Print Email Facebook Twitter Integrated Array Tomography for 3D Correlative Light and Electron Microscopy Title Integrated Array Tomography for 3D Correlative Light and Electron Microscopy Author Lane, R. (TU Delft ImPhys/Microscopy Instrumentation & Techniques; TU Delft ImPhys/Imaging Physics) Wolters, Anouk H.G. (University Medical Center Groningen) Giepmans, Ben N.G. (University Medical Center Groningen) Hoogenboom, J.P. (TU Delft ImPhys/Microscopy Instrumentation & Techniques; TU Delft ImPhys/Imaging Physics) Department ImPhys/Imaging Physics Date 2022 Abstract Volume electron microscopy (EM) of biological systems has grown exponentially in recent years due to innovative large-scale imaging approaches. As a standalone imaging method, however, large-scale EM typically has two major limitations: slow rates of acquisition and the difficulty to provide targeted biological information. We developed a 3D image acquisition and reconstruction pipeline that overcomes both of these limitations by using a widefield fluorescence microscope integrated inside of a scanning electron microscope. The workflow consists of acquiring large field of view fluorescence microscopy (FM) images, which guide to regions of interest for successive EM (integrated correlative light and electron microscopy). High precision EM-FM overlay is achieved using cathodoluminescent markers. We conduct a proof-of-concept of our integrated workflow on immunolabelled serial sections of tissues. Acquisitions are limited to regions containing biological targets, expediting total acquisition times and reducing the burden of excess data by tens or hundreds of GBs. Subject array tomographycorrelative light and electron microscopyintegrated microscopyscanning electron microscopyserial section electron microscopyvolume electron microscopy To reference this document use: http://resolver.tudelft.nl/uuid:b67121c1-9cfb-4a5d-a350-306b0b5f5c41 DOI https://doi.org/10.3389/fmolb.2021.822232 Source Frontiers in Molecular Biosciences, 8 Part of collection Institutional Repository Document type journal article Rights © 2022 R. Lane, Anouk H.G. Wolters, Ben N.G. Giepmans, J.P. Hoogenboom Files PDF fmolb_08_822232.pdf 4.5 MB Close viewer /islandora/object/uuid:b67121c1-9cfb-4a5d-a350-306b0b5f5c41/datastream/OBJ/view