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 cryo-lamella preparation through cryo-focused ion Shuffleboard 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 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 Wheelchair Cushion 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.