Fig. 2.

Expanded, antibody-stained mouse brain slice imaged by LSFM. The sample was expanded and imaged with a custom-built light-sheet microscope. In total 70 $z$-stacks with a step size of $1\mu \mathrm{m}$, covering a depth of $275\mu \mathrm{m}$ with 30% overlap were stitched to generate the image. (a) Single plane of the stitched volume at a depth of $65\mu \mathrm{m}$. The total volume shown after expansion was $3600\times 1240\times 275\mu {\mathrm{m}}^3$ (Video 1, MP4, 32 MB [URL: https://doi.org/10.1117/1.NPh.6.1.015005.1]). (b) Magnification of the region marked in (a), lateral field size $254\times 492\mu {\mathrm{m}}^2$. (c) Maximum projection of the selected region marked in (a) comprising 76 slices of the stack, lateral field size $254\times 492\mu {\mathrm{m}}^2$. Single granule cells and dendrites can well be distinguished and separated. The stack was median-filtered to remove staining artifacts before the maximum projection. (d) Segmentation and tracing of the neurites of three selected granule cells (Video 2, MP4, 19 MB [URL: https://doi.org/10.1117/1.NPh.6.1.015005.2]). Left panel: segmented GCs in the neuronal network, right panel: segmented GCs with traced neurites.