Multidimensional electron microscopy
Multidimensional electron microscopy
Electron microscopy (EM) enables the structures of cells and tissues to be visualized at the nanoscale. Though classically limited to very small fields-of-view, digitization has enabled larger areas or volumes to be imaged with nanometer resolution. Large-scale EM (‘Nanotomy’) is now routinely implemented to study the ultrastructure of cells and tissues in health and disease. However, imaging over larger scales and volumes is typically not feasible due to the low through-put of this technique.
This thesis of Peter Duinkerken explores a novel high-throughput EM method, compares it to the current state-of-the-art and demonstrates it to be up to 20 times faster than conventional EM methods. Additionally, this thesis implements large-scale energy-dispersive X-ray (EDX) imaging which reveals the elemental composition of the recorded ultrastructure. Both large-scale EM and EDX were implemented in a study of human brain tissue in which a novel phenotype of myelin was identified and described.
This study additionally demonstrated the need for a more automated approach towards the analysis of the large amounts of data that result from large-scale EM. It was demonstrated that the hyperspectral nature of EDX imaging can be utilized to automate the classification and segmentation of cellular ultrastructure, of which the spectral discernability could be further enhanced through the use of stains that contain atoms that are not abundant in the human body. Together these findings provide an enriched toolbox for biologists to study cells and tissues in health and disease.