Clarke, Margaret, Silverman, Philip (2010) CIL:7324, Escherichia coli, prokaryotic cell. CIL. Dataset

This is one of four related images of F-pili (conjugative pili encoded by the F plasmid of E. coli) dynamics by live-cell imaging. There are 2 movie (.avi) files and 2 files (.tiff) from the movies. The movies show that F-pili normally undergo cycles of extension and retraction in the absence of any obvious triggering event such as contact with a recipient cell.

Retraction of an F-pilus. E. coli strain HfrH expressing the red fluorescent marker DsRed Express were mixed with R17 bacteriophage labeled with Alexa 488 (green); R17 bacteriophage (NCBITaxon:12026) bind to the sides of F-pili.

Samples were imaged by using a Zeiss LSM 510 confocal microscope with a Plan-Apochromat 63 × 1.4 N.A. DIC objective. Alexa-488 was excited with the 488-nm line of an argon laser with a 505–530-nm filter for emission, and DsRed was excited with the 543-nm line of a HeNe laser, with a 560-nm long pass filter for emission. An HFT UV/488/543 beam splitter was used. Images were collected in a single focal plane at 2 s intervals and accompanying movies (.avi) are playing at 10fps.

本数据集收录了与大肠杆菌F质粒编码的性菌毛（F-pili）动态变化相关的四幅图像之一。其中包含两个电影文件（.avi格式）以及由电影生成的两个文件（.tiff格式）。电影展示了在没有任何明显触发事件（如与受体细胞的接触）的情况下，F-pili通常会经历伸长和收缩的循环过程。F-pili的收缩过程如下：大肠杆菌菌株HfrH表达红色荧光标记DsRed Express与标记为Alexa 488（绿色）的R17噬菌体混合；R17噬菌体（NCBITaxon:12026）绑定在F-pili的侧面。样本使用蔡司LSM 510共聚焦显微镜以及63倍1.4数值孔径的Plan-Apochromat微分干涉（DIC）物镜进行成像。Alexa-488荧光素激发使用氩激光的488nm波段，并使用505-530nm滤光片进行发射；DsRed激发使用氦氖激光的543nm波段，并使用560nm的长通滤光片进行发射。采用了HFT UV/488/543光束分离器。图像以2秒间隔在单一焦平面进行采集，并伴随的影片以每秒10帧的速率播放。