Serial Block-Face Scanning Electron Microscopy
Encyclopedia
Serial block-face scanning electron microscopy (SBFSEM) is a method to generate high resolution three-dimensional images from biological samples such as brain tissue. It consists of an ultramicrotome mounted inside the vacuum chamber of a scanning electron microscope
. The surface of the block of resin-embedded fixed tissue (not the cut sections!) is imaged via detection of back-scattered electrons. After each section, the tissue block is raised by a tiny amount (less than 50 nm) to stay in focus. The machine can acquire many thousands of images in perfect alignment in an automated fashion. SBFSEM has been invented in 2004 by Winfried Denk
at the Max-Planck-Institute in Heidelberg and is commercially available from Gatan Inc.
of the very large (~TByte) datasets generated by SBFSEM.
As the microtome is able to cut many types of samples this technique is starting to catch on in many other areas ranging from cell and developmental biology to materials science.
Scanning electron microscope
A scanning electron microscope is a type of electron microscope that images a sample by scanning it with a high-energy beam of electrons in a raster scan pattern...
. The surface of the block of resin-embedded fixed tissue (not the cut sections!) is imaged via detection of back-scattered electrons. After each section, the tissue block is raised by a tiny amount (less than 50 nm) to stay in focus. The machine can acquire many thousands of images in perfect alignment in an automated fashion. SBFSEM has been invented in 2004 by Winfried Denk
Winfried Denk
Winfried Denk is a German physicist and neurobiologist, director of the Max-Plack-Institute for Medical Research in Heidelberg. He is noted for being the first to implement two-photon microscopy while a postdoctoral fellow in Watt W. Webb's lab in 1990...
at the Max-Planck-Institute in Heidelberg and is commercially available from Gatan Inc.
Applications
One of the first applications of SBFSEM is to analyze the wiring diagram of small regions of the brain. The resolution is sufficient to trace even the thinnest axons and to identify synapses. As a result of the extremely large datasets that a technique like this can produce one of the biggest challenges is to develop algorithms for automatic segmentationSegmentation
Segmentation may mean:*Market segmentation, in economics and marketingBiology*A process of morphogenesis that divides a metazoan body into a series of semi-repetitive segments*Segmentation , a series of semi-repetitive segments...
of the very large (~TByte) datasets generated by SBFSEM.
As the microtome is able to cut many types of samples this technique is starting to catch on in many other areas ranging from cell and developmental biology to materials science.
External links
- http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.0020329 Original Publication in PloS Biology
- http://www.gatan.com/3View Gatan's 3View
- http://ccdb.ucsd.edu/sand/main?stype=lite&keyword=serial%20block%20face&event=display&Submit=Go&start=1 Cell Centered Data Base, SBFSEM datasets