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  Targeted 2D histology and ultrastructural bone analysis based on 3D microCT anatomical locations

Moreno-Jiménez, I., Garske, D., Lahr, C., Hutmacher, D., & Cipitria, A. (2021). Targeted 2D histology and ultrastructural bone analysis based on 3D microCT anatomical locations. MethodsX, 8: 101480. doi:10.1016/j.mex.2021.101480.

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 Creators:
Moreno-Jiménez, Inés1, Author           
Garske, Daniela1, Author           
Lahr, C.A., Author
Hutmacher, D.W., Author
Cipitria, Amaia1, Author           
Affiliations:
1Amaia Cipitria, Biomaterialien, Max Planck Institute of Colloids and Interfaces, Max Planck Society, ou_2489692              

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Free keywords: Mineralised tissue, Bone ultrastructure, Bone extracellular matrix, Controlled-angle microtome sectioning, Immunohistochemistry (IHC), Second harmonic generation (SHG), Confocal laser scanning microscopy (CLSM), Lacunar-canalicular network (LCN)
 Abstract: Histological processing of mineralised tissue (e.g. bone) allows examining the anatomy of cells and tissues as well as the material properties of the tissue. However, resin-embedding offers limited control over the specimen position for cutting. Moreover, specific anatomical planes (coronal, sagittal) or defined landmarks are often missed with standard microtome sectioning. Here we describe a method to precisely locate a specific anatomical 2D plane or any anatomical feature of interest (e.g. bone lesions, newly formed bone, etc) using 3D micro computed tomography (microCT), and to expose it using controlled-angle microtome cutting. The resulting sections and corresponding specimen's block surface offer correlative information of the same anatomical location, which can then be imaged using multi-scale microscopy. Moreover, this method can be combined with immunohistochemistry (IHC) to further identify any component of the bone microenvironment (cells, extracellular matrix, proteins, etc) and guide subsequent in-depth analysis. Overall, this method allows to: • Cut your specimens in a consistent position and precise manner using microCT-based controlled-angle microtome sectioning. • Locate and expose a specific anatomical plane (coronal, sagittal plane) or any other anatomical landmarks of interest based on microCT. • Identify any cell or tissue markers based on IHC to guide further in-depth examination of those regions of interest.

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Language(s): eng - English
 Dates: 2021-08-092021
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Type: -
 Identifiers: DOI: 10.1016/j.mex.2021.101480
BibTex Citekey: MORENOJIMENEZ2021101480
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Title: MethodsX
  Abbreviation : MethodsX
Source Genre: Journal
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Publ. Info: Amsterdam, The Netherlands : Elsevier BV
Pages: - Volume / Issue: 8 Sequence Number: 101480 Start / End Page: - Identifier: ISSN: 2215-0161