Bioreactor for selectively controlling the molecular diffusion between fluids

• US 7,514,256 B2
• Filed: 02/11/2005
• Issued: 04/07/2009
• Est. Priority Date: 02/11/2005
• Status: Active Grant

• Current Assignee: Emilio Barbera-Guillem
• Original Assignee: Emilio Barbera-Guillem
• Inventors: Barbera-Guillem, Emilio
• Primary Examiner(s): Beisner; William H
• Assistant Examiner(s): HOBBS, MICHAEL L
• Application Number:
  US11/056,725
• Publication Number:
  US 20060180529A1
• Time in Patent Office: 1,516 Days
• Field of Search: 435/286.6, 435/293.1, 435/294.1, 422/82
• US Class Current: 435/293.1
• CPC Class Codes: B01L 2200/0684 Venting, avoiding backpressure, avoid gas bubbles
• CPC Class Codes: B01L 2200/141 Preventing contamination, tampering
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/044 pierceable, e.g. films, membranes
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/0874 Three dimensional network
• CPC Class Codes: B01L 2400/0472 Diffusion
• CPC Class Codes: B01L 2400/0688 surface tension valves, capillary stop, capillary break
• CPC Class Codes: B01L 2400/0694 vents used to stop and induce flow, backpressure valves
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• 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: C12M 23/16 Microfluidic devices; Capillary tubes integrated microfluidic structures B01L3/5027; microreactors B01J19/0093
• CPC Class Codes: C12M 29/04 Filters; Permeable or porous membranes or plates, e.g. dialysis

Abstract

A diffusion controlling bioreactor that selectively controls the molecular diffusion between fluids through a microchannel in fluid communication with a reaction reservoir. The length and cross-sectional area are selected to obtain a predetermined rate of molecular diffusion between fluids. When the fluids are liquids, flow through the microchannel is laminar and the capillary action of the microchannel and fluid is such that the fluid does not flow into the reaction reservoir unless the pressure of the fluid is increased by an external source, thereby minimizing contamination of the bioreactor. The instant invention may also utilize at least one microchannel and reagent reservoir to regulate, rather than prevent, the passage of various molecules into the bioreactor. A pressure equalizing vent operating on similar principles to the microchannel may have a structure configured to minimize the chances of fluid leakage from the bioreactor, even if the bioreactor is turned in various directions.

Claims

1. A diffusion controlling bioreactor that selectively controls the molecular diffusion between a first fluid and a second fluid, comprising:

a reaction reservoir having at least one reaction reservoir sidewall thereby defining a reaction reservoir volume, wherein the reaction reservoir initially contains the first fluid;

a microchannel having at least one portion fully enclosed by at least one sidewall, in fluid communication with the reaction reservoir, having a proximal end with a proximal end opening, a distal end with a distal end opening, a length, the at least one microchannel sidewall defining a cross-sectional area, wherein the length and cross-sectional area are selected to obtain a predetermined rate of molecular diffusion between the first fluid and the second fluid, and wherein the microchannel is configured such when the microchannel is filled with the second fluid, and the second fluid is a liquid, that flow of the second fluid through the microchannel is laminar and the capillary action of the microchannel and the second fluid is such that the second fluid does not flow into the reaction reservoir unless the pressure of the second fluid is increased by an external source; and

a pressure equalizing vent in fluid communication with the reaction reservoir and a fluidic external environment, wherein the pressure equalizing vent is capable of equalizing pressure within the reaction reservoir with that of the external environment, wherein the pressure equalization vent has a distal end with a distal end opening in communication with the external environment, a length, at least one sidewall, having a sidewall thickness, and defining a cross-sectional area, and a proximal end with a proximal end opening in communication with the reaction reservoir, and wherein the pressure equalizing vent is configured such that when it is filled with a third fluid, and the third fluid is a liquid, that flow of the third fluid through the pressure equalizing vent is laminar and the capillary action of the pressure equalizing vent and the third fluid is such that the third fluid does not flow out of the distal end opening unless the pressure of the third fluid is increased by an external source;

wherein the reaction reservoir has a maximum first dimension in a primary axis, a maximum second dimension in a secondary axis, and a maximum third dimension in a tertiary axis, and the pressure equalizing vent is a continuous microchannel configured lying wholly exterior to the reaction reservoir and having at least one component in the primary axis with a magnitude greater than the maximum first dimension, at least one component in the secondary axis with a magnitude greater than the maximum second dimension, and at least one component in the tertiary axis with a magnitude greater than the maximum third dimension.

2. The bioreactor of claim 1, wherein the distal end opening is closed by a penetrable, elastomeric, self-sealing septum covering the distal end opening.

3. The bioreactor of claim 1, further including a reagent reservoir at the distal end of the microchannel and in fluid communication with both the microchannel and the reaction reservoir, wherein the reagent reservoir has at least one reagent reservoir sidewall thereby defining a reagent reservoir volume and a reagent reservoir opening through which the second fluid enters the reagent reservoir.

4. The bioreactor of claim 3, wherein the reagent reservoir opening is closed by a penetrable, self-sealing, elastomeric septum covering the reagent reservoir opening.

5. The bioreactor of claim 1, wherein the proximal end opening further includes a section of the microchannel sidewall configured to have a reduced thickness sufficient to minimize formation of a meniscus at the proximal end opening by a fluid within the microchannel.

6. The bioreactor of claim 3, wherein the proximal end opening blocks the flow of liquid in the microchannel when the pressure of the liquid is not increased by an external source and there is a difference in viscosity between the first fluid and the second fluid, while permitting the diffusion of molecules across the proximal end opening.

7. The bioreactor of claim 1, wherein the cross-sectional area of the microchannel is less than 4 mm².

8. The bioreactor of claim 1, wherein the length of the microchannel is equal to or between 1 cm and 10 cm.

9. The bioreactor of claim 1, wherein the distal end opening of the pressure equalizing vent further includes a filter that is hydrophobic and capable of substantially preventing flow of liquid through the filter.

10. The bioreactor of claim 9, wherein the filter further includes an assembly of at least 2 layers with a first layer being adapted to prevent the passage of particles having an average size of at least approximately 80 microns, and a second layer being adapted to prevent the passage of particles having an average size of at least approximately 0.2 microns.

11. The bioreactor of claim 1, wherein the area of the distal end opening of the pressure equalizing vent further includes a section of the vent sidewall configured to have a reduced thickness sufficient to minimize formation of a meniscus at the proximal end opening by a fluid within the vent.

12. The bioreactor of claim 11, wherein the distal end opening of the pressure equalizing vent blocks the flow of liquid in the pressure equalizing vent when the pressure of the liquid is not increased by an external source and there is a difference in viscosity between the third fluid and the external environment, while permitting the diffusion of molecules across the distal end opening.

13. The bioreactor of claim 1, wherein the reaction reservoir is a hexahedron having a first longitudinal surface, a second longitudinal surface, a first lateral surface, a second lateral surface, a top surface, and a bottom surface, thereby defining a reaction reservoir length, width, and depth, and wherein the continuous microchannel has four sections substantially parallel to the length and greater in magnitude, four sections substantially parallel to the width and greater in magnitude, and two sections substantially parallel to the depth and greater in magnitude.

14. A diffusion controlling bioreactor that selectively controls the molecular diffusion between a first fluid, a second fluid, and a third fluid, comprising:

a reaction reservoir having at least one reaction reservoir sidewall thereby defining a reaction reservoir volume, wherein the reaction reservoir initially contains the first fluid;

a microchannel having at least one portion fully enclosed by at least one sidewall, in fluid communication with the reaction reservoir, having a proximal end with a proximal end opening, a distal end with a distal end opening, a length, the at least one microchannel sidewall defining a cross-sectional area, wherein the length and cross-sectional area are selected to obtain a predetermined rate of molecular diffusion between the first fluid and the second fluid, and wherein the microchannel is configured such when the microchannel is filled with the second fluid, and the second fluid is a liquid, that flow of the second fluid through the microchannel is laminar and the capillary action of the microchannel and the second fluid is such that the second fluid does not flow into the reaction reservoir unless the pressure of the second fluid is increased by an external source; and

a pressure equalizing vent in fluid communication with the reaction reservoir and a fluidic external environment, wherein the pressure equalizing vent is capable of equalizing pressure within the reaction reservoir with that of the external environment, wherein the pressure equalization vent has a distal end with a distal end opening in communication with the external environment, a length, at least one sidewall, having a sidewall thickness, and defining a cross-sectional area, and a proximal end with a proximal end opening in communication with the reaction reservoir, and wherein the pressure equalizing vent is configured such that when it is filled with a third fluid, and the third fluid is a liquid, that flow of the third fluid through the pressure equalizing vent is laminar and the capillary action of the pressure equalizing vent and the third fluid is such that the third fluid does not flow out of the distal end opening unless the pressure of the third fluid is increased by an external source; and

wherein the area of the distal end opening of the pressure equalizing vent includes a section of the vent sidewall configured to have a reduced thickness sufficient to minimize formation of a meniscus at the proximal end opening by a fluid within the vent.

15. The bioreactor of claim 14, further including a reagent reservoir at the distal end of the microchannel and in fluid communication with both the microchannel and the reaction reservoir, wherein the reagent reservoir has at least one reagent reservoir sidewall thereby defining reagent reservoir volume and a reagent reservoir opening through which the second fluid enters the reagent reservoir.

16. The bioreactor of claim 15, wherein the reagent reservoir opening is closed by a penetrable, self-sealing, elastomeric septum covering the reagent reservoir opening.

17. The bioreactor of claim 14, wherein the proximal end opening further includes a section of the microchannel sidewall configured to have a reduced thickness sufficient to minimize formation of a meniscus at the proximal end opening by a fluid within the microchannel.

18. The bioreactor of claim 17, wherein the microchannel proximal end opening blocks the flow of liquid in the microchannel when the pressure of the liquid is not increased by an external source and there is a difference in viscosity between the first fluid and the second fluid, while permitting the diffusion of molecules across the microchannel proximal end opening.

19. The bioreactor of claim 14, wherein the cross-sectional area of the microchannel is less than 4 mm².

20. The bioreactor of claim 14, wherein the length of the microchannel is equal to or between 1 cm and 10 cm.

21. The bioreactor of claim 14, wherein the vent distal end opening further includes a filter that is hydrophobic and capable of preventing flow of liquid through the filter.

22. The bioreactor of claim 21, wherein the filter further includes an assembly of at least 2 layers with a first layer being adapted to prevent the passage of particles having an average size of at least approximately 80 microns, and a second layer being adapted to prevent the passage of particles having an average size of at least approximately 0.1 microns.

23. The bioreactor of claim 14, wherein the distal end opening of the pressure equalizing vent blocks the flow of liquid in the pressure equalizing vent when the pressure of the liquid is not increased by an external source and there is a difference in viscosity between the third fluid and the external environment, while permitting the diffusion of molecules across the vent distal end opening.

24. The bioreactor of claim 14, wherein the reaction reservoir has a maximum first dimension in a primary axis, a maximum second dimension in a secondary axis, and a maximum third dimension in a tertiary axis, and the pressure equalizing vent is a continuous microchannel configured lying wholly exterior to the reaction reservoir and having at least one component in the primary axis with a magnitude greater than the maximum first dimension, at least one component in the secondary axis with a magnitude greater than the maximum second dimension, and at least third component in the tertiary axis with a magnitude greater than the maximum third dimension.

25. The bioreactor of claim 14, wherein the reaction reservoir is a hexahedron having a first longitudinal surface, a second longitudinal surface, a first lateral surface, a second lateral surface, a top surface, and a bottom surface, thereby defining a reaction reservoir length, width, and depth, and wherein the continuous microchannel has four sections substantially parallel to the length and greater in magnitude, four sections substantially parallel to the width and greater in magnitude, and two sections substantially parallel to the depth and greater in magnitude.

26. A diffusion controlling bioreactor that selectively controls the molecular diffusion between a first fluid, a second fluid, and a third fluid, comprising:

a reaction reservoir having at least one reaction reservoir sidewall thereby defining a reaction reservoir volume, wherein the reaction reservoir initially contains the first fluid;

a first microchannel having at least one portion fully enclosed by at least one sidewall, in fluid communication with the reaction reservoir, having a proximal end with a proximal end opening, a distal end with a distal end opening, a length, the at least one microchannel sidewall defining a cross-sectional area, wherein the length and cross-sectional area are selected to obtain a predetermined rate of molecular diffusion between the first fluid and the second fluid, and wherein the first microchannel is configured such when the first microchannel is filled with the second fluid, and the second fluid is a liquid, that flow of the second fluid through the first microchannel is laminar and the capillary action of the first microchannel and the second fluid is such that the second fluid does not flow into the reaction reservoir unless the pressure of the second fluid is increased by an external source, and wherein the proximal end opening of the first microchannel includes a section of the first microchannel sidewall configured to have a reduced thickness sufficient to minimize formation of a meniscus at the proximal end opening by a fluid within the first microchannel; and

a second microchannel in fluid communication with the reaction reservoir, having a proximal end with a proximal end opening, a distal end with a distal end opening, further including a reagent reservoir at the distal end of the second microchannel and in fluid communication with both the second microchannel and the reaction reservoir, wherein the reagent reservoir has at least one reagent reservoir sidewall thereby defining reagent reservoir volume and a reagent reservoir opening through which the third fluid enters the reagent reservoir, a length, at least one second microchannel sidewall, having a sidewall thickness, and defining a cross-sectional area, wherein the length and cross-sectional area are selected to obtain a predetermined rate of molecular diffusion between the first fluid and the third fluid, and wherein the second microchannel is configured such when the second microchannel is filled with the third fluid, and the third fluid is a liquid, that flow of the third fluid through the second microchannel is laminar and the capillary action of the second microchannel and the third fluid is such that the third fluid does not flow into the reaction reservoir unless the pressure of the third fluid is increased by an external source; and

wherein the proximal end opening of the first microchannel blocks the flow of liquid in the first microchannel when the pressure of the liquid is not increased by an external source and there is a difference in viscosity between the first fluid and the second fluid, while permitting the diffusion of molecules across the proximal end opening of the first microchannel.

27. The bioreactor of claim 26, wherein the reagent reservoir opening is closed by a penetrable, self-sealing, elastomeric septum covering the reagent reservoir opening.

28. The bioreactor of claim 26, wherein the proximal end opening of the second microchannel further includes a section of the second microchannel sidewall configured to have a reduced thickness sufficient to minimize formation of a meniscus at the proximal end opening by a fluid within the second microchannel.

29. The bioreactor of claim 26, wherein the proximal end opening of the second microchannel blocks the flow of liquid in the second microchannel when the pressure of the liquid is not increased by an external source and there is a difference in viscosity between the first fluid and the second fluid, while permitting the diffusion of molecules across the proximal end opening of the second microchannel.

30. The bioreactor of claim 26, wherein the length and cross-sectional area of the second microchannel are selected to achieve a predetermined rate of diffusion between the third fluid and the first fluid.

31. The bioreactor of claim 26, wherein the reaction reservoir is constructed of a material having a diffusion capacity and with sidewalls having a surface area such that the diffusion of oxygen across the sidewalls is predetermined for a predetermined unit of time.

32. The bioreactor of claim 31 wherein the oxygen diffusion across the reaction reservoir sidewalls is approximately 4.6 ml per day at atmospheric pressure and a temperature of 37°

C.