Articles having localized molecules disposed thereon and methods of producing same
Methods of producing substrates having selected active chemical regions by employing elements of the substrates in assisting the localization of active chemical groups in desired regions of the substrate. The methods may include optical, chemical and/or mechanical processes for the deposition, removal, activation and/or deactivation of chemical groups in selected regions of the substrate to provide selective active regions of the substrate.
1. A system comprising:
an array of nanoscale wells disposed through an opaque layer, each well having a bottom surface formed from a transparent layer underlying the opaque layer; wherein a plurality of the wells in the array comprise an enzyme bound to the bottom surface and located in an observation region; and wherein portions of the array other than the bottom surface of the wells are substantially free of the enzyme;
an illumination source that provides illumination to the observation regions through the transparent layer; and
an optical detector that receives electromagnetic radiation emitted through the transparent layer from the observation regions.
2. The system of claim 1 , wherein the transparent layer comprises SiO 2 .
3. The system of claim 1 , wherein the opaque layer comprises a metal or metal oxide.
4. The system of claim 1 , wherein the opaque layer comprises aluminum.
5. The system of claim 1 , wherein the opaque layer is passivated with a compound comprising one or more phosphonic acid groups.
6. The system of claim 1 , wherein the enzyme is a nucleic acid polymerase.
7. The system of claim 6 , wherein the polymerase is part of a nucleic acid polymerase/template/primer complex.
8. The system of claim 7 , wherein the wells comprise a solution comprising one or more nucleotides or nucleotide analogs.
9. The system of claim 1 , wherein the plurality of the wells in the array comprise a single active enzyme molecule bound to the bottom surface.
10. The system of claim 1 , wherein the enzyme is bound to the bottom surface through one or more silanes.
11. The system of claim 1 , wherein the enzyme is bound to the bottom surface through a binding or coupling group applied to the bottom surface.
12. The system of claim 1 , wherein the enzyme is bound to the bottom surface through biotin.
13. The system of claim 1 , wherein the enzyme is bound to the bottom surface through a biotin-PEG-silane.