Journal: PLoS ONE

Article Title: Comparison of extracellular vesicle isolation and storage methods using high-sensitivity flow cytometry

doi: 10.1371/journal.pone.0245835

Figure Lengend Snippet: (A) Dot plots of the side scatter (SSC) or fluorescence (BP530/40) expressed in units of FITC MESF versus forward scatter (FWSC) demonstrating the sensitivity of HS-FCM when measuring 100 and 200 nm fluorescent polystyrene beads. (B) Absence of detector swarming was verified using serially diluted CFDA-SE stained EVs from DC + UC isolated EVs. Additional fluorescence plots can be found in S6 Fig in . (C) A representative distribution of EV counts in different iodixanol density gradient fractions for both DC + UC (left) and exoEasy (right) isolated EVs and the corresponding blank medium controls. The results are presented as the average ± standard deviation of three technical replicates.

Article Snippet: Conversion to units of FITC MESF was performed using FITC MESF Beads (custom product manufactured by Becton Dickinson Biosciences, 2 μm FITC Quantitation Beads; R&D Lot #2352–87) corresponding to 98 704 (bead 1), 20 292 (bead 2), 10 552 (bead 3) and 4 315 (bead 4) units of FITC MESF (S1 Fig in ).

Techniques: Fluorescence, Staining, Isolation, Standard Deviation

Journal: PLoS ONE

Article Title: Comparison of extracellular vesicle isolation and storage methods using high-sensitivity flow cytometry

doi: 10.1371/journal.pone.0245835

Figure Lengend Snippet: (A) Dot plots of CFDA-SE fluorescence intensity expressed in units of FITC MESF as a function of forward scatter (FWSC) of EVs and blank medium controls with and without treatment with the detergents 0.5% (w/v) Triton X-100 or 0.5% (w/v) SDS. Representative examples of iodixanol fractions with a density of 1.10 g/ml are presented. Additional fluorescence plots of blank medium controls with and without treatment with the detergents can be found in S7 Fig in . (B) Relative EV numbers (%) measured during 30 seconds after detergent treatment compared to EVs without detergent (100%) in iodixanol fractions for DC + UC (left) and exoEasy (right) purified EVs. The results are presented as the average ± standard deviation of three biological replicates. For every biological replicate, the average was determined based on three technical replicate measurements. Statistically significant results are indicated by an asterisk (* p-value = 0.0031, ** p-value < 0.001).

Article Snippet: Conversion to units of FITC MESF was performed using FITC MESF Beads (custom product manufactured by Becton Dickinson Biosciences, 2 μm FITC Quantitation Beads; R&D Lot #2352–87) corresponding to 98 704 (bead 1), 20 292 (bead 2), 10 552 (bead 3) and 4 315 (bead 4) units of FITC MESF (S1 Fig in ).

Techniques: Fluorescence, Purification, Standard Deviation

Journal: PLoS ONE

Article Title: Comparison of extracellular vesicle isolation and storage methods using high-sensitivity flow cytometry

doi: 10.1371/journal.pone.0245835

Figure Lengend Snippet: (A) Samples were labeled with CFDA-SE, purified on a density gradient and EV counts (density of 1.10 g/ml) measured for 30 seconds. Relative number of recorded events (%) in 30 seconds measured after different times up to 28 days of storage at 4°C or -80°C in sucrose. Normalization of events was performed between independent experiments (n = 4) and are presented as the average percentage of recorded events compared to the number of events at day 0 with error bars indicating the standard error of the mean. (B) A representative number distribution of the fluorescence intensity (expressed in units of FITC MESF) after one month of storage at -80°C shows that the intensity distribution is comparable between fresh and stored samples. When samples were stored at 4°C, hardly any fluorescent events were found (not shown).

Article Snippet: Conversion to units of FITC MESF was performed using FITC MESF Beads (custom product manufactured by Becton Dickinson Biosciences, 2 μm FITC Quantitation Beads; R&D Lot #2352–87) corresponding to 98 704 (bead 1), 20 292 (bead 2), 10 552 (bead 3) and 4 315 (bead 4) units of FITC MESF (S1 Fig in ).

Techniques: Labeling, Purification, Fluorescence