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用于收集和分离贴壁细胞的SU-8聚合物“微托盘”的脉冲激光微束释放机制。

Mechanisms of pulsed laser microbeam release of SU-8 polymer "micropallets" for the collection and separation of adherent cells.

作者信息

Quinto-Su Pedro A, To'a Salazar Georgina, Sims Christopher E, Allbritton Nancy L, Venugopalan Vasan

机构信息

Department of Chemical Engineering & Materials Science, University of California, Irvine, Irvine, California 92697-2575, USA.

出版信息

Anal Chem. 2008 Jun 15;80(12):4675-9. doi: 10.1021/ac800129a. Epub 2008 May 20.

Abstract

The release of individual polymer micropallets from glass substrates using highly focused laser pulses has been demonstrated for the efficient separation, collection, and expansion of single, adherent cells from a heterogeneous cell population. Here, we use fast-frame photography to examine the mechanism and dynamics of micropallet release produced by pulsed laser microbeam irradiation at lambda = 532 nm using pulse durations ranging between 240 ps and 6 ns. The time-resolved images show the laser microbeam irradiation to result in plasma formation at the interface between the glass coverslip and the polymer micropallet. The plasma formation results in the emission of a shock wave and the ablation of material within the focal volume. Ablation products are generated at high pressure due to the confinement offered by the polymer adhesion to the glass substrate. The ablation products expand underneath the micropallet on a time scale of several hundred nanoseconds. This expansion disrupts the polymer-glass interface and accomplishes the release of the pallet from its glass substrate on the microsecond time scale (approximately 1.5 micros). Our experimental investigation demonstrates that the threshold energy for pallet release is constant (approximately 2 microJ) over a 25-fold range of pulse duration spanning the picosecond to nanosecond domain. Taken together, these results implicate that pallet release accomplished via pulsed laser microbeam irradiation is an energy-driven plasma-mediated ablation process.

摘要

利用高聚焦激光脉冲从玻璃基板上释放单个聚合物微托盘,已被证明可用于从异质细胞群体中高效分离、收集和扩增单个贴壁细胞。在此,我们使用高速摄影来研究在波长λ = 532 nm下,使用持续时间在240 ps至6 ns之间的脉冲激光微束照射产生的微托盘释放机制和动力学。时间分辨图像显示,激光微束照射导致在玻璃盖玻片与聚合物微托盘之间的界面处形成等离子体。等离子体的形成导致冲击波的发射以及焦点体积内材料的烧蚀。由于聚合物与玻璃基板的粘附提供的限制,烧蚀产物在高压下产生。烧蚀产物在几百纳秒的时间尺度内在微托盘下方膨胀。这种膨胀破坏了聚合物 - 玻璃界面,并在微秒时间尺度(约1.5微秒)上实现了托盘从其玻璃基板上的释放。我们的实验研究表明,在跨越皮秒到纳秒范围的25倍脉冲持续时间范围内,托盘释放的阈值能量是恒定的(约2微焦)。综上所述,这些结果表明,通过脉冲激光微束照射实现的托盘释放是一个能量驱动的等离子体介导的烧蚀过程。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2018/2525502/4375de227d87/nihms60688f1.jpg

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