Fig. 5

SAsLFM facilitates volumetric observation and 3D tracking under various imaging conditions. a An add-on container filled with water was mounted on the head of a craniotomy mouse to increase the refractive index mismatch during imaging. b Orthogonal MIPs of the standard deviation across all 1009 volumes for neurons in the awake mouse brain. A CNMF framework was carried out on the MIPs of the volumetric data to extract the locations of segmented neurons. The neurons were clearly resolved even at the depth of ~ 300 μm (Supplementary Movie 1). c Temporal traces of 14 selected neurons marked in b. d Reconstructed slices of a jellyfish ephyra at different depths show that SAsLFM provides stable performance across an axial range of ~ 300 μm. Cross-section profiles along the white lines were plotted as insets. e MIPs of a freely-behaving jellyfish ephyra at various time stamps record the activities of the tentacles (Supplementary Movie 2). f SAsLFM achieves 3D tracking of the 8 folded tentacles in a large volume covering ~ 2000 × 2000 × 500 μm³. The dots represent the spatial locations of each tentacle at different time stamps. g Time series of the cell dynamics of the Drosophila embryo during the late developmental stages. h MIPs of two volumes separated by a time interval of 255 s. i Semi-automated global cell tracking in the MIPs of the entire Drosophila embryo. The color wheel for motion coding is shown in the bottom left. Scale bars: 300 μm (b), 500 μm (d) and 100 μm (h)