HK1096334A - Device having a self sealing fluid port - Google Patents

Description

Device with self-sealing fluid port

Technical Field

The present invention relates to devices having self-sealing fluid ports, methods of making the devices, systems for analyzing fluids using the devices, use of the devices in analyzing fluids, and methods of analyzing using the devices.

Background

Especially in analytical laboratories, it is of great interest to perform the analysis in a convenient, safe and reliable way. A particular problem is the contamination of the reagents, samples and equipment for performing the analysis with the environment and the contamination of the environment with the reagents or samples.

Accordingly, devices for storing, transporting and processing samples and/or reagents have been proposed.

In a simple form, such a device is a tube-based single-chamber device having a generally conical or/and cylindrical form with a single fluid port, which is closed after introduction of the sample and reagents by a lid attached to the tube body by a hinge and made of the same material as the device. The cover needs to be removed in order to remove any liquid from the device, or in order to add more reagent to the device. Such a device is disclosed in EP 907083. During the analysis, those devices require handling of the lid, i.e. use of a lid manipulator designed to retrieve the lid from the storage, to fix the lid to the device and to remove the lid from the device.

In EP 724483 a tube is disclosed which is closed by a cap which can be pierced with a steel needle. However, those assemblies still require the cap to be manipulated prior to analysis, i.e., the tube to be closed with the cap after the sample and reagents are added.

Those based on tubes have this drawback: they have only one fluid port through which introduction and removal of liquid has to be performed.

More sophisticated analysis requires more sophisticated equipment. For example, US 6,537,501 discloses a cartridge comprising a microfluidic flow channel and further compartments, like a waste storage container. Furthermore, it discloses the use of a septum to close one or more inlet channels. The septum is typically a plug used to close a fluid port and is made of an elastomeric material, such as rubber. The septum is manufactured as a mass and inserted into the fluid port of the tube after it is completely produced. To introduce liquid into the barrel interior through the slit in the septum by a steel needle, the septum may be pierced by a steel needle or cannula. Conventional septa are used to close the opening if pressed into the opening. Pressure forces perpendicular to the force piercing the septum are thus continuously acting on the opening.

In US 2003/0138969 a method of introducing a fluid sample into a small diameter tube through a pipette or syringe needle introduced into the fluid inlet port is disclosed. In order to bring the pipette tip or the injection needle into close contact with the inlet port, a stopper made of rubber or silicon called an adapter is used. The adapter fits within the inlet port.

A first need is for safe, reliable storage and/or handling of fluids (mainly comprising liquids, but also gases). A second need is for a system comprising a device for receiving and treating a fluid, which has an interior, which remains closed before or/and in use or/and after use.

This situation is for example: storage of liquids for nutrition, storage of liquids for medication, storage of liquids for analysis, and storage of samples for analysis. Another use is a system that includes and uses the device for analysis.

Safe storage and/or reliable handling of fluids is based on the requirement that: the fluid cannot be contaminated by the environment, nor can the fluid contaminate the environment. In the case of devices that need to be kept closed before use, this is based on the requirement that: the device cannot be contaminated prior to use.

One object of the present invention is: a device is provided having an easily and reliably operable fluid port that can be economically manufactured.

Disclosure of Invention

A first subject of the invention is a device having a body comprising a cavity and a fluid port made of an elastomer attached to the body.

A second subject of the invention is a method of manufacturing such a device, wherein the body is made of a first rigid material and the fluid port is made of a second elastic material, the method comprising:

-a first moulding step comprising

a) Providing a first mold reflecting the external shape of the rigid body,

b) injecting said first rigid material in liquefied form into a mould,

c) waiting until said first material becomes at least partially solid,

-retaining the result of the first moulding step in a second mould reflecting the shape of the fluid port

-a second molding step comprising

a) Injecting said second elastomeric material in liquefied form in its final state into said second mold,

b) waiting until the second material becomes at least partially solid,

and

-removing the result of the moulding step from the mould after solidification of the material.

Yet another subject of the invention is: a system for analyzing a liquid comprising an instrument with a fluid actuation module comprising a rigid fluid actuator and a device according to the invention.

Furthermore, the subject of the invention is: a device manufactured according to the manufacturing method according to the present invention.

Another subject of the invention is: use of a device according to the invention in a method of analysing a liquid.

Yet another subject of the invention is: a method of analysing a fluid or a component thereof, comprising

-providing a device according to the invention,

-introducing a fluid to the device through the fluid port by piercing the fluid port and dispensing the fluid via a breach formed by the piercing, and

-determining any property or change in property of the fluid or its components.

Drawings

In fig. 1a and 1b, two embodiments of the device according to the invention are shown.

In fig. 2a and 2b, two alternative devices are shown, each in a pre-assembled and a post-assembled state.

Detailed Description

The apparatus of the present invention is useful in receiving fluids, or/and storing fluids, or/and chemically or physically treating fluids, or/and analyzing fluids.

The fluid may be a sample, reagent, diluent or process fluid or a combination thereof or a fluid derived therefrom. It may be liquid or gaseous.

The size of the apparatus according to the invention depends mainly on the amount of fluid to be held or treated in the apparatus and the kind and number of steps to be performed. In a first embodiment, the apparatus is used as a means for separating components of a fluid. The components that need to be separated will remain in the apparatus while allowing the residual liquid to exit the apparatus. In this case, the liquid volume may exceed the volume of the cavity within the device. This allows for a rather small device. For this embodiment, the total volume employed by the apparatus is preferably between 100. mu.l and 10 ml.

In a preferred second embodiment, the device will be only slightly larger than the volume of the fluid and any reagents that will react with the fluid. Since the fluid volume preferably has a volume of more than 0.1. mu.l, preferably between 0.2. mu.l and 1L, the volume of the device will exceed 200. mu.l, preferably between 200. mu.l and 1.1L, most preferably between 500. mu.l and 110 ml. The device optionally has a substantially flat structure, i.e. over its major part it has a thickness of less than 50mm, preferably between 0.2 and 10mm, and a length and width of less than 300mm, preferably between 2 and 150 mm. If a portion of the device requires a greater thickness, the portion may extend beyond the generally flat configuration.

For receiving and maintaining a fluid, the device has a body, which further comprises one or more cavities, which temporarily or continuously receive or/and maintain the fluid or a fluid derived therefrom.

The body of the device is conveniently formed from at least one relatively rigid polymer. The polymer used for the body according to the invention is preferably selected from the group of thermoplastic materials, such as polypropylene, polyethylene, polystyrene, polycarbonate and polymethylmethacrylate. The body is further preferably made of a material that can be liquefied by heating above its melting temperature and, in the molten state, can be introduced into the mold to reflect the particular shape that the body or part thereof is required to assume.

The device according to the invention comprises a chamber. Typical volumes of the chamber may range from 1. mu.l to 1L, preferably from 100. mu.l to 100 ml. The chamber may have different forms to accommodate various intended uses of the device. The cavity may be divided into a plurality of sections with channels and chambers. Preferred chambers include one or more channels and/or one or more chambers.

The use of those channels may be varied, for example

Transport of fluid between two locations (e.g. chambers) within the apparatus,

-transporting a fluid into or out of the apparatus,

-measuring the fluid flow in a measuring chamber,

-treating a fluid or treating a substance dissolved or suspended in a fluid.

The use of those chambers may be varied, for example

-storing, receiving, transporting a fluid,

-processing the fluid, for example analysing a substance in the fluid,

suitable for measuring physical or chemical properties of the fluid (e.g. for optical absorption or fluorescence measurements).

The cavity more preferably comprises two or more channels, one leading to the chamber and one leading away from the chamber. In a preferred embodiment, the cavity has a volume of less than 1L, preferably between 1. mu.l and 100 ml. Through-hole formed inside the device, especially preferably inside the device bodyRoad, preferably having less than 10mm²Preferably between 0.01 and 2mm²In the meantime. The cavity formed inside the body is preferably of a size suitable for the intended use of the procedure. The channel for transporting the fluid through the device will preferably have a smaller size than the chamber to hold the fluid or/and to perform a process, preferably a chemical reaction. For example, a chamber for separating nucleic acids from a fluid will preferably have a volume of 5 to 100. mu.l. The chamber for performing the polymerase chain reaction may have a volume between 0.1 and 500. mu.l. If combined amplification and detection is to be performed in the chamber, the chamber preferably has a volume between 0.1 and 500. mu.l. The preferred substantially flat chamber has a depth of between 10 μm and 49mm, preferably between 10 μm and 20mm, and the length and width of the chamber may be between 10 μm and 295mm, preferably between 20 μm and 145 mm. The chamber is very preferably a flat chamber with a thickness of less than 2000 μm, preferably between 50 μm and 5 mm. The thickness is most preferably between 50 μm and 1 mm. A first channel formed in the body preferably leads from an input location into the chamber and a second channel preferably leads from the chamber to an exit location on the device. If more than one chamber is provided within the body, there may be more channels, for example connecting chambers within the body. The chamber and the channel may be selectively arranged such that any liquid introduced into the chamber through the inlet channel fills the chamber before exiting the chamber through the outlet channel. Portions of the cavity may comprise an interposed material, a so-called "solid phase", which may be used for adsorption, filtration and/or reaction on the surface. The cavity is more preferably closed by a body with a fluid port and a sealing wall attached to the body.

The final device is thus usually a composite of several elements. This means that it comprises two or more separately manufactured and subsequently assembled parts, at least one part of the device comprising a 2-component injection moulded part having a rigid body and an elastomeric fluid port attached to said body. Since it has proven difficult to manufacture the body comprising the chamber suitable for chemical analysis in one piece, it is preferred that the device is made of two or more parts, which are combined to form one or more chambers.

In a highly preferred embodiment, the device comprises a first element, called "body", with fluid ports, and a second element, called "sealing wall", the body having grooves or/and channels. The rigid body provides rigidity to the device to maintain the shape of the cavity throughout manufacture and use of the device.

The thin foil or/and the body preferably has a transmission of more than 2% for electromagnetic waves having a wavelength in the range of 300 to 4000 nm.

The two parts of body and sealing wall can be joined in a known manner. In a preferred embodiment, in which the sealing wall is a thin layer and the rigid body is made of a polymer, such as polystyrene, the two parts can be combined and then sealed by welding, such as laser welding, ultrasonic welding, heat sealing or gluing. The two parts may simply be clipped or glued together. The elastomer part may serve as a sealing part between the body and the sealing wall in case the parts are clamped or stuck together.

The bonding method, the host material and the sealant wall material must be selected to fit together. For example, if the bonding method is laser welding, then the bulk material of the body and the sealing wall is the same material (e.g., polypropylene), but one of the two materials is dyed to absorb the laser energy. If the bonding method is ultrasonic welding, the two materials are generally the same. If the bonding method is heat sealing, the sealing wall is a heat sealable foil, suitable for heat sealing to the body. Such a heat sealable foil is typically a composite of several materials, wherein the layers sealing the opposite sides are capable of being sealed to the body. Typical foils suitable for bonding to polypropylene bodies, having a composite layer of aluminium or polyester and polypropylene-such sealing foils are known and commercially available. In case the sealing wall is a foil, the foil is preferably thin, having a thickness between 20 and 1000 μm, more preferably between 50 and 250 μm. For applications requiring heat transfer, the foil has a good heat transfer rate, e.g. greater than 400W/m²a/K, more preferably more than 200W/m²/K。

The device may contain further elements which are useful for the intended purpose of the device. For example, the heat transfer wall or heating element may be integrated into the enclosure wall or body. The heat transfer wall may be used to heat or/and cool a fluid contained in the apparatus.

In another embodiment, the electrodes may be incorporated into the body or the containment wall. The electrodes may be used to determine the electrochemical state of a fluid contained in the device or to initiate an electrochemical reaction within the device. In this case, the device will have suitable connectivity to the circuitry.

In another embodiment, an optical window (allowing at least partial transmission of wavelengths) may be incorporated into the body or the containment wall.

The device according to the invention is designed to hold, transport or receive a fluid. In order to introduce fluid into the cavity of the body or/and to remove fluid from the cavity of the body, the device according to the invention has one or more fluid ports. Furthermore, the device may be used to hold a fluid inside a chamber, more preferably in one or more chambers. Furthermore, the fluid contained in the device may also be transported to the outside, for example by forcing the fluid out of the cavity through a fluid port in the device. The fluid port may be the same or different than the fluid port used to introduce fluid into the device. The device preferably has two or more fluid ports, at least one of which is closed according to the invention. More preferably two or more fluid ports in the device, most preferably all fluid ports in the device, are closed using fluid ports made of an elastomer. Forcing the liquid out of the device may require maintaining pressure to force the liquid through the fluid port, or may require applying negative pressure or even vacuum to the chamber. This may be accomplished by adding a rigid fluid removal actuator through a fluid port in the device and applying a negative pressure through the actuator. One or more or even all of those openings are closed by fluid ports.

To more easily interface with a fluid actuator, such as a hollow needle, the outside of a fluid port formed from an elastomeric material may have a recess. The recess has a conical shape at least in part thereof, wherein the angle (W) shown in fig. 1 is in the range of 5 to 150 °, more preferably between 15 and 90 °.

According to the present invention, the fluid port is made of an elastomer that is formed during a 2-compound (2C) injection molding process. The elastomeric material is preferably selected from the class of thermoplastic elastomers (TPE), thermoplastic vulcanised elastomers (TPV) or Vulcanised Elastomers (VE), which are commonly used in 2C injection moulding processes. The first and second materials are selected to adhere to each other.

Several thermoplastic elastomers can be used as the final (at room temperature) elastomeric material, such as elastomeric Thermoplastic Polyolefin (TPO) or elastomeric Thermoplastic Polyurethane (TPU) or elastomeric styrene-block-copolymer (SBS, SEBS). TPV, thermoplastic vulcanizate elastomers, which are a special class of TPEs containing a crosslinked rubber phase dispersed in a thermoplastic polymer phase, may also be used. TPVs provide elastomeric characteristics like crosslinked rubbers, but are as processable as thermoplastic polymers.

For details of TPE, see british plastic association publications, for example, in http: // www.bpf.co.uk/bpdefinition/plastics materials thermal rubber TPR.cfm.

A preferred combination of materials is a combination of polypropylene as the rigid material and TPO as the elastomeric material.

The elastomer is typically chosen to match the manufacturing (2-part injection molding process) and is chosen to have some adhesion to the host material. Typical adhesion forces between the rigid body and the elastomeric fluid port are at least 0.1N, typically greater than 1N.

Thermoplastic elastomers (TPEs) have the property of liquefying when heated to a temperature above their melting point without decomposing, and solidifying when cooled to a temperature below their melting point to reflect the geometry of the mold in which they are held. A preferred thermoplastic Elastomer for use in combination with polypropylene as the rigid material is TPESantopren  8281W-35W-237 from Advanced Elastomer Systems. A preferred group of thermoplastic elastomers has a processing temperature between 180 and 220 ℃. The elastomer has a shore hardness of 0A to 100A, preferably in the range of 20A to 60A, in its final shape and at the temperature of use.

The elastomeric fluid port forms a pierceable barrier between the cavity and the environment. The elastomeric fluid port protects the cavity within the device from environmental contamination, and vice versa, fluids stored, processed, or analyzed in the device cannot contaminate the environment.

The fluid port is configured to securely close a cavity within the device, but is of a size that allows piercing of the fluid port from outside the device with a rigid fluid actuator, preferably a pipette tip or a hollow steel needle, so that after piercing, fluid can be introduced from the actuator into or out of the cavity of the device through the fluid port. The fluid port according to the present invention more preferably covers the end of one channel, if not the end, from which the fluid is freely accessible. In this case, the fluid will be introduced into or out of a cavity present within the device by the actuator, preferably through a passage within the device leading to the chamber. There are several possible geometrical schemes regarding the geometry of the inlet and fluid ports.

In a first preferred embodiment, shown in fig. 1a, where the device is substantially flat, one fluid port (4) or two fluid ports (4) are located on the flat side of the device (1). In this case, one or more fluid ports are provided on the body side of the device, i.e. through the body leading from the flat side to the cavity (3). In this way, the fluid port can be pierced from the flat side of the device.

In a first possible embodiment (fig. 1a), at least one fluid port (4) is located on one side of the body, the fluid port being made of a thermoplastic elastomer attached to said body. Here the fluid port forms a wall relative to the surroundings, in particular a wall interfacing with the fluid actuator, on one side, and a wall of the cavity on the opposite side. In the example of fig. 1a, two fluid ports are shown interfacing with a common lumen in the device.

In another embodiment (fig. 1b), the fluid port forms a wall on one side with respect to the surroundings (in particular a wall interfacing with the fluid actuator), but, as a complement to the embodiment shown in fig. 1a, it has a cavity on the opposite side (leading to a further cavity in a rigid material). In the example of fig. 1b, the cavity in the thermoplastic elastomer is a channel that leads in a direction perpendicular to the direction of the fluid actuator interface. The material forming the fluid port is still made of a thermoplastic elastomer attached to the body.

Typically, where one of the chambers or channels is located on a plane different from the plane of the other chamber or channel to which the first chamber or channel is to be connected, the two chambers may be connected by a channel which is not in the same plane as those chambers but leads from one plane to the other.

In the case where the plane of the chamber or channel is not in the same plane as the direction of the fluid actuator piercing the fluid port, the cavity preferably extends into said fluid port. Such an embodiment is illustrated in fig. 1 b.

In another option, the material from which the fluid port is made, i.e., the elastomeric material, contains a channel through which the actuator or fluid can enter the cavity. The channel is preferably located in a direction orthogonal to the movement of the actuator when piercing the port.

In another embodiment, shown in fig. 2a, the body has the shape of a cap, while the cavity (3) is partially or curvedly contained within the sealing wall (5). In the lower half of fig. 2a, the body cover is shown above a portion of the sealing wall (only a partial view of the entire device showing the fluid port portion of the device). The body further comprises a snap-in mechanism (6) and the sealing wall comprises a snap-in recess (7). In the upper half of fig. 2a, the assembled device is shown, wherein the rigid carrier is firmly connected to the sealing wall. In this case, the connection is made by form-fitting, in particular snap-in connection. As further shown in fig. 2b, a sealing edge (8) on the sealing wall (5) is provided which compresses the fluid port to provide a fluid tight connection between the body and the sealing wall.

In another embodiment shown in fig. 2b, the body is again used as a lid to close the opening in the sealing wall. This embodiment uses gluing or holding together by welding, rather than a snap-in connection. The device is assembled by placing the rigid body (2) on the sealing wall (5).

In general, the use of an elastomer formed during a 2C injection molding process as a fluid port has the advantage of: the materials may be selected to adhere to each other. The attachment between the body and the fluid port is achieved by the properties of the material of the rigid body and the material of the elastomeric fluid port. In the molten state, the elastomer may achieve a shape that adheres tightly to the body and seals any openings in the body. The shape may have protrusions and recesses that enter recesses in or around the protrusions of the body, particularly in the vicinity of the fluid port. Thus, the press fit is preferably made through a surface having an angle formed by the outer surface of the carrier and the surface of the carrier used to connect the fluid ports.

The press fit has the effect of tightly connecting the fluid port to the body such that the connection between the fluid port and the body is impervious to fluid even when a rigid fluid-carrying actuator, such as a steel needle, is used to pierce the fluid port, and after dispensing fluid into the cavity, when the actuator is removed from the fluid port.

In one embodiment, the fluid port of the present invention is not intended and is not configured to be removed from the device. The fluid port of the present invention cannot be removed from the body without seriously damaging, i.e., destroying the integrity of, the device. The intended function of this connection is to sufficiently separate the interior (cavity) of the device from the exterior of the device, mainly to avoid undesired or uncontrolled or excessive transport of material from the environment to the interior of the device and from the interior of the device to the environment. The mass transport may be pressure driven, diffusion driven or by any other mechanism, such as gravity driven (dust).

As already indicated above, the elastomer, preferably formed during 2C injection molding, allows piercing of the fluid port. The puncture results in the introduction of a rigid fluid-carrying actuator, such as a steel cannula, and through an opening in the cannula into the cavity. By allowing this passage, the fluid port is ruptured in this position, in which the actuator enters and passes through the fluid port. In another embodiment, the fluid port may be pre-pierced, but the piercing path is closed by the repulsive force of the elastomer.

Depending on the end application, the apparatus is leak-free (meaning sufficiently closed for the transport of substances into and out of the apparatus)

a) Only during the course of the storage process is it,

b) during the course of storage and use of the article,

c) during and after storage, use.

The fluid ports are typically designed to avoid uncontrolled transport of material into and out of the chamber. The thickness of the elastomeric fluid port at the entry point of the fluid actuator is in the range of 0.1mm to 40mm, more preferably in the range of 1 to 10 mm. To receive sufficient repulsion of the elastomer toward the puncture path, sufficient diameter of the elastomer is required to close the puncture path upon fluid actuation. The diameter of the elastomer must therefore be greater than 3 times the diameter of the fluid actuator at least one point of the piercing path.

A preferred embodiment of the fluid port is a self-sealing port, i.e. a port allowing self-sealing closing the space between the fluid actuator and the fluid port, so that fluid cannot escape. Furthermore, after removal of the actuator, the fluid port still closes the opening of the body even at a pressure differential of 1 bar, and the puncture path does not allow fluid to escape the device.

In a preferred embodiment, the device is a microfluidic device. As used herein, a microfluidic device has one or more channels with a cross-section greater than 0.1 μm²More preferably at 10 μm²To 10mm²In the meantime. Microfluidic devices may additionally or alternatively include one or more chambers having a specific openingThe cross section of the channel is large. The chamber of the microfluidic device may have a volume of 10nl to 50ml, more preferably between 1 μ l to 25 ml.

The device may also have fluidic or microfluidic functions. Those functions are generally known as means for physically handling the fluid in the chamber. Those may be static elements like fittings, including walls and surfaces, e.g. for fluid mixing, differentiation or combination. Other functions that the cavity may provide are optical functions. To this end, the body surrounding the cavity, preferably the chamber, is transparent to allow light to enter or/and light to escape from the cavity to the exterior of the device. The cavity preferably has dimensions that allow for the collection of a fluid in an amount sufficient to reliably detect the components contained in the fluid. Another function of the cavity may be to receive a material that reacts with the fluid. Such materials may be selected from the group of soluble or insoluble reagents, or combinations thereof, or both, even in separate portions of the cavity, or chambers. The soluble agent may be an agent that: supporting sample lysis, amplifying nucleic acids contained in the sample or a liquid derived therefrom, or providing a signal when reacting with a component of the sample to be determined. The insoluble agent may be a solid that is designed to immobilize a fluid component or a compound derived therefrom. Examples of nucleic acid-immobilizing solids are glass wool or magnetic particles, which are capable of binding nucleic acids from a turbid solution. Suitable materials are known to those of ordinary skill in the art of nucleic acid sample preparation.

To provide the function of holding, carrying, or receiving fluid, the fluid port is located in the device so that the rigid fluid actuator is accessible from outside the device. This may be accomplished by configuring the device such that none of the elements of the device interfere with the movement of the actuator in a direction to and ultimately through the fluid port. In fact, the fluid port will be located on a flat surface of the device. To improve accessibility, the apparatus may further comprise a surface that guides the actuator to a point of the fluid port at which the actuator is to form a breach of the fluid port. Such a surface may be provided on the body or on the fluid port. The guide surface shown in the figures is indicated with reference numeral 9. The guide surface most preferably has a tapered shape, tapering towards the location where the fluid port is to be pierced.

The fluid that can be received, held or transported in the device according to the invention can be any fluid that is of interest to undergo a specific treatment. The fluid is preferably a liquid. The fluid is more preferably an aqueous solution. In a preferred use of the device according to the invention, the composition of the liquid is intended to be processed or analysed. In the diagnostic device, the liquid contains the component to be determined in the analysis. Such a liquid may be selected from the group of environmental fluids like water from rivers or fluids extracted from soil, food fluids like fruit juices or extracts of plants or fruits, or fluids received from the animal or human body like blood, serum, plasma, urine, cerebrospinal fluid or lymph, or liquids derived therefrom like liquids containing components separated from the aforementioned liquids like liquids containing pure antibodies or nucleic acids. The liquid may further comprise additional components useful for analyzing the components of the liquid, or reagents for chemical reactions to be performed within the device. Those reagents may include labeled binding partners, such as labeled oligonucleotide probes or dyes.

Further subject matter of the invention are: a method of manufacturing an apparatus having a rigid body comprising a cavity and constructed of a first rigid material and a fluid port constructed of a second elastomeric material, the method comprising

-a first moulding step comprising

a) Providing a first mold reflecting the external shape of the rigid body,

b) injecting a first material in liquefied form into a mold,

c) waiting until said first material becomes at least partially solid,

-retaining the result of the first moulding step in a second mould reflecting the shape of the fluid port,

-a second molding step comprising

a) Injecting a second material in liquefied form into the first portion of the second mold,

b) waiting until the second material becomes at least partially solid,

and

-removing the result of said moulding from said mould after solidification of said material.

The apparatus is preferably an apparatus as described above.

The method essentially combines the production of a device integrated by at least a two-step molding process. For example a first rigid material from which the body is made, preferably a material selected from the group of thermoplastics. Particularly preferred are polypropylene, polyethylene, polystyrene, polycarbonate and polymethyl methacrylate. Those materials are preferred materials for forming one or more portions of the body of the device described above. In order to give the first rigid material the shape required by the device, a mould is provided to reflect the outer shape of the body. Molds for injection molding processes are generally known and used in the art. It preferably comprises a metal or ceramic mold, preferably a cavity having one or more fluid ports to fill the mold with liquefied material. The mould is preferably made of two or more parts which are joined during the moulding process and which can be separated after the material has set in the mould.

Thus, the result of the molding process can be removed from the mold. In fact, the final shape of the result of the molding process is determined by the shape of the mold. For example, the grooves in the outer surface, which are the result of the molding process, are created by protrusions in the mold cavity, which are useful for forming channels and chambers within the body. Channels through the result of, for example, the molding process of the body, such as those leading from one side of the carrier containing the chambers to the other side of the carrier having the openings, are created by rods connecting the first part of the mold and the second part of the mold.

To perform the molding process, the first final rigid material is liquefied by heating it to a temperature above the melting temperature. The material is then injected into the cavity of the mold, preferably by applying pressure and allowing air contained within the cavity to escape through a fluid port other than the fluid port through which the material was injected.

The result of the first molding step will be subjected to a second molding step using a second material. This requires that the surface of the second part of the device, i.e. the surface of the fluid port, which is the result of the first moulding step, should be connected to be accessible to the second material in liquefied form. To achieve this, there are several ways that can be utilized. In all those methods, this is preferred: the first material becomes at least partially solid. The temperature of the first material is more preferably reduced or lowered to a temperature below the melting point. This can be achieved by active or passive cooling. In a first embodiment, in order to allow the liquefied second material to reach the surface, the part of the mould covering the surface of the shape formed in the first moulding step is removed. In addition, a second die is coupled to the remainder of the first die to reflect the external shape of the fluid port. This will form a cavity within the final assembled mold, reflecting the external shape of the fluid port connected to the device body.

In a second embodiment, the result of the first molding step is removed from the second mold and introduced into the second mold, which reflects the shape of the body surrounding the portion of the fluid port to be created and the second portion reflecting the external shape of the fluid port to be molded on the result of the first molding step.

The cavity of the second mold is then filled with a liquefied second material by injection.

This may be achieved by additional moulding steps as described above, if additional components need to be added to the body or device, or after the result of the moulding process is removed from the mould and the material set.

Although the above describes a process wherein the first material is a more rigid material and the second material is a material of the fluid port, which is preferably an elastomer shaped in a 2C injection molding process, the process may be performed in alternative ways, typically changing the order of the molding steps. In an alternative process, a mold is provided that reflects the exterior shape of the fluid port, similar to that described above for the rigid material, and a second molding step is performed using the first rigid material in liquefied form in a second mold after the second material has partially or fully solidified.

Therefore, another subject of the invention is: a method of manufacturing an apparatus having a body comprising a cavity and a fluid port comprising a second elastomeric material, the method comprising

-a first moulding step comprising

a) Providing a first mold reflecting the external shape of the fluid port,

b) injecting a second material in liquefied form into the first mold,

c) waiting until said second material becomes at least partially solid,

-providing the result of said first moulding step into a second mould, a first portion of which reflects at least part of said first mould and a second portion reflects the external shape of said body,

-a second molding step comprising

a) Injecting the first material in liquefied form into the second portion of the second mold,

b) waiting until the first material becomes at least partially solid,

and

-removing the result of said moulding from said mould after solidification of said material.

In the above manufacturing method, a further assembly step may be added. For example, any cavity may be sealed in an additional step, preferably by applying a sealing wall to the result of the molding step, which wall bounds the cavity within the body, e.g. completes the shape of one or more channels or/and chambers. This can be achieved by tightly joining the sealing wall surfaces and the result of the moulding step and gluing or welding the materials together. Preferred ways of connecting the carrier and the sealing wall are laser welding, ultrasonic welding, heat sealing or gluing.

Another subject of the invention is: a system for analyzing a liquid, comprising

An instrument having a fluid transport module comprising a rigid fluid actuator, and

-a device according to the invention.

Instruments for analyzing fluids are generally known. Those include modules that are typically needed for analysis. Preferred modules for such an instrument are optics for determining an optical property or change in an optical property of a liquid, a mechanical mechanism for moving a liquid from a first location to one or more other locations, and a liquid handling module for dispensing or/and aspirating a fluid from a tube, vessel or reagent container. The system according to the invention requires a rigid fluid actuator for dispensing a fluid into or/and removing a liquid from the device according to the invention. The functions of dispensing or delivering fluid to and removing or receiving fluid from the apparatus, according to the present invention, are considered to be both active and passive operations. For example, receiving fluid from the first rigid actuator may be achieved by applying fluid under pressure to the device, thereby forcing fluid into the device, or by applying negative pressure to the chamber, so as to draw fluid into the device, whereas removing or transporting fluid from the device to the exterior may be achieved by applying pressure to the chamber, for example by drawing fluid, such as liquid or gas, through the first fluid port, or by applying negative pressure to the chamber, so as to draw fluid through the fluid port.

Rigid fluid actuators useful in the present invention are devices having three features. It must be able to pierce the fluid port of the device. This is achieved by a certain stiffness. The required stiffness is provided by manufacturing the actuator from, for example, metal. The device is preferably designed as a tip to pierce the seal. In addition, the device needs to be long enough to pass through the fluid port to the interior of the device. Therefore, the device preferably has an elongated thin shape. Third, the device must provide a channel to transport liquid into and out of the device. Therefore, a cannula or cannula-shaped or hollow needle-shaped device is preferred. The outer diameter of the fluid actuator may be in the range 0.1 to 3mm, more preferably in the range 0.2 to 1.5mm, more preferably in the range 0.3 to 0.8 mm.

The device is preferably used in an instrument. The instrument includes fluid communication with a fluid actuator device to apply positive or negative pressure to carry or receive fluid. Suitable means include a syringe pump. The position of the fluid actuator relative to the device may be controlled by an automated system.

In a highly preferred use, the apparatus further comprises complementary means for performing the analysis, such as a heating element. The heating element is positioned so that it can contact the device at a suitable location, wherein heat is available to heat the fluid within the device, preferably when the fluid is contained within a cavity within the device. An example of an instrument that includes a heating element is a thermal cycler. Commonly known thermocyclers are used to apply a profile of different temperatures to a fluid in a repetitive manner. An exemplary thermal cycler is described in EP 236069. Preferred heating elements are Peltier elements or resistive heating elements.

During a process performed within the apparatus, the instrument may further comprise a detection module for performing a detection of a property or a change in a property of the liquid. Suitable detection modules are generally known and depend on the nature or the kind of change of nature that is made during the presence of the liquid in the device. For example, if the characteristic is a change in an optical signal, such as a fluorescent signal, the detection module will comprise a light source positioned within the instrument such that the device, preferably a chamber within the device, can be illuminated and an illumination receiving unit, preferably a photosensitive cell, to receive illumination from the liquid contained within the device and to transmit an electrical signal to the evaluation unit.

If a process performed within the instrument requires connectivity of the components of the instrument, such as electrodes or heating foils within the instrument to connect to the circuitry of the instrument, such connectors are preferably provided at locations on the instrument that are located such that when the instrument is inserted into the instrument, the connectors on the instrument connect to their counterparts on the instrument.

The system according to the invention preferably additionally comprises a fluid container (e.g. for water collection) or/and one or more reagent containers.

Further subject matter of the invention are: use of the device according to the invention in a method of analysing a sample, for example in an in vitro diagnostic test. Therefore, another subject of the invention is: a method of analysing a sample or a component thereof using one or more reagents, comprising

-providing a device according to the invention,

-introducing a fluid into the device through the fluid port, wherein the fluid is selected from the sample and the reagent, and

-determining at least one property of the fluid which relates to the presence or amount of a substance present in the sample.

In one embodiment, the device already contains reagents useful for analysis, and the sample is introduced through the fluid port. In another embodiment, the device contains a sensor that is useful for determining a characteristic of the sample or a liquid derived from the sample with or without the use of a reagent.

Fluid, preferably a sample or/and reagent to be analysed, is introduced into the apparatus, preferably by a rigid fluidic actuator such as a steel cannula, through a first fluid port into a channel leading to the chamber. The chamber further comprises, at the opposite end to its first channel entry point, an exit point for a second channel leading to a second opening closed by another of the aforementioned fluid ports. When pressure is applied on the liquid within the cannula to enter the channel, there is a second cannula that pierces the second fluid port so that the pressure can escape through the second cannula. In this very simple embodiment, the liquid comprises the fluid to be analyzed and all reagents required for analyzing the components of the fluid to be detected, for example labeled binding partners for the components to be determined in the fluid. The liquid enters the cavity by applying pressure from the first cannula or by applying negative pressure to the second cannula. The detection can be started in the room when the required reaction has been carried out. This may be done by illuminating the liquid in the chamber with light of a wavelength for which one of the fluid components or reagents has a measurable absorption. Determining the light leaving the cavity, for example by fluorescence, can be used to determine the absorbance of the liquid or any change in absorbance of the liquid over time, or compared to a standard liquid.

In a very preferred embodiment of the analysis method, the component of the liquid to be analyzed is the nucleic acid to be assayed contained in the fluid, for example part of the hepatitis C virus genome. The assay reagents will comprise primers that amplify specific fragments of the nucleic acid and probes that bind the amplified fragments. A very preferred example of such a reaction is disclosed in EP 543942. To apply thermal cycling to the liquid contained in the chamber, the instrument used comprises a combined heating/cooling block in order to bring the contents of the chamber to the temperature of the curve required to amplify the nucleic acid.

The advantages of the device according to the invention are: the interface between the device and the enclosure of the device is very tight. This allows applying high pressures to the interior of the device, e.g. when the cavity within the device has very small sized channels or/and using liquids that tend to condense during transport or reaction within the microfluidic device. The manufacturing equipment has the advantages that: by using this method, a device providing a tight seal is possible. The device according to the invention can be used very advantageously in systems for analysing liquids, because due to the tightness of the seal the instrument is better protected from contamination by the fluid escaping from the device, while on the other hand the interior of the device is protected from contamination by the surrounding environment. Furthermore, the use of the device according to the invention results in an enhanced convenience of the analysis, since the analysis can be automated using an automated fluid transport by means of a rigid fluid actuator. Furthermore, especially for small devices, the manufacture becomes simpler and more reliable.

Reference numerals:

1 apparatus

2 main body

3 cavities

4 fluid port

5 sealing wall

6 snap-in projection

7 snap-in recess

8 sealing edge

9 introducer of fluid actuator

Claims (30)

1. An apparatus has a body including a cavity and a fluid port made of a thermoplastic elastomer attached to the body.

2. The apparatus of any one of the preceding claims, wherein the apparatus is a container that holds, transports, or receives a fluid.

3. The apparatus of any one of the preceding claims, wherein the cavity comprises a channel, a chamber, or a system of channels and chambers.

4. The apparatus of any of the preceding claims, wherein the fluid port has a press fit to the body.

5. The device according to any of the preceding claims, wherein the device is a microfluidic device.

6. The device according to any of the preceding claims, wherein the chamber has a fluidic or microfluidic function, as static mixing, or an optical function, as transparency for detecting light, or a part for chemical or physical processing of fluids.

7. The apparatus of any one of the preceding claims, wherein the fluid port is located within the apparatus so as to be accessible to a rigid fluid actuator from outside the apparatus.

8. The apparatus of claim 7, wherein the fluid port comprises a channel orthogonal to the fluid actuator entry direction.

9. The apparatus of any one of the preceding claims, wherein the cavity is bounded by a body and a sealing wall covering the cavity.

10. The apparatus of claim 9, wherein the sealing wall is a thin foil.

11. The apparatus of claim 10, wherein the thin foil has a thickness greater than 200W/m²Heat transfer rate of/K.

12. The apparatus of any one of claims 10 to 11, wherein the thin foil and/or body has a transmission of greater than 2% for electromagnetic waves having a wavelength in the range of 300 to 4000 nm.

13. The apparatus of any of the preceding claims, wherein the fluid port has a conical recess for guiding the fluid actuator.

14. The apparatus of any one of the preceding claims, further comprising a material for solid phase adsorption.

15. A method of manufacturing an apparatus having a rigid body constructed of a first rigid material including a cavity having an opening closed by a fluid port constructed of a second elastomeric material, the method comprising

-a first moulding step comprising

a) A first mold is provided that reflects the shape of the rigid body,

b) a first material in liquefied form is injected into a first mold,

c) waiting until said first material becomes at least partially solid,

providing the result of said first molding step into a second mold, a first portion of which reflects the outer shape of said fluid port and a second portion of which reflects at least part of the first mold,

-a second molding step comprising

a) Injecting a second material in liquefied form into the first portion of the second mold,

b) waiting until the second material becomes at least partially solid,

and

-removing the result of the moulding step from the mould after solidification of the material.

16. The method of claim 13, wherein the fluid port is located within the apparatus so as to be accessible to a rigid fluid carrying actuator from outside the apparatus.

17. The method of claim 13, wherein the fluid port is a septum attached to the first rigid material.

18. A method of manufacturing an apparatus having a body constructed of a first rigid material including a chamber having an opening closed by a fluid port constructed of a second elastomeric material, the method comprising

-a first moulding step comprising

a) Providing a first mold reflecting the external shape of the fluid port,

b) injecting a second material in liquefied form into the first mold,

c) waiting until said second material becomes at least partially solid,

-providing the result of said first moulding step into a second mould, a first portion of which reflects at least part of said first mould and a second portion reflects the external shape of said body,

-a second molding step comprising

a) Injecting the first material in liquefied form into the second portion of the second mold,

b) waiting until the first material becomes at least partially solid,

and

-removing the result of said moulding from said mould after solidification of said material.

19. The method of any of claims 13-16, wherein the second material is an elastomer that is shaped in a 2C injection molding process.

20. The method according to claim 17, wherein the first material is selected from the group of thermoplastics, such as polypropylene, polyethylene, polystyrene, polycarbonate and polymethylmethacrylate.

21. The method of any one of claims 15 to 20, further comprising the step of sealing the cavity of the body with a sealing wall.

22. The method of claim 21, wherein the sealing is accomplished by laser welding, ultrasonic welding, heat sealing, or gluing.

23. A system for handling a fluid, comprising

An instrument having a fluid actuation module comprising a rigid fluid actuator, and

-a device according to any one of claims 1 to 22.

24. The system of claim 23, further comprising at least one reagent.

25. The system of any one of claims 23 to 24, wherein the rigid fluid actuator is a hollow needle that is movable relative to the device from at least one position in which the fluid actuator is not fluidly connected to the cavity to a second position in which the fluid actuator is fluidly connected to the cavity by piercing through the fluid port.

26. The system of any one of claims 23 to 25, wherein the instrument further comprises a device for performing an analysis.

27. The system of any one of claims 23 to 26, further comprising a heating element, preferably a thermocycler.

28. The system of any one of claims 23 to 27, wherein the instrument further comprises a detection module.

29. Use of the apparatus according to any one of claims 1 to 14 in a method of analysing a fluid.

30. A method of analyzing a sample or component thereof using one or more reagents, comprising

-providing a device according to any one of claims 1 to 12 or a system according to any one of claims 23 to 28,

-introducing a fluid into the device through the fluid port, wherein the fluid is a sample or/and a reagent to be analyzed, and

-determining at least one property of the fluid which relates to the presence or amount of a substance present in the sample.