Department of Biomedical Sciences, University of Foggia, Foggia, Italy.
Mitochondrion. 2011 Sep;11(5):764-73. doi: 10.1016/j.mito.2011.06.014. Epub 2011 Jul 6.
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.
功能生化测试是诊断与线粒体相关疾病的金标准。然而，由于患者组织的生物样本可用性有限，可筛选的酶活性数量也受到严重限制。在这项研究中，我们开发了一种荧光探针辅助显微镜检测方案，能够在单个细胞器分辨率水平上评估固定在玻璃表面上的线粒体产生 ΔΨ(m)的能力。与其他方法相比，该方法的优势在于，它可以减少所需的生物样本量，在短时间内测试氧化磷酸化系统所有成分的功能活性，而不会降低准确性。此外，还可以评估给定酶活性在整个线粒体群体中的分布情况，从而测量呼吸链功能障碍的功能异质性水平。
