Apparatus and method for clamping microfluidic devices

Abstract

The apparatus (1) suitable for clamping at least one microfluidic device (10) comprises a fluid-tight chamber (20) having a fluid inlet (24) configured to accommodate the microfluidic device (10) to be clamped by compressing at least one deformable portion of the microfluidic device (10) under the action of the pressure of a clamping fluid in the chamber (20), and a perfusion fluid management system (8) configured to regulate the pressure of the perfusion fluid in the microfluidic device (10) during the clamping operation so that the pressure of the clamping fluid in the chamber (20) is strictly higher than the pressure of the perfusion fluid in the microfluidic device (10).

Description

Technical Field

[0001] The present invention relates to an apparatus and a method for clamping at least one microfluidic device. Background Art

[0002] In the field of microfluidics, it is known to clamp microfluidic devices using chemical adhesion or mechanical systems such as bolts, C-clamps, rigid plates with magnets or shafts and levers. Chemical adhesion methods are limited in terms of compatible materials and allowable pressure ranges. Mechanical systems rely on precise, robust geometry and adjustment to achieve uniform clamping pressure, and thus uniform sealing. Summary of the Invention Problems to be Solved by the Invention

[0003] In response to these drawbacks, the present invention more specifically intends to improve these disadvantages by proposing an apparatus and a method for clamping at least one microfluidic device, which can ensure uniform clamping force over the entire surface of the microfluidic device, and thus uniform sealing, using a simple structure of the apparatus. The apparatus and method of the present invention further provide access to the microfluidic device, for example for monitoring purposes, and enable collectively clamping a plurality of microfluidic devices as needed, optionally with high-density microfluidic devices. Means for Solving the Problems

[0004] For this purpose, the subject matter of the present invention is an apparatus for clamping at least one microfluidic device, wherein a fluid-tight chamber having a fluid inlet, the fluid-tight chamber being configured to accommodate a microfluidic device that is clamped by compressing at least one deformable portion of the microfluidic device under the action of pressure of a clamping fluid within the chamber; - A perfusion fluid management system configured to adjust the pressure of the perfusion fluid within a microfluidic device so that the pressure of the clamping fluid in the chamber is strictly higher than the pressure of the perfusion fluid within the microfluidic device during the clamping operation. It is a device equipped with [a certain feature].

[0005] According to one embodiment, the perfusion fluid management system comprises at least one pressure controller. By using a pressure controller rather than a positive displacement pump or other flow generator with insufficient pressure control, the flow of the perfusion fluid is reliably stabilized, and the control of the pressure of the perfusion fluid within each microfluidic device, which is key to clamping, is improved. In particular, an electronic pressure controller, which is a pressure generator controlled by an electronic feedback loop, enables better instantaneous control of the pressure of the perfusion fluid.

[0006] According to one embodiment, the apparatus comprises a clamp fluid management system configured to adjust the pressure of the clamp fluid in a chamber, and a control unit configured to drive both the clamp fluid management system and the perfusion fluid management system so that the pressure of the clamp fluid in the chamber is strictly higher than the pressure of the perfusion fluid in the microfluidic device during clamping. Such embodiments, in which the control unit is configured to drive both the clamp fluid management system and the perfusion fluid management system, enable adjustment of the clamp of the microfluidic device according to its perfusion state, thereby ensuring efficient clamping in any operating state. The control unit may include several control modules that work together.

[0007] Therefore, a particular embodiment of the present invention is an apparatus for clamping at least one microfluidic device, -A fluid-sealed chamber having a fluid inlet, configured to house a microfluidic device that is clamped by compressing at least one deformable portion of the microfluidic device under the pressure of a clamping fluid within the chamber, - A perfusion fluid management system configured to adjust the pressure of the perfusion fluid in a microfluidic device such that the pressure of the clamp fluid in the chamber is strictly higher than the pressure of the perfusion fluid in the microfluidic device during clamping operation, the perfusion fluid management system (8) comprising at least one pressure controller, - A clamp fluid management system configured to adjust the pressure of the clamp fluid in a chamber, comprising a pressure source connected to the fluid inlet of the chamber via a duct, - A control unit configured to drive both the clamp fluid management system and the perfusion fluid management system so that during clamping operation, the pressure of the clamp fluid in the chamber is strictly higher than the pressure of the perfusion fluid in the microfluidic device. It is a device equipped with [a certain feature].

[0008] Within the framework of the present invention, a microfluidic device may be a single microfluidic chip or a stack of microfluidic chips. A microfluidic chip is typically 0.5 mm 2 The microfluidic chip comprises an internal channel having the following cross-sectional area. The microfluidic chip can be monolithic, and the channel is formed within the material constituting the chip. In a variant form, the microfluidic chip may comprise a back plate and a cover plate defining the channel between them. In this case, each of the back plate and cover plate may be a rigid plate (e.g., made of glass, or a rigid polymer such as polycarbonate, poly(methyl methacrylate) (PMMA), or cyclic olefin copolymer (COC)) or an elastomer plate (e.g., made of silicone). If both the back plate and cover plate of the microfluidic chip are rigid plates, the microfluidic chip may comprise an elastomer seal (e.g., made of polydimethylsiloxane) between the back plate and the cover plate.

[0009] In any of the above configurations, the microfluidic chip may deform due to the pressure difference between the inside and outside of the channel. In the case of a monolithic microfluidic chip, excessive pressure in the channel may cause an increase in the channel volume and deformation of the material constituting the chip, which is likely to lead to the appearance of cracks or passages that are likely to cause material rupture and leakage. In the case of a microfluidic chip that includes several components, which may be rigid and / or elastomer components, excessive pressure in the channel may cause deformation of the components of the microfluidic chip and their relative displacement, which is also likely to lead to the appearance of passages that are likely to cause leakage. In any of these cases, the microfluidic chip can be clamped by compressing at least one deformable part that is deformable under the influence of excessive pressure in the channel, which is the material constituting the chip in the case of a monolithic microfluidic chip, or at least one rigid or elastomer component in the case of a multi-component chip, under the action of the pressure of the clamping fluid in the chamber (i.e., the channel of the microfluidic chip can be closed).

[0010] Within the framework of the present invention, the clamp fluid contained within the chamber may be a gas, a liquid, or a combination thereof. The perfusion fluid circulating within the microfluidic device may be a gas, a liquid, a gel-like or semi-gel-like fluid, or a combination thereof. Examples of perfusion fluids include, for example, gas mixtures, aqueous particle or cell suspensions, non-aqueous particle suspensions, multiphase liquids, aqueous or non-aqueous solutions, and gel-like or semi-gel-like particle or cell suspensions. Several perfusion fluids may circulate within the microfluidic device, in which case multiple perfusion lines may be advantageously used to handle the circulation of different perfusion fluids independently of each other.

[0011] The apparatus of the present invention enables the clamping of microfluidic devices housed in a chamber or each microfluidic device under the action of the pressure of the clamping fluid in the chamber by uniform and omnidirectional compression of at least one deformable part of the microfluidic device, thereby ensuring optimal clamping uniformity. Therefore, it is possible to prevent leakage or damage within the microfluidic device even under high operating perfusion pressure or when a pressure difference or pressure gradient exists within the channels of the microfluidic device.

[0012] A significant advantage of the present invention is that the apparatus further allows for the clamping of multiple microfluidic devices together within the same chamber. The net clamping force applied to each microfluidic device present in the chamber, i.e., the pressure difference between the pressure of the clamping fluid in the chamber and the pressure of the perfusion fluid in the microfluidic device, can be easily controlled. In the case of microfluidic devices comprising an elastomer backplate and / or elastomer coverplate, the clamping pressure is also advantageous in that it can reduce the pressure difference between the inside and outside of the microfluidic device during use, thereby reducing deformation of the elastomer material and limiting fluctuations in the channel volume of the microfluidic device.

[0013] According to one embodiment, the pressure difference between the clamp fluid in the chamber and the perfusion fluid in the microfluidic device is maintained at 0.05 bar or higher, preferably 0.1 bar or higher. This minimum pressure difference ensures that the deformable portion(s) of the microfluidic device are sufficiently compressed, guaranteeing sealing of the microfluidic device under conventional operating conditions. In addition, such a pressure difference ensures that in the event of leakage, the flow cannot escape from the microfluidic device, which is particularly advantageous when the perfusion fluid contains harmful substances.

[0014] According to one embodiment, the control unit is configured to receive measured values ​​of the pressure of the clamp fluid in the chamber and the pressure of the perfusion fluid in the microfluidic device from a pressure sensor, and to drive the clamp fluid management system, and possibly the perfusion fluid management system, according to the received measured values. In this way, relative adjustments can be made between the pressure of the perfusion fluid in the microfluidic device and the pressure of the clamp fluid in the chamber in order to optimally seal the microfluidic device. In one embodiment, the pressure of the clamp fluid in the chamber may be maintained at a fixed value, and continuous adjustment of the pressure of the perfusion fluid in the microfluidic device may be made to optimally seal the microfluidic device. In another embodiment, continuous adjustment of the pressure of the clamp fluid in the chamber may be made according to the pressure of the perfusion fluid in the microfluidic device in order to optimally seal the microfluidic device.

[0015] According to one embodiment, the chamber is configured to house within the chamber's internal volume a plurality of microfluidic devices that are clamped together under the pressure of the clamping fluid within the chamber. In this way, the apparatus of the present invention enables the simultaneous clamping of a plurality of microfluidic devices when the pressure of the clamping fluid within the chamber is strictly higher than the pressure of the perfusing fluid within each of the microfluidic devices.

[0016] According to one embodiment, the apparatus comprises at least one active system configured to monitor the contents of a microfluidic device housed in a chamber and / or guide the contents of the microfluidic device housed in the chamber through at least one wall of the microfluidic device during a clamping operation.

[0017] According to one embodiment, the active system is an optical monitoring system configured to monitor the contents of a microfluidic device housed in a chamber through at least one wall of the microfluidic device during clamping, such as an imaging system (e.g., transmitted light imaging system, reflected light imaging system, phase imaging system, fluorescence imaging system, etc.), a spectroscopic system (e.g., FTIR, UV spectroscopic system, visible light spectroscopic system, etc.), or an interference system. The monitoring system may further be a temperature monitoring system, a calorimetry system, an electromagnetic impedance measurement system, or any other monitoring or measurement system that requires access to the vicinity of the channels of the microfluidic device.

[0018] According to one embodiment, the active system is a lithography system configured to perform lithography in the channel of a microfluidic device housed in a chamber through at least one wall of the microfluidic device during a clamping operation. The lithography system may be any type of lithography system that requires access to the vicinity of the channel of the microfluidic device, such as a visible light lithography system, a UV lithography system, an EUV lithography system, an X-ray lithography system, an electron beam lithography system, a femtosecond lithography system, a dynamic mask (e.g., a digital mirror device (DMD) or a liquid crystal dynamic mask) lithography system, a dynamic light source (e.g., an LED or a laser array) lithography system, or any combination thereof.

[0019] The clamping apparatus of the present invention enables the use of a lithography system in a chamber adjacent to the channel of the microfluidic device during the clamping operation, thereby enabling lithography operations to be performed within the microfluidic device. Performing lithography in a perfusible microfluidic device offers several advantages and capabilities, particularly enabling in-flow polymerization or stop-flow polymerization to produce fine particles with well-controlled properties at high throughput, or enabling the injection of different prepolymer mixtures, resins, developers, pigments, inhibitors, activators, or other types of reactants, thereby increasing manufacturing capabilities using lithography.

[0020] In other embodiments, the active system could be, for example, an electric field generating system used for electroporation of cells in a microfluidic device, for example an acoustic field generating system used to perform acoustic electrophoresis in a microfluidic device, for example a magnetic field generating system used to sort magnetic particles within a microfluidic device, for example an illumination system used to perform photochemistry in a microfluidic device, for example a temperature control system used to locally heat or cool a portion of a microfluidic device to perform a chemical reaction such as PCR. In this case as well, access to the entire periphery of the microfluidic device is highly advantageous.

[0021] The ability to use an active system in close proximity to the channels of the microfluidic device during the clamping operation is a significant advantage of the clamping device of the present invention over prior art mechanical clamping systems such as rigid plates with bolts, C-clamps, magnets, or shafts and levers that restrict or obstruct access to the periphery of the microfluidic device. On the other hand, in the clamping device of the present invention, access to the microfluidic device is possible all around it during the clamping operation, so an active system, which may be a monitoring system or any other type of active system, can be used as close as possible to the channels of the microfluidic device.

[0022] According to one embodiment, the apparatus comprises an imaging system configured to image the contents of the microfluidic device housed in the chamber through at least one wall of the microfluidic device. Advantageously, the at least one wall of the microfluidic device is light-transmissive in a wavelength range useful for the imaging system, such that the internal volume of the microfluidic device can be imaged using a conventional camera or another suitable optical detector.

[0023] According to one embodiment, the apparatus comprises a monitoring system configured to monitor the contents of the microfluidic device housed in the chamber during a clamping operation, and the control module is configured to drive the perfusion fluid management system in accordance with measurement values from the monitoring system. In this way, the apparatus enables monitoring of the operating conditions within the microfluidic device and adjustment of the pressure of the perfusion fluid within the microfluidic device accordingly.

[0024] According to one embodiment, the apparatus comprises a displacement system for displacing the active system and the microfluidic device housed in the chamber relative to each other, in order to position the active system in proximity to a channel of the microfluidic device during a clamping operation. The active system may be, for example, a monitoring system, a lithography system, or any other active system for applying specific guidance. According to one embodiment, the displacement system is configured to move the active system and the microfluidic device relative to each other to position them in at least one operating configuration.

[0025] According to one embodiment, the chamber is provided with a loading port for moving the microfluidic device into and out of the chamber, and the loading port is closed in a fluid-tight manner during the clamping operation. In one embodiment, a sleeve for fluid-tight penetration of at least one tube connecting the microfluidic device to the perfusion fluid management system is positioned within an opening made in a sealing surface of a door for closing the loading port.

[0026] According to one embodiment, the chamber is configured to accommodate the entire perfusion fluid management system within the internal volume of the chamber. In this case, the tube(s) connecting the microfluidic device to the perfusion fluid management system are configured to withstand the pressure of the clamping fluid within the chamber without substantially deforming, so as not to affect the circulation of fluid between the perfusion fluid management system and the microfluidic device.

[0027] According to another embodiment, the chamber is configured to accommodate only a part of the perfusion fluid management system within its internal volume, and the apparatus comprises at least one sleeve configured to be positioned within an opening in a wall of the chamber during a clamping operation to allow fluid-tight passage of at least one tube connecting the microfluidic device to the perfusion fluid management system.

[0028] According to one embodiment, the sleeve comprises at least one bore configured to receive a tube connecting the microfluidic device to the perfusion fluid management system, the bore extending between an inner end of the sleeve intended to face the internal volume of the chamber and an outer end of the sleeve intended to face the outside of the chamber, and being fluid-tightly closed around the tube. In one embodiment, the sleeve is overmolded onto the tube. In another embodiment, the sleeve is openable by reversible deformation to allow access to the bore in an open configuration of the sleeve, and the bore is fluid-tightly closed around the tube when the sleeve is closed and positioned within the opening in the wall of the chamber.

[0029] According to one embodiment, the sleeve is a sealing member configured to fluid-tightly seal the loading port of the chamber by being arranged particularly at the joint between the edge of the loading port and the door for closing the loading port.

[0030] According to one embodiment, the chamber is configured to house only a portion of the perfusion fluid management system within its internal volume, and the device includes in the chamber wall a connecting unit which includes at least one fluid passage extending through the chamber wall, a tube connected to a microfluidic device on the side toward the internal volume of the chamber, and connectors at both ends of the fluid passage for connecting a tube connected to the perfusion fluid management system on the side toward the outside of the chamber.

[0031] According to one embodiment, the apparatus comprises at least one support within a chamber configured to house a microfluidic device to be clamped. In one embodiment, the apparatus comprises a plurality of supports (e.g., in the form of shelves, slots, rails, posts, racks, suction cups, hooks, tweezers, or magnets) arranged side-by-side and / or stacked within the chamber, configured to house a plurality of microfluidic devices. In an advantageous embodiment, the support or each support is mounted on an openable wall of the chamber, particularly a door configured to close the loading port of the chamber. In another advantageous embodiment, the support or each support is mounted on a frame structure (which may be automated) configured to be loaded into the chamber by rails, wheels, or other guiding means.

[0032] Another subject of the present invention is a method for clamping at least one microfluidic device having at least one deformable portion, - The microfluidic device is connected to a perfusion fluid management system and positioned within a chamber having a fluid inlet, - The step of sealing the chamber so that it is fluid-tight with respect to the clamped fluid, - The chamber is pressurized with clamping fluid supplied through the fluid inlet, - The microfluidic device is clamped by applying pressure to the clamp fluid and the perfusion fluid such that the pressure of the clamp fluid in the chamber is strictly higher than the pressure of the perfusion fluid in the microfluidic device, thereby compressing at least one deformable part of the microfluidic device under the action of the pressure of the clamp fluid in the chamber. This method includes [something].

[0033] According to one embodiment, the pressure of the perfusion fluid within a microfluidic device is controlled by a control module configured to receive measurements from a monitoring system that monitors the contents of the microfluidic device during clamping operations, and to drive a perfusion fluid management system in accordance with the received measurements.

[0034] According to one embodiment, multiple microfluidic devices are positioned inside a chamber and clamped together under the pressure of the clamp fluid inside the chamber by applying pressures to the clamp fluid and the perfusion fluid such that the pressure of the clamp fluid inside the chamber is strictly higher than the pressure of the perfusion fluid inside each microfluidic device.

[0035] According to one embodiment, a microfluidic device or each microfluidic device is “pre-clamped” so that its components are assembled in a sealed state within the internal volume of the microfluidic device before being introduced into the chamber of the device, thereby preventing the clamping fluid from seeping into the microfluidic device (which would adversely affect the clamp) during pressurization of the chamber by the clamping fluid. Such “pre-clamping” of a microfluidic device or each microfluidic device can be achieved, for example, by an adhesive inserted between the components of the microfluidic device, by adhesive tape covering at least a portion of the edges of the microfluidic device, or by any other suitable assembly method.

[0036] The features and advantages of the present invention will become apparent from the following description of embodiments of the apparatus and method of the present invention, which are shown with reference to the accompanying drawings as merely examples. [Brief explanation of the drawing]

[0037] [Figure 1] This is a schematic cross-sectional view of a device for clamping at least one microfluidic device according to a first embodiment of the present invention in a closed configuration of a fluid-sealed chamber. [Figure 2] This diagram is similar to Figure 1 in the open configuration of the fluid-sealed chamber. [Figure 3] This is a top view of a non-limiting example of a microfluidic device clamped in the apparatus of Figure 1, comprising a rigid backplate, a rigid coverplate, and an elastomer seal between the backplate and the coverplate, wherein the channel of the microfluidic device is defined solely by the coverplate. [Figure 4] Figure 3 is a perspective view of the microfluidic device with the channels of the microfluidic device omitted. [Figure 5] This is a cross-sectional view along plane V in Figure 4. [Figure 6a] This is a magnified view of detail VI in Figure 5. [Figure 6b] This figure is similar to Figure 6a in the clamp configuration of a microfluidic device, where the elastomer seal of the microfluidic device is elastically deformed under the pressure of the clamp fluid in the fluid-sealing chamber, but the deformation of the elastomer seal is exaggerated for illustrative purposes. [Figure 7a] This figure is similar to Figure 6a in that it shows a first modified form of the microfluidic device clamped in the apparatus of Figure 1, in which the channel of the microfluidic device is defined by both the back plate and the cover plate to form a two-stage microfluidic circuit. [Figure 7b] This figure is similar to Figure 7a in the clamp configuration of a microfluidic device where the elastomer seal is elastically deformed under the pressure of the clamp fluid within the fluid sealing chamber, but the deformation of the elastomer seal is exaggerated for illustrative purposes. [Figure 8a]This figure is similar to Figure 6a in depicting a second modified form of the microfluidic device clamped by the apparatus in Figure 1, where the elastomer seal is cut according to a pattern aligned with the channels in the backplate and coverplate of the microfluidic device. [Figure 8b] This figure is similar to Figure 8a in the clamp configuration of a microfluidic device where the elastomer seal is elastically deformed under the pressure of the clamp fluid within the fluid sealing chamber, but the deformation of the elastomer seal is exaggerated for illustrative purposes. [Figure 9a] This figure is similar to Figure 6a for a third deformation mode of the microfluidic device clamped in the apparatus of Figure 1, where the microfluidic device is monolithic, and shows a deformation state due to excess pressure of the perfusing fluid in the channel of the microfluidic device that is not offset by the clamping pressure, and the deformation of the material constituting the microfluidic device is exaggerated for illustrative purposes. [Figure 9b] This figure is similar to Figure 9a in the clamp configuration of a microfluidic device, where the constituent materials of the microfluidic device have returned to a planar configuration under the pressure of the clamp fluid in the fluid-sealed chamber. [Figure 10] This is a magnified view of detail X in Figure 1. [Figure 11] This is a cross-sectional view along line XI in Figure 10. [Figure 12] This is a cross-sectional view along line XII in Figure 11. [Figure 13] This figure is similar to Figure 10 in depicting a modified form of a sealing member that enables fluid-tight penetration of a tube connecting a perfusion fluid management system to a microfluidic device clamped within the fluid-sealed chamber of the apparatus. [Figure 14] This is a cross-sectional view along line XIV in Figure 13. [Figure 15] This is a cross-sectional view along line XV in Figure 14. [Figure 16] This is a magnified view of detail XVI in Figure 1. [Figure 17]This is a schematic cross-sectional view of a device for clamping at least one microfluidic device according to a second embodiment of the present invention in a closed configuration of a fluid-sealed chamber. [Figure 18] This is a magnified view of detail XVIII in Figure 17. [Figure 19] This is a schematic cross-sectional view of a device for clamping at least one microfluidic device according to a third embodiment of the present invention in a closed configuration of a fluid-sealed chamber. [Modes for carrying out the invention]

[0038] Figure 1 shows apparatus 1 of a first embodiment of the present invention, illustrating apparatus 1 for clamping a plurality of microfluidic devices 10 arranged within a chamber 20 of apparatus 1. In the illustrated non-limiting example, each microfluidic device 10 is a microfluidic tip comprising a backplate 11 and a coverplate 12 made of poly(methyl methacrylate) (PMMA), and an elastomer seal 13 made of polydimethylsiloxane inserted between the backplate 11 and the coverplate 12. As seen in Figures 3 to 5, the backplate 11 and the coverplate 12 define a plurality of channels 14 having meandering tracks between the backplate 11 and the coverplate 12 in order to minimize the area of ​​the microfluidic device 10 while maintaining the long length of the channels 14. Each microfluidic device 10 includes an inlet port 15 and an outlet port 16 at both ends of the meandering track, which are configured to be connected to a pair of supply lines 85, 85' to circulate perfusion fluid within the channels 14.

[0039] Each microfluidic device 10 is advantageously “pre-clamped” with adhesive tape 18, shown in Figure 5, which covers at least a portion of the edges of the microfluidic device 10, before being introduced into the chamber 20 of the apparatus 1. In this way, the components of the microfluidic device 10 are assembled in a sealed state, preventing the clamp fluid from seeping into the microfluidic device 10 during pressurization of the chamber 20 (which would adversely affect the clamp).

[0040] As not limited to Figures 6a, 7a, 8a, and 9a, the channels 14 of the microfluidic device 10 may have different contours. In the first example shown in Figure 6a, cavities are provided only in the cover plate 12 of the microfluidic device 10, and each channel 14 is formed between the elastomer seal 13 covering the back plate 11 and the cavities in the cover plate 12, forming a single-stage microfluidic circuit. In the first variant shown in Figure 7a, each channel 14 is formed between two complementary cavities provided in the back plate 11 and the cover plate 12, respectively, and the elastomer seal 13 divides the channel 14 into two overlapping compartments. Thus, in this first variant, a two-stage microfluidic circuit is formed. Figure 8a shows a second variant of the microfluidic device 10 similar to the first variant in Figure 7a, except that the lower and upper stages of the microfluidic circuit are interconnected by perforations 130 in the elastomer seal 13 corresponding to the channels 14. Figure 9a shows a third variant of the microfluidic device 10, in which the microfluidic device is monolithic and the channel 14 is formed within the material constituting the chip.

[0041] In the examples shown in Figures 1 and 2, the apparatus 1 comprises a container 2 formed by a combination of a body 21 and a cover 22. In the closed configuration of the container 2 shown in Figure 1, the body 21 and the cover 22 define a fluid-sealed chamber 20 between them, having a fluid inlet 24. The chamber 20 is configured to house a plurality of microfluidic devices 10 within its internal volume, which are clamped together under the pressure of a clamping fluid present in the chamber. More precisely, the chamber 20 is pressurized by a clamping fluid supplied through the fluid inlet 24, and the microfluidic devices 10 present in the chamber 20 are clamped by the compression of their deformable parts under the pressure P of the clamping fluid. As schematically shown in Figures 6a-6b, 7a-7b, 8a-8b, and 9a-9b, the deformable parts are the elastomer seal 13 between the backplate 11 and the cover plate 12 in the examples of Figures 6a-6b, 7a-7b, and 8a-8b, and the material constituting the monolithic chip in the example of Figures 9a-9b.

[0042] In one implementation of the device 1, the clamp fluid is a gas, such as pressurized air. In another implementation of the device 1, the clamp fluid is a combination of heat transfer fluids (e.g., water or oil) contained within the body 21 of the container 2 to partially fill the internal volume of the chamber 20, for example, about 80% of its internal volume, with the remaining portion of the internal volume of the chamber 20 being filled with pressurized air supplied through the fluid inlet 24. As shown in Figures 1 and 2, a heat exchanger 27 is provided at the bottom of the body 21 to allow heating and / or cooling of the clamp fluid when the operation realized within the microfluidic device 10 requires a specific operating temperature.

[0043] As clearly shown in Figures 1 and 2, the apparatus 1 comprises a frame 9 that supports both the body 21 and the cover 22 of the container 2, and the cover 22 can be displaced relative to the body 21 to open the chamber 20. In the sealed configuration of the chamber 20 shown in Figure 1, the cover 22 fluidly closes the opening 25 of the body 21 to the clamping fluid, and the space between the cover 22 and the body 21 is sealed by sealing members 3, 211, 221. The cover 22 is held in a sealed configuration relative to the body 21 by fastening screws 28 that maintain the sealing members 3, 211, 221 in a compressed state. As shown in Figure 2, by removing the fastening screws 28, the cover 22 can be separated from the body 21 by moving a lifting arm 29 connected to the cover 22 upward. To guide the movement of the lifting arm 29, the frame 9 advantageously comprises an electric ball screw actuator and a guide rail 91 on which the sliding end 291 of the lifting arm 29 can slide up and down.

[0044] The structure of the body 21 and cover 22 of the container 2 is made of metal sheets of appropriate thickness, such as stainless steel, thereby making the container 2 robust and able to withstand the pressure levels required for clamping. For each of the body 21 and cover 22, the metal contacts are lined with thermal insulation material 23. In addition, the metal contacts of the cover 22 form a rack structure 26 intended to be housed within the internal volume of the body 21 when the cover 22 closes the opening 25 of the body 21. The rack structure 26 includes support elements 260 in which the microfluidic device 10 can be placed. The rack structure 26 further supports a monitoring system 5 configured to monitor the contents of the microfluidic device 10 housed in the chamber 20, and a displacement system 7 configured to displace the imaging head 51 of the monitoring system 5 and the microfluidic device 10 relative to each other within the chamber 20.

[0045] In the illustrated example, the monitoring system 5 includes an imaging head 51 that incorporates both a phase imaging system and a fluorescence imaging system. More specifically, as best shown in the enlarged view of Figure 16, the imaging head 51 has a U-shaped structure, with a first U-shaped arm supporting a phase-contrast light source 52 and a second U-shaped arm supporting an imaging arm 54 and a fluorescence imaging module 56. The phase-contrast light source 52 includes an electric light-emitting diode (LED) 521, a collimation lens 522, a mirror 523 positioned at 45° with respect to the optical path, a phase ring 524, and a condenser 525. On the side opposite the phase-contrast light source 52, the imaging arm 54 includes a multipurpose objective lens 541 suitable for both phase imaging and fluorescence microscopy, a lens 542, two mirrors 543, 544 positioned at 45° with respect to the optical path, and a camera 545. The fluorescence imaging module 56 is inserted into the imaging arm 54 and includes an excitation light source 561 (e.g., a laser), a diverging lens 562, and a dichroic mirror 563 positioned at a 45° angle to the optical path, wherein the dichroic mirror 563 is configured to reflect light from the excitation light source 561 and transmit other wavelengths.

[0046] To generate a phase-contrast image of the contents of the microfluidic device 10 housed in the chamber 20, the microfluidic device 10 is positioned within the space of the U-shaped imaging head 51 at a working distance from the condenser 525. Next, the LED 521 of the phase-contrast light source 52 is turned on, and its light is collimated by the lens 522, reflected by the mirror 523, spatially filtered by the phase ring 524, and focused toward the microfluidic device 10 by the condenser 525. The microfluidic device 10 and its contents transmit light, and a portion of the transmitted light is focused by the objective lens 541 positioned at a working distance from the microfluidic device 10. The focused light is reflected off two mirrors 543, 544, passes through a dichroic mirror 563, is then collimated by the objective lens 541, and is focused by lens 542 to form an image on the sensor surface of the camera 545.

[0047] To generate a fluorescence image of the contents of the microfluidic device 10 housed in the chamber 20, a fluorescence light source 561 is turned on, its beam is expanded by a divergent lens 562, redirected by dichroic mirrors 563 and 543, collimated by lens 542, and then focused to the focal plane by objective lens 541 within the microfluidic device 10. The light emitted by fluorescence emission from the illumination area is reflected off the two mirrors 543, 544, passes through the dichroic mirror 563, is partially focused by objective lens 541, collimated and focused by lens 542, and forms an image on the sensor surface of camera 545.

[0048] To adjust the relative position of the imaging head 51 with respect to the microfluidic device 10 monitored within the chamber 20, the apparatus 1 includes a displacement system 7 comprising several motorized ball screw actuators and associated guide rails, namely, a first guide rail 71 mounted substantially vertically on a rack structure 26 (allowing the imaging head 51 to slide up and down), a second guide rail 73 similarly mounted substantially vertically on the rack structure 26 (allowing a slider 74 to slide up and down), and a third guide rail 75 mounted substantially horizontally on the slider 74 (allowing the gripping head 76 to slide laterally). Naturally, the displacement system 7 may further include additional displacement means that enable movement from the plane, in particular, as shown in Figures 1, 2, and 16, so that the imaging head 51 can be moved relative to a large portion of the surface of the microfluidic device 10. For clarity, such lateral displacement means are not shown. In a variant (not shown), the displacement system 7 may further include a robotic arm configured to move the imaging head 51 around the microfluidic device 10.

[0049] The gripping head 76 is configured to grip the microfluidic device 10, which is initially positioned on the support element 260 of the rack structure 26, by a suction cup 78, and to displace the microfluidic device 10 toward the space of the U-shaped imaging head 51 by sliding along guide rails 73, 75. In addition, the imaging head 51 is configured to move perpendicular to the microfluidic device 10 positioned in space by sliding along the guide rail 71, and to adjust the subject so that it is imaged within the focal plane of the objective lens 541. To perform high-quality phase-contrast imaging, the distance between the phase-contrast light source 52 and the imaging arm 54 is adjusted according to the thickness and refractive characteristics of the microfluidic device 10 and its contents.

[0050] Naturally, more advanced imaging devices (e.g., those with multiple excitation sources, ultrashort shock sources, other types of wavelength filters and / or confocal capabilities) can also be used as monitoring systems 5 mounted within the chamber 20, as well as other types of active systems for providing specific guidance to the contents of the microfluidic device 10 housed within the chamber 20.

[0051] Frame 9 further supports other parts of the apparatus 1, including a clamp fluid management system 6 and a perfusion fluid management system 8. The clamp fluid management system 6 is configured to regulate the pressure of the clamp fluid in the chamber 20 using a pressure source, and the perfusion fluid management system 8 is configured to regulate the pressure of the perfusion fluid in each microfluidic device 10 present in the chamber 20 using another pressure source. In order to clamp the microfluidic devices 10, the pressure of the clamp fluid in the chamber 20 is strictly higher than the pressure of the perfusion fluid in the microfluidic devices 10. This operating state can be automatically controlled by a control unit which may include several control modules, such as control modules 61, 80, described below. Typically, the difference between the pressure of the clamp fluid in the chamber 20 and the pressure of the perfusion fluid in the microfluidic devices 10 is maintained at 0.05 bar or higher, preferably 0.1 bar or higher.

[0052] In the illustrated example, the clamp fluid management system 6 includes a pressure source 62 (here a pump) connected to the fluid inlet 24 of the chamber 20 via a duct 64. A pressure sensor 65 having a valve 63 and an air intake 66 is located in the duct 64 to regulate the flow of the clamp fluid at the output of the pressure source 62 and to measure the pressure of the clamp fluid supplied into the chamber 20, respectively. The control module 61 is configured to ensure that the pressure of the clamp fluid allows the chamber 20 to be pressurized such that the pressure difference between the internal volume of the chamber and the outside of the chamber is at least 0.5 bar, preferably at least 1 bar, and more preferably at least 3 bar.

[0053] In the illustrated example, the perfusion fluid management system 8 comprises a reactant module 81 having a plurality of reactant tanks 811-814 and an array of valves 815 at the outlets of the reactant tanks, the inlets of the reactant tanks being connected to two electronic pressure controllers 817, 818 via an array of valves 819, and two perfusion lines 82, 82', each having pressure sensors 83, 83', the array of valves 815 being configured to establish connections between one or more reactant tanks 811-814 and the perfusion lines 82, 82', and perfusion The perfusion fluid management system 8 further comprises an array of valves 84 configured to establish a connection between at least one of lines 82, 82' and at least one microfluidic device 10 positioned within a chamber 20, and supplying each microfluidic device 10 through a pair of supply lines 85, 85' that connect each perfusion line 82, 82' to an inlet port 15 and an outlet port 16 of the microfluidic device; a purge line 87 to which the perfusion lines 82, 82' are connected via their respective valves 86, 86', the purge line 87 equipped with a pressure sensor 88; and a waste tank 89. The perfusion fluid management system 8 further comprises a control module 80 that controls pressure controllers 817, 818 and valves 819, 815, 84, 86, 86' to regulate the pressure of the perfusion fluid distributed into each microfluidic device 10.

[0054] The structure of the reactant module 81, in which multiple reactant tanks 811-814 are connected to pressure controllers 817, 818 via a valve array 819, allows the number of pressure controllers 817, 818 to be reduced to the number of perfusion lines, i.e., two perfusion lines 82, 82' in the illustrated example. The presence of two perfusion lines 82, 82' is advantageous in that one line is used for feeding the microfluidic device 10 and the other line is used for discharging the microfluidic device 10, and they are connected to the reactant tanks 811-814 via a valve array 815 that allows for any configuration of the connection between the perfusion lines 82, 82' and the reactant tanks 811-814. By using pressure controllers 817, 818 to regulate the pressure of the perfusion fluid in each microfluidic device 10, rather than a positive displacement pump or other flow generator, the flow of the perfusion fluid is reliably stabilized and the control of the pressure of the perfusion fluid in each microfluidic device, which is key to clamping, is improved. In particular, electronic pressure controllers, which are pressure generators controlled by electronic feedback loops, enable better instantaneous control of the pressure of the perfusion fluid.

[0055] The illustrated embodiment of the perfusion fluid management system 8 allows for highly flexible use of reactant tanks for perfusing microfluidic devices, for example, the discharge from a microfluidic device can be collected in one of the reactant tanks and used to perfuse another microfluidic device in a later stage. When complex operations are required where mixing and cross-contamination between consecutive flows are reduced, it becomes possible to increase the number of perfusion lines to physically separate inlet and outlet flows having different functions in different perfusion lines. Individual connections of each microfluidic device 10 to perfusion lines 82, 82' via an array of valves 84 are also advantageous in that they allow for any combination of connections and provide high operational flexibility.

[0056] Preferably, perfusion lines 82, 82' are connected at one end to reactant tanks 811-814 via a valve array 815 and at the other end to a purge line 87 via electronically controlled valves 86, 86', which are connected to a relatively large waste tank 89. The electronically controlled valves 86, 86' connecting the perfusion lines 82, 82' to the purge line 87 may be double-layered using one-way check valves to avoid backflow. This configuration allows for complete flushing of the perfusion lines 82, 82' to efficiently reduce cross-contamination and mixing between solutions processed in the same perfusion line in consecutively opposite flow directions, for example. The pressure control system preferably comprises pressure sensors 83, 83' in each perfusion line 82, 82' and a pressure sensor 88 in the purge line 87, all of which are connected to a control module 80 of the system 8 having a feedback loop, which actively controls the perfusion pressure to be kept below a predetermined threshold to prevent the pressure of the perfusion fluid in each microfluidic device 10 from becoming higher than the pressure of the clamp fluid in the chamber 20, thereby causing leakage.

[0057] In this first embodiment, the chamber 20 houses only a portion of the perfusion fluid management system 8 within its internal volume. The reactant module 81, the waste tank 89, and portions of the perfusion lines 82, 82' and purge line 87, which include associated pressure sensors 83, 83', 88, are located outside the chamber 20. To allow the perfusion lines 82, 82' and purge line 87 to fluidly penetrate the walls of the chamber 20, a sealing member 3, which is one of the sealing members for sealing the space between the cover 22 and the body 21, has three holes 33 for receiving the tubes of the perfusion lines 82, 82' and purge line 87. As clearly shown in the cross-sectional view of Figure 12, each hole 33 extends between an inner end 32 of the sealing member 3 intended to face into the internal volume of the chamber 20 and an outer end 31 of the sealing member 3 intended to face outwards from the chamber 20. The sealing member 3 is advantageously frustoconical in shape, as shown in Figure 12, and the inner end 32 of the sealing member 3 has a larger surface area than the outer end 31, so that the pressure P of the clamping fluid in the chamber 20 during the clamping operation pushes the sealing member 3 outward, thereby improving the sealing performance at the position of the inclined peripheral wall 35 of the trapezoidal sealing member.

[0058] As shown in the enlarged views of Figures 10 and 11, in the sealed configuration of the chamber 20, the sealing member 3 is inserted between an inflatable O-ring 211 provided in a groove 210 of the main body 21 and a flat gasket 221 fastened to the cover 22. The inflatable O-ring 211, which can be replaced with any other seal having very high deformability, is very suitable for maintaining the deformation caused by the height of the sealing member 3 in the closed configuration of the container 2. The sealing member 3 is advantageously overmolded around the tubes of the perfusion lines 82, 82' and the purge line 87 and fastened to the flat gasket 221, so that in the open configuration of the cover 22, the microfluidic device 10 can be placed on the support elements 260 of the rack structure 26 with the fluid connection to the perfusion fluid management system 8 already established. This configuration makes it possible to address operations that require aseptic conditions and do not allow for intermittent separation of components of the system 8. To install the components of the microfluidic device 10 and system 8 without separation, valves 819, 815, 84, 86, and 86' are configured to receive tubes that are operated to block the flow of perfusion fluid. For example, all valves may be pinch valves, or, if their material is not suitable for reversible pinching, they may be thermal valves that operate by locally freezing the perfusion fluid in the channel or tube.

[0059] In the modified configurations shown in Figures 13 to 15, the sealing member 3' is not overmolded around the tubes of the perfusion lines 82, 82' and purge line 87, but is reversibly deformable to allow opening. More precisely, the sealing member 3' has three slots 34' extending from the hole 33' to allow access to the hole 33' in the open configuration of the slot 34', the hole 33' is fluid-tightly closed around the tubes of the perfusion lines 82, 82' and purge line 87 when the sealing member 3' is in a sealing configuration in the space between the cover 22 and the body 21. In this modified configuration, the sealing member 3' is fastened into the groove 220 of the cover 22, as a result, in the open configuration of the cover 22, the microfluidic device 10 can be placed on the support element 260 of the rack structure 26, and the tubes of the perfusion lines 82, 82' and purge line 87 can be inserted into the hole 33' of the sealing member 3' via the slots 34'. In the sealing configuration of the chamber 20, the sealing member 3' cooperates with the flat gasket 212 fastened to the main body 21.

[0060] A method for clamping multiple microfluidic devices 10 using apparatus 1 includes the steps described below.

[0061] First, in the open configuration of the container 2 shown in Figure 2, each of the multiple microfluidic devices 10, “pre-clamped” with adhesive tape 18, is positioned on the support elements 260 of the track structure 26 and connected to the perfusion fluid management system 8 via perfusion lines 82, 82' and purge line 87 through holes 33, 33' in the sealing members 3, 3'. Interconnecting tubes are connected to the valves 819, 815, 84, 86, 86 (which may be pinch valves or thermal valves in particular) in their operating positions.

[0062] Next, the chamber 20 is sealed to be fluid-tight with respect to the clamping fluid by displacing the cover 22 until the cover 22 contacts the main body 21 and seals the loading port 25. At this position, the fastening screws 28 are tightened to pressurize the sealing members 3, 211, 221. The inflatable O-ring 211 is pressurized at this stage. The displacement of the cover 22 is advantageously achieved automatically by sliding the sliding end 291 of the lifting arm 29 downward along the guide rail 91.

[0063] Once the chamber 20 is sealed, clamp fluid is supplied into the chamber 20 from the clamp fluid management system 6 to clamp the microfluidic devices 10 together by compressing the elastomer seal 13 under the pressure of the clamp fluid within the chamber 20. For this purpose, the clamp fluid is supplied through the fluid inlet 24 until a desired pressure of the clamp fluid, strictly higher than the pressure of the perfusing fluid in each of the microfluidic devices 10, is achieved within the chamber 20.

[0064] In one embodiment, the pressure of the clamp fluid in the chamber 20 and the pressure of the perfusion fluid in the microfluidic device 10 can be controlled by a control unit that includes both a control module 61 of the clamp fluid management system 6 and a control module 80 of the perfusion fluid management system 8. In one embodiment, this control unit is configured to receive a measurement of the pressure of the clamp fluid in the chamber 20 from a pressure sensor 65 and a measurement of the pressure of the perfusion fluid in the microfluidic device 10 from pressure sensors 83, 83', and to drive both the clamp fluid management system 6 and the perfusion fluid management system 8 according to the received pressure measurements.

[0065] In the second embodiment shown in Figures 17 and 18, elements similar to those in the first embodiment are given the same reference numerals. The clamping device 1 of the second embodiment differs from the first embodiment in that the fluid-tight penetration of the perfusion lines 82, 82' and purge line 87 through the wall of the chamber 20 is achieved not through a sealing member configured to seal the space between the cover 22 and the body 21, as in the first embodiment, but through a connecting unit 4 that is specially positioned to correspond to holes provided in the envelope of the container 2 for this purpose. The connecting unit 4 includes a casing 41 (in which a sealing resin may be poured) and three fluid passages 42 that extend through the wall of the chamber 20. Each passage 42 is provided with connectors 43 and 45 at both ends, respectively, for connecting tubes of perfusion lines 82, 82' or purge lines 87 connected to the microfluidic device 10 on the side facing the internal volume of the chamber 20, and for connecting tubes of perfusion lines 82, 82' or purge lines 87 connected to the perfusion fluid management system 8 on the side facing the outside of the chamber 20. In this embodiment, the inner walls of the fluid passages 42 are preferably made of a material that is easy to clean and sterilize, such as polytetrafluoroethylene (PTFE), glass, or stainless steel.

[0066] In the third embodiment shown in Figure 19, elements similar to those in the first embodiment have the same reference numerals. The clamping device 1 of the third embodiment differs from the first embodiment in that the entire perfusion fluid management system 8 (i.e., including the reactant module 81, the waste tank 89, and all of the perfusion lines 82, 82' and purge line 87 with associated pressure sensors 83, 83', 88) is housed within the internal volume of the chamber 20. In this third embodiment, the tube connecting the microfluidic device 10 to the perfusion fluid management system 8 is rigid enough to withstand the pressure of the clamping fluid in the chamber 20 without substantially deforming. In this way, the circulation of the perfusion fluid between the perfusion fluid management system 8 and the microfluidic device 10 is not affected by the pressurization of the chamber 20 by the clamping fluid during the clamping operation. For example, a small-diameter silicone tube having an inner diameter of about 0.8 mm and an outer diameter of about 2.4 mm is rigid enough not to deform excessively under operating conditions such as a clamping fluid pressure of 0 to 3 bar. In this third embodiment, the waste tank 89 is connected to a pressure controller or pressure generator, which differs from the previous embodiment in which the waste tank 89 is simply vented, for example, through a filter. The connection to the pressure controller or pressure generator is necessary to prevent the pressure of the perfused fluid in the waste tank 89 from becoming equal to the pressure of the clamped fluid in the chamber 20 (which could interfere with the purging operation and cause backflow).

[0067] The present invention is not limited to the examples described and illustrated.

[0068] In particular, any type of microfluidic device, especially a microfluidic device without elastomer components, can be clamped within the apparatus of the present invention, and each microfluidic device may be a stack of microfluidic chips rather than a single microfluidic chip as in the example above.

[0069] The microfluidic device clamped within the apparatus of the present invention may further comprise active elements such as internal valves, electrodes, sonotrodes, and light sources. The microfluidic device clamped within the apparatus of the present invention may also comprise sensors. The microfluidic device clamped within the apparatus of the present invention may further comprise built-in electronic equipment.

[0070] In addition, the clamp fluid can be a gas, a liquid, or a combination of both. The molecular composition of the clamp fluid can also be modified. In particular, if the clamp fluid is a mixture of gases, the proportion of each gas in the gas mixture can be monitored and controlled. For example, in embodiments in which living cells are processed, controlling the concentrations of CO2, O2, and N2 may be important if convective and / or diffusive gas molecular exchange occurs between the clamp fluid and the channels of the microfluidic device.

[0071] As mentioned above, active systems other than the imaging system described above may be used in the pressurized chamber during the clamping operation of the microfluidic device. In particular, the apparatus of the present invention may include any other type of monitoring system, such as a temperature monitoring system, a calorimetry system, an electromagnetic impedance measurement system, or any system configured to induce the contents of the microfluidic device.

[0072] When the perfusion fluid management system is located outside the chamber (especially in cases other than those exemplified above), any means of fluid-tight transport of the tubing may be used. In addition, the perfusion fluid management system may allow switching between several perfusion modes for perfusing the microfluidic device or each microfluidic device. For example, a microfluidic device with an alternative fluid circuit and three or more ports may be perfused along the alternative fluid circuit using valves configured to connect selected ports to selected flow lines.

Claims

1. A device (1) for clamping at least one microfluidic device (10) with a clamping fluid in order to seal the microfluidic device (10), A fluid-sealed chamber (20) having a fluid inlet (24), Clamp fluid management system (6), Perfusion fluid management system (8) Control unit, and Equipped with at least one active system, The fluid-sealed chamber (20) is configured to house the clamp fluid and the microfluidic device (10), and the microfluidic device (10) has at least one deformable portion (13) configured to be clamped by being compressed under the pressure of the clamp fluid in the chamber (20), The microfluidic device (10) contains a perfusion fluid, The device (1) is configured to seal the microfluidic device (10) while ensuring access to the entire area surrounding the microfluidic device (10) during the clamping operation. The clamp fluid management system (6) comprises a pressure source (62) connected to the fluid inlet (24) of the fluid sealing chamber (20) via a duct (64), and a control module (61). The clamp fluid management system (6) is configured to supply the clamp fluid from the pressure source (62) toward the fluid inlet (24) and to adjust the pressure of the clamp fluid inside the fluid sealing chamber (20) during the clamping operation. The perfusion fluid management system (8) is configured to be connected to the microfluidic device (10), The perfusion fluid management system (8) comprises at least one pressure controller (817, 818) and a control module (80), The perfusion fluid management system (8) is configured to adjust the pressure of the perfusion fluid in the microfluidic device (10) so that during the clamping operation, the pressure of the clamped fluid in the fluid sealing chamber (20) is strictly higher than the pressure of the perfusion fluid in the microfluidic device (10). The control unit comprises both the control module (61) of the clamp fluid management system (6) and the control module (80) of the perfusion fluid management system (8), and the control unit is configured to drive and manage both the clamp fluid management system (6) and the perfusion fluid management system (8). At least one active system is configured to monitor or guide the contents of the microfluidic device (10) received in the chamber (20) during the clamping operation, via at least one accessible peripheral wall of the microfluidic device (10). Device (1).

2. The clamp fluid management system (6) includes a pressure sensor (65) configured to measure the pressure inside the fluid sealing chamber (20), and The perfusion fluid management system (8) includes pressure sensors (83, 83') configured to measure the pressure within the microfluidic device (10), The apparatus (1) according to claim 1, wherein the control unit is configured to receive a measured value of the pressure of the clamp fluid in the fluid sealing chamber (20) from the pressure sensor (65) of the clamp fluid management system (6), and to receive a measured value of the pressure of the perfusion fluid in the microfluidic device (10) from the pressure sensors (83, 83') of the perfusion fluid management system (8), and is configured to drive the clamp fluid management system (6) and the perfusion fluid management system (8) according to claim 1.

3. The apparatus (1) according to claim 1 or 2, wherein the fluid sealing chamber (20) is configured to house a plurality of microfluidic devices (10) within its internal volume, which are clamped together under the pressure of the clamp fluid within the fluid sealing chamber (20).

4. The apparatus (1) according to any one of claims 1 to 3, comprising at least one monitoring system (5) configured to monitor the contents of the microfluidic device (10) housed in the fluid-sealed chamber (20) and / or to guide the contents of the microfluidic device (10) housed in the fluid-sealed chamber (20) through at least one wall of the microfluidic device (10) during the clamping operation.

5. The apparatus (1) according to claim 4, comprising: a monitoring system (5) configured to monitor the contents of the microfluidic device (10) housed in the fluid sealing chamber (20) during the clamping operation; and a control module (80) configured to drive the perfusion fluid management system (8) according to the measurements of the monitoring system (5).

6. The apparatus (1) according to any one of claims 1 to 5, wherein the fluid-sealed chamber (20) is provided with a loading port (25) for inserting and removing the microfluidic device (10) into and out of the fluid-sealed chamber (20), and the loading port (25) is fluid-tightly closed during the clamping operation.

7. The apparatus (1) according to any one of claims 1 to 6, wherein the fluid-sealed chamber (20) is configured to house only a portion of the perfusion fluid management system (8) within its internal volume, and the apparatus (1) comprises at least one sleeve (3, 3') configured to be positioned within an opening (25) in the wall of the fluid-sealed chamber (20) during the clamping operation in order to allow fluid-tight penetration of at least one tube (82, 82', 87) connecting the microfluidic device (10) to the perfusion fluid management system (8).

8. Apparatus (1) according to claim 7, wherein the sleeve (3, 3') comprises at least one hole (33, 33') configured to receive tubes (82, 82', 87) connecting the microfluidic device (10) to the perfusion fluid management system (8), the hole (33, 33') extending between an inner end (32) of the sleeve intended to be directed toward the internal volume of the fluid-seal chamber (20) and an outer end (31) of the sleeve intended to be directed toward the outside of the fluid-seal chamber (20), and is fluid-tightly closed around the tubes (82, 82', 87).

9. The fluid-sealed chamber (20) is configured to house only a portion of the perfusion fluid management system (8) within its internal volume, and the apparatus (1) comprises a connection unit (4) within the wall of the fluid-sealed chamber (20) including at least one fluid passage (42) extending through the wall of the fluid-sealed chamber (20), and connectors (43, 45) provided at both ends of the fluid passage (42) for connecting tubes (82, 82', 87) connected to the microfluidic device (10) on the side facing the internal volume of the fluid-sealed chamber (20) and tubes (82, 82', 87) connected to the perfusion fluid management system (8) on the side facing the outside of the fluid-sealed chamber (20).

10. The apparatus (1) according to any one of claims 1 to 9, comprising at least one support element (260) within the fluid-sealed chamber (20) configured to house a clamped microfluidic device (10).

11. A method for sealing and clamping at least one microfluidic device (10) in a device (1) during a clamping operation using a clamping fluid, wherein the microfluidic device (10) includes at least one deformable portion (13) and a perfusion fluid, and the method is as follows: The steps include: housing the microfluidic device (10) in a fluid-sealed chamber (20) of the apparatus (1), wherein the fluid-sealed chamber (20) has a fluid inlet (24), and the microfluidic device (10) is connected to a perfusion fluid management system (8) of the apparatus (1); The steps include sealing the fluid sealing chamber (20) so as to be fluid-tight with respect to the clamp fluid, The step of pressurizing the fluid sealing chamber (20) with clamp fluid supplied through the fluid inlet (24), The steps include: applying a pressure of the clamping fluid in the fluid-sealed chamber (20) that is strictly higher than the pressure of the perfusing fluid in the microfluidic device (10), thereby compressing and deforming the at least one deformable portion (13) under the action of the pressure of the clamping fluid in the fluid-sealed chamber (20) to clamp the microfluidic device (10); and A method that includes this.

12. The method according to claim 11, wherein the pressure of the perfusion fluid in the microfluidic device (10) is controlled by a control module (80) of the perfusion fluid management system (8) which is configured to receive a measurement value from a monitoring system (5) of the device (1) that monitors the contents of the microfluidic device (10) during the clamping operation, and to drive the perfusion fluid management system (8) in accordance with the received measurement value.

13. The method according to claim 11 or 12, wherein a plurality of microfluidic devices (10) are positioned within the fluid-sealed chamber (20) and are clamped together under the pressure of the clamp fluid in the chamber (20) by applying a pressure of the clamp fluid in the fluid-sealed chamber (20) that is strictly higher than the pressure of the perfusation fluid in each of the microfluidic devices (10).

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