RT Journal Article
SR Electronic
T1 Multi-fluorescence high-resolution episcopic microscopy (MF-HREM) for three dimensional imaging of adult murine organs
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.04.03.023978
DO 10.1101/2020.04.03.023978
A1 Claire Walsh
A1 Natalie Holroyd
A1 Eoin Finnerty
A1 Sean G. Ryan
A1 Paul W. Sweeney
A1 Rebecca J. Shipley
A1 Simon Walker-Samuel
YR 2020
UL http://biorxiv.org/content/early/2020/04/07/2020.04.03.023978.abstract
AB 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.