Microfluidic device, system and method
View Patent ↗A combination of capillary forces and gas pressure is used to control the movement of liquid samples within a microfluidic device. A liquid sample introduced to a proximal portion of a capillary channel of a microfluidic device moves by capillary action partway along the capillary channel. As the liquid sample moves, a pressure of a gas acting upon a distal gas-liquid interface of the liquid sample increases by an amount sufficient to stop further movement of the liquid sample. To initiate further movement of the liquid sample, a pump connected to a distal portion of the capillary channel decreases the pressure of the gas acting upon the distal gas-liquid interface of the liquid sample by an amount sufficient to permit the liquid sample to move by capillary action further along the capillary channel of the microfluidic device.
1. An immunoassay method for determining the presence of a target in a liquid sample, the method comprising:
(a) introducing the liquid sample to a proximal portion of a capillary flow channel;
(b) advancing the liquid sample at a first flow rate toward a distal portion of the capillary flow channel until at least a distal gas-liquid interface of the liquid sample contacts a conjugate disposed in dry form within the capillary flow channel, the conjugate comprising a binding agent having an affinity for the target;
(c) subsequently, by increasing a gas pressure differential between a proximal gas-liquid interface of the liquid sample and the distal gas-liquid interface of the liquid sample, advancing the liquid sample at a second flow rate toward the distal portion of the capillary flow channel until at least the distal gas-liquid interface contacts a detection zone within the capillary flow channel, the detection zone comprising a second binding agent having an affinity for a complex comprising the conjugate and the target, the second flow rate being slower than the first flow rate; and
(d) subsequently, by increasing the gas pressure differential between the proximal
and distal gas-liquid interfaces of the liquid sample, advancing the liquid sample a third flow rate toward the distal portion of the capillary flow channel until at least a conjugate is (a) bound to the second binding agent and/or been advanced beyond the detection zone toward the distal end of the capillary flow channel
(e) wherein any of the steps of “increasing a gas pressure differential” is performed by increasing a volume of gas in communication with the distal gas-liquid interface of the liquid sample.
2. The immunoassay method of claim 1 , wherein the capillary flow channel is disposed within a microfluidic device.
3. The immunoassay method of claim 1 , further comprising, after the step of introducing the liquid sample, advancing the liquid sample by capillary flow along the capillary flow channel until the gas pressure acting upon the distal gas-liquid interface stops the liquid sample from advancing further along the capillary flow channel.
4. The immunoassay method of claim 3 , wherein the liquid sample is stopped prior to contacting the conjugate.
5. The immunoassay method of claim 3 , wherein the liquid sample is stopped after contacting the conjugate.
6. The immunoassay method of claim 1 , further comprising providing a fluidic connection between a pump and a distal portion of the capillary flow channel.
7. The immunoassay method of claim 6 , wherein the step of providing a fluidic connection is performed prior to the step of introducing the liquid sample.
8. The immunoassay method of claim 6 , comprising terminating the fluidic connection between the pump and the distal portion of the capillary flow channel and then detecting conjugate present in the detection zone.
9. The immunoassay method of claim 8 , wherein the method comprising placing the microfluidic device in operable association with an optical reader for the microfluidic device.
10. The immunoassay method of claim 9 , wherein the step of providing the fluidic connection comprises automatically positioning a proximal opening of the pump with respect to a distal opening of the capillary flow channel.
11. The immunoassay method of claim 1 , wherein any of the steps of “increasing a gas pressure differential” is performed by actuating the pump.
12. The immunoassay method of claim 11 , wherein actuating the pump increases a volume of gas in communication with the distal gas-liquid interface of the liquid sample.
13. The immunoassay method of any of claim 12 , wherein the pump is a syringe pump.
14. The immunoassay method of claim 1 , wherein the liquid sample experiences a capillary force within the capillary flow channel and the magnitude of a force applied to the liquid sample by the “gas pressure differential” is less than about 15 times the magnitude of the capillary force, less than about 10 times the magnitude of the capillary force, or less than about 5 times the magnitude of the capillary force.
15. The immunoassay method of claim 1 further comprising the step of detecting conjugate bound to the detection zone.
16. The immunoassay method of claim 15 , wherein the step of detecting is performed while a volume of the detection zone is filled with liquid sample.
17. The immunoassay method of claim 15 , wherein the detection zone has a volume and the step of detecting is performed after removing a majority of the liquid sample from the detection zone.
18. The immunoassay method of claim 17 , wherein the step of detecting is performed while a majority of the volume of the detection zone is occupied by a gas.
19. The immunoassay method of claim 16 wherein the step of detecting is performed without first introducing a liquid other than the liquid sample into the detection zone.
20. The immunoassay method of claim 1 , wherein the liquid sample comprises a biological sample obtained from a mammal, blood, or urine.
21. The immunoassay method of claim 20 wherein the liquid sample comprises a reagent and the liquid sample is formed by combining the reagent and the biological sample.
22. The immunoassay method of claim 21 wherein the step of combining is performed prior to introducing the biological sample to the capillary flow channel.
23. The immunoassay method of claim 1 wherein the liquid sample is a filtered liquid sample formed by passing a liquid sample through a filter.
24. The immunoassay method of claim 23 , wherein the filter comprises pores and a size of the pores decreases proceeding from a proximal face of the filter toward a distal face of the filter.
25. The immunoassay method of claim 23 , wherein the filtered liquid sample comprises plasma and the step of passing the liquid sample through the filter comprises filtering red blood cells from the liquid sample.
26. The immunoassay method of claim 23 , wherein a total volume of blood is about 75 microliters or less, 50 microliters or less, 30 microliters or less, 20 microliters or less, such as about 15 microliters or less.
27. The immunoassay method of claim 1 wherein a total volume of the liquid sample is about 75 microliters or less, 50 microliters or less, 30 microliters or less, or 20 microliters or less.
28. The immunoassay method of claim 1 wherein a total volume of the liquid sample is about 15 microliters or less.
29. The immunoassay method of claim 1 wherein a total volume of the liquid sample is about 10 microliters or less.
30. An immunoassay method for determining the presence of a target in a liquid sample, the method comprising:
(a) introducing the liquid sample to a proximal portion of a capillary flow channel;
(b) advancing the liquid sample via capillary action at a first flow rate toward a distal portion of the capillary flow channel until at least a distal gas-liquid interface of the liquid sample contacts a conjugate disposed in dry form within the capillary flow channel, the conjugate comprising a binding agent having an affinity for the target, wherein the distal gas-liquid interface of the liquid sample is formed between the liquid sample and a gas enclosed in the capillary flow channel distal to the distal gas-liquid interface;
(c) subsequently, increasing the volume of the enclosed gas thereby decreasing a pressure of the enclosed gas acting on the distal gas-liquid interface and advancing the liquid sample via capillary action at a second flow rate toward the distal portion of the capillary flow channel until at least the distal gas-liquid interface contacts a detection zone within the capillary flow channel, the detection zone comprising a second binding agent having an affinity for a complex comprising the conjugate and the target, the second flow rate being slower than the first flow rate; and
(d) subsequently, increasing the volume of the enclosed gas thereby decreasing a pressure of the enclosed gas acting on the distal gas-liquid interface and advancing the liquid sample via capillary action at a third flow rate toward the distal portion of the capillary flow channel until at least a conjugate is (a) bound to the second binding agent and/or advanced beyond the detection zone toward the distal end of the capillary flow channel.