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Bennett. J Transl Genet Genom 2020;4:36-49 Journal of Translational
DOI: 10.20517/jtgg.2020.17 Genetics and Genomics

Review Open Access

Cryogenic electron paramagnetic resonance
spectroscopy of flash-frozen tissue for

characterization of mitochondrial disease

Brian Bennett

Department of Physics, Marquette University, Milwaukee, WI 53233, USA.

Correspondence to: Prof. Brian Bennett, Department of Physics, Marquette University, 1420 W. Clybourn St, Milwaukee, WI
53233, USA. E-mail: brian.bennett@mu.edu
How to cite this article: Bennett B. Cryogenic electron paramagnetic resonance spectroscopy of flash-frozen tissue for
characterization of mitochondrial disease. J Transl Genet Genom 2020;4:36-49. https://doi.org/10.20517/jtgg.2020.17
Received: 19 Feb 2020 First Decision: 6 Mar 2020 Revised: 4 Apr 2020 Accepted: 15 Apr 2020 Available online: 23 Apr 2020

Science Editor: Andrea L. Gropman Copy Editor: Jing-Wen Zhang Production Editor: Tian Zhang

Abstract
Electron paramagnetic resonance spectroscopy (EPR) is an analytical technique that, uniquely, can be used
to directly interrogate flash-frozen tissue. Quantitative information on the thermodynamic potential of the
mitochondrion to synthesize ATP, and the extent of reactive oxygen species-mediated oxidative stress on the
mitochondrion and the cell at large, can be obtained. A compromised ability to synthesize ATP and oxidative
stress are two of the characteristic sequelae of mitochondrial disease and therapeutic approaches may differ
widely depending on which of these is dominant. EPR, therefore, has a role to play in the characterization,
diagnosis, and ongoing evaluation of therapies for mitochondrial disease in human patients and model systems.
An introduction to mitochondrial disease is followed by a description of EPR, a summary of the EPR signals that
can be expected from tissue samples, sample preparation and analytical methods, and a case study in which EPR
and complementary techniques were employed on a rat model to study human mitochondrial disease.

Keywords: Mitochondrial disease, electron paramagnetic resonance, electron paramagnetic resonance spectroscopy,
mitochondria

INTRODUCTION
Mitochondria are characterized by mitochondrial respiratory chain (MRC) complexes that catalyze redox
reactions and act as electron transfer conduits during energy metabolism, driving ATP synthesis while
closely chaperoning potentially toxic one-electron redox equivalents. The MRC proteins contain a variety of
redox-active centers, including iron-sulfur clusters, heme, copper ions, and quinones. Each of these can exist

© The Author(s) 2020. Open Access This article is licensed under a Creative Commons Attribution 4.0
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