Description
A full understanding of the fine organization of cells and tissues requires their high resolution visualization in three dimensions (3D). Plastic embedded cells can be imaged with automated focused ion beam (FIB) - scanning electron microscopy (SEM). In this technique, the FIB removes a thin slice from the block, after which the exposed face is imaged with the SEM. This process is repeated until the desired volume is imaged, allowing for 3D imaging at high resolution (sub
5 nm) in x, y, and z directions. This technique is especially suited to obtain ultrastructural information of subcellular regions or even an entire cell.

Service provider
Cell Microscopy Core (CMC), University Medical Centre Utrecht

Location
Utrecht, The Netherlands

Contacts
Scientific contact: J. Klumperman  
Technical contact: G. Posthuma 

Description
The electron microscope is a type of microscope that uses a beam of accelerated electrons to create an image of the specimen. It is capable of much higher magnifications and has a greater resolving power than a light microscope, allowing it to see much smaller objects in finer detail.

Service provider I
Advanced Light Microscopy Facility (ALMF)

Location
Heidelberg, Germany

Contacts
Scientific contact: Rainer Pepperkok
Technical contact: Stefan Terjung

Service provider II
Cell Microscopy Core (CMC), University Medical Centre Utrecht

Location
Utrecht, The Netherlands

Contacts
Scientific contact: J. Klumperman  
Technical contact: G. Posthuma 

Service provider III
Scottish Oceans Institute (SOI), Universiy of St Andrews

Location
St. Andrews, UK

Contacts
Scientific contact: David Ferrier
Technical contact: Irvine Davidson

Description
In this variation of EM, specimens are chemically fixed, cryo-protected and frozen. After this treatment, the sample is hard enough to be thin-sectioned by cryo-ultramicrotomy. The cryo-sections are then thawed and exposed to probes or antibodies. This technique is very sensitive and adapted to identifying membrane compartments in cells.

Service provider
Cell Microscopy Core (CMC), University Medical Centre Utrecht

Location
Utrecht, The Netherlands

Contacts
Scientific contact: J. Klumperman  
Technical contact: G. Posthuma 

Description
In TEM tomography thick plastic sections - typically with a thickness of 300 nm - are tilted in the electron beam. For each tilt angle, 2D images are performed and the tilt series is computed to reconstruct the volume: the tomogram. Serial thick sections can be imaged the same way, which then results in ssTEM-tomograpy.

Service provider
Advanced Light Microscopy Facility (ALMF)

Location
Heidelberg, Germany

Contacts
Scientific contact: Rainer Pepperkok
Technical contact: Stefan Terjung