Limits...
A simple flow cytometry method improves the detection of phosphatidylserine-exposing extracellular vesicles.

Arraud N, Gounou C, Linares R, Brisson AR - J. Thromb. Haemost. (2014)

Bottom Line: We show that about 50× more Anx5-positive EVs are detected by FCM when detection is triggered on fluorescence as compared with light scattering.This study presents a simple method for enumerating EVs.We believe that this method is applicable in a general context and will improve our understanding of the roles of EVs in pathophysiological situations, which will open avenues for the development of EV-based diagnosis assays.

View Article: PubMed Central - PubMed

Affiliation: Molecular Imaging and NanoBioTechnology, UMR-5248-CBMN CNRS-University of Bordeaux-IPB, Pessac, France.

Show MeSH

Related in: MedlinePlus

Influence of Anx5-F* concentration on the concentrations of Anx5-positive extracellular vesicles (EVs) detected by FS and fluorescence (FL) triggering. Curves representing the concentrations of Anx5-positive EVs (expressed per μL of pure platelet-free plasma) detected by FS (dashed lines) and FL triggering (plain lines) at various Anx5-F* concentrations, for two platelet-free plasma (PFP) samples (represented by squares and circles, respectively). The concentrations of Anx5-F* range from 0 to 2.8 nM. Each point represents the mean ± SD of two independent aliquots measured in duplicate.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4359678&req=5

fig02: Influence of Anx5-F* concentration on the concentrations of Anx5-positive extracellular vesicles (EVs) detected by FS and fluorescence (FL) triggering. Curves representing the concentrations of Anx5-positive EVs (expressed per μL of pure platelet-free plasma) detected by FS (dashed lines) and FL triggering (plain lines) at various Anx5-F* concentrations, for two platelet-free plasma (PFP) samples (represented by squares and circles, respectively). The concentrations of Anx5-F* range from 0 to 2.8 nM. Each point represents the mean ± SD of two independent aliquots measured in duplicate.

Mentions: Next, we investigated the influence of Anx5-F* concentration on the detection of Anx5-positive EVs. For both FS and FL triggering, the number of Anx5-positive EVs detected was found to increase with increasing Anx5-F* concentration, until a plateau value was reached (Fig.2). In the conditions used here, namely 10 ×-diluted PFP, the plateau was reached at 0.8 nm Anx5-F*. Most experiments reported in this paper were performed with 2.8 nm Anx5-F*, therefore in saturating conditions.


A simple flow cytometry method improves the detection of phosphatidylserine-exposing extracellular vesicles.

Arraud N, Gounou C, Linares R, Brisson AR - J. Thromb. Haemost. (2014)

Influence of Anx5-F* concentration on the concentrations of Anx5-positive extracellular vesicles (EVs) detected by FS and fluorescence (FL) triggering. Curves representing the concentrations of Anx5-positive EVs (expressed per μL of pure platelet-free plasma) detected by FS (dashed lines) and FL triggering (plain lines) at various Anx5-F* concentrations, for two platelet-free plasma (PFP) samples (represented by squares and circles, respectively). The concentrations of Anx5-F* range from 0 to 2.8 nM. Each point represents the mean ± SD of two independent aliquots measured in duplicate.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4359678&req=5

fig02: Influence of Anx5-F* concentration on the concentrations of Anx5-positive extracellular vesicles (EVs) detected by FS and fluorescence (FL) triggering. Curves representing the concentrations of Anx5-positive EVs (expressed per μL of pure platelet-free plasma) detected by FS (dashed lines) and FL triggering (plain lines) at various Anx5-F* concentrations, for two platelet-free plasma (PFP) samples (represented by squares and circles, respectively). The concentrations of Anx5-F* range from 0 to 2.8 nM. Each point represents the mean ± SD of two independent aliquots measured in duplicate.
Mentions: Next, we investigated the influence of Anx5-F* concentration on the detection of Anx5-positive EVs. For both FS and FL triggering, the number of Anx5-positive EVs detected was found to increase with increasing Anx5-F* concentration, until a plateau value was reached (Fig.2). In the conditions used here, namely 10 ×-diluted PFP, the plateau was reached at 0.8 nm Anx5-F*. Most experiments reported in this paper were performed with 2.8 nm Anx5-F*, therefore in saturating conditions.

Bottom Line: We show that about 50× more Anx5-positive EVs are detected by FCM when detection is triggered on fluorescence as compared with light scattering.This study presents a simple method for enumerating EVs.We believe that this method is applicable in a general context and will improve our understanding of the roles of EVs in pathophysiological situations, which will open avenues for the development of EV-based diagnosis assays.

View Article: PubMed Central - PubMed

Affiliation: Molecular Imaging and NanoBioTechnology, UMR-5248-CBMN CNRS-University of Bordeaux-IPB, Pessac, France.

Show MeSH
Related in: MedlinePlus