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Sensors 2016, 16(7), 1065; doi:10.3390/s16071065

Microchambers with Solid-State Phosphorescent Sensor for Measuring Single Mitochondrial Respiration

1
Department of Biomedical Engineering, University of California, Irvine, CA 92697, USA
2
Center for Mitochondrial and Epigenomic Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA
3
Department of Electrical Engineering, University of California, Irvine, CA 92697, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Alexander Star
Received: 13 May 2016 / Revised: 20 June 2016 / Accepted: 5 July 2016 / Published: 9 July 2016
(This article belongs to the Section Biosensors)
View Full-Text   |   Download PDF [3283 KB, uploaded 9 July 2016]   |  

Abstract

It is now well established that, even within a single cell, multiple copies of the mitochondrial genome may be present (genetic heteroplasmy). It would be interesting to develop techniques to determine if and to what extent this genetic variation results in functional variation from one mitochondrion to the next (functional heteroplasmy). Measuring mitochondrial respiration can reveal the organelles’ functional capacity for Adenosine triphosphate (ATP) production and determine mitochondrial damage that may arise from genetic or age related defects. However, available technologies require significant quantities of mitochondria. Here, we develop a technology to assay the respiration of a single mitochondrion. Our “micro-respirometer” consists of micron sized chambers etched out of borofloat glass substrates and coated with an oxygen sensitive phosphorescent dye Pt(II) meso-tetra(pentafluorophenyl)porphine (PtTFPP) mixed with polystyrene. The chambers are sealed with a polydimethylsiloxane layer coated with oxygen impermeable Viton rubber to prevent diffusion of oxygen from the environment. As the mitochondria consume oxygen in the chamber, the phosphorescence signal increases, allowing direct determination of the respiration rate. Experiments with coupled vs. uncoupled mitochondria showed a substantial difference in respiration, confirming the validity of the microchambers as single mitochondrial respirometers. This demonstration could enable future high-throughput assays of mitochondrial respiration and benefit the study of mitochondrial functional heterogeneity, and its role in health and disease. View Full-Text
Keywords: respiration; mitochondria; oxygen consumption rate; phosphorescent probes; PtTFPP respiration; mitochondria; oxygen consumption rate; phosphorescent probes; PtTFPP
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Pham, T.D.; Wallace, D.C.; Burke, P.J. Microchambers with Solid-State Phosphorescent Sensor for Measuring Single Mitochondrial Respiration. Sensors 2016, 16, 1065.

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