In Situ Mechanical Characterization of the Cell Nucleus by Atomic Force Microscopy
In Situ Mechanical Characterization of the Cell Nucleus by Atomic Force Microscopy
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DOI：
10.1021/nn500553z
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发表时间：
2014-04-01
期刊：
影响因子：
17.1
通讯作者：
Sun, Yu
中科院分区：
文献类型：
--
作者：
Liu, Haijiao;Wen, Jun;Sun, Yu
核力学性质的研究可以为理解核动力学及其在细胞力学信号转导中的作用提供新的思路。虽然已经开发了几种方法来表征细胞核的力学性质，但直接在细胞内原位探测细胞核是具有挑战性的。在这里，一个修改后的AFM（原子力显微镜）针穿透技术被证明是机械表征细胞核原位。基于AFM压痕曲线上的两个不同段确定细胞质和细胞核刚度，并与同步共聚焦Z-堆叠显微镜重建相关。直接细胞内测量的基础上，我们发现，从成纤维细胞样细胞的分离的细胞核表现出显着较低的杨氏模量比完整的细胞核原位。我们还表明，有原位核软化的高转移性膀胱癌细胞系124相比，其转移性较低的对应RT 4。该技术有可能成为细胞内力学研究的可靠定量测量工具。
The study of nuclear mechanical properties can provide insights into nuclear dynamics and its role in cellular mechanotransduction. While several methods have been developed to characterize nuclear mechanical properties, direct intracellular probing of the nucleus in situ is challenging. Here, a modified AFM (atomic force microscopy) needle penetration technique is demonstrated to mechanically characterize cell nuclei in situ. Cytoplasmic and nuclear stiffness were determined based on two different segments on the AFM indentation curves and were correlated with simultaneous confocal Z-stack microscopy reconstructions. On the basis of direct intracellular measurement, we show that the isolated nuclei from fibroblast-like cells exhibited significantly lower Young's moduli than intact nuclei in situ. We also show that there is in situ nucleus softening in the highly metastatic bladder cancer cell line 124 when compared to its less metastatic counterpart RT4. This technique has potential to become a reliable quantitative measurement tool for intracellular mechanics studies.