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Systematic Methodological Evaluation of a Multiplex Bead-Based Flow Cytometry Assay for Detection of Extracellular Vesicle Surface Signatures

Overview of attention for article published in Frontiers in immunology, June 2018
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  • In the top 25% of all research outputs scored by Altmetric
  • Good Attention Score compared to outputs of the same age (70th percentile)
  • Good Attention Score compared to outputs of the same age and source (72nd percentile)

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9 X users

Citations

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Title
Systematic Methodological Evaluation of a Multiplex Bead-Based Flow Cytometry Assay for Detection of Extracellular Vesicle Surface Signatures
Published in
Frontiers in immunology, June 2018
DOI 10.3389/fimmu.2018.01326
Pubmed ID
Authors

Oscar P. B. Wiklander, R. Beklem Bostancioglu, Joshua A. Welsh, Antje M. Zickler, Florian Murke, Giulia Corso, Ulrika Felldin, Daniel W. Hagey, Björn Evertsson, Xiu-Ming Liang, Manuela O. Gustafsson, Dara K. Mohammad, Constanze Wiek, Helmut Hanenberg, Michel Bremer, Dhanu Gupta, Mikael Björnstedt, Bernd Giebel, Joel Z. Nordin, Jennifer C. Jones, Samir EL Andaloussi, André Görgens

Abstract

Extracellular vesicles (EVs) can be harvested from cell culture supernatants and from all body fluids. EVs can be conceptually classified based on their size and biogenesis as exosomes and microvesicles. Nowadays, it is however commonly accepted in the field that there is a much higher degree of heterogeneity within these two subgroups than previously thought. For instance, the surface marker profile of EVs is likely dependent on the cell source, the cell's activation status, and multiple other parameters. Within recent years, several new methods and assays to study EV heterogeneity in terms of surface markers have been described; most of them are being based on flow cytometry. Unfortunately, such methods generally require dedicated instrumentation, are time-consuming and demand extensive operator expertise for sample preparation, acquisition, and data analysis. In this study, we have systematically evaluated and explored the use of a multiplex bead-based flow cytometric assay which is compatible with most standard flow cytometers and facilitates a robust semi-quantitative detection of 37 different potential EV surface markers in one sample simultaneously. First, assay variability, sample stability over time, and dynamic range were assessed together with the limitations of this assay in terms of EV input quantity required for detection of differently abundant surface markers. Next, the potential effects of EV origin, sample preparation, and quality of the EV sample on the assay were evaluated. The findings indicate that this multiplex bead-based assay is generally suitable to detect, quantify, and compare EV surface signatures in various sample types, including unprocessed cell culture supernatants, cell culture-derived EVs isolated by different methods, and biological fluids. Furthermore, the use and limitations of this assay to assess heterogeneities in EV surface signatures was explored by combining different sets of detection antibodies in EV samples derived from different cell lines and subsets of rare cells. Taken together, this validated multiplex bead-based flow cytometric assay allows robust, sensitive, and reproducible detection of EV surface marker expression in various sample types in a semi-quantitative way and will be highly valuable for many researchers in the EV field in different experimental contexts.

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The data shown below were collected from the profiles of 9 X users who shared this research output. Click here to find out more about how the information was compiled.
Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 267 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 267 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 49 18%
Student > Ph. D. Student 45 17%
Student > Bachelor 28 10%
Student > Master 24 9%
Student > Doctoral Student 15 6%
Other 34 13%
Unknown 72 27%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 61 23%
Agricultural and Biological Sciences 30 11%
Medicine and Dentistry 21 8%
Immunology and Microbiology 19 7%
Engineering 9 3%
Other 43 16%
Unknown 84 31%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 6. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 27 April 2020.
All research outputs
#6,302,026
of 25,382,440 outputs
Outputs from Frontiers in immunology
#6,462
of 31,537 outputs
Outputs of similar age
#100,817
of 341,533 outputs
Outputs of similar age from Frontiers in immunology
#205
of 736 outputs
Altmetric has tracked 25,382,440 research outputs across all sources so far. Compared to these this one has done well and is in the 75th percentile: it's in the top 25% of all research outputs ever tracked by Altmetric.
So far Altmetric has tracked 31,537 research outputs from this source. They typically receive more attention than average, with a mean Attention Score of 8.4. This one has done well, scoring higher than 79% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 341,533 tracked outputs that were published within six weeks on either side of this one in any source. This one has gotten more attention than average, scoring higher than 70% of its contemporaries.
We're also able to compare this research output to 736 others from the same source and published within six weeks on either side of this one. This one has gotten more attention than average, scoring higher than 72% of its contemporaries.