Vitrification and Cryo-TEM Imaging of Nanoparticles

Liposomal, polymeric nanoparticle and extracellular vesicle solutions were vitrified following standard protocols described elsewhere [20 (link)]. Quantifoil holey carbon film grids (Orthogonal Array of 2 µm Diameter Holes—2 µm Separation, mounted on a 300 M Cu grid, #657-300-CU, Ted Pella) were vitrified in liquid ethane in Vitrobot (FEI) after a negative glow-discharged treatment of the grids and deposition of 3 µL of each sample. Cryo-transfer sample holders of type GATAN Model 626 kept the sample vitrified during electron microscopy analysis. Every sample was observed on a JEM-2100F UHR (80-200kV, JEOL Ltd. Akishima, Tokio, Japan) field emission gun (FEG) transmission electron microscope at different magnifications (8000× and 30,000×). Low-dose micrographs were recorded on a state of the art TVIPS F216 CMOS camera (2k × 2k).

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Osorio-Querejeta I., Carregal-Romero S., Ayerdi-Izquierdo A., Mäger I., Nash L.A., Wood M., Egimendia A., Betanzos M., Alberro A., Iparraguirre L., Moles L., Llarena I., Möller M., Goñi-de-Cerio F., Bijelic G., Ramos-Cabrer P., Muñoz-Culla M, & Otaegui D. (2020). MiR-219a-5p Enriched Extracellular Vesicles Induce OPC Differentiation and EAE Improvement More Efficiently Than Liposomes and Polymeric Nanoparticles. Pharmaceutics, 12(2), 186.

Publication 2020

Model 626 F216 CMOS camera

After treatment Carbon Cmos Electron microscopy Ethane Extracellular vesicle Liposomal Polymeric Transmission electron microscope

Corresponding Organization : Spanish Multiple Sclerosis Network

Other organizations : CIC biomaGUNE, Centro de Investigación Biomédica en Red de Enfermedades Respiratorias, Tecnalia, University of Oxford, University of Ottawa, Wilfrid Laurier University, GAIKER Technology Centre, Ikerbasque

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Variable analysis

independent variables

Liposomal, polymeric nanoparticle and extracellular vesicle solutions

dependent variables

Visualization and characterization of the nanoparticles and extracellular vesicles using electron microscopy

control variables

Quantifoil holey carbon film grids (Orthogonal Array of 2 µm Diameter Holes—2 µm Separation, mounted on a 300 M Cu grid, #657-300-CU, Ted Pella)
Negative glow-discharged treatment of the grids
Deposition of 3 µL of each sample
Cryo-transfer sample holders of type GATAN Model 626
JEM-2100F UHR (80-200kV, JEOL Ltd. Akishima, Tokio, Japan) field emission gun (FEG) transmission electron microscope
Low-dose micrographs recorded on a TVIPS F216 CMOS camera (2k × 2k)

positive controls

Not specified

negative controls

Not specified

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