Amira software version 6

Two samples (one male and one female) of D. mellyi were used for tomography. The samples were decapitated, and the heads were dehydrated in a series of graded ethanol 75%, 80%, 85%, 90%, 95%, and three times in 100% (30 min in each concentration). The samples were dried in a critical point dryer (Leica EM CPD300), then mounted on an Eppendorf tube and scanned using the Xradia scanner (Zeiss MicroXCT-400, IOZCAS, Beijing, China) at a magnification of 4× and image capture at an interval of 10 s for 5 h.
The dataset is stored in the Institute of Zoology, Beijing, China, and is available for access through the corresponding author.
Amira software version 6.0.1 (Thermo Fisher Scientific, Waltham, MA, USA) was used in the segmentation of different structures of the compound eyes from the image stacks obtained through scanning. The segmented materials were imported to VG Studio Max 3.1 (Volume Graphics, Heidelberg, Germany) for rendering and visualization.
The volume rendering of different structures that make up the compound eyes were performed through Amira software version 6.0.1, and the final images were assembled through Adobe Photoshop version 21.2.1 (Adobe Inc., San Jose, CA, USA).

Images were deconvolved using AutoQuant X Software (Media Cybernetics). Deconvolved images were then analysed using Amira Software version 6.0.1 (Thermo Scientific). Z-stacks were uploaded to create a 3D volume rendering of the crush site and labelled neurites. For samples containing more than one Z-stack, image stacks were aligned and merged using Amira’s merge module. The images were then rotated to position the crush site proximal to the retina on the left with regenerated neurites growing away from the crush site towards the right. The transformed data were resampled and axes swapped such that the XY plane became the cross-section along the length of the nerve. Images were cropped if needed to exclude any partial cross sections of the ends of the nerve caused by image rotation. Labelled regenerated neurites were segmented by thresholding pixels using Amira’s Segmentation Editor. The volume ratio of segmented neurites over total nerve volume was measured for each sample. Study results were then normalized and plotted as a bar graph. The ratio of segmented area over total nerve area for each cross-section was normalized and plotted as area ratio per slice. Area ratio plots were created starting from the behind the crush site proximal to the retina and following along the length of the nerve for the segmented regenerated neurites.