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Quantitative assessment of cerebral basement membranes using electron microscopy

Quantitative assessment of cerebral basement membranes using electron microscopy
Quantitative assessment of cerebral basement membranes using electron microscopy

In this chapter we describe in detail the tissue processing techniques we employ for the study of cerebral tissue by transmission electron microscopy (TEM). In particular, we explain a technique that enables quantification of changes in cerebral basement membranes at the ultrastructural level. This is significant, as age related pathological conditions affecting the brain are often accompanied by ultrastructural changes in the cerebral vasculature.Briefly, experimental mice are fixed by perfusion and their brains removed. Brains are then vibratomed into 100 μm slices with regions of interest microdissected and processed for TEM following a protocol optimized for the preservation of cerebral tissue. Changes in the thickness of cerebral basement membranes are then quantified using novel software. Some prior knowledge of general TEM specimen preparation and sectioning will be useful when performing this protocol.

Animals, Basement Membrane, Cerebral Cortex, Desiccation, Formaldehyde, Glutaral, Mice, Microscopy, Electron, Transmission, Microtomy, Tissue Embedding, Tissue Fixation, Journal Article
367-375
Humana Press
Sharp, Matthew MacGregor
ec57c53a-a10a-4b8a-94fe-03eca85ab7c3
Page, Anton
76ebbfb8-4fe3-495c-afff-1f2f34977fee
Morris, Alan
e70d7160-c9d9-4035-90b8-9006a1052551
Weller, Roy O.
4a501831-e38a-4d39-a125-d7141d6c667b
Carare, Roxana O.
0478c197-b0c1-4206-acae-54e88c8f21fa
Clausen, Björn E.
Laman, Jon D.
Sharp, Matthew MacGregor
ec57c53a-a10a-4b8a-94fe-03eca85ab7c3
Page, Anton
76ebbfb8-4fe3-495c-afff-1f2f34977fee
Morris, Alan
e70d7160-c9d9-4035-90b8-9006a1052551
Weller, Roy O.
4a501831-e38a-4d39-a125-d7141d6c667b
Carare, Roxana O.
0478c197-b0c1-4206-acae-54e88c8f21fa
Clausen, Björn E.
Laman, Jon D.

Sharp, Matthew MacGregor, Page, Anton, Morris, Alan, Weller, Roy O. and Carare, Roxana O. (2017) Quantitative assessment of cerebral basement membranes using electron microscopy. In, Clausen, Björn E. and Laman, Jon D. (eds.) Inflammation. (Methods in Molecular Biology, , (doi:10.1007/978-1-4939-6786-5_25), 1559) New York. Humana Press, pp. 367-375. (doi:10.1007/978-1-4939-6786-5_25).

Record type: Book Section

Abstract

In this chapter we describe in detail the tissue processing techniques we employ for the study of cerebral tissue by transmission electron microscopy (TEM). In particular, we explain a technique that enables quantification of changes in cerebral basement membranes at the ultrastructural level. This is significant, as age related pathological conditions affecting the brain are often accompanied by ultrastructural changes in the cerebral vasculature.Briefly, experimental mice are fixed by perfusion and their brains removed. Brains are then vibratomed into 100 μm slices with regions of interest microdissected and processed for TEM following a protocol optimized for the preservation of cerebral tissue. Changes in the thickness of cerebral basement membranes are then quantified using novel software. Some prior knowledge of general TEM specimen preparation and sectioning will be useful when performing this protocol.

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More information

e-pub ahead of print date: 7 January 2017
Published date: 2017
Keywords: Animals, Basement Membrane, Cerebral Cortex, Desiccation, Formaldehyde, Glutaral, Mice, Microscopy, Electron, Transmission, Microtomy, Tissue Embedding, Tissue Fixation, Journal Article

Identifiers

Local EPrints ID: 420420
URI: https://eprints.soton.ac.uk/id/eprint/420420
PURE UUID: c54adda5-5cce-46b8-9b5c-6ddb88da2f73
ORCID for Matthew MacGregor Sharp: ORCID iD orcid.org/0000-0002-6623-5078

Catalogue record

Date deposited: 08 May 2018 16:30
Last modified: 03 Dec 2019 01:41

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