Integrated Array Tomography for 3D Correlative Light and Electron Microscopy

Ryan Lane*, Anouk H.G. Wolters, Ben N.G. Giepmans, Jacob P. Hoogenboom

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

3 Citations (Scopus)
17 Downloads (Pure)

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.

Original languageEnglish
Article number822232
Number of pages11
JournalFrontiers in Molecular Biosciences
Volume8
DOIs
Publication statusPublished - 2022

Keywords

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

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