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 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.
Hofmann
Oliver
O
Department of Chemistry, Imperial College London, London SW7 2AY, UK.
Murray
Kirk
K
Wilkinson
Alan-Shaun
AS
Cox
Timothy
T
Manz
Andreas
A
D001407:Bacillus / Q000737:chemistry / Q000528:radiation effects*; D004269:DNA, Bacterial / Q000032:analysis; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D007834:Lasers; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D012828:Silicones / Q000737:chemistry; D013045:Species Specificity; D013171:Spores, Bacterial / Q000737:chemistry / Q000528:radiation effects*; D013500:Surface Tension; D014466:Ultraviolet Rays
D004269:DNA, Bacterial; D004129:Dimethylpolysiloxanes; D012828:Silicones; C013830:baysilon