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.附加文件4：补充信息4，内含figures.jpeg文件。流体动力学测试（首次泵动过程、流出阶段）由实验人员通过手指完成操作。补充信息4中的图1：带有真实内部凸起结构的榆蓝叶甲（Agelastica alni）受精囊模型（左列，红色虚线标注）与无凸起结构的对应模型（右列），及其相关的透明柔性模型与流体动力学测试（首次泵动过程、流出阶段）。(a-a’) 模型完全浸没于1,2-丙二醇中。(b-b’) 向体系中注入添加有苏丹红（Sudan Red）的1,2-丙二醇溶液。(c-e’) 以50帧每秒（fps）的帧率录制首次泵动循环的过程，展示不同条件下的静态图像。白色实心箭头指示远端C形臂的运动方向，黑色箭头标注模型内注入的墨斑的显著位移，每张图片标注的时间为泵动开始后的时长。(f-f’) 首次泵动循环中观测到的流出过程示意图。补充信息4中的图1（续）：带有真实内部凸起结构的榆蓝叶甲（Agelastica alni）受精囊模型（左列，红色虚线标注）与无凸起结构的对应模型（右列），及其相关的透明柔性模型与流体动力学测试（首次泵动过程、流入阶段）。(g-h’) 以50帧每秒（fps）的帧率录制首次泵动循环的过程，展示不同条件下的静态图像。白色实心箭头指示远端C形臂的运动方向，黑色箭头标注模型内流体的显著位移。(i-i’) 第二次泵动循环的流入过程。(j-j’) 前两次泵动循环中观测到的流入过程示意图，图中的浅粉色圆圈标注注入墨斑的初始位置。补充信息4中的图2：后续泵动循环中的流体动力学过程。(a-a’) 向体系中注入添加有苏丹红（Sudan Red）的1,2-丙二醇溶液。(b-b’) C形臂完成10次泵动后。(c-c’) C形臂完成20次泵动后，但凸起结构模型在第11次泵动循环时，苏丹红从模型侧面裂缝中部分泄漏（补充信息5，影片2）。不过在裂缝出现前已观测到一致的实验结果。(d-d’) C形臂完成30次泵动后，可见的未混合墨滴以黑色实心线条标注。