Device and method for the fluid connection of microfluidic containers

Abstract

The invention relates to a unidirectional fluid connection device (J, J1, J2, J3), the device (J, J1, J2, J3) comprising an intermediate fluid storage capacity (B) which is configured to temporarily store a given volume of a fluid, the device (J, J1, J2, J3) comprising an inlet channel (I) for the fluid coming into the device (J, J1, J2, J3) and an outlet channel (O) for the fluid going out of the device (J, J1, J2, J3), the device (J, J1, J2, J3) further comprising a first one-way valve (V1) which is configured to provide one-way fluid communication from the inlet channel (I) to the storage capacity (B), the device (J, J1, J2, J3) further comprising a second one-way valve (V2) which is configured to provide one-way fluid communication from the storage capacity (B) to the outlet channel (O).

Description

[0001] DESCRIPTION

[0002] TITLE: Device and method for the fluidic connection of microfluidic containers

[0003] The present invention relates to the field of microfluidic systems, and in particular to connection devices between microfluidic systems.

[0004] Current multi-organ microfluidic systems, i.e. allowing the culture of different organs, have limitations:

[0005] • Multi-organ microfluidic systems, also known as multi-organ chips, are usually produced in complex dedicated chips, which do not allow studying one part of the system in isolation before studying the system as a whole. Any change to a part requires the design of a new custom chip.

[0006] • Perfusion systems for circulating culture medium between different organs involve large quantities of tubing and external pumps that make the installation complex and difficult to implement. In addition, these systems involve large dead volumes in the pipes, which waste molecules that are expensive to produce at the pre-clinical stage.

[0007] • Multi-organ chips are currently poorly compatible with standard microfluidic system formats, making high-throughput screening impossible, despite growing market demand for systems generating large amounts of standardized and repeatable data.

[0008] Furthermore, in order to cultivate relevant endothelial barriers in microfluidic systems – which is also necessary to interface multi-organ chips with a vascular network – a certain level of directional shear stress must be applied to the cells, to mimic the friction of blood – or blood cells – on endothelial cells, via their culture medium. This shear stress will give the endothelial cells their elongated appearance and will confer the required porosity to the barrier. Providing shear stress is therefore a key element to enable the formation of blood-brain barriers (BBBs), for example.

[0009] The disadvantages of current systems for generating shear stress are:

[0010] • the perfusion systems for circulating the culture medium – and generating shear stress in the process – are the same as the multi-organ microfluidic systems mentioned above and therefore have the same limitations;

[0011] • Oscillating systems that tilt multi-organ microfluidic systems to generate gravity flow eliminate the complexities of perfusion systems but are limited by the low pressure (hence low shear stress) that gravity can provide under these conditions, and deviate from in vivo conditions by applying shear stress in both directions instead of just one direction.

[0012] The invention therefore aims to propose a solution to all or part of these problems.

[0013] To this end, the present invention relates to a one-way fluid connection device, the device comprising an intermediate fluid storage capacity configured to temporarily store a determined volume of a fluid, the device comprising an inlet channel for the fluid in the device and an outlet channel for the fluid coming from the device, the device further comprising a first one-way valve configured to ensure one-way fluid communication from the inlet channel to the storage capacity, the device further comprising a second one-way valve configured to ensure one-way fluid communication from the storage capacity to the outlet channel.

[0014] According to one embodiment, the invention comprises one or more of the following features, alone or in a technically acceptable combination.

[0015] According to one embodiment, the device is configured to form a one-way fluid connection from a first container to a second container, when the inlet channel is placed in fluid communication with the first container and when the outlet channel is placed in fluid communication with the second container.

[0016] According to these provisions, the device is configured to transfer the determined volume of fluid from the first container to the second container, when the fluidic connection from the first container to the second container is established with the device, and when the device is actuated by an actuator fluidically coupled with the intermediate fluidic storage capacity, the actuator being configured to implement at least one suction and discharge cycle, the suction and discharge cycle comprising a generation of a suction depression towards the storage capacity of the device to suck the determined volume of fluid from the first container to the intermediate storage capacity, followed by a generation of a discharge pressure in the storage capacity of the device to discharge the determined volume of fluid from the intermediate storage capacity to the second container.

[0017] According to one embodiment, the first container is a first microfluidic reservoir, and the second container is a second microfluidic reservoir.

[0018] A reservoir is said to be microfluidic if it belongs to a microfluidic chip, or if it has the dimensions of a reservoir of a microfluidic chip. A microfluidic chip is composed of at least two reservoirs connected by at least one channel.

[0019] According to one embodiment, the determined volume of fluid transferred is between 5 pl and 200 pl, for example 10 pl, or 50 pl, or 100 pl.

[0020] According to one embodiment, a distance d between the first container and the second container is between 1 mm and 20 mm, for example 4.5 mm or 9 mm, or 18 mm.

[0021] According to one embodiment, the first one-way valve is one of a ball check valve, a swing check valve, a disc check valve, a tesla valve, a membrane.

[0022] According to one embodiment, the second one-way valve is one of a ball check valve, a swing check valve, a disc check valve, a tesla valve, a membrane.

[0023] According to one embodiment, an opening pressure of the first valve, respectively of the second valve, is determined as a function of a suction pressure, respectively of a discharge pressure.

[0024] According to one embodiment, the actuator comprises a pump.

[0025] According to one embodiment, the pump comprises a mechanical piston.

[0026] According to one embodiment, the pump acts on an incompressible liquid fluid, or on a compressible gaseous fluid, to create the suction pressure and the discharge pressure. According to one embodiment, a filter is positioned at an interface between the actuator and the storage volume, to prevent the inclusion of foreign particles in the transferred fluid volume.

[0027] According to one embodiment, the intermediate storage capacity of the device is configured by means of a volume limiter, for example a piston, or a float, or a flexible membrane, closing the inlet of the compressible gaseous fluid to limit a sucked volume of the fluid by the device when the actuator is implemented by means of a pump acting on a compressible gaseous fluid.

[0028] According to one embodiment, the device further comprises a bridge configured to place the first container and the second container in bidirectional fluid communication, the bridge having a hydraulic resistance greater than a hydraulic resistance of the assembly comprising, by the inlet channel, the first valve, the storage capacity, the second valve, the outlet channel.

[0029] According to one embodiment, the device comprises a coupling interface of the storage capacity of the device, the coupling interface being an opening of the storage capacity configured to ensure a sealed fluidic coupling with a tip further in fluidic communication via a channel of the micro-fluidic chip actuator adjusted to the opening of the storage capacity, such that the fluidic coupling between the tip and the opening is sealed when the tip of the actuator is coupled to the opening of the storage capacity.

[0030] According to these provisions, the suction pressure and the discharge pressure created by the actuator are transmitted in full to the storage capacity via the sealed fluid coupling. The fluid coupling or a fluid connection is said to be sealed when the coupling at the coupling interface allows fluid communication between the two coupled entities without any part of the fluid being able to escape outside the coupling interface, so that all the fluid that flows from one coupled entity to the other coupled entity arrives in the other coupled entity.

[0031] According to one embodiment, the opening of the storage capacity includes an O-ring that provides fluid tight coupling between the actuator tip and the opening of the storage capacity when the actuator tip enters the opening of the storage capacity.

[0032] According to one embodiment, the opening of the storage capacity is shaped as a cone open towards the outside of the storage capacity, the open cone being configured to receive a corresponding closed cone positioned at the outlet of the tip of the actuator, such that when the tip of the actuator enters the opening of the storage capacity, contact between an outer surface of the closed cone of the tip and an inner surface of the open cone of the opening ensures the fluidic sealed coupling between the tip of the actuator and the opening of the storage capacity.

[0033] According to one embodiment, the opening of the storage capacity B may be in the shape of a closed cone, and the tip of the actuator A in the shape of an open cone corresponding to the closed cone of the opening of the storage capacity B, so that when the closed cone of the opening of the storage capacity B penetrates inside the open cone of the tip of the actuator A the fluid coupling between the tip of the actuator A and the opening of the storage capacity B is sealed.

[0034] According to one embodiment, a connection between the inlet channel and the first container is sealed. According to one embodiment, a connection between the outlet channel and the second container is sealed.

[0035] According to these provisions, the sealing of the connection between the inlet channel and the first container and the sealing of the connection between the outlet channel and the second container allows a fully pressure-controlled system, which no longer depends on its orientation relative to the ground or atmospheric pressure. This sealing makes it possible, for example, to envisage use without the presence of gravity (as on an orbital station) or without conventional atmospheric pressure (as in a submarine, an airplane or a module on the Moon or Mars).

[0036] According to another aspect, the invention also relates to an assembly comprising a first container and a second container and a device according to one of the embodiments described above, the inlet channel of the device being placed in fluid communication with the first container and the outlet channel being placed in fluid communication with the second container so as to create a one-way fluid connection from the first container to the second container.

[0037] According to one embodiment, the assembly according to the invention comprises one or more of the following characteristics, alone or in combination.

[0038] According to one embodiment, the first container of the assembly is a reservoir of a microfluidic chip, and the second container of the assembly is another reservoir of the microfluidic chip.

[0039] According to one embodiment, the reservoir and the other reservoir of the microfluidic chip are further in fluid communication via a channel of the microfluidic chip. According to these arrangements a single-chip closed loop is formed, such that the fluid circulates in a circular manner in the microfluidic chip when an actuator is connected to the device.

[0040] According to these provisions, an intermittent shear stress with a fixed direction is generated by the flow speed controlled by the actuator, without the need for an external recirculation device passing through a pump. This shear stress makes it possible to reproduce the natural stress experienced by the cells and allows them to organize and stretch longitudinally as in the vascular system, thus approaching their mode of operation in vivo.

[0041] According to one embodiment, the microfluidic chip further comprises a central container containing an organ to be perfused by the vascular system thus reproduced, the central container being provided with a nano-porous membrane and the channel of the microfluidic chip passing through this central container.

[0042] According to one embodiment, the channel of the microfluidic chip has a hydraulic resistance greater than a hydraulic resistance of the assembly comprising the inlet channel, the first valve, the storage capacity, the second valve, the outlet channel.

[0043] According to one embodiment, the first container is one of the reservoirs of a microfluidic chip and the second container is one of the reservoirs of another microfluidic chip.

[0044] According to one embodiment, the other reservoir among the reservoirs of the microfluidic chip, and the other reservoir among the reservoirs of the other microfluidic chip are in fluid communication via a fluid bridge, said fluid bridge being a passive fluid bridge, or a device according to one of the embodiments described above. According to these provisions, when the fluid bridge is passive, the level of the fluid is identical in all the reservoirs after a determined time after an implementation of the actuator to transfer a determined volume of fluid from the first container to the second container.

[0045] According to one embodiment, the first reservoir and / or the second reservoir belong to microfluidic chips of a set of microfluidic chips arranged according to a standard format, such as the format defined by the ANSI SLAS 4-2004 (R2012) standard, also recognized as ANSI / SBS 4-2004.

[0046] According to one embodiment, a connection between the inlet channel and the first container is sealed.

[0047] According to one embodiment, a connection between the outlet channel and the second container is sealed.

[0048] According to another aspect, the invention also relates to a network of microfluidic chips interconnected in series comprising a plurality of N microfluidic chips, N being an integer greater than or equal to 2, interconnected by another plurality of N fluidic bridges, in which the plurality of N fluidic bridges comprises P passive fluidic bridges, and Q devices according to one of the embodiments described above, where P and Q are integers greater than or equal to 1, such that P+Q=N or P+Q=N+1, in which:

[0049] - each microfluidic chip of the plurality of N microfluidic chips is connected with at least one other microfluidic chip of the plurality of N microfluidic chips, via a passive fluidic bridge, or via a device according to one of the embodiments described above so as to form an assembly according to one of the embodiments described above, - the network comprises at least one assembly according to one of the embodiments described above;

[0050] - if the network comprises an assembly according to one of the embodiments described above and another assembly according to one of the embodiments described above, one direction of a fluid connection of the entire network and another direction of another fluid connection of the other assembly of the network are identical.

[0051] According to one embodiment, a microfluidic chip of the network is connected, via a passive fluidic bridge or via a device according to one of the embodiments described above, to an input container.

[0052] According to one embodiment, a microfluidic chip of the network is connected, via a passive fluidic bridge or via a device according to one of the embodiments described above, to an output container.

[0053] In one embodiment, the plurality of microfluidic chips are arranged in a standard format, such as the format defined by ANSI SLAS 4-2004 (R2012), also recognized as ANSI / SBS 4-2004.

[0054] According to one embodiment, the fluidic bridges of the plurality of N fluidic bridges, comprising a subset of devices according to one of the embodiments described above, are arranged according to a predetermined configuration configured to correspond to a corresponding predetermined arrangement of microfluidic chips, arranged for example according to the format defined by the ANSI SLAS 4-2004 (R2012) standard, also recognized as ANSI / SPB 4-2004.

[0055] According to one embodiment, the fluid bridges of the plurality of fluid bridges arranged according to the predetermined configuration are preassembled on an assembly frame to which they are securely fixed. According to another aspect, the invention also relates to a method for creating a one-way active fluid connection from a first container to a second container, with at least one device according to one of the embodiments described above, the method comprising the following steps:

[0056] - placing the input channel of the at least one device in the first container;

[0057] - place the output channel of the at least one device in the second container.

[0058] According to another aspect, the invention also relates to a method for transferring a determined volume of a fluid with at least one device according to one of the embodiments described above, the method comprising the following steps:

[0059] - fluidly coupling to the storage capacity of the at least one device, an actuator configured to implement at least one suction and discharge cycle, the suction and discharge cycle comprising a generation of a suction depression towards the storage capacity of the at least one device to suck the determined volume of fluid from a first container towards the intermediate storage capacity, followed by a generation of a discharge pressure in the storage capacity of the at least one device to discharge the determined volume of fluid from the intermediate storage capacity towards a second container;

[0060] - implement at least one suction and discharge cycle of the actuator.

[0061] According to one implementation mode, the cycle is implemented according to a symmetrical period.

[0062] According to one mode of implementation, the cycle is implemented according to a period including a time pause during which the pressure is kept constant. According to one mode of implementation, the cycle is implemented according to an asymmetrical period.

[0063] According to one implementation mode, the cycle is implemented aperiodically.

[0064] According to one embodiment, the at least one device comprises at least two devices, and the actuator is fluidically coupled in parallel to the storage capacity of the at least two devices, so as to create the suction depression then the discharge pressure, in parallel in the storage capacity of each device of the at least two devices.

[0065] According to another aspect, the invention also relates to a method for storing in the storage capacity of a device according to one of the embodiments described above, a determined quantity of fluid, for example before or after the implementation of a method according to one of the embodiments described above, and for recovering the determined quantity of fluid, in a manner not synchronized with the storage, the method comprising the following steps: - fluidically coupling to the storage capacity of the at least one device, an actuator configured to implement at least one suction step, then a discharge step, the suction step and the discharge step not being synchronized,

[0066] - implement the suction step of the determined volume in the storage capacity of the device,

[0067] - implement, in a manner not synchronized with the previous step, the step of discharging the determined volume of fluid present in the storage capacity of the device.

[0068] For a better understanding, an embodiment and / or implementation of the invention is described with reference to the attached drawings representing, by way of non-limiting example, an embodiment or implementation respectively of a device and / or a method according to the invention. The same references in the drawings designate similar elements or elements whose functions are similar.

[0069] [Fig. 1] is a schematic presentation of a fluid connection device according to one embodiment of the invention

[0070] [Fig. 2] is a graphical representation of different functions (A, B, C, D) for controlling the pressure as a function of time generated by an actuator coupled to the fluid connection device according to the invention.

[0071] [Fig. 3] is another view of a fluid connection device according to an embodiment of the invention

[0072] [Fig. 4] is a schematic presentation of an embodiment of an actuator of a fluidic connection device according to an embodiment of the invention [Fig. 5] is a schematic presentation of an embodiment of two fluidic connection devices according to an embodiment of the invention [Fig. 6] is a schematic presentation of a coupling between an actuator and a fluidic connection device according to an embodiment of the invention [Fig. 7] is a schematic presentation of another coupling between an actuator and a fluidic connection device according to an embodiment of the invention [Fig. 8] is a schematic presentation of a coupling between a fluidic connection device and the microfluidic containers, according to an embodiment of the invention

[0073] [Fig. 9] is a schematic presentation of an embodiment of a fluidic connection device for implementing a microfluidic single-chip closed loop.

[0074] [Fig. 10] is a schematic presentation of another mode of implementation of a fluidic connection device to implement a single-chip microfluidic closed loop.

[0075] [Fig. 11] is a schematic presentation of another embodiment of a fluidic connection device for implementing a closed loop with two microfluidic chips connected via the fluidic connection device, with passive feedback. [Fig. 12] is a schematic presentation of another embodiment of a fluidic connection device for implementing a closed loop with two microfluidic chips connected via the fluidic connection device, with active feedback using a second fluidic connection device.

[0076] [Fig. 13] is a schematic presentation of another embodiment of more than two fluidic connection devices to implement an open loop with two microfluidic chips connected to each other via the fluidic connection devices and connected respectively to an input container and to an output container.

[0077] [Fig. 14] is a schematic presentation of another embodiment of more than two fluidic connection devices to implement different connections between microfluidic chips of a standard support in the format defined by the ANSI SLAS 4-2004 (R2012) standard, also recognized as ANSI / SBS 4-2004.

[0078] [Fig. 15] is a schematic presentation of another mode of implementation of a fluid connection device with an error compensation fluid bridge.

[0079] [Fig. 16] is a schematic presentation of another mode of implementation of a fluid connection device for storing a determined volume of fluid before or after implementation of a fluid connection device.

[0080] [Fig. 17] is a schematic presentation of a predetermined configuration of devices according to the invention.

[0081] [Fig. 18] is a schematic presentation of an intermediate storage capacity of the device, according to three particular embodiments of the invention represented respectively in Fig. 18a, Fig. 18b, and Fig. 18c, in which the intermediate storage capacity is configured to limit in a predetermined manner a volume of fluid transferred.

[0082] [Fig. 19] is a schematic diagram of the sequence of steps in a method for fluidically connecting two containers according to an embodiment of a fluidic connection device. [Fig. 20] is a schematic diagram of the sequence of steps in a method for transferring a determined volume of fluid according to an embodiment of at least one fluidic connection device

[0083] [Fig. 21] is a schematic diagram of the sequence of steps in a method for storing a determined volume of fluid before or after implementation of a fluid connection device.

[0084] The device according to the invention is intended to allow the interconnection of elementary microfluidic chips to form more complex systems, without adding tubes and dead volume.

[0085] For this purpose, the invention relates to an elementary device J for monodirectional fluid connection, configured to establish a fluid connection between a first container R1 and a second container R2. Figure 1 illustrates an embodiment of an elementary device J according to the invention. Figure 3 is another representation of an elementary device J according to the invention.

[0086] With reference to Figures 1 and 3, said fluid connection device J is described below. The device J comprises an intermediate fluid storage capacity B, intended to temporarily store a determined volume of a fluid taken from the first container R1, while waiting for said determined volume of fluid to be transferred into the second container R2. The device J comprises a fluid inlet channel I, intended to allow the taking of said determined volume of fluid from the first container R1. The device J also comprises an outlet channel O of said determined volume of fluid, intended to allow the evacuation of said determined volume of fluid, towards the second container R2.The device J further comprises a first one-way valve V1 configured to ensure one-way fluid communication from the inlet channel I to the storage capacity B, and a second one-way valve V2 configured to ensure one-way fluid communication from the storage capacity B to the outlet channel O. According to these provisions, the device J is configured to transfer the determined volume of fluid from the first container R1 to the second container R2, when the inlet channel is placed in fluid communication with the first container R1 and when the outlet channel O is placed in fluid communication with the second container R2.

[0087] To effect a transfer of the determined volume of fluid from the first container R1 to the second container R2, the device J is actuated by an actuator A fluidically coupled with the intermediate fluid storage capacity B. The actuator A is configured to implement at least one suction and discharge cycle, the suction and discharge cycle comprising a generation of a suction depression towards the storage capacity B of the device J to suck the determined volume of fluid from the first container R1 to the intermediate storage capacity B, followed by a generation of a discharge pressure in the storage capacity B of the device J to discharge the determined volume of fluid from the intermediate storage capacity B to the second container R2.

[0088] During the suction phase, the aspirated fluid can only come from the inlet channel I due to the unidirectional nature of the valve V2 whose non-return mechanism prevents the aspirated fluid from coming from the outlet channel O.

[0089] During the discharge phase, the discharged fluid can only be discharged towards the outlet channel O due to the unidirectional nature of the valve V1 whose non-return mechanism prevents the fluid from being discharged towards the inlet channel I.

[0090] Thus, after a complete suction then discharge cycle, the device J activated by the actuator A has transferred a determined volume of fluid from the inlet channel I to the outlet channel O, over a distance d, where d is the distance separating the inlet channel I from the outlet channel O. According to an exemplary embodiment, the first container R1 is a first microfluidic reservoir, and the second container R2 is a second microfluidic reservoir, the first microfluidic reservoir and the second microfluidic reservoir forming a microfluidic chip PU. A reservoir is said to be microfluidic if it belongs to a microfluidic chip PU or if it has dimensions typical of those of a reservoir of a microfluidic chip. A microfluidic chip is composed of at least two reservoirs connected by at least one channel.

[0091] By way of example, the determined volume of fluid transferred using the device according to the invention is representative of the microfluidic volumes moved from one microfluidic reservoir to another microfluidic reservoir, belonging to the same microfluidic chip, or to two separate microfluidic chips, of the order of a few pl to a few hundred pl, typically between 5 pl and 200 pl, for example 10 pl, 50 pl, 100 pl or 150 pl.

[0092] More particularly, a distance d between the first container and the second container is between 1 mm and 20 mm, for example 4.5 mm or 9 mm, or 18 mm. These orders of magnitude correspond in particular to the standard formats of microfluidic chip sets, as defined by the ANSI SLAS 4-2004 (R2012) standard, also recognized as ANSI / SBS 4-2004.

[0093] The first one-way valve V1 and / or the second one-way valve V2 may take the form, for example, of a ball check valve, or a swing check valve, or a disc check valve, or a Tesla valve, or a simple membrane. The non-return mechanism of the first valve V1 may be different from the non-return mechanism of the second valve V2. An opening pressure of the first valve V1, respectively of the second valve V2, is determined as a function of a suction pressure, respectively of a discharge pressure. As an example, illustrated in Figure 4, the actuator A may comprise a pump P, configured for example to act on an incompressible liquid fluid FLI, or on a compressible gaseous fluid FGC, to successively create the suction pressure then the discharge pressure.

[0094] The P pump may include a mechanical piston.

[0095] When the P pump is configured to act on a compressible gas, the volume of fluid transferred during activation of the J device will depend on the opening pressures and the compressibility of the gas used by the P pump. However, the use of a compressible gas allows activation of the J device remotely without liquid exchange with the exterior of the J device, which simplifies cleaning and avoids liquid contamination.

[0096] According to a particular embodiment shown in Figure 18, the intermediate storage capacity B of the device J is configured by means of a volume limiter L1, L2, L3, closing the inlet of the compressible gas to limit a volume of fluid sucked by the device J when the actuator A is implemented by means of a pump P acting on a compressible gas. Said volume limiter may for example be a piston L1, as shown in Figure 18a, or a float L2 as shown in Figure 18b, or a flexible membrane L3 as shown in Figure 18c.

[0097] When the pump P is configured for example to act on an incompressible liquid, the volume of fluid transferred upon activation of the device J is fixed and independent of the suction and discharge pressure, as long as these are higher than the opening pressure of the valves V1, V2; however, this requires local actuation, such as for example an integrated mechanical piston, to avoid external contamination and leak management during detachments. According to an exemplary embodiment, a filter Fl is positioned at an interface between the actuator A and the storage volume B, to avoid the inclusion of foreign particles in the volume of fluid transferred.

[0098] Several devices J, J' according to the invention can be activated in parallel by a single actuator A, as illustrated in Figure 5. According to this arrangement, the suction then discharge pressure generated by the actuator A is exerted in parallel on the storage capacity of each device J, J', so that each device J, J' simultaneously transfers a determined volume of fluid respectively from a first container R1 to a second container R2, belonging for example to a first microfluidic chip PU, and from another first container RT to another second container R2', belonging for example to another first microfluidic chip PU'.

[0099] The actuator A which controls the device(s) J, J' can sequence the suction and discharge periods in several ways over time, leading to several effects. We can consider that the actuation system can be controlled by any pressure or displacement function over time. As an example, Figure 2 shows four different profiles A, B, C, D of the evolution of the pressure PR, represented on the ordinate, as a function of the time T represented on the abscisce during the sequence of actuation cycles of the device(s) J':

[0100] A - symmetrical periodic control. This actuation generates an equivalent, symmetrical pressure variation on input I and output O. This is useful if one is seeking to generate an equivalent flow at several locations in the system.

[0101] B - Periodic control with pauses. This actuation is similar to A, but pauses in the system allow the different regions of the fluid circuit to recover a static pressure balance before being subjected to a next fluid transfer. C - Asymmetric periodic control. This actuation allows the inlet and outlet flows to be differentiated, which makes it possible to modulate the effects of the flows on microfluidic cells, for example.

[0102] D - Multi-periodic or aperiodic control. This actuation makes it possible to simulate more complex or erratic events, such as cardiac arrest or fibrillation, for example.

[0103] As illustrated in Figures 6 and 7, the intermediate storage capacity B of the device J comprises a coupling interface ICC, ICJT, configured to allow a sealed fluid coupling between the actuator A and the storage capacity B, such that the suction pressure and the discharge pressure created by the actuator A is transmitted entirely to the storage capacity B via said sealed fluid coupling. According to a particular exemplary embodiment, the coupling interface ICC, ICJT is an opening of the storage capacity B configured to receive a tip of the actuator A adjusted to the opening of the storage capacity B so as to ensure a sealed fluid coupling when the tip of the actuator A enters the opening of the storage capacity B.

[0104] Fluid coupling or a fluid connection is said to be tight when the coupling at the coupling interface allows fluid communication between the two coupled entities without any part of the fluid being able to escape outside the coupling interface, so that all the fluid that flows from one coupled entity to the other coupled entity arrives in the other coupled entity.

[0105] According to a particular embodiment of the ICC coupling interface shown schematically in Figure 6, the opening of the storage capacity B is in the shape of a cone open towards the outside of the storage capacity B, the open cone being configured to receive a corresponding closed cone positioned at the outlet of the tip of the actuator A, so that when the tip of the actuator A enters the opening of the storage capacity B, contact between an outer surface of the closed cone of the tip and an inner surface of the open cone of the opening ensures the sealed fluid coupling between the tip of the actuator A and the opening of the storage capacity B. According to another embodiment, the roles of the opening of the storage capacity B and the tip of the actuator A can be reversed, i.e.the opening of the storage capacity B may be in the form of a closed cone, and the tip of the actuator A in the form of an open cone corresponding to the closed cone of the opening of the storage capacity, so that when the closed cone of the opening of the storage capacity penetrates inside the open cone of the tip of the actuator A the fluid coupling between the tip of the actuator A and the opening of the storage capacity B is sealed.

[0106] According to another particular example of embodiment of the ICJT coupling interface shown schematically in Figure 7, the opening of the storage capacity B comprises an O-ring which ensures the sealed fluid coupling between the tip of the actuator A and the opening of the storage capacity B when the tip of the actuator A enters the opening of the storage capacity B.

[0107] The fluid connection of the device J with the first container R1 is made via the inlet channel I. According to a particular embodiment, the connection between the first container R1 and the inlet channel I can be sealed, for example using an O-ring adjusted to the dimension of the inlet channel I as illustrated in Figure 8, or via a cone-cone type interface not shown in Figure 8. However, it can also be free, i.e. not sealed, so that the pressure of the fluid in the first container is the ambient pressure; the fluid connection is then simpler to make.

[0108] Similarly, the fluid connection of the device J with the second container R2 is made via the outlet channel O. According to a particular embodiment, the connection between the second container R2 and the outlet channel O can be sealed, for example using an O-ring adjusted to the dimension of the outlet channel O as illustrated in Figure 8, or via a cone-cone type interface not shown in Figure 8. However, it can also be free, i.e. not sealed, so that the pressure of the fluid in the second container is the ambient pressure; the fluid connection is then simpler to make.

[0109] In Figure 8, the fluid connection of the device J with each of the containers R1, R2 is sealed. But, according to another exemplary embodiment not shown in Figure 8, a fluid connection with one of the containers R1, R2 may be sealed and the other fluid connection with the other container may be free. Finally, according to another exemplary embodiment also not shown in Figure 8, the fluid connection of the device J with each of the containers R1, R2 may be free.

[0110] The tightness of the connection between the inlet channel I and the first container R1, and the tightness of the connection between the outlet channel O and the second container R2 allows a fully pressure-controlled system, which no longer depends on its orientation relative to the ground or atmospheric pressure. This tightness makes it possible, for example, to envisage use without the presence of gravity (as on an orbital station) or without conventional atmospheric pressure (as in a submarine, an airplane or a module on the Moon or Mars).

[0111] According to one aspect, an exemplary embodiment of which is illustrated in Figure 1, the invention relates to an assembly E comprising a first container R1 and a second container R2 and a device J according to one of the embodiments described above, the inlet channel I of the device J being placed in fluid communication with the first container R1 and the outlet channel O being placed in fluid communication with the second container R2 so as to create a one-way fluid connection from the first container R1 to the second container R2. According to an exemplary embodiment, the first container R1 of the assembly is a reservoir of a microfluidic chip PU, and the second container R2 is another reservoir of the microfluidic chip PU.

[0112] In particular, as illustrated in Figure 9, the reservoir and the other reservoir of the PU microfluidic chip may be further in fluid communication via an AC channel of the PU microfluidic chip. Thus a single-chip closed loop is formed, so that the fluid circulates circularly in the PU microfluidic chip when an actuator is connected to the device.

[0113] This circular circulation has the advantage of subjecting the cells - for example endothelial cells - to an intermittent shear stress whose direction is fixed, without the need for an external recirculation system passing through a pump. This shear stress - generated by the flow speed which can be controlled by the actuator A - reproduces the natural stress undergone by these cells and allows them to organize and stretch longitudinally as in the vascular system, and therefore to approach their in vivo functionality dependent on this morphology. It should be noted that in this case, as illustrated in Figure 10, the device J could have to span a central reservoir RC equipped with a nano-porous membrane M, containing an organ to be perfused by the vascular system thus reproduced, said central reservoir being located between the first and second reservoirs R1, R2 of the microfluidic chip PU.

[0114] The device can also be used to connect several microfluidic chips together in a closed loop, thus constituting a multi-organ system connected by a single vascular system which shares and recirculates nutrients, toxins, pharmaceutical compounds, etc. The loop can be closed by completing the system with a passive return, such as a simple fluidic bridge whose purpose is to re-establish, through communicating vessels, a common liquid level for all the chips.Thus, according to an exemplary embodiment of an assembly E illustrated in Figure 11, the first container, into which the input interface I of the device J penetrates, is one of the two reservoirs R1, R2 of a microfluidic chip PU and the second container, into which the output interface O of the device J penetrates, is one of the two reservoirs RT, R2' of another microfluidic chip PU'; the other reservoir R1 among the two reservoirs R1, R2 of the microfluidic chip PU, and the other reservoir R2' among the two reservoirs RT, R2' of the other microfluidic chip PU' are in fluid communication via a fluid bridge PF. Said fluid bridge PF may be a passive fluid bridge, according to the embodiment shown in Figure 11. Said fluid bridge PF may also be another device J', according to one of the embodiments described above, according to another embodiment shown in Figure 12.

[0115] According to these provisions, when the fluid bridge PF is passive, the fluid level is identical in all the reservoirs after a determined time following an implementation of the actuator A to transfer a determined volume of fluid from the first container to the second container. This embodiment has the advantage of being simple, but it does not impose on the microfluidic chip PU the same flow conditions as on the other microfluidic chip PU', because the passive fluid bridge PF restores the levels at a different speed than the device J imposed for the transfer of the determined volume of fluid from the microfluidic chip PU to the other microfluidic chip PU'.

[0116] When the fluid bridge PF is active, i.e. produced with another device J' according to the invention, as shown in Figure 12, the device J transfers the determined volume of fluid from the microfluidic chip PU to the other microfluidic chip PU', and simultaneously, the other device J' transfers an identical determined volume from the other microfluidic chip PU' to the microfluidic chip PU. This double transfer can be carried out according to periodic cycles, driven by a rate of the actuator A which applies to both devices J and J' simultaneously. This system has the advantage of imposing the same flow conditions on all the microfluidic chips, which makes it possible, for example, to create the same shear stress conditions throughout the vascular system thus created. On the other hand, this embodiment introduces a weakness with regard to its sensitivity to the accuracy of the determined volumes of fluid transferred.Indeed, if the fluid volumes transferred by the two devices differ slightly - which is likely - one microfluidic chip will eventually dry out and the other microfluidic chip will overflow after a finite number of cycles. This phenomenon is amplified by the fact that several devices J are in parallel on the same actuator A and that the differences, or errors, in volumes transferred by each device accumulate.

[0117] This defect can be corrected by an embodiment of the device J comprising an error compensation means described below. According to this embodiment illustrated by way of example in Figure 15, the device J further comprises a fluid bridge PF configured to put the first container R1 and the second container R2 into bidirectional fluid communication, the bridge having a hydraulic resistance greater than a hydraulic resistance of the assembly comprising by the inlet channel I, the first valve V1, the storage capacity B, the second valve V2, the outlet channel O. Thus, the fluid bridge PF contributes to passively bringing the microfluidic chip(s) to a common fluid level by a communicating vessels effect. Thanks to the fact that the passive fluid bridge PF integrated into each device J has a hydraulic resistance greater than a hydraulic resistance of all the other components of said device J, iethe inlet channel I, the first valve V1, the storage capacity B, the second valve V2, the outlet channel O, the fluid flows in the direction imposed by the device(s) J used to form the set of interconnected microfluidic chips. Thanks to the passive fluidic bridge(s) PF integrated into each device J interconnecting the set of microfluidic chips PU, PU', any differential in height of the fluid level in the different microfluidic chips of the set considered is leveled over time. An additional solution to the passive fluidic bridge PF integrated into the device J according to the invention, to obtain an equalization of the fluid levels in the different microfluidic chips consists of introducing pauses in the actuation cycle of the devices so as to give time for the fluid levels to homogenize.

[0118] The devices J according to the invention can thus be used to interconnect microfluidic chips PU, PU' and containers according to various and modular configurations, such as configurations A1, A2, B1, B2, C, D, shown in Figure 14, by way of example. According to an embodiment of configurations A1, A2, B1, B2, C, D, of set of microfluidic chips, the microfluidic chips are arranged according to a standard format, such as the format defined by the ANSI SLAS 4-2004 (R2012) standard, also recognized as ANSI / SBS 4-2004. In particular, microfluidic chips can be arranged in 24-well SBS format plates with a well spacing of 18 mm, or in 384-well SBS format plates with a well spacing of 4.5 mm, or in 1536-well SBS format plates with a well spacing of 2.25 mm, or in 96-well or 6-well SBS format plates.

[0119] As an example, the following are shown in Figure 14, on a 96-well SBS format plate:

[0120] - two closed loops A1, A2 multi-chip microfluidics, with active return based on the use of several devices J to ensure the circulation of the fluid in one direction and in the return direction;

[0121] - two closed loops B1, B2 multi-chip microfluidics, with passive return based on the use of several devices J to ensure the circulation of the fluid in one direction, the return being based on passive fluidic bridges;

[0122] - An active open loop C configured to transfer the fluid between an inlet reservoir RE and an outlet reservoir RS, passing through a chain of microfluidic chips interconnected in series by devices J according to the invention;

[0123] - A passive open loop D configured to transfer the fluid between another inlet reservoir RE and another outlet reservoir RS, passing through another chain of microfluidic chips interconnected in series by devices J according to the invention for a part of the chain, and for another part of the chain by simple passive fluidic bridges PF;

[0124] These different configurations A1, A2, B1, B2, C, D, of set of microfluidic chips thus each constitutes a network A1, A2, B1, B2, C, D of microfluidic chips interconnected in series, comprising a plurality of N microfluidic chips, N being an integer greater than or equal to 2, interconnected by another plurality of N fluidic bridges, in which the plurality of N fluidic bridges comprises P passive fluidic bridges, and Q devices according to one of the embodiments described above, where P and Q are integers greater than or equal to 1, such that P+Q=N, or P+Q=N+1 in which:

[0125] - each microfluidic chip of the plurality of N microfluidic chips is connected with at least one other microfluidic chip of the plurality of N microfluidic chips via a passive fluidic bridge PF, or via a device J according to the invention so as to form an assembly E according to the embodiment described above,

[0126] - the network comprises at least one set E;

[0127] - if the network comprises a set E and another set E, one direction of a fluid connection of the set E of the network A1, A2, B1, B2, C, D and another direction of another fluid connection of the other set E of the network A1, A2, B1, B2, C, D are identical.

[0128] The network A1, A2, B1, B2 is closed if P+Q = N; the network C, D is open if P+Q=N+1.

[0129] In particular, a microfluidic chip PU of the open network C, D is connected, via a passive fluidic bridge PF or via a device J according to the invention, to an input container RE. More particularly, a microfluidic chip PU of the open network C, D is connected, via a passive fluidic bridge or via a device J according to the invention, to an output container RS.

[0130] Figure 13 illustrates in particular an example of an open network with two microfluidic chips PU, PU' and 3 devices J1, J2, J3 according to one of the embodiments of the invention described above, said devices being activated by a single actuator A, the network interconnecting an input container RE, and an output container RS.

[0131] In a particular example, the fluid bridges of the plurality of N fluid bridges, comprising a subset of devices according to the invention, are arranged in a predetermined configuration configured to correspond to a corresponding predetermined arrangement of microfluidic chips, arranged for example in the format defined by the ANSI SLAS 4-2004 (R2012) standard, also recognized as ANSI / SPB 4-2004. An example of a predetermined configuration is illustrated in Figure 17.

[0132] According to an exemplary embodiment, the fluid bridges of the plurality of fluid bridges arranged according to said predetermined configuration are preassembled on a BA assembly frame to which they are securely fixed, with fixed distances, for a dedicated application.

[0133] According to one aspect, the invention also relates to a method 100 for creating a monodirectional active fluid connection from a first container R1, RT, RE to a second container R2, R2', RS, with at least one device J, J1, J2, J3 one of the embodiments described above, the method comprising the following steps, illustrated schematically by figure 19:

[0134] - placing 101 the input channel I of the at least one device J, J1, J2, J3 in the first container R1; - placing 102 the output channel O of the at least one device J, J1, J2, J3 in the second container R2.

[0135] According to another aspect, the invention also relates to a method 200 for transferring a determined volume of a fluid with at least one device J, J1, J2, J3 according to one of the embodiments described above, the method comprising the following steps, illustrated schematically by figure 20:

[0136] - fluidly coupling 201 to the storage capacity B of the at least one device J, J1, J2, J3, an actuator A configured to implement at least one suction and discharge cycle, the suction and discharge cycle comprising a generation of a suction depression towards the storage capacity B of the at least one device J, J1, J2, J3 to suck the determined volume of fluid from a first container R1, RT, RE towards the intermediate storage capacity B, followed by a generation of a discharge pressure in the storage capacity B of the at least one device J, J1, J2, J3 to discharge the determined volume of fluid from the intermediate storage capacity B towards a second container R2, R2', RS;

[0137] - implement 202 at least one suction and discharge cycle of actuator A.

[0138] According to an example of implementation, the cycle is implemented according to a symmetrical period, typically represented by graph A of figure 2.

[0139] According to another example of implementation, the cycle is implemented according to a period including a time pause during which the pressure is kept constant, typically represented by graph B of figure 2.

[0140] According to another example of implementation, the cycle is implemented according to an asymmetric period, represented for example by graph C of figure 2. According to another example of implementation, the cycle is implemented in an aperiodic manner, represented for example by graph D of figure 2.

[0141] According to another aspect, the invention also relates to a method 300 for storing, in the storage capacity of a device J, J1, J2, J3 according to one of the embodiments described above, a determined quantity of fluid before or after the implementation of the method 200 for transferring a determined volume of a fluid according to one of the embodiments described above, the method 300 comprising the following steps, illustrated schematically in Figure 21:

[0142] - store 301 the determined volume in the storage capacity B of the device J, J1, J2, J3,

[0143] - implement 302 the method 200 of transferring the determined volume with the device J, J1, J2, J3

[0144] - recover 303 the determined volume of fluid present in the storage capacity B of the device J, J1, J2, J3.

[0145] Thus, the device J according to one of the embodiments described above can be used to store a quantity of fluid before or after its use in an open or closed loop circuit of microfluidic chips. The volume S of the storage capacity B, shown schematically in Figure 16, is constant, which can be used in different ways; for example, before use:

[0146] • Preparing a medium renewal of the value of Volumes B in the system. Example: if volume B is 100pL and 2 connected chips represent 100pL each, the active return circuit contains 400pL in total. Thus, changing the J devices for new devices with fresh medium amounts to doing a 50% medium change, or a single 25% change, which are fairly common values.

[0147] • Aliquoting a compound for introduction into the culture medium / vascular system of the multi-organ. Pre-clinical pharmaceutical compounds are complicated to maintain and often require freezing. Frozen aliquots in a J device would allow thawing of well-defined values ​​suitable for microfluidic chip testing.

[0148] • Storage / preparation of cells ready to inject into a microfluidic system

[0149] • Storage of lyophilized medium, which can be reconstituted with ultrapure water before use

[0150] Or, for example, after use, with recovery of volumes to be titrated / analyzed without disturbing the circuit by replacing the device.

Claims

CLAIMS 1. Device (J, J1, J2, J3) for one-way fluid connection, the device (J, J1, J2, J3) comprising an intermediate fluid storage capacity (B) configured to temporarily store a determined volume of a fluid, the device (J, J1, J2, J3) comprising an inlet channel (I) for the fluid in the device (J, J1, J2, J3) and an outlet channel (O) for the fluid coming from the device (J, J1, J2, J3), the device (J, J1, J2, J3) further comprising a first one-way valve (V1) configured to ensure one-way fluid communication from the inlet channel (I) to the storage capacity (B), the device (J, J1, J2, J3) further comprising a second one-way valve (V2) configured to ensure one-way fluid communication from the storage capacity (B) to the outlet channel (O).

2. Device (J, J1, J2, J3) according to one of claims 1, comprising a coupling interface of the storage capacity (B) of the device (J, J1, J2, J3), the coupling interface being an opening of the storage capacity (B) configured to ensure a sealed fluid coupling with a tip of an actuator (A) adjusted to the opening of the storage capacity (B), so that the fluid coupling between the tip and the opening is sealed when the tip of the actuator (A) is coupled to the opening of the storage capacity (B).

3. Assembly (E) comprising a first container (R1, RT, RE) and a second container (R2, R2', RS) and a device (J, J1, J2, J3) according to one of the preceding claims, the inlet channel (I) of the device (J, J1, J2, J3) being placed in fluid communication with the first container (R1, RT, RE) and the outlet channel (O) being placed in fluid communication with the second container (R2, R2', RS) so as to create a fluid connection one-way from the first container (R1, RT, RE) to the second container (R2, R2', RS).

4. Assembly (E) according to the preceding claim, in which the first container (R1) is a reservoir of a microfluidic chip (PU), and the second container (R2) is another reservoir of the microfluidic chip (PU).

5. Assembly (E) according to the preceding claim, in which the reservoir and the other reservoir of the microfluidic chip (PU) are further in fluid communication via a channel of the microfluidic chip (PU).

6. Assembly (E) according to the preceding claim, in which the channel of the microfluidic chip (PU) has a hydraulic resistance greater than a hydraulic resistance of the assembly comprising by the inlet channel (I), the first valve (V1), the storage capacity (B), the second valve (V2), the outlet channel (O).

7. Assembly (E) according to the claim, in which the first container (R1) is one of the reservoirs of a microfluidic chip (PU) and the second container (R2) is one of the reservoirs of another microfluidic chip (PU').

8. Assembly (E) according to claim 7, in which another reservoir among the reservoirs of the microfluidic chip (PU), and another reservoir among the reservoirs of the other microfluidic chip (PU') are in fluid communication via a fluid bridge, said fluid bridge being a passive fluid bridge, or a device (J, J1, J2, J3) according to one of claims 1 to 6.

9. Assembly (E) according to one of claims 3 to 8, in which a connection between the inlet channel (I) and the first container (R1, RT, RE) is sealed.

10. Assembly (E) according to one of claims 3 to 9, in which a connection between the outlet channel (O) and the second container (R2, R2', RS) is sealed.

11. Network (A1, A2, B1, B2, C, D) of microfluidic chips interconnected in series comprising a plurality of N microfluidic chips, N being an integer greater than or equal to 2, interconnected by another plurality of N fluidic bridges, in which the plurality of N fluidic bridges comprises P passive fluidic bridges (PF), and Q devices (J) according to one of claims 1 to 6, where P and Q are integers greater than or equal to 1, such that P+Q=N or P+Q=N+1, in which: - each microfluidic chip of the plurality of N microfluidic chips is connected with at least one other microfluidic chip of the plurality of N microfluidic chips, via a passive fluidic bridge (PF), or via a device (J) according to one of claims 1 to 6 so as to form an assembly (E) according to claim 10, - the network comprises at least one assembly (E) according to claim 10; - if the network comprises an assembly (E) according to claim 10 and another assembly (E) according to claim 10, one direction of a fluid connection of the assembly (E) of the network (A1, A2, B1, B2, C, D) and another direction of another fluid connection of the other assembly (E) of the network (A1, A2, B1, B2, C, D) are identical.

12. Method (100) for creating a one-way active fluid connection from a first container (R1, RT, RE) to a second container (R2, R2', RS), with at least one device (J, J1, J2, J3) according to one of claims 1 to 6, the method comprising the following steps: - placing (101) the input channel (I) of the at least one device (J, J1, J2, J3) in the first container (R1); - placing (102) the output channel (O) of the at least one device (J, J1, J2, J3) in the second container (R2); 13. Method (200) for transferring a determined volume of a fluid with at least one device (J, J1, J2, J3) according to one of claims 1 to 6, the method comprising the following steps: - fluidly coupling (201) to the storage capacity (B) of the at least one device (J, J1, J2, J3), an actuator (A) configured to implement at least one suction and discharge cycle, the suction and discharge cycle comprising a generation of a suction depression towards the storage capacity (B) of the at least one device (J, J1, J2, J3) to suck the determined volume of fluid from a first container (R1, RT, RE) towards the intermediate storage capacity (B), followed by a generation of a discharge pressure in the storage capacity (B) of the at least one device (J, J1, J2, J3) to discharge the determined volume of fluid from the intermediate storage capacity (B) towards a second container (R2, R2', RS); - implement (202) at least one suction and discharge cycle of the actuator (A).

14. Method (300) for storing in the storage capacity of a device (J, J1, J2, J3) according to one of claims 1 to 6, a determined quantity of fluid, and for recovering the determined quantity of fluid, in a manner not synchronized with the storage, the method (300) comprising the following steps: - fluidly coupling (201) to the storage capacity (B) of the at least one device (J, J1, J2, J3), an actuator (A) configured to implement at least one suction step (301), then a discharge step (302), the suction step (301) and the discharge step (302) not being synchronized, - implement the suction step (301) of the determined volume in the storage capacity (B) of the device (J, J1, J2, J3), - implement, in a manner not synchronized with the previous step (301), the step of discharging (302) the determined volume of fluid present in the storage capacity (B) of the device (J, J1, J2, J3).

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