Journal: Cell reports. Medicine
Article Title: Single extracellular vesicle detection assay identifies membrane-associated α-synuclein as an early-stage biomarker in Parkinson's disease.
doi: 10.1016/j.xcrm.2025.101999
Figure Lengend Snippet: Figure 1. Droplet-based microfluidic extra- cellular vesicle digital immunoassay (A) Workflow of the assay, also demonstrated in Video S1. (B) Schematic illustration of the droplet micro- fluidic device with further design details in Fig- ure S1. (C) Photograph of the microfluidic device (75 3 25 3 1.1 mm); scale bar, 2 cm. (D) Representative images of the picoliter-sized droplets; scale bar, 100 mm. (E) Size distribution of the droplets averaging 27.9 mm in diameter with further analysis in Figures S2–S4. (F) Representative fluorescence microscopy im- age of L1EVs immunocaptured from serum using DAPI-labeled beads, followed by staining with Alexa Fluor 647-labeled anti-CD9 in EV-depleted serum as a control and bead-to-serum-EV ratio of 1:10, or 1:1, or 1:0.1; scale bar, 2 mm. (G) Representative fluorescence microscopy im- age of L1EVs immunocaptured from serum using DAPI-labeled beads, followed by staining with Alexa Fluor 561-labeled anti-Tetraspanin Trio (CD9, CD63, and CD81) using a bead-to-serum- EV ratio of 1:0.1; scale bar, 2 mm.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Recombinant human FGF-8a protein Tocris Cat# 4745-F8 CHIR 99021 Tocris Cat# 4423 Human/mouse/rat BDNF recombinant protein PeproTech Cat# 450-02 Human GDNF recombinant protein PeproTech Cat# 450-10 Human TGF-beta 3 recombinant protein Thermo Fisher Scientific Cat# PHG9305 DAPT Tocris Cat# 2634 L-Ascorbic acid Sigma Cat# A4403 Dibutyryl cAMP sodium salt Sigma Cat# AD0627 Mouse laminin Sigma Cat# L2020 MES hydrate Sigma Cat# M8250 N-(3-Dimethylaminopropyl)-N0-ethylcarbodiimide hydrochloride (EDC) Sigma Cat# 03450 N-Hydroxysuccinimide (NHS) Sigma Cat# 130672 TWEEN 20 Sigma Cat# P7949 Streptavidin b-galactosidase (SbG) conjugate Thermo Fisher Scientific Cat# S931 Mineral oil Sigma Cat# M5904 TritonTM X-100 Sigma Cat# T8787 Fluorescein Di-b-D-Galactopyranoside (FDG) Thermo Fisher Scientific Cat# F1179 Glycine Sigma Cat# G7126 Tris buffer Sigma Cat# 648314 Paraformaldehyde Sigma Cat# 158127 2% uranyl acetate solution Electron Microscopy Sciences Cat# 22400-2 Polybrene Santa Cruz Biotechnology Cat# sc-134220 Critical commercial assays PierceTM BCA Protein Assay Kit Thermo Fisher Scientific Cat# 23225 U-PLEX human a-synuclein kit Meso Scale Discovery Cat# K151WKK-2 R-PLEX human CD81 (EV) assay Meso Scale Discovery Cat# K1515NR-2 Experimental models: Cell lines Human induced pluripotent stem cell (iPSC) This study N/A
Techniques: Microscopy, Labeling, Staining, Control
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