Ultrastructure and 3D elemental composition of frozen-hydrated neural tissue

The goal of this project is to generate the first combined structural and elemental 3D data of frozen-hydrated brain tissue at the nanoscale. For this, we will prepare high pressure frozen tissue preserved close to the native state and we will acquire both cryo X-ray holotomography and X-ray fluorescence (XRF) tomography on the same specimens. By mapping the intracellular 3D distribution of key elements (Fe, Ca, K, Zn, P, Mn, Cl) within neurons and astrocytes in the molecular layer of the dentate gyrus, combined with ultrastructural imaging, we can gain insight into neuron-astrocyte interactions. After X-ray cryogenic imaging, we will heavy metal stain and embed the tissue in resin for further assessment. Besides the technical advances, this work aims to enhance our understanding of neurons and astrocytes connectivity and function, contributing valuable insights to neurobiology and connectomics.