Document Type

Article

Publication Date

8-5-2021

Publication Source

BioTechniques

Abstract

Numerous imaging modules are utilized to study changes that occur during cellular processes. Besides qualitative (immunohistochemical) or semiquantitative (Western blot) approaches, direct quantitation method(s) for detecting and analyzing signal intensities for disease(s) biomarkers are lacking. Thus, there is a need to develop method(s) to quantitate specific signals and eliminate noise during live tissue imaging. An increase in reactive oxygen species (ROS) such as superoxide (O2•-) radicals results in oxidative damage of biomolecules, which leads to oxidative stress. This can be detected by dihydroethidium staining in live tissue(s), which does not rely on fixation and helps prevent stress on tissues. However, the signal-to-noise ratio is reduced in live tissue staining. We employ the Drosophila eye model of Alzheimer's disease as a proof of concept to quantitate ROS in live tissue by adapting an unbiased method. The method presented here has a potential application for other live tissue fluorescent images.

ISBN/ISSN

0736-6205, 1940-9818

Document Version

Published Version

Comments

This work is licensed under the Attribution-NonCommercial-NoDerivatives 4.0 Unported License. Permission documentation is on file.

DOI: https://doi.org/10.2144/btn-2021-0006

Publisher

FutureScience

Volume

71

Peer Reviewed

yes

Issue

2

Keywords

Alzheimer's disease, automated quantitation, confocal microscopy, dihydroethidium, Drosophila, ImageJ, live cell imaging, neurodegeneration, oxidative stress, reactive oxygen species


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