MEASUREMENT OF FAST ENZYME KINETICS VIA DROPLET MERGING

使用高速（1- 10 m/s）25- 200 μ m直径的液滴合并在 提出飞行来测量50 μ m-1 ms范围内的酶动力学。 两 使用技术产生极其可重复的液滴流， 为喷墨打印系统而开发。 两股液滴流汇合， 反应将几乎瞬间发生。 酶反应 会引起吸收、荧光或化学发光的变化， 反应中使用的酶和底物。 在第一阶段， 研究中，将评估荧光反应。 发射的光子 被聚焦在合并区域的光电倍增管检测到。 液滴流 该区域对应于液滴将 在1 ms内运行。 探测器被选通以测量约500 μ s-1 ms的发射。 (with 1 μ s分辨率），门与液滴同步 创造频率，允许成千上万滴的排放， 整合 液滴的小直径确保了快速混合。 的 喷墨技术固有的液滴形成的可重复性允许 在长时间内收集光子，从而增加 测量的灵敏度。

The use of high speed (1-10m/s) 25-200mum diameter droplets merging in flight is proposed to measure enzyme kinetics in the 50mum-1ms range. Two extremely repeatable droplet streams are produced using techniques developed for ink-jet printing systems. The two droplet streams merge and a reaction will take place almost instantaneously. The enzyme reaction causes changes in absorption, fluorescence, or chemiluminescence, depending on the enzyme and substrate used in the reaction. In Phase I of this study, chemiluminescent reactions will be evaluated. The photons emitted are detected by a photomultiplier tube focused on a region of the merged droplet stream. The region corresponds to the length a droplet would travel in 1ms. The detector is gated to measure emissions for ~500mus-1ms. (with 1mus resolution) and the gate is synchronized with the droplet creation frequency, allowing the emissions from thousands of drops to be integrated. The small diameter of the droplets insures rapid mixing. The repeatability of droplet formation inherent with ink-jet technology allows photons to be gathered over long time periods, thus increasing the sensitivity of the measurement.