Reshes Galina, Vanounou Sharon, Fishov Itzhak, Feingold Mario
Department of Physics, Ben Gurion University, Beer Sheva, Israel.
Biophys J. 2008 Jan 1;94(1):251-64. doi: 10.1529/biophysj.107.104398. Epub 2007 Aug 31.
Bacteria are the simplest living organisms. In particular, Escherichia coli has been extensively studied and it has become one of the standard model systems in microbiology. However, optical microscopy studies of single E. coli have been limited by its small size, approximately 1 x 3 microm, not much larger than the optical resolution, approximately 0.25 microm. As a result, not enough quantitative dynamical information on the life cycle of single E. coli is presently available. We suggest that, by careful analysis of images from phase contrast and fluorescence time-lapse microscopy, this limitation can be bypassed. For example, we show that applying this approach to monitoring morphogenesis in individual E. coli leads to a simple, quantitative description of this process. First, we find the time when the formation of the septum starts, tau(c). It occurs much earlier than the time when the constriction can be directly observed by phase contrast. Second, we find that the growth law of single cells is more likely bilinear/trilinear than exponential. This is further supported by the relations that hold between the corresponding growth rates. These methods could be further extended to study the dynamics of cell components, e.g., the nucleoid and the Z-ring.
细菌是最简单的生物。特别是,大肠杆菌已得到广泛研究,并成为微生物学中的标准模型系统之一。然而,对单个大肠杆菌的光学显微镜研究受到其小尺寸的限制,其大小约为1×3微米,仅略大于光学分辨率(约0.25微米)。因此,目前尚无足够关于单个大肠杆菌生命周期的定量动态信息。我们认为,通过仔细分析相差显微镜和荧光延时显微镜的图像,可以绕过这一限制。例如,我们表明将此方法应用于监测单个大肠杆菌中的形态发生可得出对该过程的简单定量描述。首先,我们找到隔膜开始形成的时间,即τ(c)。它比通过相差显微镜可直接观察到缢缩的时间要早得多。其次,我们发现单细胞的生长规律更可能是双线性/三线性而非指数型。相应生长速率之间的关系进一步支持了这一点。这些方法可进一步扩展用于研究细胞成分的动态变化,例如类核和Z环。