Students will be able to:
- Understand the theoretical considerations underlying three dimensional reconstruction
- Prepare a specimen for electron tomography
- Collect a dataset comprising a series of images taken from a sample rotated about a single axis
- Process the images to produce a three dimensional map – including alignment, reconstruction and correction for anisotropic shrinkage.
- Use computer-based tools to assist with the interpretation of the reconstruction
Syllabus
Introduction
- Properties of the electron microscope which make tomography possible
- Projection theory
- Back projection
- Missing wedge / cone
Software
- Spider
- Imod
Sample preparation
- Conventional
- Cryo
Machines
- The advantages of 1MeV
- Energy filters
- Computer control
- CCD cameras
Online Lectures
- Intro to Electron Microscopy for Biologists (EMU, UCT)
References
Books
- Electron Tomography: Three-Dimensional Imaging With The TEM. Edited by: Joachim Frank, © 1992, 400 pgs
Useful web sites
- Bram Koster at Utrecht University
- A manual by John Crum. National Center for Microscopy and Imaging Research, San Diego.
- Wolfgang Baumeister in Martinsried.
- Richard McIntosh in Boulder