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Updated: Aug 16, 2026
Using Isolated Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High throughput Microplate Respiratory Measurements
Published on: October 30, 2015
The problem of interlab variation in methods for mitochondrial disease diagnosis: enzymatic measurement of
Frank Norbert Gellerich1, Johannes A Mayr, Sebastian Reuter
1Muskellabor der Neurologischen Klinik und Poliklinik, Martin-Luther Universität Halle-Wittenberg, Julius-Kühn-Strasse 7, D-06097 Halle/Saale, Germany. frank.gellerich@medizin.uni-halle.de
Abstract:
Due to the large importance of mitochondrial function for numerous diseases the detection of respiratory chain defects for diagnostic purposes is an important task of mitochondrial medicine. For comparing the methods, standard mitochondrial homogenate prepared from bovine skeletal muscle was sent on dry ice to 14 labs of 8 countries. Activities of complexes I, I + III, II, II + III, IV (cytochrome c oxidase) and V (F0F1ATPase) as well as of citrate synthase were measured. For all enzymes the results differed at more than one order of magnitude. From eight labs we were able to compare the results with their control values for human skeletal muscle. Four labs found normal activity of cytochrome-c-oxidase whereas three labs found higher and one lab found lower activities compared to the own controls. Since all labs used different temperatures (25, 30 and 37 degrees C) in one lab the temperature dependencies were measured experimentally. The temperature correction did not much reduce the divergence of the results. It is concluded that differences in the lab protocols are the reason for the large variation of results. Since the experimental results strongly depend on the used method a strict standardization is necessary.
