Sectioning, Staining and TEM Imaging of Embedded Exosomes: A Protocol To Visualize the Structural Features of Exosomes Using TEM
Published: April 30, 2023
Abstract
Source: Jung, M. K. et al. Sample Preparation and Imaging of Exosomes by Transmission Electron Microscopy. J. Vis. Exp. (2018)
This video describes the technique for preparing a thin section of embedded exosomes. These sections can further be stained and imaged using transmission electron microscopy, or TEM, to study the structural features of exosomes.
Protocol
1. Sectioning of the Exosomes
- Prepare exosome block by embedding the purified exosome sample (obtained from the culture supernatant of HCT116 cells) in pure low viscosity embedding mixture.
- Section the exosome block with 60 nm thickness through an ultra-microtome.
- Double stain with 2% uranyl acetate for 20 min and lead citrate for 10 min.
For Reynolds lead solution, add 1.33 g of lead nitrate and 1.76 g of sodium citrate to a total of 50 mL distilled water. - Observe the grid under transmission electron microscopy at 80 kV.
- Click "Acquire" and then click "File" and "Save as" in the CCD camera system under the electron microscope at 80 kV. Follow automatic settings for the exposure time.
Disclosures
The authors have nothing to disclose.
Materials
| Ultra-microtome | Leica | UCT | |
| Uranyl acetate | EMS | 22400 | Hazardous chemical |
| Lead citrate | EMS | 17900 | |
| Transmission Electron Microscopy | Hitachi | H7600 | |
| Nickel grid | EMS | G200-Ni | |
| Copper grid | EMS | G200-Cu | |
| Transmission Electron Microscopy | JEOL | JEM2200FS |
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Cite This Article
Sectioning, Staining and TEM Imaging of Embedded Exosomes: A Protocol To Visualize the Structural Features of Exosomes Using TEM. J. Vis. Exp. (Pending Publication), e20418, doi: (2023).