QUANTITATIVE REDOX SCANNING OF TISSUE SAMPLES USING A CALIBRATION PROCEDURE

查看全文 作　　者：HE [1]N.XU;BAOHUA [1,2]WU;SHOKO [2]NIOKA;BRITTON [2]CHANCE;LIN [1]Z.LI

机构地区：[1]Molecular Imaging Laboratory,Department of Radiology University of Pennsylvania,Philadelphia PA 19104,USA;[2]Johnson Research Foundation Department of Biochemistry and Biophysics University of Pennsylvania,PA 19104,USA

出　　处：《Journal of Innovative Optical Health Sciences》索引2009年第2卷第4期,共11页

基　　金：the Susan G.Komen Foundation Grant KG081069(PI:L.Z.Li);an NIH supported research resource(P41-RR02305,PI:R.Reddy);the Network of Translational Research in Optical Imaging(NTROI)at the University of Pennsylvania(U54 CA105008,PI:W.S.El-Deiry);an NIH Grant UO1-CA105490(PI:L.A.Chodosh). 摘　　要：The fluorescence properties of reduced nicotinamide adenine dinucleotide(NADH)and oxidizedflavoproteins(Fp)including flavin adenine dinucleotide(FAD)in the respiratory chain are sensitive indicators of intracellular metabolic states and have been applied to the studies of mitochondrial function with energy-linked processes.The redox scanner,a three-dimensional(3D)low temperature imager previously developed by Chance et al.,measures the in vivo metabolicproperties of tissue samples by acquiring fluorescence images of NADH and Fp.The redox ratios,i.e.Fp/(Fp+NADH)and NADH/(Fp+NADH),provided a sensitive index of the mitochondrialredox state and were determined based on relative signal intensity ratios.Here we report thefurther development of the redox scanning technique by using a calibration method to quantifythe nominal concentration of the fluorophores in tissues.The redox scanner exhibited very goodlinear response in the range of NADH concentration between 165–1318µM and Fp between90–720µM using snap-frozen solution standards.Tissue samples such as human tumor mousexenografts and various mouse organs were redox-scanned together with adjacent NADH and Fpstandards of known concentration at liquid nitrogen temperature.The nominal NADH and Fpconcentrations as well as the redox ratios in the tissue samples were quantified by normalizing the tissue NADH and Fp fluorescence signal to that of the snap-frozen solution standards.This calibration procedure allows comparing redox images obtained at different time,independent of instrument settings.The quantitative multi-slice redox images revealed heterogeneity inmitochondrial redox state in the tissues. 关键词：NADH FLAVOPROTEIN redox ratio MITOCHONDRIA tissue heterogeneity

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QUANTITATIVE REDOX SCANNING OF TISSUE SAMPLES USING A CALIBRATION PROCEDURE

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