Transmission Electron Microscopy (TEM)

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:

Hitachi H7600 TEM

FEI Tecnai G2 Twin TEM

Sample Preparation:

(A) Biological tissue samples need to be fixed and embedded in the resin

  1. Initial fixation: Cryo-fixation or Chemical fixation using aldehyde fixatives
  2. Secondary fixation: Osmium fixation to fix lipids (Not needed for immunolabelling samples)
  3. Dehydration
  4. Infiltration/embedding in resin: Choice of resin
  5. Ultramicrotome sectioning
  6. Antibody incubation for immunolabelling
  7. Grid staining with uranyl acetate and lead citrate
  8. TEM imaging

(B) For isolated organelles, macromolecules, bacterial cells and viruses, please visit the TEM Negative Staining

Publications:

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.