Microfluidic system for controlling a concentration profile of molecules capable of stimulating a target

Patent metadata

Field	Value
Publication number	US-9404914-B2
Application number	US-201214003206-A
Country	US
Kind code	B2
Filing date	Mar 2, 2012
Priority date	Mar 4, 2011
Publication date	Aug 2, 2016
Grant date	Aug 2, 2016

Abstract

The invention relates to a microfluidic system for controlling a concentration profile of molecules capable of stimulating a target, for example formed by an assembly of living cells, this system comprising: —a microfluidic device ( 1 ) comprising at least one microfluidic channel ( 4 ) equipped with at least one inlet orifice ( 21 ) and with at least one outlet orifice ( 22 ) for at least one fluid; —at least one means for supplying the microfluidic channel ( 4 ) with at least one fluid comprising molecules capable of stimulating the target; —at least one chamber ( 8 ) or another microfluidic channel comprising a base ( 6 ) intended to receive the target; and —at least one microporous membrane ( 5 ) separating the chamber ( 8 ) or the other microfluidic channel from the microfluidic channel ( 4 ), said microporous membrane ( 5 ) being positioned away from the base ( 6 ) so that when the supply means provides the microfludic channel ( 4 ) with said at least one fluid flowing in laminar flow in contact with the microporous membrane ( 5 ), the molecules capable of stimulating the target then diffuse, after having passed through the microporous membrane ( 5 ) through the chamber ( 8 ) or said other microfluidic channel in order to finally form a stable concentration profile in this chamber ( 8 ) or this other microfluidic channel.

First claim

The invention claimed is:

1. A microfluidic system comprising:
a microfluidic device comprising at least one microfluidic channel having at least:
a first branch provided with an inlet orifice for a first fluid,
a second branch provided with an inlet orifice for a second fluid comprising molecules likely to stimulate a target;
a common branch for said fluids, provided with an outlet orifice for said fluids, the first and second branches combining in the common branch;
at least one fluid source connected to the inlet orifices of the at least one microfluidic channel for supplying the channel with said fluids;
at least one chamber or another microfluidic channel having a base configured to receive the target, wherein the target is formed by a set of living cells; and
at least one microporous membrane arranged at an interface of the common branch upstream from the outlet orifice, said microporous membrane being positioned to separate the at least one chamber or the other micro fluidic channel from the common branch at least one microfluidic channel,
said microporous membrane being impervious to said fluids and configured to prevent said fluids from going through to the other side of the membrane and to allow said molecules to diffuse therethrough;
said microporous membrane not directly contacting the base so that when the at least one fluid source supplies the at least one microfluidic channel with said fluids, the diffusion of molecules likely to stimulate the target occurs, after going through the microporous membrane, then through the at least one chamber or said other microfluidic channel, and wherein after the diffusion of the molecules likely to stimulate a reaction with the target in the chamber or the other micro fluidic channel, a concentration profile in the at least one chamber or the other microfluidic channel is generated.

2. The microfluidic system according to claim 1 , wherein the microfluidic channel comprises a cover made of a material selected from the group consisting of: glass or silicon, a non-elastomeric photocrosslinked polymer, a metal, an alloy that is an electrical conductor or semiconductor, a ceramic, quartz, sapphire, and an elastomer.

3. The system according to claim 1 , wherein said at least one inlet orifice and said at least one outlet orifice for the fluids are formed in the cover.

4. The microfluidic system according to claim 1 , wherein the microfluidic channel comprises at least one wall made of a photo-cured and/or thermally cured resin.

5. The microfluidic system according to claim 1 , wherein the microporous membrane extends transversely on the side wall of the microfluidic channel to close the bottom of said channel.

6. The microfluidic system according to claim 1 , wherein the microfluidic channel is organized on several levels, each level having at least one inlet orifice for at least one fluid.

7. The microfluidic system according to claim 1 , wherein the base of the chamber or of said other microfluidic channel is made of an optically transparent material.

8. The microfluidic system according to claim 1 , wherein the chamber or said other microfluidic channel comprises side walls made of a photo-cured and/or thermally cured resin.

9. The microfluidic system according to claim 8 , wherein the microporous membrane extends transversely between the side walls ( 7 a , 7 b ) of the at least one chamber or of said other microfluidic channel, to close a top of said at least one chamber or of said other microfluidic channel.

10. The microfluidic system according to claim 1 , wherein the microporous membrane is made of a material selected from a group consisting of: glass, polycarbonate, polyester, polyethylene terephthalate, quartz, silicon, silica and silicon carbide.

11. The microfluidic system according to claim 1 , wherein the microporous membrane comprises pores at a density between 10 3 and 10 10 pores/cm 2 .

12. The microfluidic system according to claim 1 , wherein the pores have a hydraulic diameter between 0.05 μm and 12 μm.

13. The microfluidic system according to claim 1 , wherein an optical visualization means is provided.

14. The microfluidic system according to claim 13 , wherein the optical visualizing means uses a technique selected from the group consisting of photoactivated localization microscopy and stimulated emission depletion microscopy.

15. The microfluidic system according to claim 1 , wherein the first and second fluids only differ by the concentration of stimulation molecules.

16. The microfluidic system according to claim 15 , wherein the first and second fluids only differ by the presence of stimulation molecules.

17. The microfluidic system according to claim 1 , wherein the first and second fluids flow in the common branch in a laminar regime.

18. The microfluidic system according to claim 1 , comprising a third branch provided with an inlet orifice for a third fluid comprising stimulation molecules, wherein the first, second and third branches combine in the common branch, wherein the third fluid flows in the common branch between the first and second fluids, and wherein the concentration of stimulation molecules in the third fluid is comprised between the concentration of stimulation molecules in the first fluid and the concentration of stimulation molecules in the second fluid.

19. The microfluidic system according to claim 18 , wherein the third fluid is a mixture of the first and second fluids.

20. A method for controlling a concentration profile of molecules likely to stimulate a target comprising:
providing a microfluidic system according claim 1 ;
placing the target on the base, wherein the target is formed by a set of living cells;
supplying the first branch with a first fluid;
supplying the second branch with a second fluid comprising molecules, the first and second branches combining in the common branch;
wherein, when the fluids are supplied, the diffusion of molecules likely to stimulate the target occurs, after going through the microporous membrane, through the at least one chamber or said other microfluidic channel, and wherein, after the diffusion of the molecules likely to stimulate a reaction with the target, a concentration profile in the at least one chamber or the other microfluidic channel is generated.

Assignees

• Dahan Maxime
• Morel Mathieu
• Galas Jean-Christophe
• Studer Vincent
• Bartolo Denis
• Centre Nat De La Rech Scientifique-Cnrs
• Ecole Normale Supérieure
• Univ Pierre Et Marie Curie Paris 6
• Fonds De L'Espci-Georges Charpak
• Univerité Bordeaux Segalen

Inventors

• Dahan Maxime
• Morel Mathieu
• Galas Jean-Christophe
• Studer Vincent
• Bartolo Denis

Classifications

• B01L2300/087

  Multiple sequential chambers · CPC title

• B01L2200/0652

  Sorting or classification of particles or molecules · CPC title

• G01N33/5008 Primary

  for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics · CPC title

• B01L2200/0647

  Handling flowable solids, e.g. microscopic beads, cells, particles · CPC title

• B01L3/5027

  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 · CPC title