 Hi, I'm Judith Cantell and I'm the lead author on our recently published manuscript in biotechnology and bioengineering entitled Automated Detection of Wholesale Mitochondrial Motility and its dependence on cytoarchitectural integrity. You'll later hear from Phillip Chao, co-author on the study, and David Ekman, the corresponding author of the work. Previous research in this field has shown connections between mitochondrial motility and mitochondrial function, as well as overall cellular health. It is for this reason that mitochondrial motility has become a subject of increasing study over the last few years. However, the methods used by others tend to either focus on the whole cell and lose information about individual mitochondrial tracts, or look at only a few mitochondria within the cell and fail to capture mitochondrial motility on the whole cell level. Thus, our goal in this work is to automatically track mitochondria on the whole cell level, but in a way which retain information about each individual mitochondrial tract. Our experimental setup is as follows. The culture human dermal fibroblasts onto glass bottom dishes two days before experiments. The afternoon before experiments, we had cell-like mito GFP, manufactured by cell technologies, to transfect the cells, ultimately resulting in fluorescent mitochondria. Our experiments the following day involved imaging a transfected cell every three seconds for five minutes. In our paper, we look at control cells as well as cells treated with various pharmacological agents. Once data is collected and needs to be processed, we first use image training to pre-process images. Use a combination of convolution, finite-porear transforms, and then thresholding in order to ultimately obtain movies showing white mitochondria migrating against the black background. Next, we create a custom MATLAB script in order to track mitochondria in space and time. It uses built-in object recognition functions in MATLAB, which automatically detects the mitochondrial objects in the middle. The hard part is matching the labels from frame to frame. They accomplish this by spatial analysis, assuming that a white pixel in the same location over two frames belongs to the same mitochondria. We also give new labels to mitochondria resulting in fusion or vision. Our results show that the net distance is traveled by mitochondria in a given cell or a group of identically treated cells are logged normally distributed. Given this whole cell metric for measuring mitochondrial motility, we were interested in comparing normal and altered motility. In particular, we used nocodizol to depolymerize microtubules or cytokalazine D to depolymerize microfilaments. We found that microtubule depolymerization shifted the entire log-normal distribution to the left, indicating shorter net distances traveled. Microfilament depolymerization, on the other hand, shifted the distribution to the right, indicating larger net distances traveled. Overall, these results support the idea that mitochondrial motility is characterized by a continuous log-normal distribution throughout the cell rather than by discrete categories of diffusive or motor-driven motion. This log-normal distribution, in turn, is affected by interfering with the cytoskeletal integrity of the cell. Going forward, we are interested in further characterizing the role of actin in mitochondrial motility. We have also begun to examine mitochondrial motility in cells with altered mechanics and actin cytoskeletons, with some very interesting results. Overall, we will continue to do research to study the interactions between cellular metabolic function and mechanical function. Thanks very much for listening.