Journal: Cell Reports Methods

Article Title: Reliable multiplex generation of pooled induced pluripotent stem cells

doi: 10.1016/j.crmeth.2023.100570

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-human SSEA-4 , Novus Biologicals , Cat#NBP2-26644AF488; RRID: AB_657849.

Techniques: Recombinant, Membrane, Flow Cytometry, Electron Microscopy, DNA Extraction, Sample Prep, Sequencing, Plasmid Preparation, Software

Journal: Frontiers in Immunology

Article Title: Extracellular vesicles epitopes as potential biomarker candidates in patients with traumatic spinal cord injury

doi: 10.3389/fimmu.2024.1478786

Figure Lengend Snippet: Characterization of plasma-isolated Extracellular Vesicles (EVs) (A) Representative results of Nanoparticle Tracking Analysis (NTA) conducted on EV isolates (1 in 100 dilution) from healthy volunteers, patients with traumatic Spinal Cord Injury (tSCI) at timepoints Emergency Room (ER) admission and 48 hours (h) after trauma. The mean particle size (nm) and concentration (particle/ml) are indicated in the graphs. (B) Representative Western-blot analysis of CD63, CD81, and CD9 expression in EVs collected from healthy volunteers and tSCI patients at ER admission, 24h and 48h after trauma. (C) Representative transmission electron microscopy-image of healthy volunteer EV isolate. Scale bar = 100 nm.

Article Snippet: Isolated EVs (20μg protein) from each sample (tSCI n=8, PT and healthy control groups, n=8), were first incubated with surface epitope-specific antibodies coupled with fluorescent-labeled beads (Panel A or B) and then incubated with MACSPlex Exosome Detect Reagents CD9, CD63 and CD81 (Cat. No. 130-108-813, Miltenyi Biotec), according to the manufacturer’s instructions and analyzed via flow cytometry (BD FACSCanto II, FACS DIVA Software, Heidelberg, Germany) ( ).

Techniques: Clinical Proteomics, Isolation, Concentration Assay, Western Blot, Expressing, Transmission Assay, Electron Microscopy

Journal: Frontiers in Cell and Developmental Biology

Article Title: Distinct miRNA profiles in human amniotic tissue and its vesicular and non-vesicular secretome

doi: 10.3389/fcell.2025.1692501

Figure Lengend Snippet: Characterization of hAM-derived extracellular vesicles. (A) Particle size distribution by nanoparticle tracking analysis (NTA) showed that most particles were ≤200 nm; n = 3, mean ± SEM. (B) Median values (X50) of particle size range from 138 nm to 211 nm after 72 h of incubation (x̄ = 177 in RA and x̄ = 174 in P), n = 3. The majority of particles were ≤200 nm. X50 values are indicated ±SD. (C) Fluorescence-triggered flow cytometry analysis (FT-FC) showed that 20%–35% of the EVs were positive for CD81 (x̄ = 19.55 in RA, x̄ = 24.40 in P). The expression of CD9 was less pronounced n = 2. (D) Western Blot analysis showed that CD63 and CD81 were more abundant in EVs than tissue, n = 3. (E) TEM confirmed EV morphologies and CD81 positive particles (immuno-gold particles (black dots)) in EV preparations from RA and P. scale bar = 100 nm. Abbreviations: human amniotic membrane (hAM), extracellular vesicles (EV), placental amnion (P), reflected amnion (RA), flotillin-1 (Flot-1), lysosomal associated membrane protein 1 (LAMP1), heat shock protein 70 (HSC70), tumor susceptibility gene 101 (TSG101), cluster of differentiation (CD), transmission electron microscopy (TEM).

Article Snippet: The staining of the respective antigens was performed by subsequent addition of 4 μL of 1:100 predilute PE-labelled antibodies for CD9 (Miltenyi Biotech, 130-118-865), CD63 (Miltenyi Biotech, Germany, 130-118-077), CD81 (Miltenyi Biotech, Germany, 130-118-481), and isotype IgG1 (Miltenyi Biotech, Germany, 130-113-438).

Techniques: Derivative Assay, Incubation, Fluorescence, Flow Cytometry, Expressing, Western Blot, Membrane, Transmission Assay, Electron Microscopy