (TRPS). A new technique for measuring the PSD is microfluidic resistive pulse sensing (MRPS). Because specific guidelines for measuring EVs together with other particles in biofluids with MRPS are lacking, we developed an operating procedure to reproducibly measure the PSD. The PSDs of particles in human plasma, conditioned medium of PC3 prostate cancer cell line (PC3 CM), and human urine were measured with MRPS (nCS1, Spectradyne LLC) to investigate: (i) the optimal diluent that reduces the interfacial tension of the sample while keeping EVs intact, (ii) the lower limit of detection (LoD) of particle size, (iii) the reproducibility of the PSD, (iv) the optimal dilution for measuring the PSD, and (v) the agreement in measured concentration between microfluidic cartridges with overlapping detection ranges. We found that the optimal diluent is 0.1% bovine serum albumin (w/v) in Dulbecco's phosphate-buffered saline. Based on the shape of the PSD, which is expected to follow a power-law function within the full detection range, we obtained a lower LoD of 75 nm for plasma and PC3 CM and 65 nm for urine. Normalized PSDs are reproducible (R-2 > 0.950) at dilutions between 10-100x for plasma, 5-20x for PC3 CM, and 2-4x for urine. Furthermore, sample dilution does not impact the dilution-corrected concentration when the microfluidic cartridges are operated within their specified concentration ranges. PSDs from microfluidic cartridges with overlapping detection ranges agreed well (R-2 > 0.936) and when combined the overall PSD spanned 5 orders of magnitude of measured concentration. Based on these findings, we have developed operating guidelines to reproducibly measure the PSD of EVs together with other particles in biofluids with MRPS.
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