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A method for spatially resolved local intracellular mechanochemical sensing and organelle manipulation

机译:一种空间分辨的局部细胞内机械化学传感和细胞器操纵的方法

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摘要

Because both the chemical and mechanical properties of living cells play crucial functional roles, there is a strong need for biophysical methods to address these properties simultaneously. Here we present a novel (to our knowledge) approach to measure local intracellular micromechanical and chemical properties using a hybrid magnetic chemical biosensor. We coupled a fluorescent dye, which serves as a chemical sensor, to a magnetic particle that is used for measurement of the viscoelastic environment by studying the response of the particle to magnetic force pulses. As a demonstration of the potential of this approach, we applied the method to study the process of phagocytosis, wherein cytoskeletal reorganization occurs in parallel with acidification of the phagosome. During this process, we measured the shear modulus and viscosity of the phagosomal environment concurrently with the phagosomal pH. We found that it is possible to manipulate phagocytosis by stalling the centripetal movement of the phagosome using magnetic force. Our results suggest that preventing centripetal phagosomal transport delays the onset of acidification. To our knowledge, this is the first report of manipulation of intracellular phagosomal transport without interfering with the underlying motor proteins or cytoskeletal network through biochemical methods
机译:由于活细胞的化学和机械性质都起着至关重要的功能作用,因此迫切需要同时解决这些性质的生物物理方法。在这里,我们提出了一种使用混合磁化学生物传感器测量局部细胞内微机械和化学性质的新颖方法(据我们所知)。我们通过研究磁性粒子对磁力脉冲的响应,将用作化学传感器的荧光染料与磁性粒子相结合,该磁性粒子用于测量粘弹性环境。为了证明这种方法的潜力,我们将该方法用于研究吞噬作用的过程,其中吞噬体的酸化与细胞骨架重组同时发生。在此过程中,我们与吞噬体pH值同时测量了吞噬体环境的剪切模量和粘度。我们发现有可能通过使用磁力使吞噬体的向心运动停滞来操纵吞噬作用。我们的结果表明,防止向心吞噬体运输会延迟酸化的开始。据我们所知,这是首次通过生化方法操纵细胞内吞噬体转运而不会干扰潜在的运动蛋白或细胞骨架网络的报道

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