Measuring Molecular Forces Using Calibrated Optical Tweezers in Living Cells.
Measuring Molecular Forces Using Calibrated Optical Tweezers in Living Cells.
复制标题
使用校准光镊测量活细胞中的分子力。
DOI：
10.1007/978-1-4939-6421-5_21
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Goldman,YaleE
中科院分区：
文献类型：
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作者：
Hendricks,AdamG;Goldman,YaleE
光镊有助于揭示马达蛋白产生力和对力作出反应的机制。虽然光阱主要应用于纯化的体外系统，但新兴的方法使活细胞中的测量成为可能，其中活性波动的粘弹性环境和变化的折射率使仪器的校准复杂化。在这里，我们描述的技术来校准光陷阱在活细胞中使用的正弦振荡和自发波动的强制响应，并测量内源性合奏的驱动蛋白和动力蛋白马达蛋白，因为它们在细胞中运输货物所施加的力。
Optical tweezers have been instrumental in uncovering the mechanisms motor proteins use to generate and react to force. While optical traps have primarily been applied to purified, in vitro systems, emerging methods enable measurements in living cells where the actively fluctuating, viscoelastic environment and varying refractive index complicate calibration of the instrument. Here, we describe techniques to calibrate optical traps in living cells using the forced response to sinusoidal oscillations and spontaneous fluctuations, and to measure the forces exerted by endogenous ensembles of kinesin and dynein motor proteins as they transport cargoes in the cell.