Date of Award

2018

Document Type

Open Access Thesis

Department

Biomedical Science

Sub-Department

School of Medicine

First Advisor

Robert L. Price

Abstract

Advances in tissue clearing have allowed biological and biomedical researchers to image entire segments of tissue and generate three-dimensional reconstructions, allowing for a better understanding of the structural and morphological characteristics of large tissues. This study compared three commonly utilized clearing methods and analyzed their effectiveness in increasing imaging depth for three-dimensional reconstruction of large tissue segments. Mouse heart, mouse brain, mouse colon, and embryonic chick tissues were cut into thick sections, cleared and immunolabeled with an antibody specific to each tissue type, and imaged using a confocal microscope. The results of this study concluded that most tissues cleared by an electrophoretic clearing method were able to be imaged deeper than samples cleared with solvent based clearing methods. Electrophoretic methods produced clearer samples with fewer pigments and artefacts when compared to other clearing methods, and allowed for greater imaging depth when compared to solvent-cleared and uncleared specimens. The results of this study support existing literature suggesting that electrophoretic clearing provides more replicable and accurate clearing by increasing imaging depth and could generate more precise threedimensional reconstructions through confocal imaging. Advances in imaging technology such as light sheet fluorescence microscopy and new computational reconstruction capabilities could also increase the impact and efficacy of electrophoretic tissue clearing on the study of large sections of tissue at the organ, system, or organism level.

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