A. N. Belozersky Research Institute of Physico-Chemical Biology MSU
Structural organization of mitochondria reflects their functional status and largely is an index of the cell viability. The indirect parameter to assess the functional state of mitochondria and cells is the degree of fragmentation, i. e. a ratio of long or branched mitochondrial structures to rounded mitochondria. The critical need for such evaluations requires the creation of an approach, that allows on the basis of confocal images of mitochondria stained with a fluorescent probe, to create an integral picture of the three-dimensional organization of mitochondria. In the present study, we tested three approaches to analyze the structural architecture of mitochondria under norm and fission induced by oxidative stress. We have revealed that while the most informative way of analysis is a three-dimensional reconstruction based on series of confocal images taken in Z-dimension, however, with some limitations it is plausible to use more simple algorithms of analysis, including that one that uses unitary two-dimensional images. Further improvement of these methods of image analysis will allow more comprehensive analysis of mitochondrial architecture under norm and different pathological states. It may also provide quantification of a number of mitochondrial parameters determining morpho-functional state of mitochondria primarily their absolute and relative volumes and give additional information on three-dimensional organization of mitochondriome.