Examination of laser microbeam cell lysis in a PDMS microfluidic channel using time-resolved imaging
Examination of laser microbeam cell lysis in a PDMS microfluidic channel using time-resolved imaging
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DOI：
10.1039/b715708h
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发表时间：
2008-01-01
期刊：
影响因子：
6.1
通讯作者：
Venugopalan, Vasan
中科院分区：
文献类型：
--
作者：
Quinto-Su, Pedro A.;Lai, Hsuan-Hong;Venugopalan, Vasan
我们使用时间分辨成像来检测非粘附的BAF-3细胞在微流体通道内的裂解动力学，微流体通道是由单个高聚焦540 ps持续激光脉冲在λ = 532 nm处产生的。时间分辨的光场图像显示，脉冲激光微束的传递导致激光诱导等离子体的形成，随后是激波发射和空化泡的形成。微流控通道提供的约束实质上限制了空化泡的膨胀，并导致PDMS通道壁的显著变形。为了检查细胞裂解和细胞内容物的分散，我们获得了细胞装载荧光染料过程的时间分辨荧光图像。这些荧光图像显示，通过等离子体形成和空化泡动力学，细胞裂解发生在纳秒到微秒的时间尺度上。此外，时间分辨荧光图像显示，当细胞内容物被激光诱导的空化泡的膨胀分散时，与气泡崩溃相关的流动随后将细胞内容物重新定位到一个小区域。这种脉冲激光微束照射在微流体通道中实现细胞快速裂解的能力，对细胞内容物的稀释程度最小，这对它们在芯片上的实验室应用具有重要意义。
We use time-resolved imaging to examine the lysis dynamics of non-adherent BAF-3 cells within a microfluidic channel produced by the delivery of single highly-focused 540 ps duration laser pulses at lambda = 532 nm. Time-resolved bright-field images reveal that the delivery of the pulsed laser microbeam results in the formation of a laser-induced plasma followed by shock wave emission and cavitation bubble formation. The confinement offered by the microfluidic channel constrains substantially the cavitation bubble expansion and results in significant deformation of the PDMS channel walls. To examine the cell lysis and dispersal of the cellular contents, we acquire time-resolved fluorescence images of the process in which the cells were loaded with a fluorescent dye. These fluorescence images reveal cell lysis to occur on the nanosecond to microsecond time scale by the plasma formation and cavitation bubble dynamics. Moreover, the time-resolved fluorescence images show that while the cellular contents are dispersed by the expansion of the laser-induced cavitation bubble, the flow associated with the bubble collapse subsequently re-localizes the cellular contents to a small region. This capacity of pulsed laser microbeam irradiation to achieve rapid cell lysis in microfluidic channels with minimal dilution of the cellular contents has important implications for their use in lab-on-a-chip applications.