WO2015150742A1

WO2015150742A1 - Fluid delivery

Info

Publication number: WO2015150742A1
Application number: PCT/GB2015/050905
Authority: WO
Inventors: Gary David KIPLING; Nathaniel Brown
Original assignee: The University Of Hull
Priority date: 2014-03-31
Filing date: 2015-03-26
Publication date: 2015-10-08
Also published as: GB201405808D0

Abstract

Certain embodiments of the present invention relate to apparatus and methods for providing a target liquid at a desired location. In certain embodiments, the apparatus includes a piercing member locatable with respect to a sealing membrane of a respective blister pouch that has a tip region which is locatable in an abutting relationship with the sealing membrane in a first mode of operation and through the sealing member in a further mode of operation for providing liquid from the blister pouch at a desired location. Also included in the present invention, inter alia, is a piercing plate comprising a piercing member, together with devices, for example, bio chips and microfluidic devices, which are for use in combination with the apparatus.

Description

FLUID DELIVERY

Field of the Invention

Certain embodiments of the present invention relate to apparatus and methods for providing a target liquid at a desired location. In certain embodiments, the apparatus includes a piercing member locatable with respect to a sealing membrane of a respective blister pouch that has a tip region which is locatable in an abutting relationship with the sealing membrane in a first mode of operation and through the sealing member in a further mode of operation for providing liquid from the blister pouch at a desired location. Also included in the present invention, inter alia, is a piercing plate comprising a piercing member, together with devices, for example, bio chips and microfluidic devices, which are for use in combination with the apparatus.

Background to the Invention

Microfluidic systems have recently emerged as powerful tools for bio-sensing, particularly in enriching and purifying molecules and cells in biological samples. Microfluidic technology has enabled the miniaturisation of normal sized equipment and has resulted in more readily portable devices and the sensing of increasingly small sample volumes and target analyte concentrations which have not been possible with previous technologies.

Microfluidic devices can be laboratory-based devices for use in genomics, sequencing, high- throughput screening and separations. Additionally, microfluidic devices provide the opportunity to integrate processes and measurement systems on a single technology platform leading to the concept of the "lab on a chip". Thus, microfluidic devices are also sometimes known as "bio-chips" or lab-on-chip" systems. Microfluidic technology has found applications in chemical and system biology, point-of-care analysis, high-throughput biological screening. Advantages associated with lab-on-chip include consumption of small volumes of reagents and sample, and delivery of results in a short timescale.

Microfluidic devices often comprise a substrate together with microstructures such as micropumps, micromixers, ports, microchannels and the like.

The delivery of fluids plays an important role in fields such as chemistry, microbiology and biochemistry. These fluids may include liquids or gases and may provide reagents, buffers, solvents or reactants, to chemical and/or biological processes, including processes involving microfluidic technology. A problem often associated with fluid delivery to microfluidic devices is the difficulty of delivering known volumes of reagents into the microfluidic flow system in a controlled manner at a steady rate. Existing technology often relies upon pipetting of reagents and/or samples into wells or chambers of the device. As a result, whilst microfluidic technology may have resulted in portable devices being developed, there still remains the problem of how to easily transport and deliver reagents and/or samples outside a laboratory environment. One way of storing and delivering fluids such as reagents and/or samples that has been developed is by way of a "blister pouch". Blister pouches include individually sealed units which store fluids or solids prior to delivery and have been used to store and subsequently deliver fluids to microfluidic devices. In order to deliver the fluid or solid to a target location, a user breaks the seal of the unit. The seal may be broken by a user manually applying a force to the seal or by way of an actuator applying pressure. Often, when the seal is broken, the release of the stored fluid is not always controlled and thus at least some of the stored fluid is not directed to the target location. Also there is a risk of fluid "surging" from the blister. One way of automatically providing the right reagents in the correct volume is to package the reagents into blister packages and allow the blister to release the reagents once the sample is on the chip. However, most conventional blister popper constructs lead to the blisters rupturing in the wrong place and the reagent splashing outside of the chip. This results in a waste of reagent and can lead to a reduction in accuracy if a precise amount of the fluid is required. In addition, on occasion there is a failure of the sealed unit to burst.

Certain embodiments of the present invention aim to at least partially mitigate the problems associated with the prior art. Summary of Certain Embodiments of the Invention

Certain embodiments of the present invention aim to provide a device and/or method which delivers a fluid, e.g. a liquid, in a controlled manner to a target location. It is an aim of certain embodiments of the present invention to provide a mechanism that can selectively burst a blister pouch when desired and in a controlled way but which doesn't pose a risk (in terms of causing an undesired breach) at other times. In a first aspect of the present invention, there is provided apparatus for providing a target liquid at a desired location, comprising at least one piercing member upstanding from a support surface at a height of about around 300 to 1200 microns and locatable with respect to a sealing membrane of a respective blister pouch; wherein a tip region of the piercing member is locatable in an abutting relationship with the sealing membrane in a first mode of operation and through the sealing membrane in a further mode of operation for providing liquid from the blister pouch at a desired location.

In one embodiment, the piercing member is upstanding from a support surface at a height of about around 300 to 1200 microns e.g. 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1 100, 1 150 or 1200 microns.

In one embodiment, the piercing member has a height of from about 300 to 1200 microns.

Aptly, in the first mode of operation the piercing member is locatable to deform but not pierce the sealing membrane. In one embodiment, in the first mode of operation, the piercing member does not pierce the sealing membrane when an abutment force of up to about around 1 N is applied by the piercing member to the sealing membrane. In one embodiment, in the first mode of operation, the piercing member does not pierce the sealing membrane when an abutment force of up to about around 1 N, e.g. 0.80, 0.85, 0.9, 0.95 or 1 .0N, is applied by the piercing member to the sealing membrane.

In one embodiment, the tip region comprises a rounded nose region having a generally arcuate shape with a radius of curvature of about around 50 to 1000 microns. Aptly, the radius of curvature is about around 100 to 500 microns. In one embodiment, the radius of curvature of the rounded nose region of the piercing member is approximately 100, 150, 200, 250, 300, 350, 400, 450 or 500 microns.

Thus, in certain embodiments of the present invention the piercing member has a comparatively blunt nose region, which is used to pierce the sealing membrane of the blister pouch when desired. As a result of the "blunt" nose region, when the sealing membrane is pierced, substantially all of the fluid held in the blister pouch flows over and through the piercing member, rather than, as is the case with many prior art fluid delivery systems, the fluid is released in an uncontrolled manner leading to a portion of the liquid not being directed into the chamber.

In one embodiment, the piercing member further comprises a plurality of abutment wall regions extending from the support surface to the tip region at a predetermined angle so that each wall region forms a tangent with an arc of a rounded nose region at an end of the tip region. Aptly, each abutment wall region meets the nose region at an angle of about around 45 degrees. In one embodiment, each abutment wall region meets the nose region at an angle of approximately 45 degrees e.g. 40, 41 , 42, 43, 44, 45, 46 or 47 degrees. Aptly, each abutment wall region meets the nose region at substantially the same angle. Alternatively, each abutment wall region meets the nose region at a different angle to each other abutment wall region.

In one embodiment, the piercing member provides a plurality of fluid communication passageways proximate to the tip region that each communicate liquid urged from a pierced blister pouch to at least one collection chamber under the support surface. In one embodiment, the piercing member comprises two or more fluid communication passageways proximate to the tip region, e.g. 2, 3, 4 or more. In one embodiment, the piercing member comprises four fluid communication passageways.

In an embodiment, the piercing member comprises a plurality of arms extending upwards and in an inclined manner from the support surface. Aptly, the fluid communication passageways are provided by openings in the support surface in notched regions between adjacent arms. In one embodiment, the piercing member comprises two or more upwardly extending arms e.g. 2, 3, 4, or more. In one embodiment, the piercing member comprises four upwardly extending arms. Certain embodiments of the present invention provide the advantage of a piercing member which provides openings through which the fluid, e.g. a liquid, released from the blister pouch may flow. The liquid may subsequently flow into a chamber e.g. a collection chamber as described herein below. As a result, the likelihood of the reagent being spilt and not collected in the chamber is reduced. Blister pouches are often pre-filled with a required volume of liquid and thus it follows that certain embodiments of the present invention may provide a more accurate result in a method which is based on calculations involving the known volume of the liquid in the blister pouch.

Certain embodiments of the present invention provide a piercing member which comprises a plurality of openings which allow fluid released from the blister pouch to flow into a chamber situated beneath the piercing member. The plurality of openings results in the fluid flowing downwardly in a plurality of flow streams which should enable the flow of fluid to be controlled more effectively. Aptly, the first mode of operation comprises a transportation mode during which a blister pouch is transportable in an abutting relationship with the piercing member without risk of puncture.

Certain embodiments of the present invention provide the advantage that the piercing member is capable of piercing a sealing membrane of a blister pouch when a predetermined level of force is applied to either the piercing member or the blister pouch but does not pierce the sealing member when simply in an abutting relationship with the sealing membrane. Thus, certain embodiments of the present invention provide an apparatus which is capable of being transported in an abutting relationship with a sealed blister pouch pre-filled with a reagent for example without the blister pouch being pierced and its contents released.

In one embodiment, in the further mode of operation in which the blister pouch is urged against the piercing member, the piercing member is disposed to rupture the sealing membrane at a rupture force of about around 3 to 12N. Aptly, the piercing member is disposed to rupture the sealing membrane at a force of about 3, 4, 5, 6, 7, 8, 9, 10, 1 1 or 12 N.

In one embodiment, each piercing member is manufactured from a material selected from a group comprising glass, polycarbonate or polymethyl methacrylate (PMMA), cyclic olefin copolymer (COC), polystyrene (PS), polyether ether ketone (PEEK), polytetrafluoroethylene (PTFE) and polypropylene (PP).

In a further aspect of the present invention, there is provided a piercing plate, securable to at least one blister pouch, comprising an apparatus for providing a target liquid at a desired location comprising at least one piercing member upstanding from a support surface at a height of about around 300 to 1200 microns and locatable with respect to a sealing membrane of a respective blister pouch; wherein a tip region of the piercing member is locatable in an abutting relationship with the sealing membrane in a first mode of operation and through the sealing member in a further mode of operation for providing liquid from the blister pouch at a desired location. Aptly, the apparatus is as further described herein.

In one embodiment, the piercing plate further comprises a planar upper surface, comprising the support surface, and a planar lower surface that is securable to an upper surface of a bio chip. Aptly the piercing plate further comprises an adhesive layer on the lower surface.

In one embodiment, the piercing plate further comprises at least one alignment element that locates the piercing plate at a desired position with respect to each blister pouch. Aptly, the alignment element comprises at least one edge of the piercing plate. In one embodiment, the alignment element comprises at least one locating lug or recess that mates with a complimentary alignment element of the blister pouch.

Aptly, the piercing plate further comprises one further alignment element that locates the piercing plate at a desired position with respect to a bio chip. In one embodiment, each further alignment element comprises at least one edge of the piercing plate.

Aptly, each further alignment element comprises at least one locating lug or recess that mates with a complimentary alignment element of the bio chip. In one embodiment, the piercing plate further comprises a cavity under each piercing member that is locatable in fluid communication with a collection chamber of a bio chip.

Aptly, the piercing plate has a thickness of about around 200 to 1000 microns.

In one embodiment, the piercing plate is manufactured from a material selected from glass, polycarbonate and PMMA.

In one embodiment, the plate and each piercing member are integrally formed. In one embodiment, the plate and the apparatus are integrally formed. In one embodiment, the plate is retrofittable to an underside of a blister package comprising one or more blister pouches sealed with a sealing membrane.

In a further aspect of the present invention, there is provided a blister pack, comprising a plurality of blister pouches secured together via a web and a common sealing membrane; and at least one piercing plate as described herein secured to a lower surface of the sealing membrane. In one embodiment, the blister pack may be fabricated using vacuum thermoforming of polymer films, for example. In one embodiment, the blister pack comprises a plurality of frangible lines or perforation lines in the web. In one embodiment, the frangible lines or perforation lines are provided between each blister pouch so as to enable a single blister pouch to be removed from the blister pack. In a further aspect of the present invention, there is provided a bio chip comprising an apparatus according to the first aspect of the invention as described herein.

As used herein, the term "bio chip" refers to a device which can be used to analyse samples having a volume of from about 10nl and 10ml. The terms "microfluidic device", "microfluidic assembly", "microfluidic systems" and the like may also be used herein to indicate devices, assembly and systems wherein samples for the processing and/or analysis of samples having a volume of from about 10 nl to about 10 ml. In one embodiment, the bio chip is used to analyse samples having a volume of between about 100μΙ and 2000μΙ. In one embodiment, the bio-chip is a microfluidic device.

In one embodiment, the bio chip comprises an upper chip layer comprising one or more piercing member as an integral unit. In one embodiment, the bio chip comprises an upper chip layer comprising each piercing member as an integral unit. In an embodiment, the bio chip further comprises an upper chip layer having an upper surface on which at least one piercing plate comprising said at least one piercing member is secured.

In one embodiment, the bio chip comprises a lower surface of the upper chip layer and wherein the lower surface of the upper chip layer comprises a collection chamber located beneath each piercing member and at least one channel connecting a collection chamber to a respective sample chamber. Aptly, the channel comprises a channel region that provides a serpentine pathway. Aptly, the channel comprises an inlet end and an outlet end. In one embodiment, the bio chip comprises a plurality of channels.

In one embodiment, the bio chip comprises a plurality of piercing members and a corresponding plurality of collection chambers. Aptly, a collection chamber is located beneath the piercing element. Aptly, the bio chip comprises a first collection chamber and at least one further collection chamber. In one embodiment, the first collection chamber and the further collection chamber are in fluid communication via a channel region. Aptly, the channel region comprises a T-junction region and/or a Y-junction region.

Certain embodiments of the present invention provide a bio-chip which incorporates at least one blister pouch which is integrally formed with the bio-chip. Certain embodiments of the present invention provide an advantage that fluids may be stored on the bio-chip and delivered using the piercing member when required. Thus, certain embodiments of the present invention may result in a reduction of gas exchange with stored solutions and reagent waste, as well as cost reduction. Certain embodiments of the present invention provide a portable bio-chip device, e.g. a microfluidic device, which incorporates a blister pouch pre-filled with one or more reagents.

Certain embodiments of the present invention provide the advantage that precise location of the piercing member relative to the sealed unit of the blister pouch is not required providing that the piercing member contacts at least a portion of the blister pouch.

In one embodiment, the bio chip is formed from a material selected from glass, polycarbonate, polymethyl methacrylate (PMMA), cyclic olefin copolymer (COC), polystyrene (PS), polyether ether ketone (PEEK), polytetrafluoroethylene (PTFE) and polypropylene (PP).

In one embodiment, the bio chip may be fabricated from a metal. Aptly, the chip may be formed from a metal which could be readily formed into the required shapes using a range of fabrication techniques. Aptly, the chip may be formed a technique selected from die casting, injection moulding and subtractive milling. In one embodiment, the bio chip is fabricated from a ceramic material. Aptly, the bio chip is produced using an injection moulding technique. In one embodiment, the bio chip is formed from a polymer. Aptly, the polymer is an injection moulded polymer.

Aptly, the bio chip further comprises one or more further elements. In one embodiment, the bio chip comprises one or more of the following elements: one or more micropumps, one or more microvalves, a power source e.g. a battery, and/or a sensor. In a further aspect of the present invention, there is provided a method of delivering a target fluid to a desired location, comprising locating an apparatus as described herein such that the piercing member of the apparatus abuts a sealing membrane of a respective blister pouch; wherein the fluid is located in the blister pouch; and applying a predetermined force to the sealing membrane such that the sealing membrane is pierced by the piercing member. Aptly, the method comprises applying a force of between about 3N and about 12N to the sealing membrane.

In one embodiment, the method comprises: locating a biochip comprising a collection chamber as described herein underneath the piercing member prior to the piercing of the sealing membrane; and collecting the fluid from the blister pouch in the collection chamber.

Aptly, the method further comprises locating the apparatus in an abutting relationship with the sealing membrane prior to application of a predetermined force. Further details of certain embodiments are provided below.

Detailed Description of Embodiments of the Invention

Certain embodiments of the present invention will now be described hereinafter, by way of example only, with reference to the accompanying drawings in which:

Figure 1 illustrates a bio chip and blister pouch according to certain embodiments of the present invention;

Figure 2 illustrates a bio chip according to certain embodiments of the present invention; Figure 3A illustrates an apparatus according to certain embodiments of the present invention which comprises a piercing member; Figure 3B illustrates an apparatus according to certain embodiments of the present invention which comprises a piercing member;

Figure 3C illustrates an apparatus according to certain embodiments of the present invention which comprises two piercing members;

Figure 4 illustrates a cross-sectional view of a blister pouch located above a piercing member according to certain embodiments of the present invention;

Figure 5 illustrates a cross-sectional view of a blister pouch located above a piercing member according to certain embodiments of the present invention;

Figure 6 illustrates a side view of an apparatus comprising a piercing member according to certain embodiments of the present invention; Figure 7 illustrates a cross sectional view of section A-A as shown in Figure 6 of an apparatus comprising a piercing member according to certain embodiments of the present invention;

Figure 8 illustrates a top view of an apparatus comprising a piercing member according to certain embodiments of the present invention;

Figure 9 illustrates a bottom view of an apparatus comprising a piercing member according to certain embodiments of the present invention; Figure 10 illustrates an exploded view of a blister pack and a piercing plate comprising a piercing member according to certain embodiments of the present invention;

Figure 1 1 illustrates a blister pouch located above a piercing member according to certain embodiments of the present invention; and Figure 12 illustrates a cross-sectional side view of a blister pouch located above a piercing member according to certain embodiments of the present invention;

Figure 1 illustrates an apparatus 100 for providing a target liquid at a desired location according to certain embodiments of the present invention. In certain embodiments, the desired location is situated on a device e.g. a bio-chip 1 10. The apparatus may be a distinct element from the bio-chip 1 10 or may be an integrally formed portion of the bio-chip. Further details of the bio chip which are not shown in the Figures are provided below. The bio-chip may comprise a number of other elements e.g. one or more of sensing regions, micropumps, valves, heaters, microchannels and other elements known in the art (not shown).

In one embodiment, the bio chip comprises one or more elements for controlling fluid flow thereon e.g. electroosmotic flow, vacuum pumps, electrokinetic pumps, membrane actuated pumps, centrifugal force, gas bubbles, ferrofluidic plugs, capillary action, electrohydrodynamic pumps, and/or magnetohydrodynamic pumps. Aptly, pumps for microfluidics may be governed by electrokinetic principles or by the mechanism of differential pressure.

The bio chip can be used for automating a variety of bioassay protocols, such as absorption, fluorescence, ELISA, enzyme immunoassay (EIA), light scattering, electrochemical measurement assays and chemiluminescence for testing a variety of analytes. Such analytes include for example proteins, nucleic acids, cells, receptors, and the like. The biochip can be configured and designed for whole blood, tears, sweat, serum, plasma, urine, and other biological fluid applications. In one embodiment, the bio-chip is adapted to perform DNA hybridization assays, expression analysis of mRNA, certain forms of DNA detection and/or DNA sequencing. Aptly, the bio chip may be used in one or more of the techniques described in Sensors (Basel). 2012; 12(8): 10713-10741 , incorporated herein by reference in its entirety.

Aptly, the bio chip is made of elastomers or polymers such as for example, PDMS, PMMA, polyurethane, PFPE, SIFEL and parylene. In one embodiment, the bio-chip is formed from a material selected from for example plastic, glass, silicon, polycarbonate, polycarbonate, polydimethylsiloxane, polymethyl methacrylate (PMMA), cyclic olefin copolymer (COC), polystyrene (PS), polyether ether ketone (PEEK), polytetrafluoroethylene (PTFE) and polypropylene (PP).

The bio-chip may comprise a sample region. As used herein, the term "sample region" refers to any portion of a bio-chip that provides storage, analysis, separation or that provides, conducts or is otherwise involved in any processing step on or with the sample liquid. In one embodiment, the sample region is involved in the analysis of a sample liquid or components therein, and the sample region is considered a sensor (or sensor region), for example a sensor surface.

In one embodiment, the sample is taken from a plant, e.g. a tree, a shrub and/or a flowering plant. In one embodiment, the sample is selected from a human and/or animal sample. In one embodiment, the sample is whole blood, serum and/or plasma. Aptly the sample is urine sample, a sample taken from a swab e.g. a nasal swab, a pharyngeal swab, a vaginal swab or a rectal swab. In one embodiment, the sample is taken from tears and/ or sweat. Aptly, multiple analytes or multiple samples may be detected in a very small quantity.

In one embodiment, the bio-chip comprises a sensor (not shown). As used herein, the term "sensor" refers to a device that detects a change in at least one physical property and produces a signal, or induces a change in a signal, in response to the detectable change. The manner in which the sensor detects a change may include, for example, acousto- mechanical changes, electrochemical changes, optical changes and/or electro-optical changes. For example, acousto-mechanical sensors can utilise resonant frequency, phase- shifts, piezoelectric properties. Aptly, electrochemical sensors utilise potentiometric and amperometric measurements. Aptly, optical sensors may utilise absorbance, fluorescence, luminescence and/or evanescent waves.

In one embodiment, the sensor is a biosensor and comprises at least one detection moiety attached thereto. In one embodiment, the detection moiety is selected from one or more of: an antibody; a nucleic acid, e.g. DNA, RNA, or PNA; a protein; a peptide; an aptamer; a cell; a charged molecule; a membrane; a vesicle; a chemical and/or electrochemical receptor.

The detection moiety may be immobilised directly to a surface of the bio-chip. In certain embodiments the detection moiety may be immobilised on a solid support e.g. latex beads or magnetic beads using techniques known in the art. The detection moiety may comprise a label for detection. Alternatively, the detection moiety may be directly detectable via a label free detection method.

The bio-chip of certain embodiments of the present invention may be used to detect for example the presence or absence of a target protein, a target DNA molecule and/or a target RNA molecule. The target molecule for detection may be for example a hormone, a cell surface molecule, a cardiac protein, a tumour marker and/or a small molecule. In one embodiment, the target molecule is a drug of abuse. In one embodiment, the target molecule may be a plurality of target molecules and the bio-chip is for the detection of the presence and/or absence of one or more target molecules.

Turning to Figure 1 again, the apparatus 100 comprises a piercing member 120 located on a support surface 130. The piercing member 120 is described in more detail below and illustrated in Figures 6 to 9. In one embodiment, the apparatus 100 may comprise a plurality of spaced apart piercing members 120.

The bio chip 1 10 comprises a lower chip sealing plate 140. The lower chip sealing plate 140 may be rectangular for example in shape. The chip sealing plate may have dimensions of for example a length of about 120mm and a width of about 60mm, although it will be appreciated that bio-chips of other dimensions are encompassed by certain embodiments of the present invention. The lower chip sealing plate 140 may comprise a rectangular glass plate. Other materials which may be suitable to fabricate the bio chip plates are described herein. The bio chip also comprises a chip top layer 150. The lower surface 160 of the chip top layer 150 can be bonded to the upper surface 170 of the lower chip sealing plate 140 by a suitable known method e.g. thermal bonding. The upper surface 180 of the chip top layer 150 forms an upper external surface of the bio chip and the lower surface 190 of the lower chip sealing plate 140 forms a lower external surface of the bio chip 1 10. By way of example, the chip top layer 150 may have a thickness of for example about 3mm and the lower chip sealing plate may have a thickness of 1 mm.

The bio chip 1 10 further comprises one or more channels. Figure 1 illustrates that the chip top layer comprises one or more channels 200. The chip top layer also comprises a sample chamber 210 which is in fluid communication with at least one channel. The chip top layer 150 further comprises a collection chamber 220. The collection chamber may be considered a fluid introduction chamber. In use, the collection chamber 220 is situated beneath a piercing member as described herein. The collection chamber is in fluid communication with at least one channel 200 and may provide an inlet for the channel 200.

The dimensions of the channels may differ between devices. In one embodiment, the channels may have a cross-sectional width of between about 0.1 mm and 5mm. The channels may have a depth of e.g. 20 microns and 1 mm. As shown in Figure 1 , in certain embodiments the piercing member 120 is provided on a piercing plate 230. The piercing plate 230 is secured to the chip top layer 150 by a layer of adhesive 240. Prior to application of the layer of adhesive, the piercing plate is positioned such that the piercing member is located above the collection chamber 220. The piercing plate may be about around 200 to 1000 microns thick and/or manufactured from a material e.g. glass, polycarbonate or PMMA.

Figure 1 illustrates a blister pouch 250 located above the piercing member 120. The blister pouch 250 is formed from a blister forming material. The blister pouch comprises a reservoir 260 which may be filled with a predetermined volume of liquid e.g. a reagent. The reservoir is sealed by a blister sealing membrane 270. The liquid may be a reagent such as for example a buffer, a liquid comprising one or more detection molecule and/ or a liquid comprising a sample material. The nature of the liquid may depend on the reaction being carried out on the bio chip. The blister sealing membrane 270 is formed from a blister sealing material. The blister sealing material is capable of being pierced by the piercing member 120 when brought into contact with the piercing member with a predetermined level of force. Aptly, the piercing member does not pierce the blister sealing membrane when it is located abutting the blister membrane at a level of force which is less than about 1 N.

When the reagent or other liquid stored in the blister pouch is required, force may be applied manually by a user pressing the blister pouch on a surface opposing the blister sealing material. Alternatively, force may be applied by an automated actuator. The force is aptly greater than about 3N. Figure 2 illustrates an embodiment in which two piercing members 120a, 120b are located in a bio chip 1 10. The illustrated bio chip may be generally square in shape as shown in Figure 2. In other embodiments, the bio chip may be alternative shapes e.g. rectangular, circular, hexagonal or pentagonal.

The piercing members 120a, 120b are each provided above a respective collection chamber 220a, 220b. During use, the collection chamber holds the liquid released from the blister pouch when the sealing membrane 270 is pierced by the corresponding piercing member. Thus, the collecting chamber helps to prevent liquid being released from the blister pouch over the surface of the bio-chip and thus away from the target location i.e. the channels of the bio-chip.

The bio chip 1 10 of Figure 2 includes a channel 200a which is generally linear and extends between a first piercing member 120a and a second piercing member 120b. The channel 200a includes a T-junction intersection with a further portion of the channel 200b. The further portion of the channel is serpentine and is connected to a sample chamber 210.

Figure 3 illustrates alternative embodiments of a bio chip 1 10. Figure 3A illustrates a bio chip 1 10 which comprises a linear channel 200. The linear channel is in fluid communication with the collection chamber 220 and the sample chamber 210. The linear channel 200 connects the collection chamber 220 and the sample chamber 210. The piercing member 120 located above the collection chamber 220 and is either integrally formed with the upper surface of the biochip or within a piercing plate. Figure 3B illustrates an alternative embodiment of a bio chip according to certain embodiments of the present invention. The bio chip comprises a channel 200 which follows a serpentine path between the collection chamber 220 and the sample chamber 210. The piercing member 120 is located above the collection chamber 220 and can be either integrally formed with the bio chip 1 10 or within a separate piercing plate 230 as described herein.

Figure 3C illustrates a further alternative embodiment of a bio-chip 1 10 according to certain embodiments of the present invention. Two piercing members 120a and 120b are located above two separate collection chambers 220a and 220b. One or both piercing members 120a and 120b are either integrally formed with the upper surface of the biochip or within a piercing plate 230.

The collection chambers 220a, 220b are linked by a linear channel 200a. The channel 200a is intersected by and in fluid communication with a further portion of the channel 200b at an intersection point 300. The further portion of the channel 200b is in fluid communication with the sample chamber 210. The channel has a linear portion 310 extending from the intersection point 300, a middle serpentine section 320 and an end linear section 330 which is connected to the sample chamber 210.

Figure 4 illustrates a cut through of a piercing member 120 provided on a piercing plate 230 and located above a collection chamber 220 located in a bio chip. As described above, the collection chamber 220 is provided in an upper chip plate 150 of the bio chip 1 10. The piercing plate 230 is secured to the upper surface of the upper chip plate by a layer of adhesive 240.

A blister pouch 250 is located above the piercing member 120. The blister membrane 270 of the blister pouch 250 is in contact with the piercing member 120 but since the force applied to the blister sealing membrane by the piercing member does not exceed about 1 N, the blister sealing material of the blister membrane is not punctured by the piercing member. As a result, the fluid is retained in the blister pouch 250 shown in Figure 4.

Figure 5 is an alternative view of the blister pouch 250, piercing plate 230 and bio chip shown in Figure 4.

The piercing member 120 is shown in more detail in Figures 6 to 9. The piercing member illustrated in Figures 6 to 9 may be comprised in a piercing plate or a bio chip as described herein. Figure 6 illustrates a cross sectional view of a piercing member 120. The piercing member 120 is located upstanding from a support surface 130 at a height of about around 300 to 1200 microns. As noted herein, the support surface 130 may be provided as an integral surface of the bio chip. Alternatively, the support surface 130 may be provided on a piercing plate or as an integral part of the piercing plate. The piercing member 120 comprises a tip region 600 which has a rounded nose region 610. In use, the upper surface of the rounded nose region 610 is locatable in an abutting relationship with the sealing membrane of the blister pouch and is capable of piercing the blister sealing membrane when a force is applied. Aptly, the force required to pierce the blister membrane is about 3N or more. As illustrated in Figure 6, Figure 8 and Figure 9, the piercing member 120 also comprises a plurality of abutment wall regions or angled struts 620, 630, 640 and 650 which extend upwardly from the support surface to the tip region 600. Each strut extends from the support surface 130 to the tip region at a predetermined angle so that each wall region or strut forms a tangent with an arc of the rounded nose region 610 at an end of the tip region. In the illustrated embodiment, the piercing member comprises four angled struts, however, in alternative embodiments the piercing member may comprise a different number of angled struts e.g. two, three or five.

The abutment wall regions 620, 630, 640, 650 form a cross when viewed from above with the centre of the cross being the rounded nose region 610. The abutment wall regions extend upwardly from the support surface at an angle of approximately 90°, as shown in Figure 6. The rounded nose region of the piercing member allows the piercing member to abut a blister sealing membrane without piercing it. This embodiment has the advantage that a blister pouch may be stored and e.g. transported in an abutting relationship with a piercing member which may be an integral part of a microfluidic device e.g. a bio-chip. Thus, certain embodiments of the present invention may have utility in systems which need to be easily transported e.g. lab-on-chip portable devices.

Thus, between each abutment wall region and an adjacent abutment wall region is a passageway 660, 670, 680, 690 through which fluid stored in the blister pouch can flow once the blister sealing membrane is punctured by the piercing member. The fluid can then flow into the collection chamber 220 located beneath the passageways formed by adjacent abutment wall regions. The passageways are sized such that liquid may pass through the passageway and do not pool on top of the piercing member and therefore the bio-chip. As a result, substantially all of the liquid stored in the blister pouch passes into the biochip which means that the amount of liquid used in the process taking place on the bio-chip can be accurately determined.

Figure 7 is a cross sectional view of section A-A of Figure 6 and shows the collection chamber 220 located underneath the rounded nose region 610 of the piercing member 120. Figure 9 is a bottom view of the apparatus 100 showing a lower surface 900 of for example a piercing plate or a bio chip. Figure 10 is an exploded view illustrating a blister pack 710 which comprises two blister pouches 250a, 250b which protrude from and are supported by a backing web 720. It will be appreciated that a blister pack 710 may comprise a plurality of blister pouches 250. Figure 10 also illustrates a piercing plate 230 which comprises two piercing members 120a, 120b. Each blister pouch may be filled with the same type of fluid or a different type of fluid. The backing web 720 includes one or more interconnecting regions 730 between the blister pouches. The backing web 720 may be provided with frangible lines or perforations (not shown) in the interconnecting regions between blister pouches to enable a section comprising one or more blister pouches to be "snapped-off" or removed from the blister pack.

The backing web may be formed from the same material as the blister forming material. In one embodiment, the backing web is formed from a plastic, a composite material, a film material or a combination of such materials.

The blister pack 710 may also include a sealing membrane (not shown) which extends across the backing web 720 and the otherwise open entrance of a plurality of blister pouches 250, thus sealing a plurality of blister pouches. The sealing membrane may comprise one or more perforations (not shown) which coincide with an outer perimeter surrounding an individual blister pouch. In embodiments in which the backing web comprises frangible lines or perforations, the sealing membrane may comprise corresponding frangible lines or perforations. The blister pouch may be sealed by a heat-sealing method. Alternatively, the blister pouch may be sealed by way of a cold-sealing method. Alternatively, the blister pack may comprise a plurality of sealing membranes, each sealing membrane extending across an otherwise open blister pouch thus sealing a single blister pouch. The sealing membrane may be heat sealed to an upper surface of the backing web.

In one embodiment, the blister pouch 250 is filled with a pre-determined amount of a fluid e.g. a reagent. The blister pouch may be filled with the fluid using an automated filling process. Alternatively, or in addition, the blister pouch may be filled with a liquid manually or using a semi-automated process.

Figure 10 also illustrates a layer of adhesive or adhesive material 740 which secures the blister pack to a piercing plate or to a biochip which comprises the piercing member. The embodiment illustrated in Figure 10 shows the piercing members being composed in a piercing plate.

The adhesive material 740 comprises one or more apertures 750a, 750b which are located to correspond to the location of the piercing members 120a, 120b respectively. The apertures 750a, 750b also provide a through passageway for fluid stored in the blister pouch to flow through the piercing member once the blister sealing membrane is pierced.

In the embodiment illustrated in Figure 10, the piercing member 120 is an integrally formed portion of a piercing plate 230. A piercing plate 230 can be used in conjunction with microfluidic devices of differing types. The piercing plate may be secured to a bio chip 1 10 by an adhesive element e.g. a layer of adhesive 240. Prior to being secured to the bio chip, the piercing plate will be aligned to the bio chip such that each piercing member is positioned above a collection chamber. The piercing plate can be used in combination with a variety of bio chips providing that the bio chip comprises at least one collection chamber in a position which corresponds to the positioning of at least one piercing member of the piercing plate.

Aptly, the piercing member 120 when comprised in the piercing plate 230 is upstanding from the upper surface of the piercing plate to a height of between about 300 to 1200 microns. Aptly, the piercing member 120 when comprised in the bio chip is upstanding from the upper surface of the piercing plate to a height of between about 300 to 1200 microns.

In certain embodiments, the piercing plate comprising one or more piercing members may be supplied in combination with a blister pack. The blister pack may be supplied with the piercing member abutting but not piercing the sealing membrane of a blister pouch comprised in the blister pack. The piercing member may pierce the blister sealing membrane only when a predetermined force is applied e.g. about 3N or greater.

Additionally or alternatively, the piercing plate may include at least one alignment element (not shown) which locates the piercing plate at a desired position with respect to each blister pouch. Aptly, the alignment element comprises at least one edge of the piercing plate. Alternatively, the alignment element comprises at least one locating lug or recess which mates with a complimentary alignment element of the blister pouch. Figure 1 1 illustrates an alternative view of an individual blister pouch 250. The blister pouch 250 comprises a continuous curved exterior wall 1 100 which define a cavity in which fluids are introduced and stored prior to use. If individual blister pouches are used, which are not an integral part of a blister pack, the blister pouch may comprise a lip region 1 1 10 which a user can grip to handle the blister pouch.

Figure 12 illustrates a side view of a blister pouch 250 positioned over a piercing member 120 which is in turn located above a collection chamber of a device e.g. a bio chip as described herein. The piercing member abuts the blister sealing membrane at an abutment point 1200 and deforms the membrane but since there is insufficient force applied the sealing membrane is not pierced. In certain embodiments, the apparatus comprising the piercing member may be stored and/or transported packaged such that the piercing member abuts but does not pierce the sealing membrane.

In use, a blister pouch 250 is prefilled with a predetermined volume of a liquid. As noted herein, the liquid may be any reagent for use in a process taking place on a microfluidic device e.g. a bio chip 1 10 as defined herein. An open mouth of the blister pouch is subsequently sealed by a sealing membrane 270. As described herein, the blister pouch may be one of a plurality of blister pouches in a blister pack. Alternatively, the blister pouch may be supplied as an individual pouch.

Aptly, the blister pouch is sealed in an air tight manner and as such may be suitable for long term storage of the liquid. Additionally, the blister pouch can be stored at low temperatures, e.g. in a refrigerator, if the liquid needs to be stored in this way. The provision of reagents in blister pouches in this way enable different liquids to be stored in their optimum conditions ready for use in a process carried out on a microfluidic device.

As discussed herein, a piercing member 120 is provided in an apparatus 100. The piercing member may be an integral element of a piercing plate 230 or an integral feature of a bio chip 1 10. The piercing member may be provided in an abutting relationship with the sealing membrane of the blister pouch such that it deforms but does not break the sealing membrane in a first mode of operation. Aptly, the piercing member deforms but does not pierce the sealing membrane when a force of up to about 1 N is applied.

In a second mode of operation, a predetermined amount of force is applied to the blister pouch and/or the piercing member such that the sealing membrane is punctured by the piercing member. Aptly, when the blister pouch is urged against the piercing member, the piercing member is disposed to rupture the membrane when a force of about around 3 to 12N is applied to the membrane. The force may be applied to the blister pouch or the piercing member manually by a user or by an actuator. In one embodiment, the actuator may be adapted to apply a pressure at a predetermined rate. The actuator may further comprise a control element adapted to instruct the actuator to apply a force to a plurality of blister pouches at substantially the same time or at predetermined time intervals.

Once the blister sealing membrane has been broken by the piercing member, the liquid stored in the blister pouch is released in a controlled manner and substantially all of the liquid flows over the piercing member and through the openings formed between the abutment wall regions. The liquid then collects underneath the piercing member e.g. in a collection chamber comprised in a biochip. The liquid can then be used in a process on the bio-chip.

Throughout the description and claims of this specification, the words "comprise" and "contain" and variations of them mean "including but not limited to" and they are not intended to (and do not) exclude other moieties, additives, components, integers or steps. Throughout the description and claims of this specification, the singular encompasses the plural unless the context otherwise requires. In particular, where the indefinite article is used, the specification is to be understood as contemplating plurality as well as singularity, unless the context requires otherwise.

Features, integers, characteristics or groups described in conjunction with a particular aspect, embodiment or example of the invention are to be understood to be applicable to any other aspect, embodiment or example described herein unless incompatible therewith. All of the features disclosed in this specification (including any accompanying claims, abstract and drawings), and/or all of the steps of any method or process so disclosed, may be combined in any combination, except combinations where at least some of the features and/or steps are mutually exclusive. The invention is not restricted to any details of any foregoing embodiments. The invention extends to any novel one, or novel combination, of the features disclosed in this specification (including any accompanying claims, abstract and drawings), or to any novel one, or any novel combination, of the steps of any method or process so disclosed.

The reader's attention is directed to all papers and documents which are filed concurrently with or previous to this specification in connection with this application and which are open to public inspection with this specification, and the contents of all such papers and documents are incorporated herein by reference.

Claims

Apparatus for providing a target liquid at a desired location, comprising:

at least one piercing member upstanding from a support surface at a height of about around 300 to 1200 microns and locatable with respect to a sealing membrane of a respective blister pouch; wherein

a tip region of the piercing member is locatable in an abutting relationship with the sealing membrane in a first mode of operation and through the sealing membrane in a further mode of operation for providing liquid from the blister pouch at a desired location.

The apparatus as claimed in claim 1 , further comprising:

in the first mode of operation the piercing member is locatable to deform but not pierce the sealing membrane.

The apparatus as claimed in claim 2, further comprising:

in the first mode of operation the piercing member does not pierce the sealing membrane when an abutment force of up to about around 1 N is applied by the piercing member to the sealing membrane.

The apparatus as claimed in any preceding claim, further comprising:

said tip region comprises a rounded nose region having a generally arcuate shape with a radius of curvature of about around 50 to 1000 microns.

The apparatus as claimed in claim 4 wherein the radius of curvature is about around 100 to 500 microns.

The apparatus as claimed in any preceding claim, further comprising:

said piercing member further comprises a plurality of abutment wall regions extending from the support surface to the tip region at a predetermined angle so that each wall region forms a tangent with an arc of a rounded nose region at an end of the tip region.

The apparatus as claimed in claim 6, further comprising: each abutment wall region meets the nose region at an angle of about around 45 degrees.

The apparatus as claimed in any preceding claim, further comprising:

the piercing member provides a plurality of fluid communication passageways proximate to the tip region that each communicate liquid urged from a pierced blister pouch to at least one collection chamber under the support surface.

The apparatus as claimed in claim 8 wherein the piercing member comprises a plurality of arms extending upwards and in an inclined manner from the support surface and the fluid communication passageways are provided by openings in the support surface in notched regions between adjacent arms.

The apparatus as claimed in any preceding claim wherein the first mode of operation comprises a transportation mode during which a blister pouch is transportable in an abutting relationship with said piercing member without risk of puncture.

The apparatus as claimed in any preceding claim wherein in the further mode of operation in which the blister pouch is urged against the piercing member the piercing member is disposed to rupture the membrane at a rupture force of about around 3 to 12N.

The apparatus as claimed in any preceding claim wherein each piercing member is manufactured from a material selected from a group comprising glass, polycarbonate, polymethyl methacrylate (PMMA), cyclic olefin copolymer (COC), polystyrene (PS), polyether ether ketone (PEEK), polytetrafluoroethylene (PTFE) and polypropylene (PP).

A piercing plate, securable to at least one blister pouch, comprising the apparatus as claimed in any preceding claim.

The piercing plate as claimed in claim 13, further comprising:

a planar upper surface, comprising said support surface, and a planar lower surface that is securable to an upper surface of a bio chip.

15. The piercing plate as claimed in claim 14 further comprising an adhesive layer on the lower surface.

16. The piercing plate as claimed in any one of claims 13 to 15, further comprising:

at least one alignment element that locates the piercing plate at a desired position with respect to each blister pouch.

17. The piercing plate as claimed in claim 16 wherein said alignment element comprises at least one edge of the piercing plate.

18. The piercing plate as claimed in claim 16 wherein said alignment element comprises at least one locating lug or recess that mates with a complimentary alignment element of the blister pouch. 19. The piercing plate as claimed in any one of claims 13 to 18 further comprising at least one further alignment element that locates the piercing plate at a desired position with respect to a bio chip.

20. The piercing plate as claimed in claim 19 wherein each further alignment element comprises at least one edge of the piercing plate.

21 . The piercing plate as claimed in claim 19 wherein said further alignment element comprises at least one locating lug or recess that mates with a complimentary alignment element of the bio chip.

22. The piercing plate as claimed in any one of claims 13 to 21 , further comprising:

a cavity under each piercing member that is locatable in fluid communication with a collection chamber of a bio chip. 23. The piercing plate as claimed in any one of claims 13 to 22, further comprising:

the plate is about around 200 to 1000 microns thick.

24. The piercing plate as claimed in any one of claims 13 to 23 wherein the plate is manufactured from a material selected from the list of glass, polycarbonate and PMMA. The piercing plate as claimed in any one of claims 13 to 24 wherein the plate and each piercing member are integrally formed.

The piercing plate as claimed in any one of claims 13 to 25 wherein the plate is retrofittable to an underside of a blister package comprising one or more blister pouches sealed with a sealing membrane.

27. A blister pack, comprising:

a plurality of blister pouches secured together via a web and a common sealing membrane; and

at least one piercing plate as claimed in any one of claims 13 to 26 secured to a lower surface of the sealing membrane.

28. The blister pack as claimed in claim 27, further comprising:

a plurality of frangible lines or perforation lines in the web.

29. A bio chip, comprising the apparatus as claimed in any one of claims 1 to 9.

30. The bio chip as claimed in claim 29, further comprising:

an upper chip layer comprising each piercing member as an integral unit.

31 . The bio chip as claimed in claim 29, further comprising:

an upper chip layer having an upper surface on which at least one piercing plate comprising said at least one piercing member is secured.

32. The bio chip as claimed in claim 30 or claim 31 wherein a lower surface of the upper chip layer includes a collection chamber located beneath each piercing member and at least one channel connecting a collection chamber to a respective sample chamber.

33. The bio chip as claimed in claim 32 wherein said channel comprises a channel region that provides a serpentine pathway.

34. Apparatus constructed and arranged substantially as hereinbefore described with reference to the accompanying drawings.

35. A piercing plate constructed and arranged substantially as hereinbefore described with reference to the accompanying drawings.

36. A bio chip constructed and arranged substantially as hereinbefore described with reference to the accompanying drawings.

37. A method substantially as hereinbefore described with reference to the accompanying drawings.