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Development of protocols for the first serial block-face scanning electron microscopy (SBF SEM) studies of bone tissue

Development of protocols for the first serial block-face scanning electron microscopy (SBF SEM) studies of bone tissue
Development of protocols for the first serial block-face scanning electron microscopy (SBF SEM) studies of bone tissue
There is an unmet need for a high-resolution three-dimensional (3D) technique to simultaneously image osteocytes and the matrix in which these cells reside. In serial block-face scanning electron microscopy (SBF SEM), an ultramicrotome mounted within the vacuum chamber of a microscope repeatedly sections a resin embedded block of tissue. Backscattered electron scans of the block-face provide a stack of high-resolution two-dimensional (2D) images, which can be used to visualise and quantify cells and organelles in 3D. High-resolution 3D images of biological tissues from SBF SEM have been exploited considerably to date in the neuroscience field. However, non-brain samples, in particular hard biological tissues, have appeared more challenging to image by SBF SEM due to the difficulties of sectioning and rendering the samples conductive. We have developed and propose protocols for bone tissue preparation using SBF SEM, for imaging simultaneously soft and hard bone tissue components in 3D. We review the state of the art in high resolution imaging of osteocytes, the historical perspective of SBF SEM and we present first SBF SEM proof-of-concept studies for mouse and human tissue. The application of SBF SEM to hard tissues will facilitate qualitative and quantitative 3D studies of tissue microstructure and ultrastructure in bone development, ageing and pathologies such as osteoporosis and osteoarthritis.
8756-3282
Goggin, Patricia, Mary
c2d225c4-b84d-4ead-a433-320057cb5fa9
Ho, Elaine, Ming Li
7fa9df7f-4dbf-4be4-b03f-ff79012dd44b
Gnaegi, Helmut
f5305499-8141-4197-816e-f31ee843f7b6
Searle, Stuart
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Oreffo, Richard
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Schneider, Philipp
a810f925-4808-44e4-8a4a-a51586f9d7ad
Goggin, Patricia, Mary
c2d225c4-b84d-4ead-a433-320057cb5fa9
Ho, Elaine, Ming Li
7fa9df7f-4dbf-4be4-b03f-ff79012dd44b
Gnaegi, Helmut
f5305499-8141-4197-816e-f31ee843f7b6
Searle, Stuart
18ced818-c4e0-4e1e-8898-9c4a303f10bf
Oreffo, Richard
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Schneider, Philipp
a810f925-4808-44e4-8a4a-a51586f9d7ad

Goggin, Patricia, Mary, Ho, Elaine, Ming Li, Gnaegi, Helmut, Searle, Stuart, Oreffo, Richard and Schneider, Philipp (2020) Development of protocols for the first serial block-face scanning electron microscopy (SBF SEM) studies of bone tissue. Bone, 131, [115107]. (doi:10.1016/j.bone.2019.115107).

Record type: Article

Abstract

There is an unmet need for a high-resolution three-dimensional (3D) technique to simultaneously image osteocytes and the matrix in which these cells reside. In serial block-face scanning electron microscopy (SBF SEM), an ultramicrotome mounted within the vacuum chamber of a microscope repeatedly sections a resin embedded block of tissue. Backscattered electron scans of the block-face provide a stack of high-resolution two-dimensional (2D) images, which can be used to visualise and quantify cells and organelles in 3D. High-resolution 3D images of biological tissues from SBF SEM have been exploited considerably to date in the neuroscience field. However, non-brain samples, in particular hard biological tissues, have appeared more challenging to image by SBF SEM due to the difficulties of sectioning and rendering the samples conductive. We have developed and propose protocols for bone tissue preparation using SBF SEM, for imaging simultaneously soft and hard bone tissue components in 3D. We review the state of the art in high resolution imaging of osteocytes, the historical perspective of SBF SEM and we present first SBF SEM proof-of-concept studies for mouse and human tissue. The application of SBF SEM to hard tissues will facilitate qualitative and quantitative 3D studies of tissue microstructure and ultrastructure in bone development, ageing and pathologies such as osteoporosis and osteoarthritis.

Text
BONE-D-19-00771R1-1_for_Pure - Accepted Manuscript
Restricted to Repository staff only until 24 October 2020.
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More information

Accepted/In Press date: 9 October 2019
e-pub ahead of print date: 24 October 2019
Published date: February 2020

Identifiers

Local EPrints ID: 435233
URI: http://eprints.soton.ac.uk/id/eprint/435233
ISSN: 8756-3282
PURE UUID: f7fe8397-90d0-4607-8162-dcf9b43ab496
ORCID for Richard Oreffo: ORCID iD orcid.org/0000-0001-5995-6726
ORCID for Philipp Schneider: ORCID iD orcid.org/0000-0001-7499-3576

Catalogue record

Date deposited: 28 Oct 2019 17:30
Last modified: 27 Jan 2020 13:46

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