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Controlling Brownian motion of single protein molecules and single fluorophores in aqueous buffer

机译:控制水性缓冲液中单个蛋白质分子和单个荧光团的布朗运动

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摘要

We present an Anti-Brownian Electrokinetic trap (ABEL trap) capable of trapping individual fluorescently labeled protein molecules in aqueous buffer. The ABEL trap operates by tracking the Brownian motion of a single fluorescent particle in solution, and applying a time-dependent electric field designed to induce an electrokinetic drift that cancels the Brownian motion. The trapping strength of the ABEL trap is limited by the latency of the feedback loop. In previous versions of the trap, this latency was set by the finite frame rate of the camera used for video-tracking. In the present system, the motion of the particle is tracked entirely in hardware (without a camera or image-processing software) using a rapidly rotating laser focus and lock-in detection. The feedback latency is set by the finite rate of arrival of photons. We demonstrate trapping of individual molecules of the protein GroEL in buffer, and we show confinement of single fluorophores of the dye Cy3 in water.
机译:我们提出了一种反布朗电动陷阱(ABEL陷阱),能够捕获水性缓冲液中单个荧光标记的蛋白质分子。 ABEL捕集阱通过跟踪溶液中单个荧光粒子的布朗运动,并施加时间相关的电场来工作,该电场旨在诱发抵消布朗运动的电动漂移,从而进行操作。 ABEL陷阱的陷阱强度受反馈回路的等待时间限制。在陷阱的早期版本中,此延迟是由用于视频跟踪的摄像机的有限帧速率设置的。在本系统中,使用快速旋转的激光聚焦和锁定检测,可以完全在硬件(没有相机或图像处理软件)中跟踪粒子的运动。反馈等待时间由光子的有限到达速率设置。我们展示了蛋白质GroEL的单个分子在缓冲液中的捕获,并且展示了染料Cy3在水中的单个荧光团的封闭。

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