Additional file 1.


Ca2+ signalling evoked by ATP in the acute NAc slice from the rat brain. Movie of time measurement showing ATP (100 μM) application onto a Fluo-4 AM loaded acute NAc slice (300 μm). ATP was applied for 60 s (long puff) through a glass micropipette right above the tissue surface. Image acquisition frequency was 2 s in depth of 25 μm from the slice surface. Olympus FluoView300 software collected .tiff file stacks were coloured and converted to .avi file by ImageJ 1.44 (32-bit) image processing and analysis software [80]. ATP-responsive cells were counted by using an ImageJ macro developed to average stacks before the time of ATP application and to subtract this average from every stack of the 10 minute-long recording. In this way, we got a 10-minute-long movie showing only the cells that had a fluorescence intensity change.

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Molnár et al. BMC Neuroscience 2011 12:96   doi:10.1186/1471-2202-12-96