Abstract
One recent technical innovation in neuroscience is microcircuit analysis using three-dimensional reconstructions of neural elements with a large volume Electron microscopy (EM) data set. Large-scale data sets are acquired with newly-developed electron microscope systems such as automated tape-collecting ultramicrotomy (ATUM) with scanning EM (SEM), serial block-face EM (SBEM) and focused ion beam-SEM (FIB-SEM). Currently, projects are also underway to develop computer applications for the registration and segmentation of the serially-captured electron micrographs that are suitable for analyzing large volume EM data sets thoroughly and efficiently. The analysis of large volume data sets can bring innovative research results. These recently available techniques promote our understanding of the functional architecture of the brain.
Highlights
Electron microscopy (EM) has been used in neuroscience research for more than 60 years
Assuming that 80% of the projected electrons carry a sufficient signal that could be detected by the scanning EM (SEM) detector, the simulation analysis results suggest that electrons projected with an accelerating voltage of 1.5 keV or 2 keV would interact with the 50-nm-thick tissue section most efficiently (Figure 2) (Kubota et al, 2018)
Automated image acquisition of serial ultrathin sections collected on grids with transmission EM (TEM) was introduced (Bloss et al, 2018), and the features including the collection of serial sections on a silicon wafer and manual observation with SEM are available, allowing the acquisition of large volume EM data set using (Boeckeler Instruments, Inc., Tucson, AZ, USA; JEOL Ltd., Akishima, Japan) two commonly available EM tools, namely, ultramicrotome and EM
Summary
Electron microscopy (EM) has been used in neuroscience research for more than 60 years. Some jitters are found in serial section images before alignment due to the instability of the SEM beam for charging, or deformation/expansion of the block surface by the heat during imaging/sputtering, and this makes the orthogonal xz view slightly irregular (Figure 4N) Despite these issues, the FIB-SEM has unique features that are not available with other methods as discussed in the above, and can be used to capture serial electron micrographs for various research purposes. Automated image acquisition of serial ultrathin sections collected on grids with TEM was introduced (Bloss et al, 2018), and the features including the collection of serial sections on a silicon wafer and manual observation with SEM are available, allowing the acquisition of large volume EM data set using (Boeckeler Instruments, Inc., Tucson, AZ, USA; JEOL Ltd., Akishima, Japan) two commonly available EM tools, namely, ultramicrotome and EM This likely promotes the use of 3D-EM methods. These pieces of software will undoubtedly advance brain structure analysis in neuroscience
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