High-throughput, subpixel precision analysis of bacterial morphogenesis and intracellular spatio-temporal dynamics.
High-throughput, subpixel precision analysis of bacterial morphogenesis and intracellular spatio-temporal dynamics.
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细菌形态发生和细胞内时空动力学的高通量，子像素精确分析。
DOI：
10.1111/j.1365-2958.2011.07579.x
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发表时间：
2011-05
影响因子：
3.6
通讯作者：
Jacobs-Wagner C
中科院分区：
文献类型：
--
作者：
Sliusarenko O;Heinritz J;Emonet T;Jacobs-Wagner C
细菌表现出各种形状，并依赖于其细胞内成分的复杂空间组织来进行许多细胞过程。这个组织会随着内部和外部的提示而变化。对这些时空动力学的定量、无偏见的研究需要对大型显微数据集进行自动图像分析。因此，我们开发了MicrobeTracker，这是一种多功能和高通量的图像分析程序，即使在拥挤的图像和微型克隆中也能以亚像素精度勾勒和分割细胞，从而实现细胞谱系跟踪。MicrobeTracker带有一个集成的辅助工具SpotFinder，它可以精确跟踪细胞内荧光标记分子的焦点。利用MicrobeTracker，我们发现被广泛研究的大肠杆菌Min振荡器的动力学依赖于Min蛋白浓度，揭示了振荡器内稳健性的关键限制。我们还发现，随着细胞长度的增加，振荡的Mind蛋白质的比例增加，这表明当细胞达到适当的长度时，振荡器已经进化成最有效的。MicrobeTracker还被用来发现新月弯杆菌的形态发生和细胞周期调控的新方面。通过对丝状细胞的追踪，我们发现老极点的染色体起源是大多数复制/分离事件的原因，而其他染色体尽管细胞质相连，但在很大程度上保持沉默。这种令人惊讶的与位置相关的沉默是由分裂控制的。
Bacteria display various shapes and rely on complex spatial organization of their intracellular components for many cellular processes. This organization changes in response to internal and external cues. Quantitative, unbiased study of these spatio-temporal dynamics requires automated image analysis of large microscopy datasets. We have therefore developed MicrobeTracker, a versatile and high-throughput image analysis program that outlines and segments cells with subpixel precision, even in crowded images and mini-colonies, enabling cell lineage tracking. MicrobeTracker comes with an integrated accessory tool, SpotFinder, that precisely tracks foci of fluorescently labeled molecules inside cells. Using MicrobeTracker, we discover that the dynamics of the extensively-studied Escherichia coli Min oscillator depends on Min protein concentration, unveiling critical limitations in robustness within the oscillator. We also find that the fraction of MinD proteins oscillating increases with cell length, indicating that the oscillator has evolved to be most effective when cells attain an appropriate length. MicrobeTracker was also used to uncover novel aspects of morphogenesis and cell cycle regulation in Caulobacter crescentus. By tracking filamentous cells, we show that the chromosomal origin at the old pole is responsible for most replication/separation events while the others remain largely silent despite contiguous cytoplasm. This surprising position-dependent silencing is regulated by division.
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影响因子：
11.4
作者：
Kruse, T;Moller-Jensen, J;Gerdes, K
通讯作者：
Gerdes, K
DOI：
10.1073/pnas.96.19.10661
发表时间：
1999-09-14
影响因子：
11.1
作者：
Jensen, RB;Shapiro, L
通讯作者：
Shapiro, L
DOI：
10.1073/pnas.0911708107
发表时间：
2010-04-06
影响因子：
11.1
作者：
Fischer-Friedrich, Elisabeth;Meacci, Giovanni;Kruse, Karsten
通讯作者：
Kruse, Karsten
DOI：
10.1126/science.1188658
发表时间：
2010-06-04
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Christen M;Kulasekara HD;Christen B;Kulasekara BR;Hoffman LR;Miller SI
通讯作者：
Miller SI
影响因子：
19.5
作者：
KASS, M;WITKIN, A;TERZOPOULOS, D
通讯作者：
TERZOPOULOS, D