UNC Chapel Hill, Applied Physical Sciences, Chapel Hill, North Carolina.
UNC Chapel Hill, Applied Physical Sciences, Chapel Hill, North Carolina.
Biophys J. 2021 May 4;120(9):1542-1564. doi: 10.1016/j.bpj.2021.01.042. Epub 2021 Mar 9.
Mechanical properties of the cell are important biomarkers for probing its architectural changes caused by cellular processes and/or pathologies. The development of microfluidic technologies has enabled measuring the cell's mechanical properties at high throughput so that mechanical phenotyping can be applied to large samples in reasonable timescales. These studies typically measure the stiffness of the cell as the only mechanical biomarker and do not disentangle the rheological contributions of different structural components of the cell, including the cell cortex, the interior cytoplasm and its immersed cytoskeletal structures, and the nucleus. Recent advancements in high-speed fluorescent imaging have enabled probing the deformations of the cell cortex while also tracking different intracellular components in rates applicable to microfluidic platforms. We present a, to our knowledge, novel method to decouple the mechanics of the cell cortex and the cytoplasm by analyzing the correlation between the cortical deformations that are induced by external microfluidic flows and the nucleus displacements, induced by those cortical deformations, i.e., we use the nucleus as a high-throughput microrheological probe to study the rheology of the cytoplasm, independent of the cell cortex mechanics. To demonstrate the applicability of this method, we consider a proof-of-concept model consisting of a rigid spherical nucleus centered in a spherical cell. We obtain analytical expressions for the time-dependent nucleus velocity as a function of the cell deformations when the interior cytoplasm is modeled as a viscous, viscoelastic, porous, and poroelastic material and demonstrate how the nucleus velocity can be used to characterize the linear rheology of the cytoplasm over a wide range of forces and timescales/frequencies.
细胞的力学特性是探测其因细胞过程和/或病变引起的结构变化的重要生物标志物。微流控技术的发展使得能够以高通量测量细胞的机械特性,从而使机械表型分析能够在合理的时间内应用于大样本。这些研究通常仅测量细胞的刚性作为唯一的机械生物标志物,而无法区分细胞的不同结构成分(包括细胞皮层、内部细胞质及其浸入的细胞骨架结构和核)的流变贡献。最近在高速荧光成像方面的进展使得能够探测细胞皮层的变形,同时以适用于微流控平台的速率跟踪不同的细胞内成分。我们提出了一种新颖的方法,通过分析外部微流引起的皮层变形与核位移之间的相关性,来分离细胞皮层和细胞质的力学特性,即我们使用核作为高通量微流变探针,来研究细胞质的流变特性,而不考虑细胞皮层力学特性。为了证明这种方法的适用性,我们考虑了一个由刚性球形核位于球形细胞中心的概念验证模型。当内部细胞质被建模为粘性、粘弹性、多孔和多孔弹性材料时,我们获得了随时间变化的核速度作为细胞变形的函数的解析表达式,并演示了如何使用核速度来表征细胞质的线性流变特性在广泛的力和时间尺度/频率范围内。