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Multi-fluorescence high-resolution episcopic microscopy (MF-HREM) for three dimensional imaging of adult murine organs. / Walsh, Claire; Holroyd, Natalie; Finnerty, Eoin; Ryan, Sean; Sweeney, Paul; Shipley, Rebecca; Walker-Samuel, Simon.

In: Advanced Photonics Research, 26.05.2021.

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Walsh, Claire ; Holroyd, Natalie ; Finnerty, Eoin ; Ryan, Sean ; Sweeney, Paul ; Shipley, Rebecca ; Walker-Samuel, Simon. / Multi-fluorescence high-resolution episcopic microscopy (MF-HREM) for three dimensional imaging of adult murine organs. In: Advanced Photonics Research. 2021.

Bibtex

@article{43ec51789a184728852ca29a7a166d5f,
title = "Multi-fluorescence high-resolution episcopic microscopy (MF-HREM) for three dimensional imaging of adult murine organs",
abstract = "Three-dimensional microscopy of large biological samples (greater than 0.5 cm^3) 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 mm^3 at resolutions of 2.75 mm, quantification of glomeruli volume in the adult mouse kidney, and quantification of vascular networks and white matter track orientation in adult mouse brain.",
keywords = "serial-sectioning, HREM, deconvolution, tumor, whole-mount",
author = "Claire Walsh and Natalie Holroyd and Eoin Finnerty and Sean Ryan and Paul Sweeney and Rebecca Shipley and Simon Walker-Samuel",
note = "{\textcopyright} 2021 The Authors. Advanced Photonics Research published by Wiley-VCH GmbH. This is an open access article under the terms of the Creative Commons Attribution License, https://creativecommons.org/licenses/by/4.0/",
year = "2021",
month = may,
day = "26",
doi = "10.1002/adpr.202100110",
language = "English",
journal = "Advanced Photonics Research",

}

RIS

TY - JOUR

T1 - Multi-fluorescence high-resolution episcopic microscopy (MF-HREM) for three dimensional imaging of adult murine organs

AU - Walsh, Claire

AU - Holroyd, Natalie

AU - Finnerty, Eoin

AU - Ryan, Sean

AU - Sweeney, Paul

AU - Shipley, Rebecca

AU - Walker-Samuel, Simon

N1 - © 2021 The Authors. Advanced Photonics Research published by Wiley-VCH GmbH. This is an open access article under the terms of the Creative Commons Attribution License, https://creativecommons.org/licenses/by/4.0/

PY - 2021/5/26

Y1 - 2021/5/26

N2 - Three-dimensional microscopy of large biological samples (greater than 0.5 cm^3) 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 mm^3 at resolutions of 2.75 mm, quantification of glomeruli volume in the adult mouse kidney, and quantification of vascular networks and white matter track orientation in adult mouse brain.

AB - Three-dimensional microscopy of large biological samples (greater than 0.5 cm^3) 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 mm^3 at resolutions of 2.75 mm, quantification of glomeruli volume in the adult mouse kidney, and quantification of vascular networks and white matter track orientation in adult mouse brain.

KW - serial-sectioning

KW - HREM

KW - deconvolution

KW - tumor

KW - whole-mount

U2 - 10.1002/adpr.202100110

DO - 10.1002/adpr.202100110

M3 - Article

JO - Advanced Photonics Research

JF - Advanced Photonics Research

M1 - 2100110

ER -