Challenges in Large Volume Analysis
Until now, the axial resolution in serial block face imaging (SBFI) was limited by the minimal section thickness that can be physically cut from the block-face using an in-chamber microtome. However, with a combination of SBF-SEM and Multi-Energy Deconvolution SEM (MED-SEM), our Large Volume Analysis workflow solution now enables large-volume imaging with truly isotropic 3D resolution. This is achieved by acquiring data from multiple layers of the sample in between the physical cuts using different beam energies. The combination of mechanical sectioning and optical sectioning using multi-energy deconvolution provides true isotropic resolution. Optimized detectors and low-vacuum operation ensure high-quality imaging. The acquisition software provides automation of processes ranging from low-level setup and alignment to walkaway acquisition of a complete image series. This approach includes a complete workflow from initial setup to final results, including software solutions for large area/volume analysis, LM image overlay, reconstruction, visualization, and segmentation.