Negative Stain Transmission Electron Micrographs of Haliangium ochraceum Encapsulin:Encapsulated Ferritin complex

Purified encapsulin nanocompartments were initially imaged by negative stain TEM. Continuous carbon/formvar coated copper grids (200 mesh) were glow-discharged for 30 seconds using a Pelco glow discharge system. 5 µL Enc was pipetted onto the glow-discharged grids and excess liquid was removed after 30 seconds with Whatman filter paper (grade 1, diameter 24.0 cm). The grids were washed with distilled water three times, followed by staining with 2 % uranyl acetate for 5 seconds. Grids were left to air dry and then imaged with a JEOL JEM-1400 transmission electron microscope. Images were collected with a Gatan CCD OneView camera and analyzed using FIJI.

纯化的包壳蛋白纳米区室（encapsulin nanocompartments）首先通过负染透射电子显微镜（negative stain TEM）进行成像。实验使用200目连续碳/福马尔瓦（formvar）包被铜网，借助Pelco辉光放电仪对铜网开展30秒的辉光放电处理。取5微升包壳蛋白（Enc）溶液滴加至经辉光放电处理的铜网上，30秒后采用Whatman 1号滤纸（直径24.0 cm）吸除多余液体。随后用蒸馏水对铜网洗涤三次，再以2%乙酸铀酰溶液染色5秒。将铜网自然晾干后，使用JEOL JEM-1400型透射电子显微镜完成成像，图像通过Gatan CCD OneView相机采集，并借助FIJI软件进行分析。