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Proceedings Paper

Fluorescence spectroscopy and cryoimaging of rat lung tissue mitochondrial redox state
Author(s): R. Sepehr; S. Audi; K. Staniszewski; S. Maleki; M. Ranji
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Paper Abstract

The objective of this study was to demonstrate the utility of optical cryoimaging and fluorometry to evaluate tissue redox state of the mitochondrial metabolic coenzymes NADH (Nicotinamide Adenine Dinucleotide) and FAD (Flavin Adenine Dinucleotide) in intact rat lungs. The ratio (NADH/FAD), referred to as mitochondrial redox ratio (RR), is a measure of the lung tissue mitochondrial redox state. Isolated rat lungs were connected to a ventilation-perfused system. Surface NADH and FAD fluorescence signals were acquired before and after lung perfusion in the absence (control perfusate) or presence of potassium cyanide (KCN, complex IV inhibitor) to reduce the mitochondrial respiratory chain (state 5 respiration). Another group of lungs were perfused with control perfusate or KCN-containing perfusate as above, after which the lungs were deflated and frozen rapidly for subsequent 3D cryoimaging. Results demonstrate that lung treatment with KCN increased lung surface NADH signal by 22%, decreased FAD signal by 8%, and as result increased RR by 31% as compared to control perfusate (baseline) values. Cryoimaging results also show that KCN increased mean lung tissue NADH signal by 37%, decreased mean FAD signal by 4%, and increased mean RR by 47%. These results demonstrate the utility of these optical techniques to evaluate the effect of pulmonary oxidative stress on tissue mitochondrial redox state in intact lungs.

Paper Details

Date Published: 8 June 2011
PDF: 6 pages
Proc. SPIE 8087, Clinical and Biomedical Spectroscopy and Imaging II, 80870A (8 June 2011); doi: 10.1117/12.890019
Show Author Affiliations
R. Sepehr, Univ. of Wisconsin-Milwaukee (United States)
S. Audi, Marquette Univ. (United States)
Zablocki VA Medical Ctr. (United States)
K. Staniszewski, Univ. of Wisconsin-Milwaukee (United States)
S. Maleki, Univ. of Wisconsin-Milwaukee (United States)
M. Ranji, Univ. of Wisconsin-Milwaukee (United States)


Published in SPIE Proceedings Vol. 8087:
Clinical and Biomedical Spectroscopy and Imaging II
Nirmala Ramanujam; Jürgen Popp, Editor(s)

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