Use of computer-assisted motion analysis for quantitative measurements of swimming behavior in peritrichously flagellated bacteria
Use of computer-assisted motion analysis for quantitative measurements of swimming behavior in peritrichously flagellated bacteria
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DOI：
10.1006/abio.1996.0086
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发表时间：
1996-03-01
影响因子：
2.9
通讯作者：
Amsler, CD
中科院分区：
文献类型：
--
作者：
Amsler, CD
开发了一种鉴定单个细菌翻滚的测定法，从而允许快速定量测量自由游动细菌中的翻滚频率。翻滚频率由细胞调节以实现趋化性。趋化性信号转导途径中的突变通常具有改变的翻滚频率的表型。本试验旨在定量测量稳态翻滚频率，以比较突变菌株表型。它是使用大肠杆菌开发的，但应适用于其他具有围毛鞭毛模式的物种，如鼠伤寒沙门氏菌。翻滚由方向变化率和游泳速度的参数的组合来定义，其中方向的快速变化定义翻滚的开始，并且增加的游泳速度定义翻滚的结束。这些参数以前已经被证明是相关的翻滚一般，但不用于识别离散翻滚事件。通过将其结果与人工肉眼观察结果进行比较，验证了计算机分析。该试验旨在对与野生型相似的游泳模式最敏感，以解决部分功能突变引起的细微变化。它还可以定量检测极端行为表型，并且可以在必要时进行修改以增加任一极端的分辨率。(C)出版社：Academic Press，Inc.
An assay was developed which identifies individual bacterial tumbles and so allows rapid, quantitative measurements of tumble frequency in free-swimming bacteria. Tumble frequency is modulated by cells to enable chemotaxis. Mutations in the chemotaxis signal transduction pathway typically have phenotypes of altered tumble frequency. The purpose of this assay is to quantitatively measure steady-state tumble frequency to enable comparisons of mutant strain phenotypes. It was developed using Escherichia coli but should be applicable to other species with a peritrichous flagellation pattern, such as Salmonella typhimurium. Tumbles are defined by a combination of the parameters rate of change of direction and swimming speed, with a rapid change of direction defining the beginning of a tumble and increased swimming speed defining the end. These parameters have previously been shown to be correlated with tumbles in general but not used to identify discrete tumble events. The computer assay was validated by comparing its results with manual observations by eye. The assay was intended to be most sensitive to swimming patterns similar to wild type so as to resolve subtle changes which would result from partial-function mutations. It quantitatively detects extreme behavioral phenotypes as well and can be modified to increase resolution at either extreme if necessary. (C) 1996 Academic Press, Inc.