Microfluidic device for rapid (<15 min) automated microarray hybridization.
BACKGROUND
Current hybridization protocols on microarrays are slow and need skilled personnel. Microfluidics is an emerging science that enables the processing of minute volumes of liquids to perform chemical, biochemical, or enzymatic analyzes. The merging of microfluidics and microarray technologies constitutes an elegant solution that will automate and speed up microarray hybridization.


METHODS
We developed a microfluidic flow cell consisting of a network of chambers and channels molded into a polydimethylsiloxane substrate. The substrate was aligned and reversibly bound to the microarray printed on a standard glass slide to form a functional microfluidic unit. The microfluidic units were placed on an engraved, disc-shaped support fixed on a rotational device. Centrifugal forces drove the sample and buffers directly onto the microarray surface.


RESULTS
This microfluidic system increased the hybridization signal by approximately 10fold compared with a passive system that made use of 10 times more sample. By means of a 15-min automated hybridization process, performed at room temperature, we demonstrated the discrimination of 4 clinically relevant Staphylococcus species that differ by as little as a single-nucleotide polymorphism. This process included hybridization, washing, rinsing, and drying steps and did not require any purification of target nucleic acids. This platform was sensitive enough to detect 10 PCR-amplified bacterial genomes.


CONCLUSIONS
This removable microfluidic system for performing microarray hybridization on glass slides is promising for molecular diagnostics and gene profiling.
Peytavi
Régis
R
Centre de Recherche en Infectiologie de l'Université Laval, Centre Hospitalier Universitaire de Québec (Pavillon CHUL), Sainte-Foy, Québec, Canada.
Raymond
Frédéric R
FR
Gagné
Dominic
D
Picard
François J
FJ
Jia
Guangyao
G
Zoval
Jim
J
Madou
Marc
M
Boissinot
Karel
K
Boissinot
Maurice
M
Bissonnette
Luc
L
Ouellette
Marc
M
Bergeron
Michel G
MG
D004247:DNA / Q000737:chemistry*; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods*; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013210:Staphylococcus / Q000145:classification / Q000235:genetics; D013499:Surface Properties
D004129:Dimethylpolysiloxanes; D004247:DNA