Fig. 3: Practical use of descSPIM in neuroscience.

a An example of propidium iodide (PI)-stained whole Thy1-YFP-H mouse brain imaging with full FOV (FF) mode and flat-field correction (FFC) adoption. PI and YFP were excited with 561 nm and 488 nm laser beams, respectively. Four image-stack tiles were collected from a single direction (angle 0°). Using the θ stage, a second set of four image tiles from the opposite direction (angle 180°) were then obtained. All the image tiles were stitched, aligned, and fused with BigStither and ANTs software. See Supplementary Fig. 13 for the procedure. Voxel size: 3.45 × 3.45 × 10 µm³. b Magnified images of YFP-expressing neurons within the dataset. Signal elongation along the axial direction was more modest in somas (10–20 µm in diameter) than in neurites (approximately 1 µm in diameter), consistent with the light sheet thickness (approximately 25 µm). Quantitative data is shown in Supplementary Fig. 14. The original grayscale 8-bit maps were pseudo-colored with a Blue, Yellow or Green Fire Blue look-up table. The imaging experiment was performed twice with nearly identical results.