Laser induced disruption of bacterial spores on a microchip

We report on the development of a laser based spore disruption method. Bacillus globigii spores were mixed with a laser light absorbing matrix and co-crystallized into 200-microm-wide and 20-microm-deep nanovials formed in a polydimethylsiloxane (PDMS) target plate. Surface tension effects were expl...

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Bibliographic Details
Published in:Lab on a chip 2005-01, Vol.5 (4), p.374-377
Main Authors: Hofmann, Oliver, Murray, Kirk, Wilkinson, Alan-Shaun, Cox, Timothy, Manz, Andreas
Format: Article
Language:English
Subjects:
Bacillus - chemistry
Bacillus - radiation effects
Dimethylpolysiloxanes - chemistry
DNA, Bacterial - analysis
Lasers
Oligonucleotide Array Sequence Analysis - methods
Polymerase Chain Reaction - methods
Silicones - chemistry
Species Specificity
Spores, Bacterial - chemistry
Spores, Bacterial - radiation effects
Surface Tension
Ultraviolet Rays
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Summary:We report on the development of a laser based spore disruption method. Bacillus globigii spores were mixed with a laser light absorbing matrix and co-crystallized into 200-microm-wide and 20-microm-deep nanovials formed in a polydimethylsiloxane (PDMS) target plate. Surface tension effects were exploited to effect up to 125-fold spore enrichment. When the target zones were illuminated at atmospheric pressure with pulsed UV-laser light at fluences below 20 mJ cm(-2) a change in spore morphology was observed within seconds. Post illumination PCR analysis suggests the release of endogenous DNA indicative of spore disruption. For laser fluences above 20 mJ cm(-2), desorption of spores and fragments was also observed even without a matrix being employed. Desorbed material was collected in a PDMS flowcell attached to the target plate during laser illumination. This opens up a route towards the direct extraction of released DNA in an integrated spore disruption-PCR amplification microchip device.
ISSN:1473-0197
1473-0189
DOI:10.1039/b418663j