Evaporation management in digital microfluidic devices

• US 10,695,762 B2
• Filed: 06/06/2016
• Issued: 06/30/2020
• Est. Priority Date: 06/05/2015
• Status: Active Grant

• Current Assignee: Miroculus Inc.
• Original Assignee: Miroculus Inc.
• Inventors: Jebrail, Mais, Renzi, Ronald Francis, Branda, Steven
• Primary Examiner(s): Noguerola, Alexander S
• Application Number:
  US15/579,239
• Publication Number:
  US 20180178217A1
• Time in Patent Office: 1,485 Days
• CPC Class Codes: B01L 2200/142 Preventing evaporation
• CPC Class Codes: B01L 2200/143 Quality control, feedback systems
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/1805 Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2400/0427 Electrowetting
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502784 specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: B01L 3/502792 for moving individual droplets on a plate, e.g. by locally altering surface tension
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones

Abstract

Described herein are digital microfluidic (DMF) devices and corresponding methods for managing reagent solution evaporation during a reaction. Reactions on the DMF devices described here are performed in an air or gas matrix. The DMF devices include a means for performing reactions at different temperatures. To address the issue of evaporation of the reaction droplet especially when the reaction is performed at higher temperatures, a means for introducing a replenishing droplet has been incorporated into the DMF device. A replenishing droplet is introduced every time when it has been determined that the reaction droplet has fallen below a threshold volume. Detection and monitoring of the reaction droplet may be through visual, optical, fluorescence, colorimetric, and/or electrical means.

Claims

1. A method of replenishing a reaction droplet on an air-matrix digital microfluidic (DMF) apparatus to correct for evaporation, the method comprising:

monitoring a reaction droplet in an air gap region of the air-matrix DMF apparatus to determine when a volume of the reaction droplet falls below a threshold, wherein the reaction droplet comprises a solvent and reaction reagents;

introducing a replenishing droplet from an aperture through one of two plates of the air-matrix DMF apparatus forming the air gap region, wherein the replenishing droplet consists of solvent; and

combining the replenishing droplet with the reaction droplet after the volume of the reaction droplet falls beneath the threshold.

2. The method of claim 1, wherein combining comprises moving the replenishing droplet, the reaction droplet, or both the replacement droplet and the reaction droplet by applying energy to electrodes adjacent to the replacement droplet, the reaction droplet or both the replacement droplet and the reaction droplet.

3. The method of claim 1, wherein monitoring comprises determining a change in size of the reaction droplet.

4. The method of claim 1, wherein monitoring comprises detecting a change in an electrical property due to the reduction in volume of the reaction droplet.

5. The method of claim 1, wherein monitoring comprises detecting a capacitance change at an electrode adjacent and above or beneath the reaction droplet.

6. The method of claim 1, wherein monitoring comprises determining a change in size of the reaction droplet based relative to two or more actuation electrodes of the air-matrix DMF apparatus.

7. The method of claim 1, wherein the threshold level for triggering reagent replenishment is a loss of reaction droplet volume of 30% or more.

8. The method of claim 1, further comprising heating the reaction droplet in a thermal zone of the air gap region of the air-matrix DMF apparatus.

9. The method of claim 1, wherein introducing the replenishing droplet comprises introducing a replenishing droplet having a volume of between 10% and 50% the volume of the reaction droplet.

10. A method of replenishing a reaction droplet on an air-matrix digital microfluidic (DMF) apparatus to correct for evaporation, the method comprising:

optically monitoring the reaction droplet in an air gap region of the air-matrix DMF apparatus to determine when a volume of the reaction droplet falls below a threshold, wherein the reaction droplet comprises a solvent and reaction reagents;

introducing a replenishing droplet into the air gap region of the air-matrix DMF, wherein the replenishing droplet consists of solvent; and

combining the replenishing droplet with the reaction droplet after the volume of the reaction droplet falls beneath the threshold.

11. A method of replenishing a reaction droplet on an air-matrix digital microfluidic (DMF) apparatus to correct for evaporation, the method comprising:

monitoring a reaction droplet in an air gap region of the air-matrix DMF apparatus to determine when a volume of the reaction droplet falls below a threshold, wherein the reaction droplet comprises a solvent and reaction reagents;

introducing the replenishing droplet from an aperture through one of two plates of the air-matrix DMF apparatus forming the air gap region, wherein the replenishing droplet consists of solvent;

adjusting a temperature the replenishing droplet to a temperature the reaction droplet; and

combining the replenishing droplet with the reaction droplet when the temperature of the replenishing droplet matches the temperature of the reaction droplet, after the volume of the reaction droplet falls beneath the threshold.

12. The method of claim 11, wherein adjusting the replenishing droplet temperature comprises holding the replenishing droplet at a region that is adjacent to a reaction droplet and in thermal communication with a region beneath the reaction droplet.

13. The method of claim 11, wherein adjusting the replenishing droplet temperature comprises holding the replenishing droplet at a thermal zone and adjusting a temperature of the thermal zone to match the temperature of the reaction droplet.

14. The method of claim 11, wherein combining comprises moving the replenishing droplet, the reaction droplet, or both the replacement droplet and the reaction droplet by applying energy to electrodes adjacent to the replacement droplet, the reaction droplet or both the replacement droplet and the reaction droplet.

15. The method of claim 11, wherein monitoring comprises determining a change in size of the reaction droplet.

16. The method of claim 11, wherein monitoring comprises detecting a change in an electrical property due to the reduction in volume of the reaction droplet.

17. The method of claim 11, wherein monitoring comprises detecting a capacitance change at an electrode adjacent and above or beneath the reaction droplet.

18. The method of claim 11, wherein monitoring comprises determining a change in size of the reaction droplet based relative to two or more actuation electrodes of the air-matrix DMF apparatus.

19. The method of claim 11, wherein the threshold level for triggering reagent replenishment is a loss of reaction droplet volume of 30% or more.

20. The method of claim 11, further comprising heating the reaction droplet in a thermal zone of the air gap region of the air-matrix DMF apparatus.

21. The method of claim 11, wherein introducing the replenishing droplet comprises introducing a replenishing droplet having a volume of between 10% and 50% of the volume of the reaction droplet.

22. A method of replenishing a reaction droplet on an air-matrix digital microfluidic (DMF) apparatus to correct for evaporation, the method comprising:

optically monitoring the reaction droplet in an air gap region of the air-matrix DMF apparatus to determine when a volume of the reaction droplet falls below a threshold, wherein the reaction droplet comprises a solvent and reaction reagents;

introducing a replenishing droplet into the air gap region of the air-matrix DMF, wherein the replenishing droplet consists of solvent;

adjusting a temperature of the replenishing droplet to a temperature of the reaction droplet; and

combining the replenishing droplet with the reaction droplet when the temperature of the replenishing droplet matches the temperature of the reaction droplet, after the volume of the reaction droplet falls beneath the threshold.

23. A method of replenishing a reaction droplet on an air-matrix digital microfluidic (DMF) apparatus to correct for evaporation, the method comprising:

monitoring a reaction droplet in an air gap region of the air-matrix DMF apparatus to determine when a volume of the reaction droplet falls below 30% of an initial volume, wherein the reaction droplet comprises a solvent and reaction reagents;

introducing a replenishing droplet into the air gap region of the air-matrix DMF through an aperture in one or two plates forming the air gap region, wherein the replenishing droplet consists of solvent;

moving the replenishing droplet to a region adjacent to the reaction droplet;

adjusting a temperature of the replenishing droplet to a temperature of the reaction droplet; and

combining the replenishing droplet with the reaction droplet when the temperature of the replenishing droplet matches the temperature of the reaction droplet, after the volume of the reaction droplet falls beneath a threshold.