Margaret Clarke, Lucinda Maddera (2010) CIL:7654, Dictyostelium discoideum, cell by organism, eukaryotic cell, amoeboid cell, Eukaryotic Protist. CIL. Dataset

A movie of phagocytosis of bacteria by Dictyostelium discoideum amoebae. These Dictyostelium cells are expressing GFP-2FYVE, which binds to phosphatidylinositol 3-phosphate [PI(3)P], and mRFP-LimEdelta, which binds to actin filaments. The bacteria are E. coli that had been incubated with an inhibitor of cytokinesis, producing elongated forms. Phosphoinositides help to confer specific identities to compartments along the endocytic pathway. PI(3)P identifies early endosomes. It associates with phagosomes ~1 minute after uptake and persists as acidification occurs and digestion begins. Note that the long bacteria do not bind the green PI(3)P marker until after they have been fully internalized. The changing morphology of the long bacteria within the labeled phagosomes reveals the action of digestive enzymes. Three successive time series of 100 frames were collected in a single focal plane using a Zeiss LSM510 confocal microscope with a 63x DIC objective lens. The frames are separated by 3.94 seconds. There was a space of 6 seconds between the first two time series and a space of 9 seconds between the second and third time series. The scaling (pixel size) was 0.10 x 0.10 microns. The images were exported as TIFs using Zeiss LSM 5 Image Examiner and saved both as a TIF stack (available at CIL:7653) and as an AVI movie using NIH ImageJ. The movie spans 20 minutes; it plays at 7.5 fps, 30 times actual speed. For related experiments and further details, see Clarke et al., PLoS ONE 5, e8585 (2010).

一帧展示枯草杆菌被发网粘菌（Dictyostelium discoideum）阿米巴吞噬的动态电影。这些发网粘菌细胞表达绿色荧光蛋白GFP-2FYVE，该蛋白与磷脂酰肌醇3-磷酸[PI(3)P]结合，以及红色荧光蛋白mRFP-LimEdelta，该蛋白与肌动蛋白丝结合。所使用的细菌是大肠杆菌，经过细胞分裂抑制剂的处理，形成细长的形态。磷脂酰肌醇有助于在胞吞途径中的各个区室赋予特定的身份。PI(3)P识别早期内体。它在吞噬体形成后大约1分钟内与之结合，并在酸化发生和消化开始时持续存在。需要注意的是，长形细菌在完全内化后才与绿色PI(3)P标记结合。在标记的吞噬体内长形细菌形态的变化揭示了消化酶的作用。使用蔡司LSM510共聚焦显微镜和63倍DIC物镜，在同一焦平面上收集了三个连续的时间序列，每个序列包含100帧。帧与帧之间的间隔为3.94秒。前两个时间序列之间相隔6秒，第二个和第三个时间序列之间相隔9秒。缩放（像素大小）为0.10 x 0.10微米。图像使用蔡司LSM 5 Image Examiner导出为TIF格式，并保存为TIF堆栈（可在CIL:7653处获得）以及使用NIH ImageJ制作的AVI电影。该电影时长20分钟，播放速度为7.5帧每秒，实际速度的30倍。有关相关实验和更详细的信息，请参阅Clarke等人发表于PLoS ONE 5，e8585（2010）的文章。