Journal: eLife

Article Title: Expansion-assisted selective plane illumination microscopy for nanoscale imaging of centimeter-scale tissues

doi: 10.7554/elife.91979

Figure Lengend Snippet: Figure 5. Expansion and imaging of a large volume of macaque brain. A 1 cm×1 cm×1.5 cm block of macaque primary motor cortex was expanded (3×) and imaged on the ExA-SPIM (Videos 7–9). Corticospinal neurons were transduced by injecting tdTomato-expressing retro-AAV into the spinal cord. (a–b) Maximum intensity projections of the imaged volume pseudo-colored by depth. The axes descriptors in (a) indicate the 5×3 tiling used to image this volume. (c–f) Fine axonal and dendritic structures including descending axons, collaterals, and dendritic spines are clearly discernible in the images throughout the entire volume. See Video 10. Images are displayed as maximum intensity projections with the following thicknesses: (a) 23 mm, (b) 45 mm, (c–f) 1 mm. Inset scale bars correspond to 10 µm post tissue expansion unless otherwise specified.

Article Snippet: Spinal injection procedure and motor cortex collection To retrogradely label corticospinal neurons in the hand- wrist and trunk regions of primary motor cortex in a pigtail macaque, we injected a retro AAV vector (rAAV2- CAG- tdTomato; Addgene plasmid #59462 packaged in- house, titer of 1.88×1013) into the left lateral funiculus and ventrolateral part of the gray matter in the C6/C7 spinal segments.

Techniques: Imaging, Blocking Assay, Expressing