| Autor: |
Comfort N; Department of Environmental Health Sciences, Columbia University Mailman School of Public Health; nicole.comfort@columbia.edu., Cai K; Department of Medicine, Naomi Berrie Diabetes Center, Columbia University., Bloomquist TR; Department of Environmental Health Sciences, Columbia University Mailman School of Public Health., Strait MD; Department of Environmental Health Sciences, Columbia University Mailman School of Public Health., Ferrante AW Jr; Department of Medicine, Naomi Berrie Diabetes Center, Columbia University., Baccarelli AA; Department of Environmental Health Sciences, Columbia University Mailman School of Public Health; ab4303@cumc.columbia.edu. |
| Jazyk: |
angličtina |
| Zdroj: |
Journal of visualized experiments : JoVE [J Vis Exp] 2021 Mar 28 (169). Date of Electronic Publication: 2021 Mar 28. |
| DOI: |
10.3791/62447 |
| Abstrakt: |
The physiological and pathophysiological roles of extracellular vesicles (EVs) have become increasingly recognized, making the EV field a quickly evolving area of research. There are many different methods for EV isolation, each with distinct advantages and disadvantages that affect the downstream yield and purity of EVs. Thus, characterizing the EV prep isolated from a given source by a chosen method is important for interpretation of downstream results and comparison of results across laboratories. Various methods exist for determining the size and quantity of EVs, which can be altered by disease states or in response to external conditions. Nanoparticle tracking analysis (NTA) is one of the prominent technologies used for high-throughput analysis of individual EVs. Here, we present a detailed protocol for quantification and size determination of EVs isolated from mouse perigonadal adipose tissue and human plasma using a breakthrough technology for NTA representing major advances in the field. The results demonstrate that this method can deliver reproducible and valid total particle concentration and size distribution data for EVs isolated from different sources using different methods, as confirmed by transmission electron microscopy. The adaptation of this instrument for NTA will address the need for standardization in NTA methods to increase rigor and reproducibility in EV research. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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