title: Functional imaging of membrane potential at the single mitochondrion level: possible application for diagnosis of human diseases.
abstract: Functional biochemical tests are the gold standard for the diagnosis of mitochondria-related diseases. However, the availability of the biological samples from patients' tissues represents a severe limitation to the number of screenable enzymatic activities. In this study we developed a fluorescent probe-assisted microscopy protocol enabling to assess the ΔΨ(m)-generating capacity by mitochondria immobilized on a glass surface at the single organelle resolution-level. The advantage of this assay over others is to scale-down the amount of the biological sample required to test in a short time the functional activity of all the components of the oxidative phosphorylation system without loss of accuracy. Furthermore, the distribution of a given enzymatic activity can also be evaluated within the mitochondrial population enabling to measure the level of functional heterogeneity of the respiratory chain dysfunction.
authors.affiliation: Department of Biomedical Sciences, University of Foggia, Foggia, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Quarato
authors.forename: Claudia
authors.initials: C
authors.lastname: Piccoli
authors.forename: Rosella
authors.initials: R
authors.lastname: Scrima
authors.forename: Nazzareno
authors.initials: N
authors.lastname: Capitanio
mesh_terms: D000165:Acridine Orange / Q000031:analogs & derivatives; D000818:Animals; D004396:Coloring Agents; D045222:Electron Transport Chain Complex Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D057075:Enzyme Assays / Q000379:methods; D006801:Humans; D057230:Limit of Detection; D008297:Male; D053078:Membrane Potential, Mitochondrial; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal / Q000379:methods; D008930:Mitochondria, Liver / Q000378:metabolism*; D028361:Mitochondrial Diseases / Q000175:diagnosis*; D010084:Oxidation-Reduction; D010101:Oxygen Consumption; D014475:Uncoupling Agents / Q000737:chemistry
chemical_list: D004396:Coloring Agents; D045222:Electron Transport Chain Complex Proteins; D014475:Uncoupling Agents; C056258:N(10)-nonylacridine orange; D000165:Acridine Orange