*4.8. Calcium Event Analysis*

Calcium event detection in astrocytes was performed by a previously developed algorithm [16]. Input data for the algorithm consist of two image series with the same dimensionality that are obtained from the same microscope: the astrocyte activity and images of a cell-free medium. The software pipeline includes the following steps:


Initially, the entire image plane is considered, and events are identified as spatiotemporally connected areas with significant activity. Next, the watershed method is applied to the long-exposure calcium intensity image to discriminate the areas corresponding to individual astrocytes. Finally, calcium events are segmented according to astrocyte regions. Further analysis utilizes the obtained individual astrocyte calcium events and filtered intensity signals.
