通过使用四足点扩散函数的三维定位显微镜观察细胞中的活染色质动力学。
Observation of live chromatin dynamics in cells via 3D localization microscopy using Tetrapod point spread functions.
作者信息
Shechtman Yoav, Gustavsson Anna-Karin, Petrov Petar N, Dultz Elisa, Lee Maurice Y, Weis Karsten, Moerner W E
机构信息
Department of Chemistry, Stanford University, 375 North-South Mall, Stanford, California 94305, USA.
Currently with the Department of Biomedical Engineering, Technion-Israel Institute of Technology, Haifa, 32000 Israel.
出版信息
Biomed Opt Express. 2017 Nov 21;8(12):5735-5748. doi: 10.1364/BOE.8.005735. eCollection 2017 Dec 1.
Abstract
We report the observation of chromatin dynamics in living budding yeast () cells, in three-dimensions (3D). Using dual color localization microscopy and employing a Tetrapod point spread function, we analyze the spatio-temporal dynamics of two fluorescently labeled DNA loci surrounding the locus. From the measured trajectories, we obtain different dynamical characteristics in terms of inter-loci distance and temporal variance; when the locus is activated, the 3D inter-loci distance and temporal variance increase compared to the inactive state. These changes are visible in spite of the large thermally- and biologically-driven heterogeneity in the relative motion of the two loci. Our observations are consistent with current euchromatin vs. heterochromatin models.
摘要
我们报告了在活的出芽酵母（）细胞中对染色质动力学进行的三维（3D）观察。使用双色定位显微镜并采用四脚架点扩散函数，我们分析了围绕基因座的两个荧光标记DNA位点的时空动力学。从测量的轨迹中，我们在基因座间距离和时间方差方面获得了不同的动力学特征；当基因座被激活时，与非激活状态相比，三维基因座间距离和时间方差增加。尽管两个位点的相对运动存在由热和生物驱动的巨大异质性，但这些变化仍然可见。我们的观察结果与当前的常染色质与异染色质模型一致。
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