2007 NSTI Nanotechnology Conference and Trade Show - Nanotech 2007 - 10th Annual

Automated 3D cortex image data acquisition

B.H. Lich and G. Knott
FEI Electron Optics, NL

Keywords:
automation imaging synaps cortex

Abstract:
The study of the 3D architecture of cortex tissue requires the creation of image data sets with sufficient detail in all directions. The detail level required to produce meaningful data requires imaging quality that allows identification of synaptic regions with their characteristic neurotransmitter carrying vesicles. One way to provide a data set is to apply microtomes and TEM imaging, however, this process requires a lot of manual input and control – therefore it is time consuming and labor intensive. Another, novel way to create a data set of the cortex, is to apply a Focused Ion Beam (FIB) based slicing with successive in-situ SEM based high-resolution imaging. The process for 3D imaging of the cortex is easy to describe: 1. Prepare the sample and create a FIB aligned edge at the volume of interest. 2. Define milling conditions and patterning definitions for the proper volume. 3. Select an imaging detector, to be applied after the slicing process and define the imaging conditions. 4. Run the successive slicing and viewing process in an automated way. For this indicated process, the parameter settings for the cortex data set with some conditions and limitations have been studied in more detail

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