Fluidics systems for sequential delivery of reagents
The invention provides a passive fluidics circuit for directing different fluids to a common volume, such as a reaction chamber or flow cell, without intermixing or cross contamination. The direction and rate of flow through junctions, nodes and passages of the fluidics circuit are controlled by the states of upstream valves (e.g. opened or closed), differential fluid pressures at circuit inlets or upstream reservoirs, flow path resistances, and the like. Free diffusion or leakage of fluids from unselected inlets into the common outlet or other inlets at junctions or nodes is prevented by the flow of the selected inlet fluid, a portion of which sweeps by the inlets of unselected fluids and exits the fluidics circuit by waste ports, thereby creating a barrier against undesired intermixing with the outlet flow through leakage or diffusion.
1. An apparatus comprising:
a reagent source;
one or more wash solution sources;
a flowcell including a fluidics interface member mounted on and sealingly attached to a housing, the flowcell defining a plurality of separate flow chambers, each flow chamber having an inlet and an outlet, the inlet in fluid communication with the reagent source and the outlet connected to waste, each flow chamber having interior walls formed by attachment of the fluidics interface member to the housing;
the reagent source to provide a nucleotide solution to the flowcell; and
for each flow chamber of the plurality of separate flow chambers:
a passage in fluid communication with the inlet;
a junction in fluid communication with the passage, the reagent source, and a wash solution source of the one or more wash solution sources;
a valve to control passage of a wash solution to the junction; and
a fluidic circuit comprising:
a plurality of fluid inlets;
a waste port;
the reagent source;
a node in fluid communication with the reagent source; and
a plurality of fluid passages, each fluid passage of the plurality of fluid passages including a junction in unique fluid communication with a fluid inlet of the plurality of fluid inlets, each fluid passage of the plurality of fluid passages providing a distinct fluid pathway between the node and the waste port.
2. The apparatus of claim 1 , wherein, for the each flow chamber, the outlet is disposed diagonally opposite the inlet.
3. The apparatus of claim 1 , further comprising a reference electrode in electrical communication with an inlet of the wash solution source.
4. The apparatus of claim 1 , wherein, when the valve is open, the nucleotide solution is blocked from entering the passage and flow chamber.
5. The apparatus of claim 1 , wherein, when the valve is closed, the nucleotide solution is permitted to enter the passage and flow chamber.
6. The apparatus of claim 1 , further comprising a second passage to waste in fluid communication with the reagent source and the wash solution source and bypassing the flow chamber.
7. The apparatus of claim 1 , wherein the plurality of fluid passages is free of valves.
8. The apparatus of claim 1 , wherein each distinct fluid pathway is free of impermeable barriers.
9. The apparatus of claim 1 , further comprising a wash fluid inlet in fluid communication with the reagent source opposite the node.
10. The apparatus of claim 9 , further comprising a reference electrode in electrical communication with the wash fluid inlet.
11. The apparatus of claim 9 , wherein the wash fluid inlet includes a branch passage connected to the fluid outlet, the reference electrode disposed in the branch passage.
12. The apparatus of claim 9 , further comprising a reference electrode having a continuous electrolyte pathway through the reagent source to the each flow chamber.
13. The apparatus of claim 1 , wherein each reagent of the plurality of reagents is stored in a respective reservoir.
14. The apparatus of claim 1 , wherein each fluid passage of the plurality of fluid passages includes a first segment extending from the junction to the node and a second segment extending from the junction away from the node, wherein the second segment has greater flow resistance than the first segment.
15. An apparatus comprising:
a reagent source;
one or more wash solution sources;
a flowcell including a fluidics interface member mounted on and sealingly attached to a housing, the flowcell defining a plurality of separate flow chambers, each flow chamber having an inlet and an outlet, the inlet in fluid communication with the reagent source and the outlet connected to waste, each flow chamber having interior walls formed by attachment of the fluidics interface member to the housing;
the reagent source to provide a nucleotide solution to the flowcell; and
for each flow chamber of the plurality of separate flow chambers:
a passage in fluid communication with the inlet;
a junction in fluid communication with the passage, the reagent source, and a wash solution source of the one or more wash solution sources;
a valve to control passage of a wash solution to the junction; and
a fluidic circuit comprising:
a node having a fluid outlet;
a plurality of fluid inlets, each fluid inlet of the plurality of fluid inlets uniquely associated with a respective reagent source of a plurality of reagent sources, the plurality of reagent sources comprising the reagent source;
at least one waste port;
a plurality of passages, each passage of the plurality of passages providing a distinct fluid pathway between the node and the at least one waste port, each passage of the plurality of passages including a junction to a respective inlet passage providing fluid communication between the each passage and a respective fluid inlet of the plurality of fluid inlets;
an outlet passage in fluid communication with the node through the fluid outlet and in fluid communication with the reagent source; and
a wash fluid inlet in fluid communication with the outlet passage.
16. The apparatus of claim 15 , wherein each reagent of the plurality of reagents is stored in a respective reservoir.
17. The apparatus of claim 15 , wherein each passage includes a first segment an inner leg extending from the junction to the node and a second segment an outer leg extending from the junction away from the node toward the at least one waste port.
18. The apparatus of claim 15 , wherein, for the each flow chamber, the outlet is disposed diagonally opposite the inlet.
19. An apparatus comprising:
a reagent source;
one or more wash solution sources;
a flowcell including a fluidics interface member mounted on and sealingly attached to a housing, the flowcell defining a plurality of separate flow chambers, each flow chamber having an inlet and an outlet, the inlet in fluid communication with the reagent source and the outlet connected to waste, each flow chamber having interior walls formed by attachment of the fluidics interface member to the housing;
the reagent source to provide a nucleotide solution to the flowcell; and
for each flow chamber of the plurality of separate flow chambers:
a passage in fluid communication with the inlet;
a junction in fluid communication with the passage, the reagent source, and a wash solution source of the one or more wash solution sources;
a valve to control passage of a wash solution to the junction; and
a fluidic circuit comprising:
a fluidics node having an outlet in fluid communication with the reagent source;
a plurality of fluid inlets;
at least one waste port; and
a plurality of passages, each fluid inlet of the plurality of fluid inlets in fluid communication with a unique passage of the plurality of passages, the at least one waste port in fluid communication with the fluidics node by each passage of the plurality of passages without intervening valves, the plurality of passages having a fluid resistance selected so that when a fluid is selected to flow solely through a single fluid inlet of the plurality of fluid inlets a first portion of the fluid flows to the at least one waste port via an associated passage uniquely associated with the single fluid inlet without entering the fluidics node and a second portion of the fluid flows via the associated passage to the fluidics node, a part of the second portion exits the fluidics node through the outlet and the remainder of the second portion exits the fluidics node to the at least one waste port through the plurality of passages other than the associated passage uniquely associated with the single fluid inlet, another fluid entering one of the plurality of passages from an unselected fluid inlet of the plurality of fluid inlets is directed to the at least one waste port without entering the fluidics node.
20. The apparatus of claim 19 , wherein said passages of said plurality are radially disposed about said outlet.