Fluidic connectors and microfluidic systems

• US 8,802,445 B2
• Filed: 02/12/2013
• Issued: 08/12/2014
• Est. Priority Date: 05/04/2007
• Status: Active Grant

• Current Assignee: Opko Diagnostics, LLC (Opko Health, Inc.)
• Original Assignee: Opko Diagnostics, LLC (Opko Health, Inc.)
• Inventors: Linder, Vincent, Steinmiller, David, Sia, Samual K.
• Primary Examiner(s): Sines, Brian J
• Application Number:
  US13/765,042
• Publication Number:
  US 20130157286A1
• Time in Patent Office: 546 Days
• Field of Search: 422/50, 422/68.1, 422/502, 422/503, 422/504, 422/509, 436/43, 436/63, 436/174, 436/180
• US Class Current: 436/180
• CPC Class Codes: B01L 2200/025 Align devices or objects to ensure defined positions relative to each other
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/021 Identification, e.g. bar codes
• CPC Class Codes: B01L 2300/041 Connecting closures to device or container
• CPC Class Codes: B01L 2300/0627 Sensor or part of a sensor is integrated
• CPC Class Codes: B01L 2300/0672 Integrated piercing tool
• CPC Class Codes: B01L 2300/0838 Capillaries
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/12 Specific details about materials
• CPC Class Codes: B01L 2400/049 vacuum
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: B01L 3/563 Joints or fittings in general F16L; Separable fluid transfer means to transfer fluids between at least two containers, e.g. connectors
• CPC Class Codes: B01L 3/565 Seals in general F16L
• CPC Class Codes: C12Q 1/6834 : Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: G01F 22/00 : Methods or apparatus for measuring volume of fluids or fluent solid material, not otherwise provided for
• CPC Class Codes: G01N 21/6428 : Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 33/5302 : Apparatus specially adapted for immunological test procedures
• CPC Class Codes: G01N 33/5304 : Reaction vessels, e.g. agglutination plates for solid-phase systems G01N33/543
• CPC Class Codes: G01N 33/5306 : Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
• CPC Class Codes: G01N 33/54366 : Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/545 : Synthetic resin
• CPC Class Codes: Y10T 403/22 : with fluid pressure responsive component
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/25 : including sample preparation
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

Fluidic connectors, methods, and devices for performing analyses (e.g., immunoassays) in microfluidic systems are provided. In some embodiments, a fluidic connector having a fluid path is used to connect two independent channels formed in a substrate so as to allow fluid communication between the two independent channels. One or both of the independent channels may be pre-filled with reagents (e.g., antibody solutions, washing buffers and amplification reagents), which can be used to perform the analysis. These reagents may be stored in the channels of the substrate for long periods amounts of time (e.g., 1 year) prior to use.

Claims

1. A method comprising:

providing a microfluidic system comprising a first microfluidic channel including at least one inlet and one outlet and a second microfluidic channel including at least one inlet and one outlet,

wherein the first microfluidic channel contains a first reagent disposed therein,

wherein the second microfluidic channel contains a second reagent disposed therein, and

wherein a seal covers the inlet of the first microfluidic channel and a seal covers the outlet of the first microfluidic channel so as to store the first reagent in the first microfluidic channel; and

connecting a fluidic connector comprising a fluid path to the microfluidic system to cause fluid communication between the first and second microfluidic channels,

wherein upon connection, the fluid path of the fluidic connector connects to an inlet or an outlet of the first microfluidic channel to allow fluid communication between the fluid path and the first microfluidic channel, and the fluid path connects to an inlet or an outlet of the second microfluidic channel to allow fluid communication between the fluid path and the second microfluidic channel,

wherein the first and second microfluidic channels are not in fluid communication with one another absent connection via the fluidic connector.

2. A method as in claim 1, wherein the microfluidic system comprises a reaction area in fluid communication with the second microfluidic channel prior to the connecting step.

3. A method as in claim 1, wherein the fluid path includes a fluid path inlet and a fluid path outlet, and wherein upon connection, the fluid path inlet connects to the outlet of the first microfluidic channel to allow fluid communication between the fluid path and the first microfluidic channel, and the fluid path outlet connects to the inlet of the second microfluidic channel to allow fluid communication between the fluid path and the second microfluidic channel.

4. A method as in claim 1, wherein the first reagent disposed in the first microfluidic channel is a liquid reagent.

5. A method as in claim 1, wherein the second reagent disposed in the second microfluidic channel is a substantially dry reagent.

6. A method as in claim 1, wherein the first microfluidic channel comprises a third reagent disposed therein.

7. A method as in claim 6, wherein the third reagent is a liquid reagent.

8. A method as in claim 7, wherein the first and third reagents are separated by a fluid that is immiscible with said first and third reagents.

9. A method as in claim 8, wherein the fluid immiscible with the first and third reagents is a gas.

10. A method as in claim 9, wherein at least one of the first and third reagents is a rinse solution.

11. A method as in claim 1, wherein the microfluidic system comprises a seal covering the inlet of the second microfluidic channel and a seal covering the outlet of the second microfluidic channel.

12. A method as in claim 2, wherein the second reagent is disposed in the reaction area of the second microfluidic channel, and wherein the second reagent is a substantially dry reagent.

13. A method as in claim 12, wherein the second reagent comprises at least one of an antibody or an antigen.

14. A method as in claim 12, wherein the second reagent is adsorbed to a surface of the second microfluidic channel.

15. A method as in claim 2, wherein the reaction area comprises at least one meandering channel region.

16. A method as in claim 15, wherein the reaction area comprises at least two meandering channel regions connected in series.

17. A method as in claim 16, wherein each of the at least two meandering channel regions comprises a chemical and/or biological species disposed therein that can undergo a chemical and/or biological reaction.

18. A method as in claim 17, wherein the chemical and/or biological reaction is a binding event between at least two binding partners, and wherein at least one of the binding partners comprises an antibody.

19. A method as in claim 2, wherein the reaction area comprises at least one cross-sectional dimension of less than 100 microns.

20. A method as in claim 1, wherein the microfluidic system comprises a substrate comprising more than one substrate layers that are mated to one another.

21. A method as in claim 1, wherein the fluidic connector comprises a substantially planar substrate comprising a channel formed in at least a portion of the substantially planar substrate.

22. A method as in claim 1, wherein the fluidic connector comprises a monolithic device comprising a channel formed in at least a portion of the monolithic device.

23. A method as in claim 1, wherein the fluidic connector comprises a tube.

24. A method as in claim 1, wherein the fluidic connector comprises a fluid path having a volume between 1 μ

L and 1000 μ

L.

25. A method as in claim 1, wherein the fluid path of the fluidic connector comprises a reagent disposed therein.

26. A method as in claim 25, wherein the reagent disposed in the fluid path of the fluidic connector is a substantially dry reagent that coats at least a portion of the fluid path.

27. A method as in claim 25, wherein the reagent is an anti-coagulant.

28. A method as in claim 1, wherein the fluidic connector comprises a volume control element that is constructed and arranged to introduce a controlled volume of a fluid sample from a biological entity into the fluid path.

29. A method as in claim 28, wherein the volume control element is constructed and arranged to allow a fluid sample to fill the entire volume of the fluid path.

30. A method as in claim 28, wherein the volume control element is constructed and arranged to allow a fluid sample to fill a portion, but not all, of the volume of the fluid path.

31. A method as in claim 1, wherein the microfluidic system comprises a substrate comprising the first and second microfluidic channels formed therein.

32. A method as in claim 1, comprising introducing a sample into the fluidic connector prior to the connection step.

33. A method as in claim 32, wherein the fluidic connector contains a binding component disposed therein, the method comprising binding at least a portion of the sample with the binding component prior to the connecting step.

34. A method as in claim 32, comprising applying a vacuum to an outlet of the microfluidic system and flowing at least the sample and the first reagent in series across an analysis region of the second microfluidic channel.