“…However, fast 2D and 3D imaging of large spine assemblies and spiny dendritic segments in awake, running, and behaving animals has remained an important challenge because locomotion can more than double firing rate in most neurons (Niell and Stryker, 2010, Fu et al., 2014). Moreover, the majority of the top-down and bottom-up input integration occurs in complex, distant apical and basal dendritic segments separated by distances of several hundred micrometers in 3D (Schiller et al., 2000, Magee and Johnston, 2005, Larkum et al., 2009, Smith et al., 2013, Phillips, 2015). The maximal, over 1,000 μm z-scanning range of AO microscopy (Katona et al., 2012), which was limited during in vivo measurements with GECIs to about 650 μm by the maximal power of the currently available lasers, already permitted the simultaneous measurement of these apical and basal dendritic segments of layer II/III neurons and dendritic segments of layer V neurons in a range of over 500 μm during running periods, or in different behavioral experiments (Harvey et al., 2012, Hangya et al., 2015) where motion-induced fluorescence transients had similar amplitude and kinetics as behavior-related Ca 2+ transients without motion compensation.…”