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 system for nucleic acid sample processing, comprising:
a substrate comprising a first region and a second region different from said first region, wherein said first region or said second region comprises at least 10,000 indexed locations;
a first source comprising a first set of nucleic acid molecules, wherein said first region of said substrate comprises a first set of indexed locations comprising nucleic acid molecules of said first set of nucleic acid molecules;
a second source comprising a second set of nucleic acid molecules, wherein said second source is different than said first source, wherein said second region of said substrate comprises a second set of indexed locations comprising nucleic acid molecules of said second set of nucleic acid molecules;
a detector configured to detect signals from said first set of indexed locations that are indicative of at least one nucleotide having been incorporated into a growing nucleic acid strand that is complementary to a nucleic acid molecule of said first set of nucleic acid molecules; and
one or more processors, individually or in combination, configured to (i) receive said signals from said detector and (ii) associate each nucleic acid molecule of said first set of indexed locations with said first source or (iii) associate each nucleic acid molecule of said second set of indexed locations with said second source.
2 . The system of claim 1 , wherein said one or more processors are operatively coupled to said first source and said second source.
3 . The system of claim 2 , wherein said one or more processors are individually or collectively programmed to direct a fluid flow unit to dispense a solution from said first source or said second source to said substrate.
4 . The system of claim 1 , wherein said detector comprises an optical imaging unit that is operatively coupled to said one or more processors.
5 . The system of claim 4 , wherein said optical imaging unit is configured to detect (i) signals from said first region to determine a first set of sequences of said nucleic acid molecules of said first set of indexed locations and (ii) signals from said second region to determine a second set of sequences of said nucleic acid molecules of said second set of indexed locations.
6 . The system of claim 5 , wherein said one or more processors are configured, individually or in combination, (1) to associate said first set of sequences to said first source using said first set of indexed locations and (2) to associate said second set of sequences to said second source using said second set of indexed locations.
7 . The system of claim 6 , wherein said first set of sequences is associated with said first source and said second set of sequences is associated with said second source in absence of determining a barcode sequence of said first set of nucleic acid molecules and said second set of nucleic acid molecules.
8 . The system of claim 1 , wherein each nucleic acid molecule of said first set of nucleic acid molecules and said second set of nucleic acid molecules comprises a synthetic sequence of no more than 6 bases in length, and wherein a subset of nucleic acid molecules originating from a same source each comprises a common synthetic sequence, which common synthetic sequence is different from synthetic sequences of another subset of nucleic acid molecules originating from a different source.
9 . The system of claim 1 , wherein said substrate comprises a plurality of binders, wherein a binder of said plurality of binders is capable of immobilizing both said nucleic acid molecules of said first set of nucleic acid molecules and said nucleic acid molecules of said second set of nucleic acid molecules, wherein said nucleic acid molecules of said first set of nucleic acid molecules are immobilized to said first set of indexed locations via a first set of binders of said plurality of binders, and wherein said nucleic acid molecules of said second set of nucleic acid molecules are immobilized to said second set of indexed locations via a second set of binders of said plurality of binders different from said first set of binders.
10 . The system of claim 9 , wherein said binder is configured to immobilize said first set of nucleic acid molecules or said second set of nucleic acid molecules using one or more interactions selected from a group consisting of: hydrophilic interactions, hydrophobic interactions, electrostatic interactions, and hybridization interactions.
11 . The system of claim 1 , wherein said first region and said second region are adjacent and in contact on said substrate.
12 . The system of claim 1 , wherein said substrate comprises (1) a plurality of first regions, including said first region, configured to receive nucleic acid molecules of said first source and (2) a plurality of second regions, including said second region, configured to receive nucleic acid molecules of said second source, which plurality of first regions and plurality of second regions are arranged on said substrate such that a given first region of said plurality of first regions is not in contact with another first region of said plurality of first regions.
13 . The system of claim 12 , wherein said plurality of first regions and said plurality of second regions are arranged as alternating regions on said substrate.
14 . The system of claim 12 , wherein said plurality of first regions and said plurality of second regions are arranged as radially alternating regions on said substrate.
15 . The system of claim 12 , wherein (i) said plurality of first regions are chemically distinct from said plurality of second regions, or (ii) said plurality of first regions and said plurality of second regions are separated by barriers.
16 . The system of claim 1 , wherein said first region and said second region are of different sizes or comprise different numbers of individually addressable locations on said substrate.
17 . The system of claim 1 , further comprising, a first loading fluid comprising said first set of nucleic acid molecules and a second loading fluid comprising said second set of nucleic acid molecules, wherein said first loading fluid and said second loading fluid are immiscible.
18 . The system of claim 1 , further comprising a magnetic source configured to direct said first set of nucleic acid molecules or said second set of nucleic acid molecules to said substrate.
19 . The system of claim 1 , wherein said first region and said second region are separated by a barrier on said substrate, and wherein said barrier comprises a viscous solution, polyethylene glycol (PEG), a fluid that is immiscible with a loading solution comprising said first set of nucleic acid molecules and said second set of nucleic acid molecules, an air knife, an injection molded guide, or a hydrophobic region.
20 . The system of claim 1 , wherein said substrate is substantially planar.
21 . The system of claim 1 , wherein said substrate is patterned.
22 . The system of claim 1 , wherein said substrate is rotatable about a rotational axis.
23 . The system of claim 22 , wherein said first set of indexed locations and said second set of indexed locations have different radial distances from said rotational axis.
24 . The system of claim 1 , wherein said nucleic acid molecules of said first set of nucleic acid molecules have sequences that are different from sequences of said nucleic acid molecules of said second set of nucleic acid molecules.
25 . The system of claim 1 , wherein said nucleic acid molecules of said first set of nucleic acid molecules have the same sequences as said nucleic acid molecules of said second set of nucleic acid molecules.
26 . The system of claim 1 , wherein said first set of nucleic acid molecules is attached to a plurality of beads, which plurality of beads is immobilized adjacent to said substrate at said first set of indexed locations.
27 . The system of claim 26 , wherein a bead of said plurality of beads comprises a plurality of nucleic acid molecules attached thereto, and wherein said plurality of nucleic acid molecules comprises amplification products derived from a template nucleic acid molecule.
28 . The system of claim 27 , wherein said first set of indexed locations comprises at least 100 individually addressable locations, and wherein said second set of indexed locations comprises at least 100 individually addressable locations.
29 . The system of claim 28 , wherein said first set of indexed locations and said second set of indexed locations comprise at least about 5,000,000,000 individually addressable locations.
30 . The system of claim 28 , wherein said first set of nucleic acid molecules is attached to a first plurality of beads and said second set of nucleic acid molecules is attached to a second plurality of beads, and wherein said first plurality of beads and said second plurality of beads are associated to said at least 100 individually addressable locations of said first set of indexed locations or said at least 100 individually addressable locations of said second set of indexed locations.