biostudies-literatureKiss DSKK-UKNKBKK-PhDOTKA417-29https://www.ebi.ac.uk/biostudies/studies/S-EPMC4887559biostudies-literatureUnknown3In order to measure the activity of neuronal mitochondria, a representative proof of neuronal processes, physiologically relevant mitochondrial samples need to be gained as simply as possible. Existing methods are, however, either for tissue samples of large size and/or homogenous microstructures only, or are not tested for mitochondrial function measurements. In the present article we describe a gradient fractionation method to isolate viable and well-coupled mitochondria from relatively heterogeneous histological microstructures such as the hypothalamus. With this new method, we are able to isolate a sufficient amount of functional mitochondria for determination of respiratory activity, in a short period of time, using affordable equipment. •Verified by electron microscopy, our method seMethodsXPreparation of purified perikaryal and synaptosomal mitochondrial fractions from relatively small hypothalamic brain samples.PMC488755915263104982120211593072186115613Toth ISterczer AKiss DSJocsak GBartha TZsarnovszky AFrenyo LVfalsePreparation of purified perikaryal and synaptosomal mitochondrial fractions from relatively small hypothalamic brain samples.In order to measure the activity of neuronal mitochondria, a representative proof of neuronal processes, physiologically relevant mitochondrial samples need to be gained as simply as possible. Existing methods are, however, either for tissue samples of large size and/or homogenous microstructures only, or are not tested for mitochondrial function measurements. In the present article we describe a gradient fractionation method to isolate viable and well-coupled mitochondria from relatively heterogeneous histological microstructures such as the hypothalamus. With this new method, we are able to isolate a sufficient amount of functional mitochondria for determination of respiratory activity, in a short period of time, using affordable equipment. •Verified by electron microscopy, our method se2016-01-01T00:00:00Z20162025-06-01T01:57:22.535Z2025-06-01T01:57:22.535ZS-EPMC48875592728453310.1016/j.mex.2016.05.004