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 A. Holroyd
A1 Eoin Finnerty
A1 Sean G. Ryan
A1 Paul W. Sweeney
A1 Rebecca J. Shipley
A1 Simon Walker-Samuel
YR 2021
UL http://biorxiv.org/content/early/2021/02/23/2020.04.03.023978.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, sample size and method complexity. A simple robust instrument with the potential to perform large volume 3D imaging currently exists in the form of the Optical HREM, however the development of the instrument to date is limited to single fluorescent wavelength imaging with non-specific eosin staining. This work presents developments to realize the potential of the HREM to become Multi-fluorescent High Resolution Episcopic Microscopy (MF-HREM).MF-HREM is a serial-sectioning and block-facing wide-field fluorescence imaging technique, which does not require tissue clearing or optical sectioning. Multiple developments are detailed in sample preparation and image post-processing to enable multiple specific stains in large samples, and show how these enable segmentation and quantification of the data. The application of MF-HREM is demonstrated in a variety of biological contexts: 3D imaging of whole tumor vascular networks and tumor cell invasion in xenograft tumors up to 7.5 mm3 at resolutions of 2.75 μm, quantification of glomeruli volume in the adult mouse kidney, and quantification of vascular networks and white matter track orientation in adult mouse brain.Competing Interest StatementThe authors have declared no competing interest.