Integrated Array Tomography for 3D Correlative Light and Electron Microscopy

Integrated Array Tomography for 3D Correlative Light and Electron Microscopy

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)

ImPhys/Imaging Physics

2022

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.

array tomography
correlative light and electron microscopy
integrated microscopy
scanning electron microscopy
serial section electron microscopy
volume electron microscopy

http://resolver.tudelft.nl/uuid:b67121c1-9cfb-4a5d-a350-306b0b5f5c41

https://doi.org/10.3389/fmolb.2021.822232

Frontiers in Molecular Biosciences, 8

Institutional Repository

journal article

© 2022 R. Lane, Anouk H.G. Wolters, Ben N.G. Giepmans, J.P. Hoogenboom