Microfluidic device comprising members for retaining objects within capillary traps

Abstract

The invention relates to a microfluidic device (1) comprising capillary traps (5) each provided with a microcavity (6) formed on a wall of a microchannel (2) for circulating a fluid and with a retaining member (7) extending in the microchannel (2) opposite the microcavity (6) of said capillary trap (5) so as to retain objects previously trapped in said microcavity (6) when said objects move from said microcavity (6) toward said retaining member (7), for example due to a change in the flow direction of the fluid in the microchannel (2).

Description

[0001] Description

[0002] Title: Microfluidic device comprising object retention organs within capillary traps

[0003] Technical field

[0004] The invention relates to the field of microfluidics and “lab-on-chip” systems.

[0005] The invention is of particular interest, in no way limiting, for the study of biological macromolecules by crystallography.

[0006] State of the prior art

[0007] Microfluidics can be defined as the study of flows and their implementation in microchannel networks whose dimensions are of the order of a micrometer. At this scale, capillary forces predominate over gravity forces.

[0008] Typically, a microfluidic device includes capillary traps designed to trap objects such as crystals or biological cells for analysis. These traps typically consist of cavities formed along microchannels through which a liquid is moved, with the objects suspended in this liquid.

[0009] An example of a microfluidic device and method for crystal analysis is described in the article by Lyubimov et al, entitled "Capture and X-ray diffraction studies of protein microcrystals in a microfluidic trap array" and published in Acta Crystallographica D71, pages 928-940, in 2015.

[0010] The introduction of the liquid into the microchannels is typically carried out using an injection tube which is disconnected after filling the device, causing a liquid suction phenomenon which tends to force the objects out of the capillary traps.

[0011] There is therefore a need to improve the capture of objects for analysis.

[0012] Statement of the invention

[0013] To this end, the subject of the invention is a microfluidic device comprising: - a support comprising a microchannel configured to allow a flow of a fluid from an inlet of the microchannel to an outlet of the microchannel,

[0014] - capillary traps succeeding one another between the inlet and the outlet of the microchannel, each of the capillary traps comprising a microcavity formed on a wall of the microchannel so as to open into the microchannel in order to trap one or more objects suspended in the fluid when the latter flows in the microchannel.

[0015] According to the invention, each of the capillary traps comprises a retention member extending into the microchannel opposite the microcavity of this capillary trap.

[0016] Such a retention member is thus configured to allow retention of at least one of the object(s) previously trapped in this microcavity in the event of movement of this object from this microcavity to this retention member.

[0017] The invention thus makes it possible to reduce the escape of objects trapped in the microcavities, particularly when the fluid changes direction of circulation, for example when removing an injection tube.

[0018] The invention makes it possible in particular to improve the reproducibility of the analysis results.

[0019] In the present description, a microchannel is a channel whose diameter or largest transverse dimension is of the order of a micrometer, i.e., a dimension less than 1 mm and greater than or equal to 1000 nm. Similarly, it is considered that a microcavity is a cavity whose diameter or largest dimension is of the order of a micrometer, i.e., a dimension less than 1 mm and greater than or equal to 1000 nm.

[0020] In a non-limiting manner, the objects can be crystals, cells or even particles whose size is also of the order of a micrometer, that is to say less than 1 mm and greater than or equal to 1000 nm.

[0021] The fluid may be liquid or gaseous. In one embodiment, for each of the capillary traps, the retention member comprises a retention surface.

[0022] The retention surface is a surface of the retention member that extends opposite the microcavity of the corresponding capillary trap.

[0023] The retention surface is preferably concave or hollow.

[0024] The retention surface and / or more generally the retention member may have an asymmetrical profile.

[0025] In one embodiment, for each of the capillary traps, the retention member comprises a first edge delimiting an upstream end of the retention surface and a second edge delimiting a downstream end of the retention surface, the first edge being located at a first distance from the microcavity of this capillary trap, the second edge being located at a second distance from the microcavity of this capillary trap, the first distance being greater than the second distance.

[0026] The terms "upstream" and "downstream" are defined here with reference to a direction of flow of the fluid in the microchannel, for example when the fluid is introduced therein.

[0027] A different distance from the microcavity between the first edge and the second edge makes it possible to promote the capture of objects when the fluid flows in a direction going from the inlet to the outlet of the microchannel and / or to promote their retention when the fluid changes direction.

[0028] Without limitation, the second distance may be greater than 1.2 times the first distance, for example equal to or close to 1.5 times the first distance.

[0029] By way of non-limiting example, the first distance may be 10 pm and the second distance 15 pm.

[0030] In one embodiment, for each of the capillary traps, the retention member extends along a flow direction of the microchannel, having a width less than or equal to a width of the microcavity of this capillary trap. In a non-limiting manner, the length of the retention member may be less than 0.8 times the width of the microcavity, for example equal to or close to 0.7 times the width of the microcavity.

[0031] By way of non-limiting example, the width of the microcavity may be 40 pm and the length of the retention member 28 pm.

[0032] In one embodiment, said microchannel is a primary microchannel, each of the capillary traps comprising a secondary microchannel opening into the microcavity of this capillary trap.

[0033] A secondary microchannel improves the capture of objects within the corresponding capillary trap.

[0034] According to an alternative embodiment, for each of the capillary traps, the secondary microchannel comprises:

[0035] - a first end through which it opens into the microcavity of this capillary trap which opens into a section of the primary microchannel and

[0036] - a second end through which it opens into another section of the primary microchannel.

[0037] Such an arrangement makes it possible to improve the efficiency of the capillary traps while reducing the compactness of the device.

[0038] In one embodiment, the device comprises an injection member such as a tube configured to introduce said fluid into the microchannel and means for connecting / disconnecting this injection member.

[0039] In other words, the device of the invention may comprise conventional fluid injection means.

[0040] The carrier may be a chip of a lab-on-a-chip type system.

[0041] The invention also relates to a method for analyzing objects comprising a step of injecting a fluid into the microchannel of a microfluidic device as defined above, said objects being suspended in the fluid thus injected. This method may comprise a step of optical detection of the objects retained by the capillary traps of the microfluidic device, for example by irradiating the objects with X-rays.

[0042] Other advantages and characteristics of the invention will appear on reading the detailed, non-limiting description which follows.

[0043] Brief description of the drawings

[0044] The following detailed description refers to the attached drawings in which:

[0045] - Figure 1 is a schematic view of a microfluidic device according to the invention, this device comprising a chip supporting a microfluidic network;

[0046] - Figure 2 is an enlargement of a part of the microfluidic network of the device of Figure 1, showing more precisely secondary microchannels belonging to capillary traps fluidically connected to a primary microchannel;

[0047] - Figure 3 is a further magnification of a portion of the microfluidic network of Figure 2, showing more precisely two adjacent capillary traps, each of these capillary traps comprising a microcavity, a secondary microchannel and a retention member.

[0048] Detailed description of embodiments

[0049] Figure 1 shows a non-limiting example of a microfluidic device 1 according to the invention.

[0050] The device 1 comprises a support 10 forming a chip of a laboratory-on-chip system, generally called “lab-on-chip” in English.

[0051] The support 10 comprises a microfluidic network forming a microchannel 2, called primary, having an inlet 3 and an outlet 4.

[0052] With reference to Figure 2, the microchannel 2 comprises sections 201-219 defining a serpentine flow trajectory or direction, making it possible in particular to increase the compactness of the device 1. The section 201, partially shown in Figure 2, comprises one end forming the inlet 3 of the microchannel 2 and another end connected to the section 202. Similarly, the section 219, partially shown in Figure 2, comprises one end forming the outlet 4 of the microchannel 2 and another end connected to the section 218.

[0053] In this example, sections 202, 204, 206, 208, 210, 212, 214, 216 and 218 extend parallel to each other and sections 203, 205, 207, 209, 211, 213, 215 and 217 respectively connect sections 202 and 204, 204 and 206, 206 and 208, 208 and 210, 210 and 212, 212 and 214, 214 and 216, 216 and 218 to each other.

[0054] Sections 203, 209 and 215 on the one hand and 205, 211 and 217 on the other hand define a width DI of the microfluidic network.

[0055] In this example, sections 204, 210 and 216 extend over the entire width DI of the microfluidic network while sections 202, 206, 208, 212, 214 and 218 extend over a distance corresponding to approximately half the width DI of the microfluidic network.

[0056] Considering a median axis passing between sections 203, 209 and 215 on the one hand and sections 205, 211 and 217 on the other hand, sections 206, 212 and 218 extend mainly on a first side of this median axis (on the left in Figure 2) and sections 202, 208 and 214 extend mainly on a second side of this median axis (on the right in Figure 2).

[0057] Such a geometry presents advantages in particular in terms of arrangement of capillary traps 5 of the device 1, described further below, and operation of the device 1.

[0058] In this example, microchannel 2 has a diameter of 200 pm and defines an overall volume of approximately 3.2 nL.

[0059] In a manner known per se, the microchannel 2 makes it possible to convey a fluid from the inlet 3 to the outlet 4, introduced into the microchannel 2 by an injection member (not shown) such as a tube removably connected to the inlet 3 of the microchannel 2. In this example, the fluid is a liquid in which objects are suspended.

[0060] Without limitation, the liquid may typically form a saline medium having a pH between five and nine.

[0061] The objects suspended in the liquid are in this example crystals having a size of the order of a micrometer, that is to say less than 1 mm and greater than or equal to 1000 nm, typically an average size between 10 pm and 15 pm.

[0062] This description applies by analogy to an implementation of the device 1 with another fluid, for example a gaseous fluid, and / or objects other than crystals.

[0063] Figure 3 shows two capillary traps 5 of the device 1.

[0064] Each of these capillary traps 5 comprises a microcavity 6, a retention member 7 and a microchannel 8, called a secondary microchannel.

[0065] In this example, the secondary microchannel 8 has a diameter of 10 pm.

[0066] The capillary trap 5 located on the right of Figure 3 will now be described.

[0067] The microcavity 6 is formed on a wall of the primary microchannel 2 so as to open into the primary microchannel 2 through an opening of width D2, the width D2 being considered according to the flow direction.

[0068] In this example, the microcavity 6 has a hemispherical shape so that the aforementioned opening is circular. The width D2 thus corresponds to an opening diameter of the microcavity 6.

[0069] The secondary microchannel s comprises a first end 8A through which it opens into the microcavity 6 and a second end 8B through which it opens into the primary microchannel 2.

[0070] In this particular example, this secondary microchannel 8 opens via its second end 8B into the section 204 of the primary microchannel 2 and via its first end 8A into the microcavity 6 which opens into the section 202 of the primary microchannel 2 (see figures 2 and 3). Still with reference to the capillary trap 5 located on the right of figure 3, the retention member 7 extends into the primary microchannel 2, in this case into the section 202, opposite the microcavity 6 of this capillary trap 5.

[0071] The retention member 7 comprises a surface 71, called retention, which extends opposite the microcavity 6, an opposite surface 72, an upstream surface 73 and a downstream surface 74.

[0072] The terms "upstream" and "downstream" refer to a flow direction SI of the liquid in the primary microchannel 2 when it moves from the inlet 3 to the outlet 4.

[0073] The surfaces 71-74 of the retention member 7 are separated from each other by edges 75-78 forming ridges in this example. The edge 75 delimits the surfaces 71 and 73, the edge 76 delimits the surfaces 72 and 73, the edge 77 delimits the surfaces 72 and 74 and the edge 78 delimits the surfaces 71 and 74.

[0074] The surfaces 71 and 72 define a thickness of the retention member 7.

[0075] According to the flow direction, the retention member 7 has a width D3 which corresponds in this example to a distance between the edges 75 and 77, the edge 75 delimiting an upstream end of both the retention surface 71 and the retention member 7, the edge 77 delimiting a downstream end of both the opposite surface 72 and the retention member 7.

[0076] The width D3 of the retention member 7 is less than the width D2 of the microcavity 6. In this particular example, D2 is equal to 40 pm and D3 is equal to 28 pm.

[0077] The retention member 7 is located at a distance from the microcavity 6.

[0078] More precisely, the edge 75 is located at a distance D4 from the microcavity 6, that is to say from said opening of the microcavity 6, while the edge 78, which delimits a downstream end of the retention surface 71, is located at a distance D5 from the microcavity 6, that is to say from said opening of the microcavity 6.

[0079] The distance D4 is here greater than the distance D5. In this example, D4 ​​is equal to 15 pm and D5 is equal to 10 pm. The retention member 7 has an asymmetrical geometry. In particular, the thickness of the retention member 7 is smaller at its upstream end than at its downstream end.

[0080] In this example, the retention surface 71 is concave and the opposing surface 72 is convex.

[0081] Such a geometry and such dimensions of the retention member 7 make it possible to both reduce disturbances to the flow of liquid in the microchannel 2 while contributing to the capture and retention of objects within this capillary trap 5.

[0082] The foregoing description applies by analogy to the capillary trap 5 located on the left of Figure 3 and to each of the other capillary traps 5 shown in Figure 2.

[0083] With reference to figure 2, the different capillary traps 5 of the device 1 follow one another between the inlet 3 and the outlet 4 of the primary microchannel 2 in series of 30 adjacent capillary traps 5.

[0084] A first series of capillary traps 5 follow one another along the section 202 in the sense that the microcavity 6 of each of these capillary traps 5 opens into the section 202 and the retention member 7 of each of these capillary traps 5 extends into the section 202, the secondary microchannel s of each of these capillary traps 5 opening through its second end 8B into the section 204.

[0085] Similarly, a second series of capillary traps 5 follow one another along the section 204, their microcavity 6 opening into the section 204, their retention member 7 extending into the section 202, their secondary microchannel 8 opening through their second end 8B into the section 206. A third series of capillary traps 5 follow one another along the section 208, their microcavity 6 opening into the section 208, their retention member 7 extending into the section 208, their secondary microchannel 8 opening through their second end 8B into the section 210. A fourth series of capillary traps 5 follow one another along the section 210, their microcavity 6 opening into the section 210, their retention member 7 extending into the section 210, their secondary microchannel 8 opening at their second end 8B into section 212.A fifth series of capillary traps 5 follow one another along the section 214, their microcavity 6 opening into the section 214, their retention member 7 extending into the section 214, their secondary microchannel 8 opening through their second end 8B into the section 216. A sixth series of capillary traps 5 follow one another along the section 216, their microcavity 6 opening into the section 216, their retention member 7 extending into the section 216, their secondary microchannel 8 opening through their second end 8B into the section 218.

[0086] The microfluidic network of the device 1 is thus formed by the primary microchannel 2 and by all the secondary microchannels 8 of the capillary traps 5.

[0087] When the liquid is injected into the microchannel 2 through its inlet 3, the different capillary traps 5 each make it possible to retain within the microcavity 6 one or more objects suspended in the liquid. Typically, the objects enter a trap 5 through the space extending between the microcavity 6 and the upstream end of the corresponding retention member 7.

[0088] The capture of objects is facilitated in particular by the resistance of the different parts of the microfluidic network to the flow of the liquid, which evolves as the capture progresses. More precisely, the secondary microchannels 8 having a lower hydraulic resistance than that of the primary microchannel 2, objects located at a certain position along the flow direction will tend to move towards a secondary microchannel 8 near which they are located so as to lodge in the microcavity 6 of the corresponding capillary trap 5. The objects thus captured tend to block this secondary microchannel 8 locally increasing the hydraulic resistance, so that uncaptured objects tend to move towards capillary traps 5 further downstream.

[0089] When the injection tube is removed from the device 1, a suction phenomenon occurs, typically resulting in a change in the direction of flow of the liquid, causing it to move in a direction S2 from the outlet 4 to the inlet 3 of the primary microchannel 2, or resulting in back-and-forth movements of the liquid within this microchannel 2. Typically, the removal of the injection tube produces a vacuum, corresponding to a volume of approximately 5 pL in this example, which sucks the liquid located in the microfluidic network out of it, this suction vacuum volume being in this case much greater than the overall volume of the liquid filling the microchannel 2, i.e. approximately 3.2 nL in this example. This typically tends to cause the objects to come out of the microcavities 6 in which they were trapped, and to move at least some of them towards the retention members 7 given the arrangement of the latter opposite the microcavities 6.The geometry of the retention members 7, in particular in this example the concavity of the retention surface 71 and the arrangement of the upstream edge 75, contribute to retaining these objects in the traps 5. An analysis can then be carried out, for example by optical detection of the objects thus retained in the capillary traps 5.

[0090] Many variants can be made to the microfluidic device 1 described above, in particular in terms of geometry and / or dimension of the microchannel 2, of the capillary traps 5 and / or in particular of the retention members 7, for example depending on the type of objects to be analyzed and / or the nature of the transport fluid used. Thus, in a variant not shown, said retention surface can have a non-curved hollow shape. For another example, the position and / or the orientation of the retention members can be different from those shown in FIG. 3.

Claims

Demands 1. Microfluidic device (1) comprising: - a support (10) comprising a microchannel (2) configured to allow the flow of a fluid from an inlet (3) of the microchannel (2) to an outlet (4) of the microchannel (2), - capillary traps (5) following one another between the inlet (3) and outlet (4) of the microchannel (2), each of the capillary traps (5) comprising a microcavity (6) formed on a wall of the microchannel (2) so as to open into the microchannel (2) in order to trap one or more objects suspended in the fluid when it flows into the microchannel (2), characterized in that each of the capillary traps (5) comprises a retention element (7) extending into the microchannel (2) opposite the microcavity (6) of this capillary trap (5).

2. Device (1) according to claim 1, wherein, for each of the capillary traps (5), the retention member (7) comprises a concave retention surface (71).

3. Device (1) according to claim 2, wherein, for each of the capillary traps (5), the retention member (7) comprises a first edge (75) delimiting an upstream end of the retention surface (71) and a second edge (78) delimiting a downstream end of the retention surface (71), the first edge (75) being located at a first distance (D4) from the microcavity (6) of this capillary trap (5), the second edge (78) being located at a second distance (D5) from the microcavity (6) of this capillary trap (5), the first distance (D4) being greater than the second distance (D5).

4. Device (1) according to any one of claims 1 to 3, wherein, for each of the capillary traps (5), the retention member (7) extends along a flow direction of the microchannel (2) with a width (D3) less than or equal to a width (D2) of the microcavity (6) of this capillary trap (5).

5. Device (1) according to any one of claims 1 to 4, wherein said microchannel (2) is a primary microchannel, each of the capillary traps (5) comprising a secondary microchannel (8) opening into the microcavity (6) of this capillary trap (5).

6. Device (1) according to claim s, wherein, for each of the capillary traps (5), the secondary microchannel (8) comprises: - a first end (8A) through which it opens into the microcavity (6) of this capillary trap (5) which opens into a section (202) of the primary microchannel (2) and - a second end (8B) through which it opens into another section (204) of the primary microchannel (2).

7. Device (1) according to any one of claims 1 to 6, comprising an injection member such as a tube configured to introduce said fluid into the microchannel (2) and means for connecting / disconnecting this injection member.

8. Device (1) according to any one of claims 1 to 7, wherein the support (10) is a chip of a lab-on-a-chip type system.

9. Method for analyzing objects comprising a step of injecting a fluid into the microchannel (2) of a microfluidic device (1) according to any one of claims 1 to 8, said objects being suspended in the fluid thus injected.

10. Method according to claim 9, comprising an optical detection step of objects retained by the capillary traps (5) of the microfluidic device (1).

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