PIE-scope, integrated cryo-correlative light and FIB/SEM microscopy

PIE-scope, integrated cryo-correlative light and FIB/SEM microscopy

Blog Article

Cryo-electron tomography (cryo-ET) is emerging as a revolutionary method for resolving the structure of macromolecular complexes in situ.However, sample preparation for in situ Cryo-ET is labour-intensive and can require both Toy Abacuses cryo-lamella preparation through cryo-focused ion beam (FIB) milling and correlative light microscopy to ensure that the event of interest is present in the lamella.Here, we present an integrated cryo-FIB and light microscope setup called the Photon Ion Electron microscope (PIE-scope) that enables direct and rapid isolation of cellular regions containing protein complexes shoes of interest.Specifically, we demonstrate the versatility of PIE-scope by preparing targeted cryo-lamellae from subcellular compartments of neurons from transgenic Caenorhabditis elegans and Drosophila melanogaster expressing fluorescent proteins.We designed PIE-scope to enable retrofitting of existing microscopes, which will increase the throughput and accuracy on projects requiring correlative microscopy to target protein complexes.

This new approach will make cryo-correlative workflow safer and more accessible.