Journal: bioRxiv

Article Title: Enhanced Workflow for Urinary Extracellular Vesicle Isolation Using Membrane-Sensing Peptides

doi: 10.64898/2026.05.21.726982

Figure Lengend Snippet: Comparative analysis of tetraspanin expression and co-localization in isolated uEVss. A) MACSPlex analysis in pre-cleared urine samples (yellow bars) and in MSP-isolated uEVss (purple bars). Data were normalized to the median fluorescence intensity of all detectable markers; B) CD9 Median Fluorescence Intensity in the small EV isolated measured using HS-FCM technology comparing dUC and MSP; C) Schematic representation of the single-tetraspanin SiMoA assay; corresponding plots show the average enzyme per bead (AEB) signal for individual tetraspanins (CD9, CD63, and CD81) in uEVss isolated by dUC, SEC, and MSP-functionalized beads (N = 5). B) Sketch of EV-labeling with mAbs anti-tetraspanin for F-NTA analysis; percentage of events that colocalized for two different tetraspanin simultaneously.

Article Snippet: For each sample, 3 x 10 8 particles (calculated by NTA quantification) were incubated for 15 minutes protected from light with 0,75 μg/ml of Phycoerythrin (PE)-conjugated anti-CD9 antibody (Cat. No. 130-124-758, Miltenyi Biotec), reaching a total volume of 100 μl with filtered (0,1 μm pores) PBS.

Techniques: Expressing, Isolation, Fluorescence, Labeling

Journal: Alzheimer's Research & Therapy

Article Title: Stress-induced alteration of small extracellular vesicles drives amyloid-beta sequestration and exacerbates Alzheimer’s disease pathogenesis

doi: 10.1186/s13195-026-02028-1

Figure Lengend Snippet: Graphical representation of workflow A . In-vitro assessment of amyloid-β (Unaggregated) and EV association. Imaging was performed at three key stages: Control (Without any stresses), Sonication only, and Sonication and agitation combined. TEM micrograph shows morphological characterization of different Aβ aggregated structures formed in the presence of EVs ( B ), Scale bar= 100 nm; CFM images for respective groups ( C )and Colocalization analysis ( D ) showed maximum PCC in the post-sonication group (PCC = 0.73). Colour yellow is the merged signal of EVs (Green) and Aβ (red). Scale bar= 20 μm. PCC between CD9-Alexa 488 EVs and AβSA-Alexa 647. Control: 0.033 ± 0.005; Sonicated: 0.780 ± 0.046; Sonicated & Agitated: 0.499 ± 0.066 ( n = 3). One-way ANOVA: p < 0.0001 (E)

Article Snippet: sEVs and Aβ42 suspension was labeled using Human CD9 Alexa Fluor ® 488-conjugated Antibody (R&D systems, FAB1880G) for sEVs and Alexa Fluor ® 647 Anti-beta Amyloid 1–42 antibody [mOC64] (Abcam, ab300742) in final concentration of 2%(v/v) and 0.5%(v/v) of the total suspension respectively. sEVs and the aggregate suspension were incubated for 2 h with both antibodies at 25 °C before mounting on labolene-cleaned glass slides (Blue Star) and covered with 18 mm;10Gms glass covers (Blue Star), and were imaged on Zeiss LSM980 system using the 40X objective.

Techniques: In Vitro, Imaging, Control, Sonication

Journal: Alzheimer's Research & Therapy

Article Title: Stress-induced alteration of small extracellular vesicles drives amyloid-beta sequestration and exacerbates Alzheimer’s disease pathogenesis

doi: 10.1186/s13195-026-02028-1

Figure Lengend Snippet: Comparison of interaction between EV and small amyloid-β aggregates ( A ) and big amyloid-β aggregates/Fibrils ( B ) at low temperature 4℃. The TEM, Scale bar= 100 nm;, and CFM micrograph shows the EVs sequestering the SA group as opposed to no association between BA/Fibril B . Graphical representation of workflow C . CFM image showing Alexa-Fluor-488 CD9 (Green) and Alexa-Fluor-647Amyloid-β (Red) signals of: EVs only; Aβ; and EVs and Aβ together D . Scale bar= 20 μm. Fluorescence intensity per cell was measured as Mean Fluorescence intensity/Area of ROI. Each condition included 3 cells per dish with a total of n = 9 cells per group. Statistical analysis using one-way ANOVA (Kruskal–Wallis test) indicated a significant difference ( p < 0.001) E . Cell viability ( F ) 10,000cells/well were seeded and included 4 wells per plate with a total of n = 4. Statistical analysis using one-way ANOVA and multiple comparisons showing a highly significant effect ( p < 0.00001)

Article Snippet: sEVs and Aβ42 suspension was labeled using Human CD9 Alexa Fluor ® 488-conjugated Antibody (R&D systems, FAB1880G) for sEVs and Alexa Fluor ® 647 Anti-beta Amyloid 1–42 antibody [mOC64] (Abcam, ab300742) in final concentration of 2%(v/v) and 0.5%(v/v) of the total suspension respectively. sEVs and the aggregate suspension were incubated for 2 h with both antibodies at 25 °C before mounting on labolene-cleaned glass slides (Blue Star) and covered with 18 mm;10Gms glass covers (Blue Star), and were imaged on Zeiss LSM980 system using the 40X objective.

Techniques: Comparison, Fluorescence

Journal: Alzheimer's Research & Therapy

Article Title: Stress-induced alteration of small extracellular vesicles drives amyloid-beta sequestration and exacerbates Alzheimer’s disease pathogenesis

doi: 10.1186/s13195-026-02028-1

Figure Lengend Snippet: Graphical representation of workflow A . Circulating sEVs derived from Control, MCI and AD patient does not show colocalization signal B . Co-incubation with Aβ SA was performed only for control and AD EVs. Controls sEVs incubated with small amyloid-β aggregates C . AD sEVs, when incubated with small amyloid-β aggregates, colocalize (White arrows) at low temperature 4℃ D . Colour Yellow is the colocalised signal for sEVs (Green)and Aβ (Red). Scale bar= 20 μm. PCC ( E ) between CD9–Alexa Fluor 488 EVs and AβSA-Alexa Fluor 647, calculated using Costes thresholding (FIJI/JACoP). Control EVs: 0.033 ± 0.005; AD EVs: 0.780 ± 0.046 ( n = 3). One-way ANOVA: p < 0.0001. Percentage of AβSA associated with EVs ( F ), Control EVs: 2.87 ± 0.90%; AD EVs: 40.37 ± 1.98% (( n = 3; incubation maintained at 4 °C)). Unpaired t-test: p < 0.0001. AD ( n = 9), MCI ( n = 3), and non-demented controls ( n = 10)

Article Snippet: sEVs and Aβ42 suspension was labeled using Human CD9 Alexa Fluor ® 488-conjugated Antibody (R&D systems, FAB1880G) for sEVs and Alexa Fluor ® 647 Anti-beta Amyloid 1–42 antibody [mOC64] (Abcam, ab300742) in final concentration of 2%(v/v) and 0.5%(v/v) of the total suspension respectively. sEVs and the aggregate suspension were incubated for 2 h with both antibodies at 25 °C before mounting on labolene-cleaned glass slides (Blue Star) and covered with 18 mm;10Gms glass covers (Blue Star), and were imaged on Zeiss LSM980 system using the 40X objective.

Techniques: Derivative Assay, Control, Incubation