Quantitative motion analysis of subchromosomal foci in living cells using four-dimensional microscopy
Quantitative motion analysis of subchromosomal foci in living cells using four-dimensional microscopy
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DOI：
10.1016/s0006-3495(99)77119-5
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发表时间：
1999-11-01
影响因子：
3.4
通讯作者：
Cremer, C
中科院分区：
文献类型：
--
作者：
Bornfleth, H;Edelmann, P;Cremer, C
在四维时空图像中，研究了活体人细胞核内亚染色体焦点和整个染色体区域的运动。在显微注射后，通过将Cy3-dUTP掺入两种不同细胞类型的核DNA中，实现了亚染色体LOOK的可视化，随后标记的细胞核的分离导致在非标记染色质的背景上仅存在少数标记的染色体区域(Zink等人，1998)。哼。吉内。102.241-251年)。这一过程在给定的细胞核中产生了许多不同的信号。使用单粒子跟踪和统计方法对四维时空图像中的运动进行分析，以检测焦点相对于染色体区域的质心可能的定向运动。分析的准确性是使用模拟数据集进行测试的，这些数据集紧密地反映了实验设置，并使用了已知尺寸的微粒。分析工具对实验数据的应用表明，位于不同染色体上的病灶之间的相互扩散样运动比区域内更明显。在观察到的时间范围内，单个病灶的运动可以最好地用随机扩散过程来描述。对染色体区域内多个焦点的联合定向运动的统计检验表明，在区域内，焦点偶尔会从随机运动切换到定向运动。
The motion of subchromosomal foci and of whole chromosome territories in live human cell nuclei was investigated in four-dimensional space-time images. Visualization of subchromosomal fool was achieved by incorporating Cy3-dUTP into the nuclear DNA of two different cell types after microinjection, A subsequent segregation of the labeled cell nuclei led to the presence of only a few labeled chromosome territories on a background of nonlabeled chromatin (Zink et al., 1998. Hum. Genet. 102.241-251). This procedure yielded many distinct signals in a given cell nucleus. Motion analysis in four-dimensional space-time images was performed using single-particle tracking and a statistical approach to the detection of a possible directional motion of foci relative to the center of mass of a chromosome territory. The accuracy of the analysis was tested using simulated data sets that closely mirrored the experimental setup and using microparticles of known size. Application of the analysis tools to experimental data showed that mutual diffusion-like movements between foci located on different chromosomes were more pronounced than inside the territories. In the time range observed, movements of individual foci could best be described by a random diffusion process. The statistical test for joint directed motion of several foci inside chromosome territories revealed that foci occasionally switched from random to directional motion inside the territories.