STARDUST: A pipeline for the unbiased analysis of astrocyte regional calcium dynamics

Calcium imaging has become a popular way to probe astrocyte activity, but few techniques holistically capture discrete calcium signals occurring across the astrocyte domain. Here, we introduce STARDUST, a pipeline for the spatio-temporal analysis of regional dynamics and unbiased sorting of transients from fluorescence recordings of astrocytes. We describe steps for installing software, detecting active pixel patches, obtaining region of activity (ROA) maps, and extracting time series from ROAs. We then detail procedures for extracting signal features using custom-made code. [Display omitted] •STARDUST is a pipeline for the unbiased analysis of astrocyte calcium activity•Steps for capturing regional calcium activity in cellular micro-domains (ROAs)•Guidance on performing signal detection and features extraction in all ROAs•Instructions on accessing the spatial and temporal properties of ROAs Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Calcium imaging has become a popular way to probe astrocyte activity, but few techniques holistically capture discrete calcium signals occurring across the astrocyte domain. Here, we introduce STARDUST, a pipeline for the spatio-temporal analysis of regional dynamics and unbiased sorting of transients from fluorescence recordings of astrocytes. We describe steps for installing software, detecting active pixel patches, obtaining region of activity (ROA) maps, and extracting time series from ROAs. We then detail procedures for extracting signal features using custom-made code.