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.
Hitachi H7600 TEM
FEI Tecnai G2 Twin TEM
(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)
- 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
Deeg, CM., Chow, CET., and Suttle, CA. (2018) The kinetoplastid-infecting Bodo saltans virus (BsV), a window into the most abundant giant viruses in the sea. eLIFE 7: e33014
Shumborski, SJ., Samuels, AL., and Bird, DA. (2016) Fine structure of the Arabidopsis stem cuticle: effects of fixation and changes over development. Planta 244(4): 843-851.