Module 3

Maximum participants: 6

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

+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