High spatial resolution readout of 3-D metabolic organ structure: an automated, low-temperature redox ratio-scanning instrument.

Intraorgan compartmentation of metabolic processes plays an important role in the understanding of the physiological function of the integrated organ under normal as well as under pathological conditions. We describe here a technique by which 3-D information on tissue redox state may be obtained by means of automated scanning of surface fluorescence. The instrument allows for serial scanning of frozen tissue. A typical scan of a tissue volume of 3 X 3 X 2 mm at a linear resolution of 50 micron and a spatial resolution of ca. 3 X 10(-7) ml includes 144,000 single-point measurements of pyridine nucleotide and flavoprotein fluorescence within the tissue block. The scanning process is fully computerized and programs have been developed which allow 2- or 3-D reconstruction of the data in terms of "redox ratio models," exemplified here by a 3-D model of a spreading depression wave in the cerebral cortex of a gerbil.

[1]  F. Jöbsis,et al.  Oxidative and Glycolytic Recovery Metabolism in Muscle Fluorometric observations on their relative contributions , 1967 .

[2]  J. Williamson.  GLYCOLYTIC CONTROL MECHANISMS. I. INHIBITION OF GLYCOLYSIS BY ACETATE AND PYRUVATE IN THE ISOLATED, PERFUSED RAT HEART. , 1965, The Journal of biological chemistry.

[3]  Britton Chance,et al.  In vivo one-dimensional imaging of phosphorus metabolites by phosphorus-31 nuclear magnetic resonance. , 1983 .

[4]  H. Poulsen,et al.  Evaluation of a freeze-clamping technique designed for two- and three-dimensional metabolic studies of rat liver in vivo. Quenching efficiency and effect of clamping on tissue morphology. , 1980, Analytical biochemistry.

[5]  K. Jungermann,et al.  Functional heterogeneity of rat liver parenchyma and of isolated hepatocytes , 1975, FEBS letters.

[6]  B. Quistorff,et al.  Digitonin perfusion of rat liver. A new approach in the study of intra-acinar and intracellular compartmentation in the liver. , 1985, The Biochemical journal.

[7]  J. Lemasters,et al.  A non‐invasive method to study metabolic events within sublobular regions of hemoglobin‐free perfused liver , 1980, FEBS letters.

[8]  A. Gjedde,et al.  Blood‐Brain Glucose Transfer in Spreading Depression , 1981, Journal of neurochemistry.

[9]  K. Jungermann,et al.  Regulation of hepatic metabolism : intra- and intercellular compartmentation , 1986 .

[10]  B Chance,et al.  In vivo measurement of pyridine nucleotide fluorescence from cat brain cortex. , 1976, Journal of applied physiology.

[11]  E. Ogata,et al.  Microfluorometry of oxidation-reduction state of the rat kidney in situ. , 1971, Journal of applied physiology.

[12]  D. Häussinger.  Hepatocyte heterogeneity in glutamine and ammonia metabolism and the role of an intercellular glutamine cycle during ureogenesis in perfused rat liver. , 1983, European journal of biochemistry.

[13]  B Chance,et al.  Oxidation-reduction properties of the mitochondrial flavoprotein chain. , 1968, Biochemical and biophysical research communications.

[14]  B. Quistorff.  A mechanical device for the rapid removal and freezing of liver or brain tissue from unanaesthetized and nonparalyzed rats. , 1975, Analytical biochemistry.

[15]  A. Gjedde,et al.  Relationship between local changes in cortical blood flow and extracellular K+ during spreading depression. , 1980, Acta physiologica Scandinavica.

[16]  B. Chance,et al.  On the fluorescence of NAD(P)H in whole-cell preparations of tumours and normal tissues. , 1970, European journal of biochemistry.

[17]  A. Kovách.  Oxygen transport to tissue , 1981 .

[18]  B. Chance,et al.  Ischemic areas in perfused rat hearts: measurement by NADH fluorescence photography. , 1976, Science.

[19]  B. Chance,et al.  Flavin and pyridine nucleotide oxidation-reduction changes in perfused rat liver. I. Anoxia and subcellular localization of fluorescent flavoproteins. , 1969, The Journal of biological chemistry.

[20]  B. Chance,et al.  Initiation of endochondral calcification is related to changes in the redox state of hypertrophic chondrocytes. , 1982, Science.

[21]  B Chance,et al.  A versatile time-sharing multichannel spectrophotometer, reflectometer, and fluorometer. , 1975, Analytical biochemistry.

[22]  F. Jöbsis,et al.  Changes in Fluorescence in a Frog Sartorius Muscle Following a Twitch , 1959, Nature.

[23]  M. Reivich,et al.  THE [14C]DEOXYGLUCOSE METHOD FOR THE MEASUREMENT OF LOCAL CEREBRAL GLUCOSE UTILIZATION: THEORY, PROCEDURE, AND NORMAL VALUES IN THE CONSCIOUS AND ANESTHETIZED ALBINO RAT 1 , 1977, Journal of neurochemistry.

[24]  B. Chance,et al.  Oxidation-reduction ratio studies of mitochondria in freeze-trapped samples. NADH and flavoprotein fluorescence signals. , 1979, The Journal of biological chemistry.

[25]  B Chance,et al.  Simple techniques for freeze clamping and for cutting and milling of frozen tissue at low temperature for the purpose of two- or three-dimensional metabolic studies in vivo. , 1980, Analytical biochemistry.

[26]  J. B. Chapman,et al.  Fluorometric Studies of Oxidative Metabolism in Isolated Papillary Muscle of the Rabbit , 1972, The Journal of general physiology.