Transmission electron microscopy (TEM) is an electron microscopy technique in which a wide beam of electrons passes through a thin slice of specimen (<100 nm) to form an image. For biological samples, it is used to examine cell ultrastructure and subcellular localization of proteins/molecules using immnolabelling techniques. Other TEM applications are Cryo-TEM and TEM Tomography.
Equipment:
FEI Tecnai Spirit 120kV TEM
FEI Tecnai G2 Twin 200kV TEM
Sample Preparation:
(A) Biological tissue samples need to be fixed and embedded in the resin
- Initial fixation: Cryo-fixation or Chemical fixation using aldehyde fixatives
- Secondary fixation: Osmium fixation to fix lipids (Not needed for immunolabelling samples)
- Dehydration
- Infiltration/embedding in resin: Choice of resin
- Ultramicrotome sectioning
- Antibody incubation for immunolabelling
- Grid staining with uranyl acetate and lead citrate
- TEM imaging
(B) For isolated organelles, macromolecules, bacterial cells and viruses, please visit the TEM Negative Staining
Publications:
Golla, K., Paul, M., Lengyell, T. C., Simpson, E. M., Falet, H., & Kim, H. (2023). A novel association between platelet filamin A and soluble N-ethylmaleimide sensitive factor attachment proteins regulates granule secretion. Research and Practice in Thrombosis and Haemostasis, 7(4), 100019. https://doi.org/10.1016/j.rpth.2022.100019
Leung, J., Strong, C., Badior, K. E., Robertson, M., Wu, X., Meledeo, M. A., Kang, E., Paul, M., Sato, Y., Harashima, H., Cap, A. P., Devine, D. v., Jan, E., Cullis, P. R., & Kastrup, C. J. (2023). Genetically engineered transfusable platelets using mRNA lipid nanoparticles. Science Advances, 9(48), eadi0508. https://doi.org/10.1126/sciadv.adi0508
Mriga, D., Duo, C., Till, M., & Vanessa J., A. (2022). DLG5 and Cadherins Are Key to Peripheral Glia Integrity. bioRxiv. https://doi.org/10.1101/2022.11.01.514384