Serial block-face scanning electron microscopy: Difference between revisions
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'''Serial Block-Face Scanning Electron Microscopy ''' (SBFSEM) is a method to generate high resolution three-dimensional images from biological samples such as brain tissue.<ref>Denk W, Horstmann H (2004) Serial Block-Face Scanning Electron Microscopy to Reconstruct Three-Dimensional Tissue Nanostructure. PLoS Biol 2(11): e329. doi:10.1371/journal.pbio.0020329</ref> 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. |
'''Serial Block-Face Scanning Electron Microscopy ''' (SBFSEM) is a method to generate high resolution three-dimensional images from biological samples such as brain tissue.<ref>Denk W, Horstmann H (2004) Serial Block-Face Scanning Electron Microscopy to Reconstruct Three-Dimensional Tissue Nanostructure. PLoS Biol 2(11): e329. doi:10.1371/journal.pbio.0020329</ref> 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 [http://www.gatan.com/products/sem_products/products/3View_landing.php]. |
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== External links == |
== External links == |
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* [http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.0020329] Original Publication in PloS Biology |
* [http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.0020329] Original Publication in PloS Biology |
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* [http://www.gatan.com/products/sem_products/products/3View_landing.php] Gatan's 3View |
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Revision as of 22:24, 27 August 2010
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Serial Block-Face Scanning Electron Microscopy (SBFSEM) is a method to generate high resolution three-dimensional images from biological samples such as brain tissue.[1] 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 [1].
Applications
SBFSEM will be used 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. The biggest challenge is to develop algorithms for automatic segmentation of the very large (~TByte) datasets generated by SBFSEM.
References
- ^ Denk W, Horstmann H (2004) Serial Block-Face Scanning Electron Microscopy to Reconstruct Three-Dimensional Tissue Nanostructure. PLoS Biol 2(11): e329. doi:10.1371/journal.pbio.0020329