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Evaluation of mitochondrial activity in living cells using fluorescence methods
Fedorov, Vasilii ; Mézl, Martin (referee) ; Čmiel, Vratislav (advisor)
Monitoring mitochondrial dynamics can help in the therapeutic treatment of serious diseases and slowing down the aging process. This work was parted into a theoretical part and a practical part. In the theoretical part, research was carried out in the field of mitochondria and methods of activity evaluation. In the practical part in the Python environment, an automatic algorithm for evaluating mitochondrial activity based on the analysis of morphological signs was implemented. The algorithm included image preprocessing, localization of mitochondria, segmentation, and calculation of morphological features such as number of mitochondria, total area, average area, circularity index, eccentricity index. We were able to get and perform an analysis on 16 microscopic images and confirm the success of an algorithm that could be used for drug discovery and testing purposes.
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Mitochondria morphology and membrane potential under stress conditions
Ezati, Masoumeh ; Zumberg, Inna ; Cmiel, Vratislav
Mitochondria, organelles found in the cytoplasm of almost all eukaryotic cells, generate large quantities of energy in the form of adenosine triphosphate (ATP). In addition to producing energy, mitochondria store calcium for cell signaling activities and play an important role in maintenance of ionic balance. Mitochondria, however, are highly sensitive to any kind of stress in which they mainly response by disturbance of respiration, reactive oxygen species (ROS) production and release of cytochrome c into the cytoplasm. Osmotic stress, in particular, generates ROS that degrade lipids, proteins, and DNA. High levels of salt concentration can cause an imbalance in cellular ion homeostasis that results in ion toxicity and osmotic stress. This study aims to investigate possible effects of KCl and NaCl on Bone marrow-derived (MSCs) mitochondria membrane potential (MMP) and morphology. The results indicated that KCl and NaCl of salt concentration can cause an imbalance in cellular ion homeostasis that results in ion toxicity and osmotic stress. This study aims to investigate possible effects of KCl and NaCl on MSCs mitochondria membrane potential (MMP) and morphology. The results indicated that KCl and NaCl decreased the potential and changed the morphology of mitochondria membrane compared to cells growing in normal condition.
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