On Friday, February 17th at 13:00 CET, Nico Sommerdijk, Department of Medical Biosciences, Electron Microscopy Center, Radboudumc Technology Center Microscopy, Radboudumc, Nijmegen-NL delivers a lecture on “3D Correlative Live and Cryogenic imaging of Biological Tissues combining Raman, Light and Electron Microscopy” as part of the Volume EM Series, brought to you by the Volume EM community and Euro-BioImaging.
Imaging biological tissues at different length scales (e.g. correlative light and electron microscopy (CLEM)) has shown its strength in the last decades, by combining dynamics and high-resolution imaging with fluorescent-LM and EM, respectively. For EM, ultrastructure preservation is essential and is traditionally done with chemical fixation and resin embedding. However, these chemical agents can induce artefacts. Instead, physical fixation is shown to be a powerful method allowing for high resolution imaging of samples in the hydrated native stage. Cryo-CLEM is already a well-established method on 2D cell culture, but is unexplored for volume imaging on 3D samples such as tissues and organoids, which are more biological relevant.
Nico Sommerdijk will present a new 3D cryoCLEM workflow for tissue imaging connecting live-cell fluorescence with volume cryo-FIB/SEM. To connect the two modalities, cryo-fluorescence was used to localize the live-cell imaged region of interest in the high-pressure frozen carriers/samples. For high accurate localization of LM to FIB/SEM, he and his group developed a tailor-made finderTOP, which leaves a grid pattern on the ice surface that can be imaged with both modalities. This approach allows high resolution live-cell imaging to be combined with 3D cryo-FIB/SEM volume imaging and eventually cryo-lamellae lift-outs can be made for cryoTEM. In addition he will demonstrate the integration of Raman microscopy in their workflow to provide (bio)chemical information correlated with the 3D structural information of the tissue.
Nico Sommerdijk and his colleagues are currently exploring this workflow to study matrix development in bone organoids-on-chip, focusing on the organization and mineralization of collagen.
Friday, February 17th at 13:00 CET
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