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Imaging in acute slices

WS Weida Shen
LN Ljiljana Nikolic
CM Claire Meunier
FP Frank Pfrieger
EA Etienne Audinat
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Astrocytes were loaded with Calcium indicator Rhod-2 AM (9 µM) at room temperature (24 °C) for 1 h with 0.02% Pluronic F-127 (In vitrogen) and 0.6% DMSO (Sigma) in aCSF. Slices were then incubated at least 30 mins in aCSF to wash out the excess of dye before transferring to the imaging chamber. During experiments, slices were perfused with aCSF containing TTX (0.5 µM) and GBZ (10 µM). Fluorescence images were acquired with a custom-built two-photon laser scanning microscope. Details on imaging acquisition and analysis of Ca2+ signals are provided in Supplementary Information.

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