Integrated Array Tomography for 3D Correlative Light and Electron Microscopy.

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.
Competing Interests: RL, AHGW, and BNGG declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The integrated microscope used in this study is a product of Delmic B.V. of which JPH is a co-founder and shareholder.
(Copyright © 2022 Lane, Wolters, Giepmans and Hoogenboom.)