Pinhole zero-mode waveguides
Devices and methods useful for sequencing and characterizing single nucleic acid molecules involve large arrays of nanopore zero-mode waveguides (NZMWs). The methods and devices are made possible by fabrication of nanoporous membranes of appropriate porosity for use in nucleotide sequencing. The fabrication methods eliminate the need for serial nanopore formation and make possible the mass production of highly efficient DNA and RNA single molecule sequencing devices.
1. A nanoporous membrane comprising a plurality of nanopores through the material; wherein the nanopores have a diameter in a range from about 0.3 nm to about 15 nm and provide ion conductive pathways across the membrane, the membrane having a specific conductance of about 10000 S/m 2 .
2. The nanoporous membrane of claim 1 comprising a material selected from the group consisting of aluminum oxide, hafnium oxide, silicon dioxide, and titanium oxide.
3. The nanoporous membrane of claim 1 having a thickness in the range from about 2 nm to about 50 nm.
4. The nanoporous membrane of claim 1 , wherein the ion conductive pathways are in the form of straight or tortuous channels through the membrane.
5. The nanoporous membrane of claim 1 formed by a process comprising reactive ion etching, wet chemical etching, atomic layer deposition, or molecular layer deposition.
6. A device comprising a zero-mode waveguide, the device comprising first and second fluid chambers separated by a nanoporous membrane of claim 1 , wherein the first and second fluid chambers and the membrane form said zero-mode waveguide, and wherein the membrane comprises a plurality of nanopores within said zero-mode waveguide that provide ion conductance pathways across the membrane.
7. The zero-mode waveguide device of claim 6 , wherein the first fluid chamber has a width in the range from about 60 to about 200 nm and a depth from about 50 to about 150 nm, and is open at a top end to provide access to a fluid in the fluid chamber.
8. A zero-mode waveguide device comprising a plurality of first fluid chambers disposed in a two-dimensional array on a single chip and a single common second fluid chamber, the first and second fluid chambers separated by a nanoporous membrane of claim 1 and form a plurality of zero-mode waveguides disposed in the two-dimensional array, wherein the membrane of each waveguide of said plurality of waveguides comprises a plurality of nanopores that provide ion conductance pathways across the membrane.
9. The zero-mode waveguide device of claim 8 , further comprising a first electrode disposed in each first chamber, a second electrode disposed in the second chamber, and a voltage source configured for providing a user-defined voltage between said first and second electrodes.
10. A system comprising the zero-mode waveguide device of claim 9 , a fluorescence microscope, an image acquisition device, a processor, and a memory.
11. A filter or filtration system comprising the membrane of claim 1 .