Illumination of integrated analytical systems
An analytical device including an optically opaque cladding, a sequencing layer including a substrate disposed below the cladding, and a waveguide assembly for receiving optical illumination and introducing illumination into the device. The illumination may be received from a top, a side edge, and a bottom of the device. The waveguide assembly may include a nanoscale aperture disposed in the substrate and extending through the cladding. The aperture defines a reaction cell for receiving a set of reactants. In various aspects, the device includes a sensor element and the illumination pathway is through the sensor element. Waveguides and illumination devices, such as plasmonic illumination devices, are also disclosed. Methods for forming and operating the devices are also disclosed.
1. An analytical device comprising:
a fluidic layer comprising components of an analytical reaction;
a chip comprising an array of nanoscale apertures extending through an opaque layer to a transparent layer below the opaque layer, the nanoscale apertures comprising reaction cells in fluidic communication with the fluidic layer, wherein the reaction cells comprise fluorescent components; and
light deflecting elements below the nanoscale apertures including semitransparent light directing elements which feed light to one or more nanoscale apertures above the semitransparent light directing elements, thereby providing illumination light to the nanoscale apertures,
wherein the nanoscale apertures comprise zero mode waveguides.
2. The analytical device of claim 1
wherein the semitransparent light directing elements are embedded in the transparent layer.
3. The analytical device of claim 1 wherein the semitransparent light directing elements comprise metal or semiconductor mirrors, dichroics, prisms, Bragg scatterers, acousto-optic devices, or electro-optic devices.
4. The analytical device of claim 1 wherein the analytical reaction comprises nucleic acid sequencing, and the fluorescent components comprise fluorescently labeled nucleotides.
5. The analytical device of claim 1 further comprising a gap in the opaque layer providing for the illumination light to pass through the gap to the light deflecting elements.
6. The analytical device of claim 5 wherein the illumination light is directed to the light directing elements by a light pipe.
7. The analytical device of claim 1 wherein the opaque layer comprises a metal.
8. The analytical device of claim 1 further comprising a detector disposed below the nanoscale apertures for detecting light emitted by the fluorescent components.
9. The analytical device of claim 8 further comprising integrated directional optical elements for directing the emitted light to one or more sensor elements on the detector.
10. The analytical device of claim 8 further comprising a filter layer having different filters for multicolor distinction by the detector.
11. The analytical device of claim 1 wherein the device comprises multiple light pipes that allow illumination light to pass through a layer of nanoscale apertures.
12. The analytical device of claim 11 wherein the device comprises between about 100,000 and 1 million nanoscale apertures.
13. A method for monitoring an analytical reaction comprising:
providing an analytical device of claim 8 ;
providing illumination light to the nanoscale apertures via the light deflection elements; and
detecting light emitted from the fluorescent components, thereby monitoring the analytical reaction.
14. The method of claim 13 wherein the semitransparent light directing elements comprise metal or semiconductor mirrors, dichroics, prisms, Bragg scatterers, acousto-optic devices, or electro-optic devices.
15. The method of claim 13 wherein the analytical reaction comprises nucleic acid sequencing, and the fluorescent components comprise fluorescently labeled nucleotides.
16. The method of claim 13 wherein the analytical device further comprises a gap in the opaque layer providing for illumination light to pass through the gap into the light pipe.
17. An analytical system comprising:
an analytical device of claim 8 ;
an illumination system providing the illumination light to the light deflection elements; and
a processing system for processing electrical signals from the detector.
18. The analytical system of claim 17 further comprising a fluidic delivery system for delivering fluid to the fluidic layer.
19. The analytical system of claim 17 wherein the semitransparent light directing elements comprise metal or semiconductor mirrors, dichroics, prisms, Bragg scatterers, acousto-optic devices, or electro-optic devices.