Additional file 4 of 3D printed spermathecae as experimental models to understand sperm dynamics in leaf beetles

Additional file 4 Supplementary information 4, figures.jpeg. The fluid dynamics tests (first pumping movement, outflow) were performed by the experimenter’s fingers. Supplementary information 4, Fig. 1. Transparent flexible models and fluid dynamics tests (first pumping movement, outflow) using the Agelastica alni spermatheca models with a realistic internal protuberance (left column, highlighted with red dashed lines) and without the protuberance (right column). (a-a’) The models fully immersed in 1,2-propanediol. (b-b’) Sudan Red in 1,2-propanediol was injected. (c-e’) The first pumping cycle filmed at 50 fps, and still images at different conditions are exhibited. The solid white arrows show the direction of the distal C-arm movement. The black arrows indicate a marked shift of the injected ink spots in the models. The times listed in each image are the time after pumping started. (f-f’) Schemes of the observed outflow in the first pumping cycle. Supplementary information 4, Fig. 1 (cont.). Transparent flexible models and fluid dynamics tests (first pumping, inflow) using the Agelastica alni spermatheca models with a realistic internal protuberance (left column, highlighted with red dashed lines) and without the protuberance (right column). (g-h’) The first pumping cycle filmed at 50 fps, with still images shown under different conditions. The solid white arrows show directions of the distal C-arm movement. The black arrows show a marked shift of the fluid in the models. (i-i’) The inflow at the second pumping cycle. (j-j’) Schemes of the observed inflow during the first two pumping cycles. The light-pink circles in the schemes indicate the original locations of the injected ink. Supplementary information 4, Fig. 2. Fluid dynamics in subsequent pumping cycles. (a-a’) Sudan Red in 1,2-propanediol was injected. (b-b’) After the C-arms were pumped ten times. (c-c’) After the C-arms were pumped 20 times, but in the protuberance model at the 11th pumping cycle, Sudan Red partly leaked (Supplementary Information 5, Movie 2) from a lateral crack. However, similar results were observed before the lateral crack occurred. (d-d’) After the C-arms were pumped 30 times. The visible unmixed ink drops are marked with solid black lines.