Marcel Mettlen, Sandra L. Schmid (2010) CIL:10105, Homo sapiens, epithelial cell, retinal pigmented epithelial cell. CIL. Dataset

Human retinal pigmented epithelial (RPE) cells labeled for clathrin-coated pits (green), focal adhesions (red) and nuclei (blue). RPE cells stabily expressing 'clathrin light chain a' tagged with EGFP (EGFP-LCa) were plated directly on glass coverslides. After 72 hrs, cells were fixed for 2 min in a 1:1 mixture of 2% paraformaldehyde/0.5% Triton X-100, followed by a 30 min fixation in 4% paraformaldehyde. Cells were then stained with mouse monoclonal antibodies to paxillin, detected with goat anti-mouse Alexa 568. Nuclei were stained with DAPI. Image was collected on an Olympus IX71 inverted microscope coupled to a Hamamatsu ORCA-ER digital camera using a 60x 1.42 NA Plan Apo N objective (Olympus). The exposure times were 200, 500 and 70 msec for the red, green and blue channel, respectively. A xenon-arc lamp (Lambda LS stand-alone illuminator; Sutter Instrument) was used with adequat filters (Chroma). Bar: 5µm

人视网膜色素上皮（RPE）细胞被标记为网格蛋白包被的凹陷（绿色）、焦点粘附（红色）和细胞核（蓝色）。表达‘网格蛋白轻链a’（EGFP-LCa）的RPE细胞稳定地被直接培养在玻璃载玻片上。经过72小时后，细胞在2%多聚甲醛与0.5%Triton X-100的1:1混合液中固定2分钟，随后在4%多聚甲醛中固定30分钟。接着，使用针对paxillin的小鼠单克隆抗体进行染色，并使用山羊抗小鼠Alexa 568进行检测。细胞核则用DAPI染色。图像是在Olympus IX71倒置显微镜上，通过连接Hamamatsu ORCA-ER数字相机使用60x 1.42 NA Plan Apo N物镜（Olympus）采集的。曝光时间分别为红色通道200毫秒，绿色通道500毫秒和蓝色通道70毫秒。使用氙弧灯（Lambda LS独立照明器；Sutter Instrument）以及适当的滤光片（Chroma）进行照明。标尺：5微米。