Multi-oscillation microrheology via acoustic force spectroscopy enables frequency-dependent measurements on endothelial cells at high-throughput.
Multi-oscillation microrheology via acoustic force spectroscopy enables frequency-dependent measurements on endothelial cells at high-throughput.
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DOI：
10.1039/d0lc01135e
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发表时间：
2021-05-18
期刊：
影响因子：
6.1
通讯作者：
Betz T
中科院分区：
文献类型：
--
作者：
Nguyen A;Brandt M;Muenker TM;Betz T
主动微流变学是确定细胞和组织力学特性的主要方法之一，这些粘弹性特性的建模与许多相互竞争的方法存在激烈的争论。大多数主动微流变学的实验方法，如光学镊子或基于原子力显微镜的方法依赖于单细胞测量，因此受到低通量的影响。在这里，我们提出了一种利用声学力对细胞进行频率相关微流变学的新方法，该方法允许并行地对几个细胞进行多路测量。声学力谱（AFS）用于对附着在微流控芯片内培养的人脐静脉内皮细胞（HUVEC）上的颗粒产生pN-nN范围内的多振荡力。虽然AFS是作为一种单分子技术来测量生物分子的机械化学特性，但我们利用AFS来测量暴露在不同条件下（如流动剪切应力或药物注射）的细胞的动态粘弹性特性。通过控制力和测量颗粒的位置，可以在几个小时内连续测量复合剪切模量G*(ω)。得到的幂律剪切模量与分数黏弹性模型一致。在我们的实验中，我们证实了通过细胞松弛素b干扰肌动蛋白细胞骨架后剪切模量的降低，这种影响在冲洗药物后是可逆的。此外，我们还提供了使用新方法AFS作为测量工具的关键信息，展示了它的功能和局限性，我们发现使用AFS进行粘弹性测量，彻底的校准和仔细的数据分析是至关重要的，为此我们提供了协议和指南。我们提出了一种在高通量和流体流动下内皮细胞单层频率依赖性微流变学的新方法。
Active microrheology is one of the main methods to determine the mechanical properties of cells and tissue, and the modelling of these viscoelastic properties is under heavy debate with many competing approaches. Most experimental methods of active microrheology such as optical tweezers or atomic force microscopy based approaches rely on single cell measurements, and thus suffer from a low throughput. Here, we present a novel method for frequency-dependent microrheology on cells using acoustic forces which allows multiplexed measurements of several cells in parallel. Acoustic force spectroscopy (AFS) is used to generate multi-oscillatory forces in the range of pN–nN on particles attached to primary human umbilical vein endothelial cells (HUVEC) cultivated inside a microfluidic chip. While the AFS was introduced as a single-molecule technique to measure mechanochemical properties of biomolecules, we exploit the AFS to measure the dynamic viscoelastic properties of cells exposed to different conditions, such as flow shear stresses or drug injections. By controlling the force and measuring the position of the particle, the complex shear modulus G*(ω) can be measured continuously over several hours. The resulting power-law shear moduli are consistent with fractional viscoelastic models. In our experiments we confirm a decrease in shear modulus after perturbing the actin cytoskeleton via cytochalasin B. This effect was reversible after washing out the drug. Additionally, we include critical information for the usage of the new method AFS as a measurement tool showing its capabilities and limitations and we find that for performing viscoelastic measurements with the AFS, a thorough calibration and careful data analysis is crucial, for which we provide protocols and guidelines. We present a novel method for frequency-dependent microrheology on endothelial cell monolayer at high throughput and fluid flow.
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影响因子：
10
作者：
Hinderliter PM;Minard KR;Orr G;Chrisler WB;Thrall BD;Pounds JG;Teeguarden JG
通讯作者：
Teeguarden JG
DOI：
10.6028/jres.097.013
发表时间：
1992-05
影响因子：
1.5
作者：
Jones FE;Harris GL
通讯作者：
Harris GL
影响因子：
3.4
作者：
Kollmannsberger, Philip;Mierke, Claudia Tanja;Fabry, Ben
通讯作者：
Fabry, Ben
DOI：
10.1021/i260061a012
发表时间：
1977-01-01
期刊：
INDUSTRIAL & ENGINEERING CHEMISTRY PROCESS DESIGN AND DEVELOPMENT
影响因子：
--
作者：
GOLETZ, E;TASSIOS, D
通讯作者：
TASSIOS, D
影响因子：
3.4
作者：
Alcaraz, J;Buscemi, L;Navajas, D
通讯作者：
Navajas, D