Methods, devices, and systems for analyte detection and analysis
Provided are systems and methods for analyte detection and analysis. A system can comprise an open substrate. The open substrate may be configured to rotate or otherwise move. The open substrate can comprise an array of individually addressable locations, with analytes immobilized thereto. The substrate may be spatially indexed to identify nucleic acid molecules from one or more sources, and/or sequences thereof, with the respective one or more sources. A solution comprising a plurality of probes may be directed across the array to couple at least one of the plurality of probes with at least one of the analytes to form a bound probe. A detector can be configured to detect a signal from the bound probe via scanning of the substrate while minimizing temperature fluctuations of the substrate or optical aberrations caused by bubbles.
1. A method for reagent dispensing, comprising:
(a) dispensing a first reagent to a first region of a surface of a substrate, wherein said surface comprises a second region comprising an analyte coupled thereto, wherein said second region is different from said first region;
(b) dispensing a second reagent to said first region such that said first reagent mixes with said second reagent to form a mixture, wherein said first region is a location of initial contact between said second reagent and said substrate; and
(c) causing said mixture to move from said first region to said second region and be in contact with said analyte.
2. The method of claim 1 , wherein (b) comprises forming a substantially homogenous mixture.
3. The method of claim 1 , wherein said first reagent and said second reagent are delivered substantially simultaneously to said first region.
4. The method of claim 1 , wherein said second reagent is delivered to said first region subsequent to delivery of said first reagent to said first region.
5. The method of claim 1 , wherein said first reagent is dispensed to said surface via a first nozzle and wherein said second reagent is dispensed to said surface via a second nozzle different from said first nozzle.
6. The method of claim 1 , wherein said first region is more radially proximal to a central axis of said surface than said second region.
7. The method of claim 1 , wherein (c) comprises rotating said surface relative to an axis of rotation to facilitate radially outward motion of said mixture on said surface.
8. The method of claim 7 , wherein, subsequent to (c), said surface comprises a layer of film of said mixture having a predetermined thickness.
9. The method of claim 1 , wherein said analyte is a nucleic acid molecule.
10. The method of claim 1 , wherein said analyte is coupled to a bead, wherein said bead is immobilized to said surface.
11. The method of claim 10 , wherein said surface comprises an array of a plurality of individually addressable locations, wherein said bead is coupled to an individually addressable location of said plurality of individually addressable locations.
12. The method of claim 11 , wherein a plurality of beads is coupled to said plurality of individually addressable locations, each bead of said plurality of beads comprising a respective analyte coupled thereto.
13. The method of claim 12 , wherein said plurality of beads comprises a reference bead, wherein said reference bead is configured to fluoresce during all imaging events of said surface.
14. The method of claim 11 , wherein said plurality of individually addressable locations comprise at least 50,000,000,000 individually addressable locations.
15. The method of claim 10 , wherein said bead comprises a plurality of analytes coupled thereto.
16. The method of claim 15 , wherein said plurality of analytes comprises a plurality of nucleic acid molecules having sequence identity.
17. The method of claim 1 , wherein during (a) or (b) said surface is rotating relative to an axis of rotation at a rotational frequency of no more than 5 revolutions per minute (rpm).
18. The method of claim 1 , wherein during (a) or (b) said surface is stationary.
19. The method of claim 1 , wherein said first reagent comprises a nucleotide solution.
20. The method of claim 1 , wherein said second reagent comprises a cleaving reagent.
21. The method of claim 1 , further comprising, subsequent to (c), detecting a signal from said surface.
22. The method of claim 1 , wherein said surface is substantially planar.
23. The method of claim 1 , wherein said surface is patterned.
24. The method of claim 23 , wherein said surface is patterned via surface chemistry.