Development of New Objective Lens with Double Cylinders

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We have developed a florescence microscope by using objective lens with the double cylinders. Since this objective enabled us to detect the time course of fluorescence at different wavelengths simultaneously, we performed the following two experiments, 1) In adrenocortical cells and C6 glioma cells, we observed calcium signaling and mitochondrial membrane potentials. When the intracellular calcium concentration shows a local and rapid increase near mitochondria, the mitochondria show the transient and sudden depolarization, leading to the release of intramitochondrial calcium. When the calcium concentration increases gradually near the mitochondria, however, mitochondria show the uptake of calcium ion and decrease the calcium concentration at cytosol. 2) The changes in mitochondrial membrane potential were detected during the mitosis. The stage of mitosis was identified with Hoechst 33342. The mitochondrial membrane potential decreased during mitosis. On the other hand, the membrane potential recovered after the mitosis. This results might suggest that the intracellular ATP concentration could decrease during mitosis and that it could increase after the mitosis.