CIL:13468, Drosophila melanogaster, garland cell. In Cell Image Library

Transmission electron micrograph of Drosophila nephrocyte garland cell from ema[1] mutant. ema mutant garland cells are filled with very large electron-lucent alpha and electron-dense alpha and beta vacuoles, many of which are 5–10 microns in diameter, whereas the largest vacuoles in wild type are ∼2 microns in diameter. In wild type, both types of vacuoles usually contain a single aggregate of electron-dense granular material that appears to be attached to the limiting membrane. However, the large vacuolar compartments in the mutant contain numerous electron-dense granular structures, suggesting they arise from homo-/heterotypic fusions of alpha and beta vacuoles. Samples for TEM were fixed for 1 h at 4°C in 2% paraformaldehyde, 2% glutaraldehyde, and 1% tannic acid in 0.1 M cacodylic acid buffer, pH 7.2. Then, the samples were postfixed in 1% OsO4 in 0.1 M cacodylic acid buffer, pH 7.2, for 1 h at room temperature, stained en bloc with 1% uranyl acetate, dehydrated in a grade series of ethanol and propylene oxide, and embedded in epon resin (Electron Microscopy Sciences). Blocks were sectioned in an ultramicrotome (RMC Products) at ∼70-nm thickness with a Delaware Diamond knife and post-stained for 1 h in Reynolds lead citrate and uranyl acetate. Electron micrographs were taken on a transmission electron microscope (H-7500; Hitachi) using an AMT camera system. Mag: 4000x. Image corresponds to Figure 2B, right ema[1] panel in Kim et al. J Cell Biol. 188: 717-734. 2010. High magnification image taken from this micrograph is in CIL# 13469.本图像为ema[1]突变体果蝇肾细胞花环细胞的透射电子显微照片。ema突变体的花环细胞内充满了体积巨大的电子透亮α囊泡与电子致密α、β囊泡，其中多数囊泡直径为5~10微米；而野生型个体中最大的囊泡直径仅约2微米。在野生型样本中，两类囊泡通常均含有单个电子致密颗粒物质聚集体，且该聚集体似乎附着于囊泡的界膜之上。但突变体中的大型囊泡腔室却含有大量电子致密颗粒结构，提示这些大型囊泡源自α与β囊泡的同型/异型融合。透射电镜样本的固定流程如下：于4℃下在含2%多聚甲醛、2%戊二醛与1%单宁酸的0.1 M 二甲胂酸缓冲液（pH 7.2）中固定1小时。随后将样本置于室温，在含1%四氧化锇的0.1 M 二甲胂酸缓冲液（pH 7.2）中后固定1小时；再以1%醋酸铀进行整体染色，经梯度乙醇与环氧丙烷脱水后，使用环氧树脂（Electron Microscopy Sciences公司）进行包埋。使用超薄切片机（RMC Products公司）与Delaware钻石刀将包埋块切成约70 nm厚的切片，随后用雷诺柠檬酸铅与醋酸铀进行复染色1小时。采用AMT成像系统在透射电子显微镜（日立H-7500）下采集电子显微照片，放大倍数为4000×。本图像对应Kim等人2010年发表于《Journal of Cell Biology》第188卷第717-734页的论文中图2B的右侧ema[1]组图。从该显微照片截取的高倍放大图像收录于CIL#13469。