Multi-capillary optical waveguides for DNA sequencing.
Cylindrical capillaries can be used as optical elements in a waveguide, where refraction will confine an appropriately focused light beam to pass through the interiors of successive capillaries in a flat parallel array. Such a capillary waveguide allows efficient illumination of samples in multiple capillaries with relatively little laser power. Analytical expressions derived under paraxial and thin-lens approximations provide guidance in selecting the capillary sizes and the refractive indices that will produce the waveguiding effect, but accurate predictions require exact ray tracing. Small reflective losses as the light passes through the capillary surfaces cause cumulative intensity decreases, but the resulting lack of uniformity can be compensated to a considerable extent by illuminating the capillary array from both sides. A 12-capillary waveguide illuminated from both sides in air has a difference of less than 10% from the strongest to the weakest illumination. By increasing the refractive index of both the external medium and the contents of the capillaries, a 96-capillary waveguide for DNA sequencing could be produced that would also provide nearly uniform illumination. A 12-capillary, bi-directionally illuminated waveguide system for DNA sequencing has been constructed. The two focused laser beams are delivered by integrated fiber optic transmitters (IFOTs), and fluorescence is collected by a set of optical fibers whose spacing exactly matches that of the capillaries in the waveguide. The system is easy to align and provides sensitive detection of fluorescence with minimal cross-talk between channels.
Biology Department, Brookhaven National Laboratory, Upton, NY 11973, USA.
M A
MA
Quesada
H S
HS
Dhadwal
D
D
Fisk
F W
FW
Studier
D019075:Electrophoresis, Capillary / Q000379:methods; D008029:Lighting; D008432:Mathematical Computing; D008958:Models, Molecular; D017422:Sequence Analysis, DNA / Q000379:methods