Improved DiOlistic labelling technique for neurons in situ: Detailed visualisation of dendritic spines and concurrent histochemical-detection in fixed tissue

Bevan, Ryan J. ORCID: https://orcid.org/0000-0002-2557-2887, Cimaglia, Gloria, Morgan, James E. ORCID: https://orcid.org/0000-0002-8920-1065 and Taylor, Philip R. ORCID: https://orcid.org/0000-0003-0163-1421 2024. Improved DiOlistic labelling technique for neurons in situ: Detailed visualisation of dendritic spines and concurrent histochemical-detection in fixed tissue. Methods 229 , pp. 82-93. 10.1016/j.ymeth.2024.06.009

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Abstract

DiOlistic labelling is a robust, unbiased ballistic method that utilises lipophilic dyes to morphologically label neurons. While its efficacy on freshly dissected tissue specimens is well-documented, applying DiOlistic labelling to stored, fixed brain tissue and its use in polychromatic multi-marker studies poses significant technical challenges. Here, we present an improved, step-by-step protocol for DiOlistic labelling of dendrites and dendritic spines in fixed mouse tissue. Our protocol encompasses the five key stages: Tissue Preparation, Dye Bullet Preparation, DiOlistic Labelling, Confocal Imaging, and Image Analysis. This method ensures reliable and consistent labelling of dendritic spines in fixed mouse tissue, combined with increased throughput of samples and multi-parameter staining and visualisation of tissue, thereby offering a valuable approach for neuroscientific research.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine Optometry and Vision Sciences
Publisher:	Elsevier
ISSN:	1046-2023
Date of First Compliant Deposit:	24 June 2024
Date of Acceptance:	20 June 2024
Last Modified:	10 Jul 2024 10:30
URI:	https://orca.cardiff.ac.uk/id/eprint/170084

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