High-speed single molecule imaging of membrane proteins in rat basophilic leukaemia cells.

A high-speed fluorescence microscope operating at a 490 Hz frame rate was used to image two different membrane proteins- the high-affinity IgE receptor FcεRI, a transmembrane protein, and an outer-leaflet GPI-anchored protein. The IgE receptor was imaged via IgE labeled with Janelia Fluor 646 and the GPI-anchored protein was imaged using a GPI-GFP fusion protein and an Atto647N labeled anti-GFP nanobody. Data was collected for both proteins in untreated cells and cells that had actin stabilized by phalloidin.

本数据集采用帧率为490赫兹的高速荧光显微镜，对两种不同的膜蛋白开展成像实验：其一为高亲和力免疫球蛋白E（IgE）受体FcεRI，属于跨膜蛋白；其二为细胞膜外层脂单层的糖基磷脂酰肌醇（GPI）锚定蛋白。其中，FcεRI的成像通过经Janelia Fluor 646标记的IgE完成；GPI锚定蛋白则借助GPI-GFP融合蛋白与经Atto647N标记的抗GFP纳米抗体实现成像。实验分别采集了未处理细胞，以及经鬼笔环肽稳定肌动蛋白的细胞中两种靶蛋白的成像数据。