Authors: B. Lich, J. Greiser, P. Faber, G. Knott
Affilation: FEI Company, Netherlands
Pages: 472 - 474
Keywords: SEM FIM automated serial section
The study of the 3D architecture of cortex requires the creation of image datasets 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 dataset 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 way to create a dataset 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 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. After investigating the requirements for obtaining the resolution level that is required for the data acquisition process we have now focused on the challenges associated with increasing the investigated volume, while maintaining the required resolution.