Selection of compartmentalized screening method
View Patent ↗The present invention provides novel microfluidic substrates and methods that are useful for performing biological, chemical and diagnostic assays. The substrates can include a plurality of electrically addressable, channel bearing fluidic modules integrally arranged such that a continuous channel is provided for flow of immiscible fluids.
1. A method for analyzing an amplification reaction product, the method comprising:
providing a plurality of droplets separated by an oil in a reservoir that is external to a microfluidic device, wherein a plurality of the droplets comprise nucleic acid molecules and reagents in an aqueous fluid for a nucleic acid amplification reaction;
conducting the nucleic acid amplification reaction simultaneously in the droplets while the droplets are in the reservoir;
subsequent to the conducting step, moving the droplets from the reservoir into a channel of the microfluidic device; and
flowing the droplets in the microfluidic device past an optical detector, wherein an optical signal in at least one of the droplets is detected, the optical signal indicating presence of a nucleic acid amplification product in at least one of the at least one droplets.
2. The method accordingly to claim 1 , wherein the droplets and the immiscible fluid have different densities.
3. The method according to claim 2 , wherein the density differences assist droplet collection by separating at least a portion of the immiscible fluid from the droplets.
4. The method according to claim 1 , wherein the droplets are monodisperse within the channel.
5. The method according to claim 1 , wherein the oil is a fluorinated oil.
6. The method according to claim 1 , wherein the oil comprises a surfactant.
7. The method according to claim 6 , wherein the surfactant is a fluorosurfactant.
8. The method according to claim 1 , wherein the amplification reaction is a polymerase chain reaction.
9. The method according to claim 1 , wherein after detecting, the method further comprising sorting the droplets.
10. The method according to claim 9 , further comprising collecting at least one of the sorted droplets.
11. The method according to claim 1 , wherein the optical detector comprises a laser.
12. The method according to claim 1 , wherein the moving step further comprises spacing the droplets.
13. The method according to claim 12 , wherein spacing comprises adding an immiscible fluid to the channel so as to increase spacing between droplets prior to the droplets moving past the detector.
14. The method according to claim 1 , wherein the droplets pass through the channel and are detected by the detector one at a time.
15. The method according to claim 14 , wherein the width of the channel is arranged to position the droplets so that they are detected by the detector one at a time.
16. The method according to claim 1 , wherein the at least one droplet is a merged, fused, or coalesced droplet.
17. The method according to claim 1 , further comprising the step of merging a reagent into the droplets prior to detecting the reaction product.
18. The method according to claim 1 , wherein the reservoir is in fluid communication with the channel.
19. The method according to claim 1 , wherein said moving step comprises aspirating.
20. The method according to claim 1 , wherein said providing step comprises aspirating.
21. The method of claim 1 , wherein the reservoir is a syringe, cartridge, vial, tube, or well plate.
22. The method of claim 1 , wherein the reservoir is a collection module.
23. The method of claim 22 , wherein the collection module is on a microfabricated substrate.