Pooled SEC fractions from the first EV containing peak (F1/S1) were dialysed against 10 mM HEPES, 140 mM NaCl, pH 7.4 using Amicon®Ultra-0.5 ml centrifugal filters, 10kDa (Merck Millipore) to remove phosphate. Electron microscopy samples were prepared as previously described.[41 (link)] Samples were applied undiluted onto previously glow discharged home-made carbon coated copper grids and incubated for 10 s. Subsequently specimens were stained three times with 1% uranyl formate solution. Raw micrographs were acquired automatically at 67,000 magnification on specimen level using the Leginon system [42 (link)] operated on a Tecnai Spirit BioTWIN TEM (Thermo Fisher Scientific, Hillsboro, OR, USA) at 80kv acceleration voltage and a Tietz 416 CMOS detector (TVIPS, Martinsried, Germany).
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Electron Microscopy of Extracellular Vesicles
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Corresponding Organization : Aarhus University
Other organizations : Arla Foods (Denmark)
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Variable analysis
independent variables
- Dialysis of pooled SEC fractions from the first EV containing peak (F1/S1) against 10 mM HEPES, 140 mM NaCl, pH 7.4 using Amicon®Ultra-0.5 ml centrifugal filters, 10kDa
- Electron microscopy samples
- Samples were prepared as previously described [41]
- Samples were applied undiluted onto previously glow discharged home-made carbon coated copper grids and incubated for 10 s
- Specimens were stained three times with 1% uranyl formate solution
- Raw micrographs were acquired automatically at 67,000 magnification on specimen level using the Leginon system [42] operated on a Tecnai Spirit BioTWIN TEM at 80kv acceleration voltage and a Tietz 416 CMOS detector
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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