Microfluidic genetic analysis with an integrated a-Si:H detector.
We have developed an integrated hydrogenated amorphous silicon (a-Si:H) fluorescence detector for microfluidic genetic analysis. It consists of a half-ball lens, a ZnS/YF3 multilayer optical interference filter with a pinhole, and an annular a-Si:H PIN photodiode allowing the laser excitation to pass up through the central aperture in the photodiode and the filter. Microfluidic separations of multiplex PCR products generated from methicillin-resistant/sensitive Staphylococcus aureus (MRSA/MSSA) DNA on microfluidic capillary electrophoresis (CE) devices are successfully detected with the integrated detector. Similarly, multiplex PCR amplicons from the kanamycin resistant and K12 serotype-specific genes of E. coli cells are detected. The direct detection of multiplex PCR amplicons indicates that the fluorescence detector can be successfully coupled with current microfluidic PCR-CE platforms. This work establishes that the integrated a-Si:H detector provides relevant limits of detection for point-of-care genetic and pathogen analysis with microfluidic devices.
Kamei
T
T
Department of Chemistry, University of California, Berkeley, CA 94720, USA. toshi-kamei@aist.go.jp
Toriello
N M
NM
Lagally
E T
ET
Blazej
R G
RG
Scherer
J R
JR
Street
R A
RA
Mathies
R A
RA
D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics*; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006865:Hydrogenation; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods; D017422:Sequence Analysis, DNA / Q000295:instrumentation* / Q000379:methods; D012825:Silicon; D013050:Spectrometry, Fluorescence / Q000295:instrumentation* / Q000379:methods; D018511:Systems Integration; D014159:Transducers
D004269:DNA, Bacterial; D012825:Silicon