Electron Tomography

Electron tomorgraphy (ET) is a technique that retrieves three-dimensional (3D) structural information from a tilt series of two-dimensional (2D) projections that is imaged at several different angles. This method provides nanometer resolution from thick samples (100 nm to 1 mm) to provide more information about the depth of the sample.

While the electron beam is stationary, the specimen holder can rotate and the specimen can be tilted at 1 – 2° angular increments over a range of ± 70 degree about a fixed axis. To prevent radiation damage to the sample, the cumulative electron dose must be kept within tolerable limits.

Equipment and Software:

FEI Tecnai G2 Twin TEM – to collect a tilt series of 2D projections

Inspect 3D software – to reconstruct 3D volume

Amira software – to align the tilt series and perform modeling

Sample Preparation:

Regular TEM sample preparation

Section thickness is in the range of 100 nm – 1 mm and thicker than normal sections


Optimizing Data Collection

– Staining conditions

– Pre-irradiation needed (sections may shrink due to the beam intensity)

Image Data Set Alignment

– Calculation of alignment shifts

– Adjustment of tilt axis


Asghari, P., Scriven, D. R., Ng, M., Panwar, P., Chou, K. C., van Petegem, F. and Moore, E. D. (2020). Cardiac ryanodine receptor distribution is dynamic and changed by auxiliary proteins and post-translational modification. eLife9, e51602.

Baldelli, A., Trivanovic, U., and Rogak, SK. (2019) Electron tomography of soot for validation of 2-D image processing and observation of new structural features. Aerosol Science and Technology, 1-8.

Lyon, K., Adams, A., Piva, M., Asghari, P., Moore, ED., and Vogl, AW. (2017) Ca2+ signaling machinery is present at intercellular junctions and structures associated with junction turnover in rat Sertoli cells. Biology of Reproduction 96 (6): 1288-1302.