Astrocytes are a major type of glial cell in the mammalian brain, essentially regulating neuronal development and function.Quantitative imaging represents an important approach to study astrocytic signaling in neural circuits.Focusing on astrocytic Ca2+ activity, a key pathway implicated in astrocye-neuron interaction, we here report a strategy combining fast light sheet fluorescence microscopy (LSFM) and correlative screening-based time series analysis, to map activity domains in astrocytes in living mammalian nerve tissue.Light sheet of micron-scale thickness destiny 2 beyond light steam key enables wide-field optical sectioning to image astrocytes in acute mouse brain slices.Using both chemical and genetically encoded Ca2+ indicators, we demonstrate the complementary advantages of LSFM in mapping Ca2+ domains in astrocyte populations as compared to epifluorescence and two-photon microscopy.
Our approach then revealed distinct kinetics of Ca2+ signals between cortical and hypothalamic astrocytes in resting conditions and following the activation of adrenergic G protein coupled receptor (GPCR).This observation highlights the activity heterogeneity across regionally distinct astrocyte populations, and indicates the potential of fr5945 our method for investigating dynamic signals in astrocytes.