Characterization and use of laser-based lysis for cell analysis on-chip
Characterization and use of laser-based lysis for cell analysis on-chip
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DOI：
10.1098/rsif.2008.0177.focus
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发表时间：
2008-10-06
影响因子：
3.9
通讯作者：
Allbritton, Nancy L.
中科院分区：
文献类型：
--
作者：
Lai, Hsuan-Hong;Quinto-Su, Pedro A.;Allbritton, Nancy L.
我们演示了使用脉冲激光微束进行细胞裂解，然后在微流体装置中对细胞分析物进行电泳分离。测量了微通道内的脉冲能量和激光焦点对等离子体形成阈值的影响。光束穿过的聚（二甲基硅氧烷）（PDMS）层的厚度是阈值能量的关键决定因素。激光束的有效光路长度 L-eff 可用于预测不同微通道位置处光学击穿的阈值。基于激光的细胞裂解的一个主要优点是裂解区域非常有限（小于 5 毫米）。第二个优点是快速的细胞裂解时间（约微秒）。这些功能使得细胞中的两种分析物（荧光素和俄勒冈绿）能够在发生细胞裂解的通道中进行电泳分离。细胞分析物分离的分辨率和效率与标准相似，证明了在单细胞分析中使用脉冲激光微束的可行性。
We demonstrate the use of a pulsed laser microbeam for cell lysis followed by electrophoretic separation of cellular analytes in a microfluidic device. The influence of pulse energy and laser focal point within the microchannel on the threshold for plasma formation was measured. The thickness of the poly(dimethylsiloxane) (PDMS) layer through which the beam travelled was a critical determinant of the threshold energy. An effective optical path length, L-eff, for the laser beam can be used to predict the threshold for optical breakdown at different microchannel locations. A key benefit of laser-based cell lysis is the very limited zone (less than 5 mm) of lysis. A second asset is the rapid cell lysis times (approx. microseconds). These features enable two analytes, fluorescein and Oregon Green, from a cell to be electrophoretically separated in the channel in which cell lysis occurred. The resolution and efficiency of the separation of the cellular analytes are similar to those of standards demonstrating the feasibility of using a pulsed laser microbeam in single-cell analysis.