Module 3

Sample Preparation for 2D and 3D Electron Microscopy (University of Zurich, Campus Irchel)

In this module, you will learn how to prepare biological specimens for 2D and 3D electron microscopy techniques such as serial-section transmission and scanning electron microscopy, focused ion beam and serial block-face scanning electron microscopy. To preserve the sample ultra-structure, chemical and cryo-fixation techniques (high-pressure freezing, freeze-substitution) will be used. Subsequently, resin-embedded samples will be sectioned by ultramicrotomy and imaged by transmission, scanning and focused ion beam scanning electron microscopy. The cellular ultra-structure in the resulting micrographs will be reviewed in context to preparation. Samples will be provided.

Contact:

Dr. Andres Kaech
Center for Microscopy and Image Analysis
University of Zurich
andres.kaech@zmb.uzh.ch
+41 44 634 26 65

Learning outcomes:

  • Participants can process biological samples for imaging the subcellular ultra-structure by electron microscopy
  • Participants know the advantages and disadvantages of different preparation steps and procedures
  • Participants know the advantages and disadvantages of the different 2D/3D electron microscopy techniques and can select the optimal approach for their research

Practical activities:

  • Room temperature fixation and cryo fixation of biological specimens, dehydration, resin embedding, ultrathin sectioning (ultramicrotomy), contrast enhancement
  • Imaging sections in the transmission electron microscope

Prerequisites:

  • Have a need to apply the trained skills/one of the techniques for your research question at least within a year after the course