Examination of laser microbeam cell lysis in a PDMS microfluidic channel using time-resolved imaging.
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.
Quinto-Su
Pedro A
PA
Department of Chemical Engineering & Materials Science, University of California, Irvine, CA 92697, USA.
Lai
Hsuan-Hong
HH
Yoon
Helen H
HH
Sims
Christopher E
CE
Allbritton
Nancy L
NL
Venugopalan
Vasan
V
D002460:Cell Line; D007834:Lasers; D044085:Microfluidics / Q000295:instrumentation*; D008856:Microscopy, Fluorescence