Microindentation of fresh soft biological tissue: A rapid tissue sectioning and mounting protocol
Clíona M. McCarthy, Kevin L. McKevitt, Sinéad A. Connolly, Isabel Andersson, Fiona C. Leahy, Siobhan Egan, Michael A. Moloney, Eamon G. Kavanagh, Colin Peirce, Eoghan M. Cunnane, Kieran D. McGourty, Michael T. Walsh, John J. E. Mulvihill
TL;DR
This paper introduces a new method for quickly and effectively preparing fresh biological tissue for microindentation, using a mixture of agarose and gelatin.
Contribution
The novel contribution is an optimized mounting protocol using 2.5% agarose and 1.5% gelatin for microindentation of fresh tissue.
Findings
Embedding tissue in a 2.5% agarose and 1.5% gelatin mixture is the most favorable method for microindentation.
The protocol enables rapid processing of fresh tissue slices without significant degradation.
The method is applicable to a variety of tissue types.
Abstract
Microindentation of fresh biological tissues is necessary for the creation of 3D biomimetic models that accurately represent the native extracellular matrix microenvironment. However, tissue must first be precisely sectioned into slices. Challenges exist in the preparation of fresh tissue slices, as they can tear easily and must be processed rapidly in order to mitigate tissue degradation. In this study, we propose an optimised mounting condition for microindentation and demonstrate that embedding tissue in a mixture of 2.5% agarose and 1.5% gelatin is the most favourable method of tissue slice mounting for microindentation. This protocol allows for rapid processing of fresh biological tissue and is applicable to a variety of tissue types.
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Taxonomy
TopicsTissue Engineering and Regenerative Medicine · Bone Tissue Engineering Materials · Additive Manufacturing and 3D Printing Technologies