University of Hertfordshire

From the same journal

By the same authors

  • Claire Walsh
  • Natalie Holroyd
  • Eoin Finnerty
  • Sean Ryan
  • Paul Sweeney
  • Rebecca Shipley
  • Simon Walker-Samuel
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Original languageEnglish
JournalElife
DOIs
Publication statusSubmitted - 7 Apr 2020

Abstract

Three-dimensional microscopy of large biological samples (>0.5 cm3) is transforming biological research. Many existing techniques require trade-offs between image resolution and sample size, require clearing or use optical sectioning. These factors complicate the implementation of large volume 3D imaging. Here we present Multi-fluorescent High Resolution Episcopic Microscopy (MF-HREM) which allows 3D imaging of large samples without the need for clearing or optical sectioning. MF-HREM uses serial-sectioning and block-facing wide-field fluorescence, without the need for tissue clearing or optical sectioning. We detail developments in sample processing including stain penetration, resin embedding and imaging. In addition, we describe image post-processing methods needed to segment and further quantify these data. Finally, we demonstrate the wide applicability of MF-HREM by: 1) quantifying adult mouse glomeruli. 2) identifying injected cells and vascular networks in tumour xenograft models; 3) quantifying vascular networks and white matter track orientation in mouse brain.

ID: 20849680