SEC-EV peak fractions were pooled and concentrated as previously described. TEI-EV samples were used without further concentration. Holey Carbon 200 mesh Grids (Electron Microscopy Supplies) were glow-discharged (15 sec O2/H2 1:1) with a Solaris plasma cleaner (Gatan, Pleasanton, CA, U.S.). Samples were prepared with a Leica EM GP plunge-freezer. Sample grids were prepared by adding 4 µL of sample within an environmental chamber (20°C, 90% relative humidity). Excess sample was blotted away on filter paper prior to plunging into liquid ethane. Samples were stored and transferred in liquid nitrogen in a Gatan model 914 cryo transfer holder. Samples were imaged at −170°C in a JEOL 2100 Plus Transmission Electron Microscope (JEOL, Tokyo, Japan) at 200 kV, using a Minimum Dose System software. Images were acquired with an Orius SC 100 camera (Gatan) at 30 k magnification over 5-sec exposure time.
2100 plus transmission electron microscope
Manufactured by JEOL
Sourced in Japan
The JEOL 2100 Plus Transmission Electron Microscope is a high-performance, versatile instrument designed for advanced materials characterization. It features a LaB6 electron source, a robust and stable column, and a high-resolution imaging system. The JEOL 2100 Plus provides researchers and scientists with a powerful tool for studying the structure and composition of a wide range of materials at the nanoscale.
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2 protocols using 2100 plus transmission electron microscope
1
Cryo-TEM Imaging of Extracellular Vesicles
2
High-Resolution Ultrastructural Imaging
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Samples were prepared by fixing plant material by high-pressure freezing using a Leica HPM100, followed by freeze substitution with 100% ethanol (Leica EM Auto Freeze substitution) and infiltration with LRWhite resin (Agar Scientific). Ultra-thin sections were prepared with a Leica EM UCT ultramicrotome (section thickness: 90 nm) and were collected on pioloform/carbon-coated nickel grids (Agar Scientific) and stained with 2.5% uranyl acetate and Reynolds lead citrate (Reynolds, 1963 (link)). Ultrathin sections were imaged using a JEOL-2100Plus transmission electron microscope (JEOL, Japan) equipped with a Gatan OneView IS camera (Gatan, USA).
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