Liquid treatment apparatus

Abstract

The present invention relates to a liquid treatment device comprising: a fluid layer comprising: a plurality of channels; and an opening in a surface of the fluid layer, where the opening extends through at least a portion of a thickness of the fluid layer to one of the plurality of channels; a rigid layer including an actuatable portion; and a reservoir capsule positioned between the fluid layer and the actuatable portion, where the reservoir capsule comprises: a body defining a volume of stored liquid, the body comprising a first deformable portion, a main reservoir portion, and a restriction portion connecting the first deformable portion to the main reservoir portion; and a sealing layer configured to seal the volume; wherein the reservoir capsule is positioned over the opening such that a portion of the sealing layer covers the opening; wherein the actuatable portion comprises a protrusion extending toward the reservoir capsule, and wherein the actuatable portion is actuatable from a first position in which the protrusion does not deform the first deformable portion to a second position in which the protrusion deforms the first deformable portion into contact with a portion of the sealing layer to cause a rupture of the portion of the sealing layer, and wherein when the actuatable portion is in the second position, the first deformable portion deforms the second deformable portion into contact with the portion of the sealing layer to cause the rupture of the portion of the sealing layer. The protrusion does not deform the restricting portion.

Description

TECHNICAL FIELD

[0001] The present disclosure relates to a liquid handling device, a liquid handling system, and a method of moving a liquid in a liquid handling device. BACKGROUND

[0002] Point-of-care diagnostic devices are commonly used to perform diagnostic tests, such as immunoassays, on biological samples, such as whole blood, serum, or plasma. To perform such diagnostic tests, the biological sample needs to be transferred to the diagnostic device. The diagnostic device is then inserted into an analyzer device (or instrument) that controls the movement of fluids (e.g., biological sample, reagents, buffer solutions, etc.) within the diagnostic device and performs measurements of biological markers in order to conduct the diagnostic test.

[0003] Point-of-care testing provides diagnostic tests to patients conveniently and quickly, allowing for better and faster clinical decisions. However, it is challenging to integrate diagnostic tests into point-of-care devices or systems. Preparing a sample for an immunoassay can require mixing multiple solutions and reagents, as well as precise control of volumes and mixing times. Moreover, the device should ideally be automated to avoid the need for medical professionals.

[0004] Therefore, there is a need for improved liquid handling devices capable of performing liquid handling operations used in point-of-care diagnostic tests. SUMMARY

[0005] This section introduces concepts more fully described in the detailed description. It should not be used to identify the fundamental features of the claimed subject matter nor should it be used to limit the scope of the claimed subject matter. The inclusion of multiple statements in a single paragraph does not imply that there is a structural or functional relationship between the statements.

[0006] According to a first aspect of the present disclosure, there is provided a liquid handling device comprising: a fluidic layer comprising: a plurality of channels; and an opening in a surface of the fluidic layer, wherein the opening extends through at least a portion of a thickness of the fluidic layer to one of the plurality of channels; a rigid layer comprising an actuatable portion; and a reservoir capsule positioned between the fluidic layer and the actuatable portion, wherein the reservoir capsule comprises: a body defining a volume in which a liquid is stored, the body comprising a first deformable portion, a main reservoir portion, and a restriction portion connecting the first deformable portion to the main reservoir portion; and a sealing layer configured to seal the volume; wherein the reservoir capsule is positioned over the opening such that a portion of the sealing layer covers the opening; wherein the actuatable portion comprises a protrusion extending towards the reservoir capsule, wherein the actuatable portion is actuatable from a first position in which the protrusion does not deform the first deformable portion to a second position in which the protrusion deforms the first deformable portion into contact with the portion of the sealing layer to cause the portion of the sealing layer to rupture, wherein the protrusion does not deform the restriction portion when the actuatable portion is in the second position.

[0007] The protrusion can be unable to pass into the opening.

[0008] The reservoir capsule can be positioned over the opening such that the first deformable portion and the restriction portion are positioned at least partially over the opening. The opening can have an oblong shape.

[0009] The first deformable portion can comprise a first region and a second region, wherein a first distance between the first region and the sealing layer is greater than a second distance between the second region and the sealing layer. The first region and the second region can be concentric such that the first region surrounds the second region.

[0010] A width of the opening can be less than a width of the protrusion.

[0011] The protrusion can comprise an end portion that engages the first deformable portion of the reservoir capsule. The end portion of the protrusion can be flat. The end portion of the protrusion can have a cross-section of a main sector shape.

[0012] The protrusion can comprise a groove that defines a cross-section of the end portion of the protrusion. The groove can be aligned with the restriction portion of the reservoir capsule. The groove can define a main sector shape of the cross-section of the end portion of the protrusion.

[0013] The cross-section of the end portion of the protrusion can comprise one or more outwardly extending protruding ribs. The one or more outwardly extending protruding ribs can extend from a curved portion of the main sector shape.

[0014] The protrusion can be a first protrusion, and the actuatable portion can include a second protrusion extending toward the liquid storage capsule. The body of the liquid storage capsule can include a second deformable portion. The opening can be a first opening, and the fluid layer can include a second opening extending through at least a portion of a thickness of the fluid layer to one of the plurality of channels. The portion of the sealing layer can be a first portion of the sealing layer, and the liquid storage capsule can be positioned above the second opening such that a second portion of the sealing layer covers the second opening. In the first position, the second protrusion can not deform the second deformable portion. In the second position, the second protrusion can deform the second deformable portion to contact the second portion of the sealing layer, causing the second portion of the sealing layer to rupture.

[0015] The liquid storage capsule can be a first liquid storage capsule, and the liquid handling device can include a second liquid storage capsule. The second liquid storage capsule can include a body defining a volume in which liquid is stored, the body including a first deformable portion and a second deformable portion, and a sealing layer configured to seal the volume. The actuatable portion can include a third protrusion extending toward the first deformable portion of the second liquid storage capsule and a fourth protrusion extending toward the second deformable portion of the second liquid storage capsule. The fluid layer can include a third opening extending through at least a portion of a thickness of the fluid layer to one of the plurality of channels and a fourth opening extending through at least a portion of a thickness of the fluid layer to one of the plurality of channels. The second liquid storage capsule can be positioned above the third opening and the fourth opening such that a first portion of the sealing layer of the second liquid storage capsule covers the third opening and a second portion of the sealing layer of the second liquid storage capsule covers the fourth opening. In the first position, the third protrusion can not deform the first deformable portion of the body of the second liquid storage capsule, and the fourth protrusion can not deform the second deformable portion of the body of the second liquid storage capsule. In the second position, the third protrusion can deform the first deformable portion of the body of the second liquid storage capsule into contact with the first portion of the sealing layer of the second liquid storage capsule to cause the first portion of the sealing layer of the second liquid storage capsule to rupture. In the second position, the fourth protrusion can deform the second deformable portion of the body of the second liquid storage capsule into contact with the second portion of the sealing layer of the second liquid storage capsule to cause the second portion of the sealing layer of the second liquid storage capsule to rupture.

[0016] According to a second aspect of the present disclosure, there is provided a liquid handling device comprising: a fluidic layer comprising: a plurality of channels; and an opening in a surface of the fluidic layer, wherein the opening extends through at least a portion of a thickness of the fluidic layer to one of the plurality of channels; a rigid layer comprising an actuatable portion; and a reservoir capsule positioned between the fluidic layer and the actuatable portion, wherein the reservoir capsule comprises: a body defining a volume in which a liquid is stored; and a sealing layer configured to seal the volume; wherein the reservoir capsule is positioned over the opening such that a portion of the sealing layer covers the opening; wherein the actuatable portion comprises a protrusion extending towards the reservoir capsule, wherein the actuatable portion is actuatable from a first position in which the protrusion does not deform the body to a second position in which the protrusion deforms the body into contact with the portion of the sealing layer to cause the portion of the sealing layer to rupture; and wherein the protrusion is unable to pass into the opening.

[0017] The opening can have an oblong shape.

[0018] The body of the reservoir capsule can comprise a first deformable portion configured to be deformed by the protrusion when the actuatable portion is in the second position. The body can further comprise a main reservoir portion and a restriction portion connecting the first deformable portion to the main reservoir portion, wherein the reservoir capsule is positioned over the opening such that the first deformable portion and the restriction portion are positioned at least partially over the opening.

[0019] The first deformable portion can comprise a first region and a second region, wherein a first distance between the first region and the sealing layer is greater than a second distance between the second region and the sealing layer. The first region and the second region are concentric such that the first region encloses the second region.

[0020] The width of the opening can be less than the width of the protrusion.

[0021] The protrusion can comprise an end portion that engages the first deformable portion of the reservoir capsule. The end portion of the protrusion can be flat. The end portion of the protrusion can have a cross-section of a main sector shape.

[0022] The protrusion can comprise a groove defining a cross-section of the end portion of the protrusion. The groove can be aligned with the restriction portion of the reservoir capsule. The groove can define a main sector shape of the cross-section of the end portion of the protrusion.

[0023] The cross-section of the end portion of the protrusion can comprise one or more outwardly extending protruding ribs. The one or more outwardly extending protruding ribs can extend from a curved portion of the main sector shape.

[0024] The protrusion can be a first protrusion, and the actuatable portion can include a second protrusion extending toward the liquid reservoir capsule. The opening can be a first opening, and the fluid layer can include a second opening extending through at least a portion of a thickness of the fluid layer to one of the plurality of channels. The portion of the sealing layer can be a first portion of the sealing layer, and the liquid reservoir capsule can be positioned above the second opening such that a second portion of the sealing layer covers the second opening. In the first position, the second protrusion can not deform the body of the liquid reservoir capsule. In the second position, the second protrusion can deform the body to contact the second portion of the sealing layer, causing the second portion of the sealing layer to rupture.

[0025] The liquid reservoir capsule can be a first liquid reservoir capsule, and the liquid handling device can include a second liquid reservoir capsule comprising: a body defining a volume storing a liquid; and a sealing layer configured to seal the volume. The actuatable portion can include: a third protrusion extending toward the second liquid reservoir capsule; and a fourth protrusion extending toward the second liquid reservoir capsule. The fluid layer can include: a third opening extending through at least a portion of a thickness of the fluid layer to one of the plurality of channels; and a fourth opening extending through at least a portion of a thickness of the fluid layer to one of the plurality of channels. The second liquid reservoir capsule can be positioned above the third opening and the fourth opening such that a first portion of the sealing layer of the second liquid reservoir capsule covers the third opening and a second portion of the sealing layer of the second liquid reservoir capsule covers the fourth opening. In the first position, the third protrusion can not deform the body of the second liquid reservoir capsule and the fourth protrusion can not deform the body of the second liquid reservoir capsule. In the second position, the third protrusion can deform the body of the second liquid reservoir capsule to contact the first portion of the sealing layer of the second liquid reservoir capsule, causing the first portion of the sealing layer of the second liquid reservoir capsule to rupture. In the second position, the fourth protrusion can deform the body of the second liquid reservoir capsule to contact the second portion of the sealing layer of the second liquid reservoir capsule, causing the second portion of the sealing layer of the second liquid reservoir capsule to rupture.

[0026] According to a third aspect of the disclosure, there is provided a liquid handling device comprising: a fluid layer comprising a chamber, wherein the chamber comprises: an opening at an upper end of the chamber; and a sealing layer configured to seal the opening of the chamber; wherein the chamber comprises one or more protrusions, wherein each of the one or more protrusions extends from an inner wall of the chamber; wherein the one or more protrusions prevent a reagent ball from contacting the sealing layer when the reagent ball is contained in the chamber.

[0027] The opening can be configured to allow a reagent ball to be inserted into the chamber.

[0028] The one or more protrusions are elastically deformable. The fluid layer is formed from a thermoplastic elastomer.

[0029] Each of the one or more protrusions can extend from an inner wall of the chamber at the upper end of the chamber. The opening can be partially defined by each of the one or more protrusions.

[0030] The fluid layer can include a protrusion extending from an upper surface of the fluid layer, wherein the protrusion partially defines the chamber.

[0031] The chamber can further include a fluid inlet below the one or more protrusions and an air outlet in the inner wall, wherein a first maximum distance between the air outlet and the upper end base is less than or equal to a second maximum distance between the one or more protrusions and the upper end.

[0032] Each of the one or more protrusions can include a first protrusion portion extending from an inner wall of the chamber and a second protrusion portion extending from a distal end of the first protrusion portion, wherein the second protrusion portion extends away from the upper end of the chamber.

[0033] According to a fourth aspect of the disclosure, there is provided a liquid handling device comprising: a first reservoir and a second reservoir, wherein each of the first reservoir and the second reservoir is configured to be opened by creating an inlet and an outlet in the reservoir; and a fluid layer comprising: a network of channels; and a pneumatic port configured to receive positive pressure; wherein the network of channels comprises: a first reservoir inlet channel configured to provide a fluid connection between the pneumatic port and the inlet in the first reservoir when the first reservoir is opened; and a second reservoir inlet channel configured to provide a fluid connection between the pneumatic port and the inlet in the second reservoir when the second reservoir is opened.

[0034] The liquid handling device can further comprise: a first reservoir valve configured to control flow of liquid from the outlet of the first reservoir; and a second reservoir valve configured to control flow of liquid from the outlet of the second reservoir.

[0035] The network of channels can further comprise: a first reservoir outlet channel configured to be in fluid communication with the outlet in the first reservoir when the first reservoir is opened, wherein the first reservoir valve is on the first reservoir outlet channel.

[0036] The network of channels can further comprise: a second reservoir outlet channel configured to be in fluid communication with the outlet in the second reservoir when the second reservoir is opened, wherein the second reservoir valve is on the second reservoir outlet channel.

[0037] The liquid treatment device can further include a well portion, where the well portion is defined at least in part by an opening extending through at least a portion of a thickness of the fluid layer, where the well portion is in fluid communication with the pneumatic port, the first reservoir inlet channel, and the second reservoir inlet channel.

[0038] The opening, the first reservoir inlet channel, and the second reservoir inlet channel can each be disposed in an upper surface of the fluid layer. A depth of the opening can be greater than depths of the first reservoir inlet channel and the second reservoir inlet channel.

[0039] The liquid treatment device can further include a lower portion including a slot in an upper surface of the lower portion. The fluid layer can be positioned on top of the lower portion. The well portion can be defined at least in part by the opening in the fluid layer and the slot in the lower portion.

[0040] The opening in the fluid layer can extend through a thickness of the fluid layer.

[0041] The pneumatic port can extend through a thickness of the fluid layer, where the pneumatic port is in fluid communication with the slot via a connection channel in the lower portion.

[0042] The well portion can include an absorbent material.

[0043] Each of the first reservoir inlet channel and the second reservoir inlet channel can include: one or more first channel regions; and a second channel region, where a first depth of the one or more first channel regions is less than a second depth of the second channel region.

[0044] The second depth can be equal to a thickness of the fluid layer. A first width of the one or more first channel regions can be less than a second width of the second channel region.

[0045] The liquid treatment device can further include a third reservoir, where the third reservoir is configured to be opened by forming an inlet and an outlet in the reservoir, and where the network of channels further includes a third reservoir inlet channel configured to provide a fluid connection between the pneumatic port and the inlet in the third reservoir when the third reservoir is opened.

[0046] The pneumatic port can be a first pneumatic port. The liquid treatment device can further include a second pneumatic port configured to be in fluid communication with an outlet in the first reservoir.

[0047] The second pneumatic port can be configured to be in fluid communication with an outlet in the second reservoir.

[0048] The liquid treatment device can further include a third pneumatic port configured to be in fluid communication with an outlet in the second reservoir.

[0049] According to a fifth aspect of the disclosure, there is provided a method of controlling liquid flow in a liquid handling device, the method comprising: creating an inlet and an outlet in a first reservoir of the liquid handling device to create a fluid connection between a pneumatic port of the liquid handling device and the inlet in the first reservoir; creating an inlet and an outlet in a second reservoir of the liquid handling device to create a fluid connection between the pneumatic port and the inlet in the second reservoir; dispensing liquid from the outlet in the first reservoir by applying a positive pressure via the pneumatic port; and dispensing liquid from the outlet in the second reservoir by applying a positive pressure via the pneumatic port.

[0050] Dispensing liquid from the outlet in the first reservoir can comprise opening a first reservoir valve configured to control flow of liquid from the outlet in the first reservoir.

[0051] Dispensing liquid from the outlet in the second reservoir can comprise opening a second reservoir valve configured to control flow of liquid from the outlet in the second reservoir.

[0052] The method can further comprise: creating an inlet and an outlet in a third reservoir of the liquid handling device to create a fluid connection between the pneumatic port and the inlet of the third reservoir; and dispensing liquid from the outlet in the third reservoir by applying a positive pressure via the pneumatic port.

[0053] The pneumatic port can be a first pneumatic port. The liquid handling device can comprise a second pneumatic port. Dispensing liquid from the outlet in the first reservoir can comprise: opening one or more valves of the liquid handling device to establish a fluid connection between the outlet in the first reservoir and the second pneumatic port; and venting the second pneumatic port during the application of the positive pressure via the first pneumatic port.

[0054] Dispensing liquid from the outlet in the second reservoir can comprise: opening one or more valves of the liquid handling device to establish a fluid connection between the outlet in the second reservoir and the second pneumatic port; and venting the second pneumatic port during the application of the positive pressure via the first pneumatic port.

[0055] The liquid handling device can comprise a third pneumatic port. Dispensing liquid from the outlet in the second reservoir can comprise: opening one or more valves of the liquid handling device to establish a fluid connection between the outlet in the second reservoir and the third pneumatic port; and venting the third pneumatic port during the application of the positive pressure via the first pneumatic port.

[0056] According to a sixth aspect of the present disclosure, there is provided a liquid handling device comprising: a first chamber in fluid communication with a second chamber; a first pneumatic port in fluid communication with one of the first chamber and the second chamber; a second pneumatic port in fluid communication with the other of the first chamber and the second chamber; a chamber inlet conduit configured to allow liquid to flow into the first chamber; a chamber outlet conduit configured to allow liquid to flow out of the second chamber; and a third pneumatic port in fluid communication with a first liquid reagent capsule, wherein the first liquid reagent capsule is in selective fluid communication with the first chamber or the second chamber when the first liquid reagent capsule is opened.

[0057] As used herein, the term "selectively in fluid communication" between A and B means that A can be brought into fluid communication with B by opening one or more valves so as to provide a fluid connection between A and B through components of the fluid network.

[0058] A chamber inlet valve can be used to control the flow of liquid through the chamber inlet conduit. A chamber outlet valve can be used to control the flow of liquid through the chamber outlet conduit. A first liquid reagent capsule valve can be used to control the flow of liquid from the first liquid reagent capsule.

[0059] The first pneumatic port can be in permanent fluid communication with the first chamber. The second pneumatic port can be in permanent fluid communication with the second chamber.

[0060] The liquid handling device can further comprise: a first flow cell in fluid communication with the second chamber via the chamber outlet conduit; a second flow cell in fluid communication with the first flow cell; and a bypass conduit configured to allow liquid to bypass the second flow cell after flowing through the first flow cell. A bypass conduit valve can be used to control the flow of liquid through the bypass conduit.

[0061] The first flow cell is in fluid communication with a waste liquid chamber through a first flow cell outlet conduit. A bypass conduit valve can be used to control the flow of liquid into the waste liquid chamber via the first flow cell outlet conduit.

[0062] The waste liquid chamber can be permanently vented. The second flow cell can be in fluid communication with the waste liquid chamber via a second flow cell outlet conduit. A second flow cell outlet valve can be used to control the flow of liquid into the waste liquid chamber via the second flow cell outlet conduit.

[0063] The third pneumatic port can be in fluid communication with a second liquid reagent capsule. A second liquid reagent capsule valve can be used to control the flow of liquid from the second liquid reagent capsule. The second liquid reagent capsule can be in selective fluid communication with the second flow cell when the second liquid reagent capsule is opened.

[0064] The second liquid reagent capsule can be in fluid communication with the second flow cell via a second flow cell outlet conduit when the second liquid reagent capsule is opened.

[0065] The third pneumatic port can be in fluid communication with a third liquid reagent capsule. A third liquid reagent capsule valve can be used to control liquid flow from the third liquid reagent capsule. The third liquid reagent capsule can be in selective fluid communication with the first chamber when the third liquid reagent capsule is opened. Additionally or alternatively, the third liquid reagent capsule can be in selective fluid communication with the first flow cell when the third liquid reagent capsule is opened.

[0066] The liquid handling device can further include a sample inlet chamber in fluid communication with the chamber inlet conduit. A sample inlet valve can be used to control liquid flow from the sample inlet chamber. The sample inlet chamber can be permanently vented.

[0067] The liquid handling device can further include a sample flow cell in fluid communication with the sample inlet chamber via a sample flow cell inlet conduit. A sample flow cell valve can be used to control liquid flow into the sample flow cell.

[0068] The sample flow cell can be in fluid communication with a sample flow cell waste chamber via a sample flow cell outlet conduit. A sample flow cell valve can be used to control liquid flow into the sample flow cell waste chamber. The sample flow cell waste chamber can be in fluid communication with a second pneumatic port.

[0069] The liquid handling device can further include a third chamber, wherein the third chamber is in fluid communication with the first pneumatic port. The first pneumatic port can be in permanent fluid communication with the third chamber. A third chamber valve can be used to control liquid flow into the third chamber. The third chamber can be in fluid communication with the first liquid reagent capsule. The third chamber can contain a dry reagent.

[0070] The first liquid reagent capsule can be in fluid communication with the first chamber via a prime conduit. A prime conduit valve can be used to control liquid flow through the prime conduit. The prime conduit can be in fluid communication with the sample flow cell inlet conduit via the prime conduit valve.

[0071] The third pneumatic port can be in fluid communication with an air / wash solution supply conduit. An air supply valve can be used to control air flow through the air / wash solution supply conduit. The air / wash solution supply conduit can be in fluid communication with the chamber outlet conduit.

[0072] One or more of the first chamber and the second chamber can contain a dry reagent.

[0073] According to a seventh aspect of the disclosure, there is provided a method of metering a liquid in a channel of a liquid handling device, the method comprising: applying a first pressure to move a first liquid from a first chamber through a first fluid path comprising two junctions; applying a second pressure to move a fluid through a second fluid path comprising a portion of the first fluid path between the two junctions, such that the fluid moves a volume of the first liquid between the two junctions.

[0074] The first pressure can be applied using a first pneumatic port of the liquid handling device, wherein the first pneumatic port is in selective fluid communication with the first chamber. The second pressure can be applied using a second pneumatic port of the liquid handling device, wherein the second pneumatic port is in selective fluid communication with the second chamber.

[0075] The second pressure can be applied to dispense the fluid and the first liquid to a third chamber.

[0076] The first pressure can be applied to aspirate the first liquid from the first chamber through the first fluid path. The first chamber can be permanently vented. Alternatively, the first pressure can be applied to dispense the first liquid from the first chamber through the first fluid path.

[0077] The fluid can be a second liquid. The second pressure can be applied to dispense the second liquid from the second chamber through the second fluid path.

[0078] Alternatively, the fluid can be air. The second pressure can be applied to supply air to the second fluid path through an air supply conduit.

[0079] The method can further comprise metering a plurality of volumes of the first liquid by repeating one or more times: applying a first pressure to move a first liquid from a first chamber through a first fluid path comprising two junctions; applying a second pressure to move a fluid through a second fluid path comprising a portion of the first fluid path between the two junctions, such that the fluid moves a volume of the first liquid between the two junctions.

[0080] According to an eighth aspect of the disclosure, there is provided a method of priming a fluidic circuit of a liquid handling device, the method comprising: applying a first pressure to move a liquid from a chamber into a first channel; continuing to apply the first pressure to move the liquid until a meniscus of the liquid has moved past a junction on the first channel, wherein the junction connects the first channel to a second channel; and applying a second pressure to move liquid from the chamber into the second channel via the first channel and the junction.

[0081] Applying the first pressure can comprise applying a negative pressure to move the liquid. The chamber can be permanently vented.

[0082] Applying the second pressure can include applying negative pressure to move the liquid.

[0083] Applying the first pressure can include applying negative pressure to a first pneumatic port of the liquid handling device. Applying the second pressure can include applying negative pressure to a second pneumatic port of the liquid handling device.

[0084] Applying the first pressure can include applying positive pressure to move the liquid. Applying the first pressure can include applying positive pressure to a first pneumatic port of the liquid handling device while venting a second pneumatic port of the liquid handling device.

[0085] Applying the second pressure can include applying positive pressure to move the liquid. Applying the second pressure can include applying positive pressure to a first pneumatic port of the liquid handling device. Applying the second pressure can include applying positive pressure to a first pneumatic port of the liquid handling device while venting a third pneumatic port of the liquid handling device.

[0086] According to a ninth aspect of the disclosure, there is provided a method of diluting a sample using a liquid handling device, the method comprising: applying negative pressure to draw a volume of a first liquid into a first flow cell of a liquid handling device; applying negative pressure to draw a volume of the first liquid into a first chamber of the liquid handling device; applying positive pressure to dispense a volume of a second liquid into the first chamber; repeating one or more times the following steps: applying positive pressure to dispense the first liquid and the second liquid from the first chamber to a second chamber; and applying negative pressure to draw the first liquid and the second liquid from the second chamber to the first chamber; applying positive pressure to move a mixture comprising the first liquid and the second liquid from the first chamber or the second chamber to the second flow cell.

[0087] The method can further comprise measuring a detection signal in the first flow cell. The method can further comprise measuring a detection signal in the second flow cell.

[0088] The volume of the first liquid drawn into the first chamber can be a metered volume of the first liquid. The volume of the second liquid dispensed into the first chamber can be a metered volume of the second liquid.

[0089] According to a tenth aspect of the present disclosure, there is provided a method of resuspending dry reagents using a liquid handling device, the method comprising: applying a first pressure to transfer a metered volume of a first liquid into a first chamber to resuspend one or more first dry reagents, wherein resuspension of the one or more first dry reagents forms a first reagent; applying a second pressure to transfer a metered volume of the first liquid into a second chamber to resuspend one or more second dry reagents, wherein resuspension of the one or more second dry reagents forms a second reagent; applying a third pressure to move the first reagent through at least one flow cell; applying a fourth pressure to move the second reagent through the at least one flow cell; and measuring a detection signal in the at least one flow cell.

[0090] The volume of the first liquid transferred into the first chamber can be a metered volume of the first liquid. The volume of the first liquid transferred into the second chamber can be a metered volume of the first liquid.

[0091] Advantages associated with features of the aspects described above are set forth in the detailed description that follows. BRIEF DESCRIPTION OF DRAWINGS

[0092] Particular embodiments are described below by way of example only and with reference to the accompanying drawings, in which:

[0093] FIG. 1 is an isometric view of a liquid handling device.

[0094] FIG. 2 is FIG. 1 is an isometric view of a liquid handling device, wherein labels of the liquid handling device have been removed.

[0095] FIG. 3 is an exploded view showing components of a liquid handling device in FIG. 1

[0096] FIG. 4 is an exploded view of components shown in FIG. 3 assembled at a first assembly stage.

[0097] FIG. 5 is an exploded view of components shown in FIG. 3 assembled at a second assembly stage.

[0098] FIG. 6 is an exploded view of components shown in FIG. 3 assembled at a third assembly stage.

[0099] FIG. 7 is a top view of a liquid handling device, wherein labels of the liquid handling device have been removed. FIG. 1

[0100] is a top view of a liquid handling device, wherein labels of the liquid handling device have been removed.​FIG. 8 is a top view showing the fluid layer of the liquid handling device of FIG. 1

[0101] FIG. 9A is a top view showing the fluid layer of the liquid handling device of FIG. 8

[0102] FIG. 9B is a top view showing the fluid layer of the liquid handling device of FIG. 8

[0103] FIG. 10 is a top view showing the fluid layer of the liquid handling device of FIG. 1

[0104] FIG. 11 FIG. 10

[0105] FIG. 12 FIG. 10

[0106] FIG. 13 FIG. 12

[0107] FIG. 14 FIG. 1

[0108] FIG. 15 FIG. 14

[0109] FIG. 16 FIG. 14

[0110] FIG. 17 FIG. 15

[0111] FIG. 18 FIG. 1

[0112] FIG. 19 FIG. 18 FIG. 10

[0113] FIG. 20 FIG. 7 ​​​​​​​​​​​​​​​​​​​​​​​​​

[0114] FIG. 21 is FIG. 20 an enlarged sectional view of the region S shown.

[0115] FIG. 22 is a schematic view showing the protrusion, the deformable portion of the reservoir capsule, and the opening in the fluid layer.

[0116] FIG. 23 is FIG. 1 an isometric view of a second portion of the liquid handling device shown.

[0117] FIG. 24 is FIG. 23 a top view of the second portion shown.

[0118] FIG. 25 is FIG. 3 an exploded view of certain components shown.

[0119] FIG. 26 is FIG. 1 a top view of the fluid layer of the liquid handling device shown.

[0120] FIG. 27 is FIG. 26 a bottom view of the fluid layer shown.

[0121] FIG. 28 is a schematic view of a fluid network defined by channels in the fluid layer shown by FIG. 26 and FIG. 27 .

[0122] FIG. 29 is FIG. 28 another schematic view of the fluid network shown.

[0123] FIG. 30 is FIG. 26 a side view of the fluid layer shown.

[0124] FIG. 31 is FIG. 26 a sectional view of the fluid layer shown.

[0125] FIG. 32 is FIG. 25 a top view of the components assembled.

[0126] FIG. 33 is a sectional view through line Z-Z in FIG. 32 .

[0127] FIG. 34 is FIG. 33 an enlarged sectional view of the region T shown.

[0128] FIG. 35This is a flowchart of a method for controlling the flow of liquid in a liquid handling device.

[0129] FIG. 36 This is a flowchart of a method for metering liquid in the channel of a liquid handling device.

[0130] FIG. 37 This is a flowchart of a method for the fluid circuit of a pre-filled liquid treatment device.

[0131] FIG. 38 This is a flowchart of a method for diluting samples using a liquid handling device.

[0132] FIG. 39 This is a flowchart of a method for resuspending and drying reagents using a liquid handling device.

[0133] FIG. 40 Is using FIG. 1 The flowchart shows the method for performing the first test using the liquid processing device shown.

[0134] FIG. 41 Is using FIG. 1 The flowchart shows the method for performing a second test using the liquid processing device shown.

[0135] FIG. 42 Is using FIG. 1 The flowchart shows the method for performing a third test using the liquid processing device shown.

[0136] FIG. 43 Is using FIG. 1 The flowchart shows the method for performing the fourth test using the liquid processing device shown. Detailed Implementation

[0137] The various embodiments of this disclosure are explained below with specific reference to microfluidic cartridges used to perform diagnostic tests. However, it should be understood that the embodiments described herein can be applied to microfluidic cartridges for other purposes. As used herein, the term "microfluidic cartridge" means a device having a conduit with a width or depth of less than 1 mm. It should also be understood that the embodiments described herein are not limited to microfluidics and can be applied to liquid handling devices of various sizes for various purposes.

[0138] FIG. 1 This is an isometric view of a liquid handling device in the form of a diagnostic cartridge 100 (e.g., a microfluidic cartridge). FIG. 2 An isometric view of box 100 is shown, with label 110 removed. From FIG. 3 As can be seen from the exploded view shown, box 100 includes several components.

[0139] In particular, the cartridge 100 includes a first portion 200 and a second portion 500, each portion formed of a rigid material. In use (i.e., when the cartridge 100 is in the orientation shown FIG. 1 The first portion 200 is an upper portion and the second portion 500 is a lower portion, in the orientation shown. The first portion 200 and the second portion 500 together define a housing of the cartridge 100. In particular, the first portion 200 includes a rigid face 250 that defines an upper surface of the cartridge 100. Likewise, the second portion 500 includes a rigid face 570 that defines a lower surface of the cartridge 100 (e.g., as shown in FIG. 15 The first portion 200 and the second portion 500 together define a housing of the cartridge 100. In particular, the first portion 200 includes a rigid face 250 that defines an upper surface of the cartridge 100. Likewise, the second portion 500 includes a rigid face 570 that defines a lower surface of the cartridge 100 (e.g., as shown in FIG. 3 As can be seen, the first portion 200 also includes a sidewall 252 connected to the rigid face 250 and the second portion 500 also includes a sidewall 572 connected to the rigid face 570. The sidewall 252 of the first portion 200 and the sidewall 572 of the second portion 500 cooperate together to define a sidewall of the cartridge 100.

[0140] The cartridge 100 also includes a fluid layer 300 disposed within the housing defined by the first portion 200 and the second portion 500. In particular, the fluid layer 300 is disposed between the rigid face 250 of the first portion 200 and the rigid face 570 of the second portion 500. Thus, the fluid layer 300 is disposed between a first rigid layer in the form of the rigid face 250 and a second rigid layer in the form of the rigid face 570. The fluid layer 300 is formed of an elastomeric material, such as a thermoplastic elastomer (TPE), such as a silicone-based TPE or styrene-ethylene-butylene-styrene (SEBS); polydimethylsiloxane (PDMS); or liquid silicone rubber (LSR).

[0141] As described in greater detail below, the first surface 308 of the fluid layer 300 includes a plurality of valve regions 302 (e.g., as shown in FIG. 25 and FIG. 26 The cartridge 100 is housed in an analyzer device that includes an actuator that applies a force to the valve regions 302 of the fluid layer 300 to close one or more conduits 600 within the cartridge 100 (as best shown in FIG. 16 and FIG. 21 The properties of the material used for the fluid layer 300 depend on the available force that can be applied to the valve regions 302 of the fluid layer 300 by the actuator. Two important properties are the hardness of the material and the relaxation time of the material (i.e., the time it takes for the material to return to its original form after being deformed). Examples of suitable materials include the elastomeric materials listed above. In some embodiments, the fluid layer 300 can be a medical grade material to prevent the fluid layer 300 from reacting with reagents used in diagnostic tests or assays.

[0142] As described in greater detail below, the fluid layer 300 includes a network of channels 304 (as shown in FIG. 26 andFIG. 27 The cartridge 100 also includes a fluid network comprising a plurality of conduits 600 defined at least in part by the network of channels 304 in the fluid layer 300. In particular, the conduits 600 are defined by: (i) the network of channels 304 in the fluid layer 300; (ii) the first sealing layer 400 (shown in FIG. 3 configured to seal the channels 304 in the second surface 310 of the fluid layer 300; and (iii) the second sealing layer 410 configured to seal the channels 304 in the first surface 308 of the fluid layer.

[0143] Providing the channels 304 in the elastomeric fluid layer 300 provides improved sealing of the fluid layer regardless of the joining process used to seal the network of channels 304 (e.g. pressure sensitive tape, laser welding, etc.). This is because the elastomeric fluid layer 300 acts as a compliant layer when it is sealed to another layer (e.g. the sealing layer 400). Furthermore, using an elastomeric material for the fluid layer 300 means that the channels 304 can be compressed so as to close the respective conduits 600. This means that the channels 304 and valves (i.e. valve regions 302) can be implemented with a single layer, thereby providing a simplified cartridge construction.

[0144] Returning to FIG. 3 the exploded view shown in FIG. 4, it can be seen that the cartridge 100 also includes: a label 110 arranged to cover at least a portion of the rigid face 250 of the first portion 200; a plurality of reagent capsules 120 disposed within the cartridge 100 between the fluid layer 300 and the first face 250; sealing bands 130a and 130b arranged to seal the chambers 332 in the fluid layer (where the chambers 332 are best shown in FIG. 25 FIG. 3); a sealing band 180 for attaching the reagent capsules 120 to the second sealing layer 410; a sensor strip 150 comprising a plurality of sensors each in fluid communication with a respective measurement chamber 610 defined by a respective aperture 454 in the first sealing layer 400; and a plurality of absorbent waste pads 160 each arranged to fit in a corresponding waste chamber 508a, 508b or slot 514a, 514b (where the waste chambers 508 and slots 514 are best shown in FIG. 24 FIG. 3) disposed in the second portion 500.

[0145] As FIG. 1 and FIG. 2The first portion 200 comprises a container in the form of a round box 202 configured to house a portion of a reservoir container, such as a blood collection tube (e.g. a Vacutainer (RTM) blood collection tube produced by Dickinson and Company, Franklin Lakes, New Jersey, USA). The blood collection tube typically contains a volume of liquid (e.g. blood) and a headspace containing a volume of gas. The round box 202 can be a round box as described in PCT / EP2022 / 087658, the entire contents of which are incorporated herein by reference, and the round box can further comprise an actuatable liquid extraction mechanism as described in PCT / EP2022 / 087658. Alternatively, the actuatable liquid extraction mechanism can be any of the actuatable liquid extraction mechanisms described in PCT / EP2022 / 071433, the entire contents of which are incorporated herein by reference.

[0146] FIG. 3 Also shown is a plurality of reservoir capsules 120 arranged within the box 100. In particular, the reservoir capsules 120 are sealed to the fluid layer 300 using a sealing tape 180. As FIG. 8 As shown in Figures 8 and 9, each reservoir chamber 120 comprises an inlet chamber 122, a main chamber 124 (also referred to herein as a reservoir chamber) storing a liquid such as a liquid reagent, and an outlet chamber 126. A sealing layer 136 (e.g. a sealing foil) is used to seal the chambers 122, 124, 126 of each reservoir capsule 120. The capsule 120 further comprises a first restriction 123 between the inlet chamber 122 and the reservoir chamber 124 and a second restriction 125 between the reservoir chamber 124 and the outlet chamber 126. Each of the first and second restrictions 123, 125 is provided in the form of a neck (or narrow region) between the reservoir chamber 124 and the inlet / outlet chamber. The inlet chamber 122 is in fluid communication with the reservoir chamber 124 via the first restriction 123. The outlet chamber 126 is in fluid communication with the reservoir chamber 124 via the second restriction 125. The reservoir capsule 120 can be a reservoir capsule as described in PCT / EP2022 / 058804, the entire contents of which are incorporated herein by reference.

[0147] FIG. 3 The reservoir capsules 120 shown comprise one larger reservoir capsule (i.e. a first reservoir capsule 120a) and two smaller reservoir capsules (i.e. a second reservoir capsule 120b and a third reservoir capsule 120c). Each of the smaller reservoir capsules 120b, 120c is arranged perpendicular to the larger reservoir capsule 120a, such that the smaller reservoir capsules 120b, 120c are parallel to each other.

[0148] As explained in more detail below, each reagent capsule 120 is positioned above two openings 350 in the fluid layer 300. When force is applied to the inlet chamber 122 and the outlet chamber 126 of the reagent cartridge 120, the material of the reagent cartridge 120 deforms into each opening 350. When sufficient force is applied, the deformation of the reagent capsule 120 in the opening 350 causes the seal layer (e.g., foil) used to seal the capsule 120 to break.

[0149] FIG. 4 A first assembly stage of the cartridge 100 is shown. First, the absorbent waste pad 160 is positioned in the waste chamber 508 and the slot 514 disposed in the second portion 500. Next, the sensor strip 150 is positioned at one end of the second portion 500. The fluid layer assembly (including the fluid layer 300, the first seal layer 400, and the second seal layer 410) is then positioned within the second portion 500. FIG. 5 A second assembly stage is shown, in which the liquid reagent capsules 120 and the seal strip 180 are attached to the fluid layer assembly such that the liquid reagent capsules 120 are positioned above the openings 350 in the fluid layer 300. FIG. 6 A third assembly stage is shown, in which the first portion 200 is coupled to the second portion 500 to form the housing of the cartridge.

[0150] FIG. 7 is a top view of the cartridge 100 with the label 110 removed. As FIG. 7 shown, the first portion 200 includes an actuatable portion 240 (e.g., an actuatable platform). The actuatable portion 240 is positioned above the reagent capsules 120. The actuatable portion 240 is actuatable from a first position in which the actuatable portion 240 does not deform the reagent capsules 120 to a second position in which the actuatable portion 240 deforms the reagent capsules 120. The actuatable portion 240 is actuatable relative to the rigid face 250 of the first portion 200 and can be actuatable in a direction that is orthogonal to the rigid face 250 of the first portion 200. The actuatable portion 240 is rigid. As FIG. 7 shown, the actuatable portion 240 is U-shaped such that it can be deformed toward each reagent capsule 120. The U-shape of the actuatable portion 240 also allows the actuatable portion 240 to be deformed around the pneumatic port 312 in the fluid layer 300 and the protrusion 330 extending from the first surface 308 of the fluid layer 300 (also shown in FIG. 25 FIG. 8

[0151] FIG. 9A is shown in more detail. As FIG. 9A ​​As shown, each reservoir cartridge 120 includes a capsule body 134 that defines the shape of the reservoir chamber 124, the inlet chamber 122, the outlet chamber 126, and the first and second restrictions 123, 125. The capsule body 134 includes an aperture through which the capsule 120 is filled, as FIG. 9B is best shown.

[0152] The capsule 120 also includes a seal layer 136 that seals the aperture, as FIG. 9B is shown. The seal layer 136 is attached to a seal band 180. As FIG. 9A is shown, the seal band 180 includes a first set of openings 184 that have the same size and shape as the openings 350 in the fluid layer 300 in which the reservoir capsules 120 are positioned. The size and shape of these openings 350 (and the corresponding first set of openings 184 in the seal band 180) are further described below. FIG. 9A It is also shown that the seal band 180 includes a second set of openings 186 that allow access to the valve region 302 defined by the fluid layer 300.

[0153] As best shown in FIG. 17 , the inlet chamber 122 and the outlet chamber 126 each include a first region 127a, 127b and a second region 128a, 128b (also referred to herein as “recesses”) in the top surface of the chamber 122, 126. For each chamber 122, 126, the first region 127 and the second region 128 are concentric, such that the first region 127 surrounds the second region 128. The distance between the first region 127 and the seal layer 136 is greater than the distance between the second region 128 and the seal layer 136. In other words, the second region 128 provides a recess in the top surface of the chamber 122, 126, where the recess is surrounded by the first region 127 of the chamber 122, 126.

[0154] FIG. 9B An alternative construction of the reservoir capsules is shown, in which a single portion 134a defines the capsule body 134 of all three reservoir capsules 120. In this alternative example, a single seal layer 136a is used to seal the aperture in the reservoir capsules 120. The single seal layer 136a includes an opening 138 that allows access to the valve region 302 defined by the fluid layer 300.

[0155] FIG. 10 is an isometric top view of the first portion 200. As FIG. 10 shown, the top face (i.e., the outer surface) of the actuatable portion 240 is flat (or includes multiple flat regions). The flat surface of the actuatable portion 240 allows the actuatable portion 240 to be easily moved from the first position to the second position without requiring a specially shaped actuator to move the actuatable portion 240.

[0156] AsFIG. 10 and FIG. 11 The first portion 200 comprises a plurality of elastically deformable members 246 (in the example shown in FIG. 10 and FIG. 11 In the example shown, four elastically deformable members 246 are shown. The provision of a plurality of elastically deformable members 246 allows the actuatable portion 240 to be actuated in a vertical direction, meaning that the actuatable portion 240 is parallel to the base (i.e. the sealing layer) of each reservoir capsule 120 when the actuatable portion 240 is in the first portion and when the actuatable portion 240 is in the second position. The vertical movement of the actuatable portion 240 allows substantially the same force to be applied to the recess 128 of a particular reservoir capsule 120.

[0157] In particular, as shown in FIG. 11 a first one of the elastically deformable members 246 is connected to a first edge 256a of the actuatable portion 240; a second one of the elastically deformable members 246 is connected to a second edge 256b of the actuatable portion 240 opposite the first edge 256a; a third one of the elastically deformable members 246 is also connected to the first edge 256a of the actuatable portion 240 but is spaced apart from the connection point of the first one of the elastically deformable members 246; and a fourth one of the elastically deformable members 246 is connected to a third edge 256c of the actuatable portion 240 which is also opposite the first edge 256a.

[0158] As shown in FIG. 12 the underside of the actuatable portion 240 comprises three pairs of protrusions 242 (shown in cross-section in FIG. 13 Each pair of protrusions 242 extends towards the reservoir capsules 120 and is aligned with the inlet chamber 122 and the outlet chamber 126 of one of the reservoir capsules 120.

[0159] When the actuatable portion 240 is moved to the second position, one protrusion in each pair of protrusions 242 deforms the inlet chamber 122, while the other protrusion in each pair deforms the outlet chamber 126. Specifically, when the actuatable portion is moved from the first position to the second position, the inlet chamber 122 and the outlet chamber 126 are forced into contact with the sealing layer 136 by the deformation of the protrusions 242. When the actuatable portion 240 reaches the second position, the force exerted by the recess 128 onto the sealing layer 136 is high enough to cause the sealing layer 136 to rupture. The rupture of the sealing layer 136 provides an inlet to the inlet chamber 122 (in the sealing layer 136) and an outlet from the outlet chamber 126 (in the sealing layer 136). By providing an inlet and an outlet in the liquid storage capsule 120, air will be supplied to the inlet (via the fluid layer 300) to force liquid to flow out through the outlet (to the fluid layer 300). In alternative embodiments, the liquid storage capsule 120 can not include a recess 128, in which case the protrusions 242 can deform a portion of the inlet chamber 122 and the outlet chamber 126 of each liquid storage capsule 120 (e.g. a flat or domed upper surface of the inlet chamber 122 and the outlet chamber 126) such that a portion of the capsule body 134 is forced into contact with the sealing layer 136.

[0160] When creating an opening in the liquid storage capsule 120 in this way, there can be three challenges. Firstly, there is a risk that the recess 128a displacement will block the opening in the fluid layer 300 to provide air, or that the recess 128b displacement will block the opening in the fluid layer 300 to allow liquid from the capsule 120 to flow. Secondly, if too much force is applied by the external actuator that actuates the actuatable portion 240, there is a risk that one or more of the protrusions 242 will be forced through both the capsule body 134 and the sealing layer 136. Thirdly, there is a risk that the protrusions 242 deform the restricting portions 123, 125 and prevent air from entering the liquid storage chamber 124 and / or prevent liquid from leaving the liquid storage chamber 124. Without addressing these challenges, precise actuator control is required to ensure that the opening is not blocked and that the capsule body 134 is not ruptured.

[0161] As described in more detail below, the protrusions 242 are sized and shaped such that they cannot pass into the opening 350 in the fluid layer covered by the liquid storage capsule 120. This means that the downward displacement of the protrusions 242 is limited by the second sealing layer 410 and the first surface 308 of the fluid layer 300. Therefore, the protrusions 242 cannot be forced through both the capsule body 134 and the sealing layer 136.

[0162] Furthermore, as described in more detail below, the opening 350 covered by the reservoir capsule 120 is positioned such that the inlet chamber 122 and the restrictor 123 of the reservoir capsule 120 are at least partially positioned above one opening 350, while the outlet chamber 126 and the restrictor 125 are at least partially positioned above the other opening 350. As explained below, the protrusion 242 deforms the inlet chamber 122 and the outlet chamber 126, but does not deform the restrictors 123, 125. Therefore, even if the recess 128 is pressed into the opening 350, a portion of the opening remains unblocked because it is located below one of the restrictors 123, 125. Positioning each restrictor 123, 125 partially above one of the openings 350 also means that when force is applied to the top of the restrictors 123, 125, the fluid layer 300 provides a smaller reaction force, thereby reducing the likelihood that the restrictors 123, 125 will be compressed during actuation of the actuable part 240.

[0163] Furthermore, the protrusion 242 and the reservoir capsule 120 are sized and positioned in relation to each other so that when the actuating part 240 is in the second position, the protrusion 242 will not deform the restricting parts 123, 125. Therefore, the risk of blockage within the reservoir capsule 120 is reduced.

[0164] like FIG. 19 As shown, each protrusion 242 includes a groove 243 defining the cross-section of the end of the protrusion 242. The groove 243 of each protrusion 242 is aligned with the corresponding limiting portions 123, 125 of the reservoir capsule 120. FIG. 19 Four protrusions 242 are shown, each having a main fan-shaped end cross-section, and two additional protrusions 242 having trapezoidal end cross-sections with grooves 243 in the longer parallel sides of the trapezoids. The grooves 243 in the end cross-sections of each protrusion 242 reduce the risk of deformation of the limiting portions 123, 125 of the reservoir capsule 120 when the inlet chamber 122 and outlet chamber 126 are deformed.

[0165] For a protrusion 242 having a main fan-shaped end cross-section, the end cross-section also includes a plurality of outwardly extending protruding ribs 245 extending from the curved portion of the main fan shape. The outwardly extending protruding ribs 245 serve to maximize the total width of the protrusion 242 (thus preventing the protrusion 242 from penetrating the opening 350), while minimizing the total cross-sectional area of ​​the end of the protrusion 242 (because the larger the area of ​​the end of the protrusion 242, the greater the force required to deform the capsule body 134). For a protrusion 242 having a trapezoidal end cross-section, the periphery of the trapezoid prevents the protrusion 242 from penetrating the opening 350. In an alternative example, the trapezoidal protrusion 242 may also be provided with outwardly extending protruding ribs 245.

[0166] FIG. 13is a cross-section through the first portion 200, which shows the resiliently deformable members 246 in more detail. As FIG. 13 shown, the underside of the actuable portion 240 also comprises recessed regions 244. In particular, the actuable portion 240 comprises three recessed regions, which can be seen in FIG. 16 and FIG. 17 Each recessed region 244 is provided between two protrusions 242. Each recessed region 244 is configured to accommodate the main chamber 124 of its corresponding liquid reservoir capsule 120 when the actuable portion 240 is in the second position. This means that the main chamber 124 is not deformed by the actuable portion 240 when the actuable portion 240 is in the second position.

[0167] Assuming that the protrusions 242 extend from a single actuable portion 240, actuation of the actuable portion 240 into the second position causes deformation of each of the plurality of capsules 120 simultaneously. Thus, a single movement of the actuable portion 240 can be used to open all of the capsules 120 within the cartridge 100.

[0168] FIG. 13 Also shown is a resiliently deformable member 246 coupling the actuable portion 240 to the first portion 200. The resiliently deformable member 246 is configured to bias the actuable portion 240 away from the second position (i.e. towards the first position). Thus, the resiliently deformable member 246 forces the actuable portion away from engagement with the inlet chamber 122 and the outlet chamber 126 of the liquid reservoir capsule 120.

[0169] Each resiliently deformable member 246 has a curved (in particular, U-shaped) profile, which allows the resiliently deformable member 246 to undergo elastic deformation during movement of the actuable portion 240 into the second position. The resiliently deformable member 246 is formed from the same material as the actuable portion 240 and the first portion 200. In other words, the resiliently deformable member 246 is integral with the first portion 200 and the actuable portion 240, and each is provided in the form of an elastic living hinge. This allows the manufacture of the first portion 200, including the actuable portion 240 and the resiliently deformable member 246, to be simplified (e.g. by injection moulding).

[0170] FIG. 11 and FIG. 12 Also shown is a rigid face 250 comprising a plurality of apertures 254. The plurality of apertures 254 comprises an aperture 254a aligned with the valve region 302 of the fluid layer 300, an aperture 254b aligned with the first pneumatic port 312a of the fluid layer 300, and apertures 254c providing openings for the second pneumatic port 312b and the third pneumatic port 312c of the fluid layer 300, as well as a protrusion 330 extending from the first surface 308 of the fluid layer 300 (e.g. as shown in FIG. 25 Port 312 and protrusion 330 are described with reference to FIG. 30 .

[0171] The aperture 254a in the rigid face 254 allows the valve region 302 of the fluid layer 300 to be accessed by an external valve actuator. Likewise, the apertures 254b and 254c allow the pneumatic ports 312 of the fluid layer 300 to be accessed by external pneumatic actuators.

[0172] FIGS. 15-17 The protrusion 242 is shown aligned with the inlet chamber 122 and the outlet chamber 126 of the reservoir capsule 120 when the actuatable portion 240 is in the first position. It will be appreciated that when a force is applied to the actuatable portion 240 to move the actuatable portion 240 towards its second position, the protrusion 242 engages with the inlet chamber 122 and the outlet chamber 126 and causes the inlet chamber 122 and the outlet chamber 126 of the reservoir capsule 120 to deform to force the recess 128 into contact with the sealing layer 136. In particular, the portion of the sealing layer 136 on the underside of the inlet chamber 122 deforms to a first opening 350 and the portion of the sealing layer 136 on the underside of the outlet chamber 126 deforms to a second opening 350. The deformation of the inlet chamber 122 and the outlet chamber 126 causes the material on the underside of the inlet chamber 122 and the outlet chamber 126 to rupture, which means that an opening is formed in each of the inlet chamber 122 and the outlet chamber 126. The top surface of the capsule 120 is formed from a material that is able to plastically deform in order to allow the recess 128 to deform.

[0173] FIG. 15 and FIG. 17 One of the recessed regions 244 of the main chamber 124 containing the second reservoir capsule 120b is shown. FIG. 15 and FIG. 17 It is also shown that the second reservoir capsule 120b and the openings 350 it covers are sized and positioned relative to each other such that the inlet chamber 122 and the restriction 123 of the second reservoir capsule 120b are positioned partially over one of the openings 350 and the outlet chamber 126 and the restriction 125 are positioned partially over the other of the openings 350. FIG. 16 The recessed regions 244 of the main chamber 124 containing the first reservoir capsule 120a are shown, which can also be seen in FIG. 15 . FIG. 16 It is also shown that the first reservoir capsule 120a and the openings 350 it covers are sized and positioned relative to each other such that the inlet chamber 122 and the restriction 123 of the first reservoir capsule 120a are positioned partially over one of the openings 350 and the outlet chamber 126 and the restriction 125 are positioned partially over the other of the openings 350.

[0174] FIG. 18 The positioning of the reservoir capsules 120 relative to the openings 350 in the fluid layer 300 is shown in a partially transparent view. As FIG. 18As shown, the openings 350 and the reservoir capsules 120 are sized and positioned relative to each other such that each opening 350 extends partially under the restriction 123, 125 of one of the reservoir capsules 120. In other words, the openings 350 and the reservoir capsules 120 are sized and positioned relative to each other such that, for each reservoir capsule 120, the inlet chamber 122 and the restriction 123 of the reservoir capsule 120 are partially positioned over the first opening 350 covered by the reservoir capsule 120, while the outlet chamber 126 and the restriction 125 of each reservoir capsule 120 are partially positioned over the second opening 350 covered by the reservoir capsule 120. FIG. 18 It is also shown that each opening 350 has an oblong shape. This oblong shape allows the opening 350 to extend under the inlet 122 or outlet chamber 126 and the adjoining restriction 123, 125, while minimizing the width of the opening 350. For comparison, a circular opening extending under the inlet chamber or outlet chamber and the adjoining restriction would have to be large enough to prevent the occlusion of the opening by the capsule body 134, in particular the recess 128, but would not prevent the protrusion 242 from piercing both the capsule body 134 and the sealing layer 136 of the reservoir capsule 120.

[0175] In FIG. 19 , the cross-section of the end of the protrusion 242 covers FIG. 18 . FIG. 19 It is shown that the width of each opening 350, i.e. the width of the oblong shape, is smaller than the width of the respective protrusion 242 aligned with the opening 350. In other words, the footprint of each protrusion 242 extends beyond the footprint of the corresponding opening 350 aligned therewith. In other words, the perimeter of each protrusion 242 extends beyond the perimeter of the respective opening 350 aligned therewith. This means, as shown, that each protrusion 242 cannot penetrate into the corresponding opening 350 aligned therewith. FIG. 19 FIG. 19 It is also shown that the groove 243 of each protrusion 242 is aligned with the restriction 123, 125 that partially covers the opening 350 aligned with the protrusion 242. In other words, the groove 243 of each protrusion 242 is aligned with a center line that passes through: (i) the recess 128 of the inlet chamber or outlet chamber 122, 126 aligned with the protrusion 242, and (ii) the restriction 123, 125 adjoining the inlet chamber 122 or outlet chamber 126 aligned with the protrusion 242.

[0176] FIG. 20 is a cross-section through the inlet chamber 124 of each smaller reservoir capsule 120b, 120c. As FIG. 20 and FIG. 21 ​As shown in the enlarged view of FIG. 2B, the width of each protrusion 242 is greater than the width of the corresponding opening 350 aligned therewith, meaning that no part of the protrusion 242 can pass into the corresponding opening 350 aligned therewith. In addition, FIG. 20 and FIG. 21 It is shown that the end of each protrusion 242 is flat. Providing a flat end of each protrusion 242 means that no part of the protrusion 242 can pass into the opening 350 aligned with the protrusion 242. The flat end of the protrusion 242 also reduces the risk of piercing the capsule body 134 when the inlet chamber 122 and the outlet chamber 126 of the liquid storage capsule 120 are deformed.

[0177] FIG. 22 The protrusion 242, the inlet chamber 122 or the outlet chamber 126 of the liquid storage chamber 120, and the opening 350 are shown schematically from the same position (i.e. looking through the width of the opening 350) as shown in FIG. 2B. FIG. 20 and FIG. 21 The protrusion 242, the inlet chamber 122 or the outlet chamber 126 of the liquid storage chamber 120, and the opening 350 are shown schematically from the same position (i.e. looking through the width of the opening 350) as shown in FIG. 2B. FIG. 22 Three distances are shown in FIG. 2B. A first distance, labelled "C", is the gap between the end of the protrusion 242 and the inlet chamber 122 or the outlet chamber 126 (specifically, the first region 127 of the inlet chamber 122 or the outlet chamber 126). A second distance, labelled "D1", is the displacement of the capsule body 134 (specifically, the recess 128 of the inlet chamber 122 or the outlet chamber 126) after the protrusion contacts the first region 127 of the inlet chamber 122 or the outlet chamber 126 without piercing the seal layer 126. A third distance, labelled "D2", is the displacement of the capsule body 134 (specifically, the recess 128 of the inlet chamber 122 or the outlet chamber 126) during piercing of the seal layer 126. From FIG. 22 It can be seen that D2 is limited by the protrusion 242 not being able to pass into the opening 350, which reduces the risk of the protrusion 242 piercing the capsule body 134.

[0178] To prevent early piercing of the liquid storage capsule 120 during manufacture and handling, distances C and D1 are implemented. The width of the protrusion 242 is also greater than the width of the second portion 128 (or recess), thereby reducing the likelihood of the protrusion 242 passing into the recess 128 and, in turn, reducing the likelihood of the protrusion 242 piercing both the capsule body 134 and the seal layer 136.

[0179] FIG. 23 An isometric view of the second portion 500 is shown in FIG. 3A, while FIG. 24A top view of the second portion 500 is shown. The second portion 500 includes a sample waste chamber defined by a first well 504 in the upper surface of the second portion 500. The sample waste chamber receives excess sample during the receipt of a sample in the cartridge 100. The sample waste chamber is in fluid communication with a permanent vent 506, which is provided as a hole in the second portion 500. Thus, the sample inlet chamber 236 (shown) in the first portion 200 for receiving a sample is permanently vented through the permanent vent 506. FIG. 12

[0180] The second portion 500 also includes two additional waste chambers (a first waste chamber 508a and a second waste chamber 508b), each provided as a recess in the inner surface of the rigid face 570. The first waste chamber 508 is in fluid communication with a first slot 514a in the inner surface of the rigid face 570. The second waste chamber is in fluid communication with the first well 504 and is thus permanently vented via the permanent vent 506.

[0181] The second portion 500 also includes a first slot 514a in the inner surface of the rigid face 570 in the form of a circular recess, a second slot 514b in the inner surface of the rigid face 570 in the form of a circular recess, and a third slot 514c in the inner surface of the rigid face 570 in the form of a circular recess with integral fingers. Each slot 514 provides a fluid connection between a pneumatic port 312 in the fluid layer 300 and one or more channels 304 in the fluid layer 300, as described in more detail below. In particular, as described with reference to FIG. 26 the plurality of channels 304 are fluidly connected to the slots 514 through holes in the sealing layer 400.

[0182] The second portion 500 also includes a first pneumatic port support region 516a provided adjacent to the first slot 514a, a second pneumatic port support region 516b provided adjacent to the second slot 514b, and a third pneumatic port support region 516c provided adjacent to the third slot 514c. When the cartridge 100 is assembled, each pneumatic port support 516 is disposed beneath a corresponding pneumatic port 312 of the fluid layer 300.

[0183] The first pneumatic port support region 516a includes a connection channel 518a connecting the first pneumatic port 312a to the first slot 514a. Likewise, the second pneumatic port support region 516b includes a connection channel 518b connecting the second pneumatic port 312b to the second slot 514b. Furthermore, the third pneumatic port support region 516c includes a connection channel 518c connecting the third pneumatic port 312c to the third slot 514c. When the cartridge 100 is assembled, there is a fluid connection between the opening 316 of each pneumatic port 312 and the connection channel 518 of the corresponding pneumatic port support 516 (via the sealing layer 400).​

[0184] The port support region 516 is in contact with the underside of the seal layer 400 for sealing the channels 304 in the second surface 310 of the fluid layer 300. This means that when a force is applied to the pneumatic port 312 of the fluid layer 300, the contact between the seal layer 400 and the port support 516 provides a counter force against the applied force, thereby preventing the seal layer 400 from deforming downwards. This helps the pneumatic port 312 to form a seal with the pneumatic actuator.

[0185] The slot 514 in the second portion 500 prevents liquid from reaching the pneumatic port 312 of the pneumatic actuator connected to the analyser device. Therefore, the slot 514 prevents liquid from reaching the analyser device, particularly during the aspiration of liquid. Such liquid can contaminate or damage the analyser device. Pneumatic pressure is provided via the connection channel 518 of the port support 516. The connection channel 518 is disposed above the bottom of the slot 514, which means that any liquid aspirated from the channels 304 in the fluid layer 300 collects in the bottom of the slot 514 and does not reach the connection channel 518. Therefore, any liquid aspirated from the channels 304 in the fluid layer 300 is not aspirated through the connection channel 518 and into the analyser via the pneumatic port 312.

[0186] The fluid communication between the first waste liquid chamber 508a and the first slot 514a allows the venting state of the waste liquid chamber 508 to be controlled via the first pneumatic port 312a (i.e. by the analyser using the containment cartridge 100 venting or not venting the first pneumatic port 312a).

[0187] The second portion 500 also comprises a plurality of valve support regions 524 (indicated by dashed circles in FIG. 24 In the example shown in FIG. 23 and FIG. 24 twelve valve support regions 524 are shown. Each valve support region 524 is aligned with a corresponding valve region 302 in the fluid layer 300. Each valve support region 524 is in contact with the underside of the seal layer 400 for sealing the channels 304 in the second surface 310 of the fluid layer 300. This means that when a force is applied to the valve region 302 of the fluid layer 300 by an actuator of the analyser, the valve region 302 between the actuator and the corresponding valve support region 524 is compressed. Therefore, the waste liquid chambers 508a, 508b do not extend under the valve region 302 of the fluid layer 300.

[0188] The second portion 500 also comprises a plurality of capsule support regions 526 (again indicated by dashed squares in FIG. 24 In the example shown in FIG. 23 and FIG. 27In the example shown, six capsule support regions 526 are shown. Each capsule support region 526 is aligned with a corresponding opening 350 in the fluid layer 300, which is aligned with an inlet chamber 122 or an outlet chamber 126 of one of the plurality of capsules 120. Each capsule support region 526 contacts the underside of the sealing layer 400, which is used to seal the channels 304 in the second surface 310 of the fluid layer 300. This means that, when a force is applied to the inlet or outlet chamber 122, 126 of one of the plurality of capsules 120 by the corresponding protrusion 242 of the actuatable portion 240, the second surface 410 of the fluid layer 300 is prevented from being displaced downwards by the respective capsule support region 526. As a result, the waste liquid chambers 508a, 508b do not extend below the openings 350 in the fluid layer 300.

[0189] FIG. 25 An exploded view of the fluid layer 300, the first sealing layer 400 and the second sealing layer 410 is shown. The first sealing layer 400 is arranged to seal the channels 304 in the second surface 310 of the fluid layer 300, while the second sealing layer 410 is arranged to seal the channels 304 in the first surface 308 of the fluid layer 300.

[0190] FIG. 25 A plurality of valve regions 302 of the cartridge are shown, each valve region 302 being provided in the form of a recess in the fluid layer 300. This means that the fluid layer 300 has a reduced thickness in each valve region 302. By providing a region of reduced thickness aligned with the corresponding channel 304, the force required to deform the valve region 302 and close the corresponding channel 304 is reduced.

[0191] FIG. 25 A protrusion 330 extending from the first surface 308 of the fluid layer 300 is also shown. The protrusion 330 comprises openings defining a plurality of chambers 332, as described in more detail with reference to FIG. 25 and FIG. 26 The implementation of the protrusion 330 extending from the first surface 308 of the fluid layer 300 means that the volume of the chambers 332 defined by the protrusion 330 is not limited by the thickness of the fluid layer 300 between its first surface 308 and its second surface 310. The fluid layer 300 also comprises a plurality of openings 350 extending through the thickness of the fluid layer 300.

[0192] As FIG. 25 shown, the fluid layer 300 also comprises a plurality of pneumatic ports 312 (for example, three pneumatic ports 312a, 312b and 312c as shown in FIG. 26 Each pneumatic port 312 comprises a protrusion 314 extending from the first surface 308.

[0193] Each port 312 also includes an opening 316 that extends through the protrusion 314 and at least part of the thickness of the fluid layer. In FIG. 26 and FIG. 27 In the example shown, the openings 316 extend through the entire thickness of the fluid layer 300.

[0194] Accordingly, the fluid layer 300 includes two sets of openings: a plurality of first openings 350 disposed directly below the reservoir capsules 120 and aligned with the protrusions 242 on the actuatable portion 240, and a plurality of second openings 316, each extending through a corresponding protrusion 314 of the pneumatic port 312. These sets of openings allow for communication between the network of channels 304 in the fluid layer 300 and other fluidic components of the cartridge 100 (e.g., the capsules 120 and the pneumatic port 312).

[0195] All of the components of the fluid layer 300 described above are integrally formed with the fluid layer 300, meaning that they are all formed from the same elastomeric material as the fluid layer 300. More specifically, the protrusions 330 and the protrusions 314 are both integrally formed with the fluid layer 300 and are formed from the same elastomeric material as the fluid layer 300.

[0196] The first seal layer 400 includes a plurality of apertures 454, each of which partially defines a corresponding measurement chamber 610 of the cartridge 100 and is accessed via two channels 304 in the fluid layer 300 that provide an inlet and an outlet to the measurement chamber 610.

[0197] FIG. 26 A top view of the fluid layer 300 is shown, which shows the first surface 308. FIG. 26 The locations of the valve regions 302, the ports 312, the protrusions 330 (including the chambers 332a-c, as shown in FIG. 27 The openings 350 and the openings 316 extend through the thickness of the fluid layer 300, as explained above.

[0198] Each valve region 302 allows the flow of fluid through one conduit 600 of the cartridge 100 to be controlled. Each valve is defined by a valve region 302 (each disposed over a corresponding channel 304) and one of the seal layer 400 that seals the corresponding channel 304. To close a valve, a force is applied to the valve region 302 to press the corresponding channel 304 against the seal layer 400. Closing the valve prevents fluid flow through the corresponding conduit 600. To open the valve, the force applied to the valve region 302 to close the valve is retracted. Opening the valve allows fluid to flow through the corresponding conduit 600.

[0199] Fluid flow through the conduit 600 is controlled by applying variable pressures to the pneumatic ports 312. As described in more detail below, each pneumatic port 312 can: (i) receive positive pressure through a corresponding pneumatic actuator; (ii) receive negative pressure through a corresponding pneumatic actuator; (iii) be vented (i.e. open to atmospheric pressure) through a corresponding pneumatic actuator; or (iv) be closed (i.e. not vented), meaning that the pneumatic port 312 is disconnected from the pneumatic actuator. In case (iv), there is no airflow through the pneumatic port 312.

[0200] The plurality of channels 304 in the fluid layer extend from a point in the fluid layer over one of the slots 514 in the second portion 500. This means, for example, that when a positive pneumatic pressure is applied to the first pneumatic port 312a, pressurised air flows through the corresponding opening 316 in the first pneumatic port 312a, through the corresponding hole in the seal layer 400, through the channel 518a of the first port support region 516a, through the slot 514a and into one of the channels 304. Negative pressure is applied in the same way, but with the airflow in the opposite direction.

[0201] This arrangement reduces the risk of liquid being drawn into the pneumatic actuator (for example, during the application of negative pressure to draw liquid). This is because any liquid drawn through the channels 304 falls to the bottom of the slot 514a under the action of gravity. As FIG. 23 and FIG. 24 shown, the channels 518a are located on the flat upper surface of the port support regions 516a, meaning that the channels 518a are disposed above the base of the slots 514a. Therefore, any liquid that collects in the slots 514a is not drawn through the channels 518a and into the pneumatic actuator via the first pneumatic port 312a.

[0202] As FIG. 25 shown, two protrusions 330 extend from the first surface 308 of the fluid layer 300. The first protrusion 330a comprises a hole that partially defines a first chamber 332a and a second chamber 332b. The second protrusion 330b comprises a hole that partially defines a third chamber 332c. In particular, the holes in the first and second protrusions 330 define the inner walls of the chambers 332, and therefore the height of the chambers 332. The holes defined by the protrusions 330 extend through the entire thickness of the fluid layer 300 in the region of the protrusions 330 (for example, shown in a lower side view of the fluid layer 300, as FIG. 27 shown). This means that the base of each chamber 332 is defined by the first seal layer 400 (for example, shown in FIG. 25 ), while the top of each chamber 332 is defined by the seal band 130 (specifically, the first seal band 130a seals the first chamber 332a and the second chamber 332b defined by the first protrusion 330a, while the second seal band 130b seals the third chamber 332c defined by the second protrusion 330b).FIG. 26 and FIG. 27 It is also shown that each chamber 332 includes a protrusion 338 extending from an inner wall 340 (see FIG. 31 ) of the chamber 332, as described in detail below with reference to FIGS. 31-34 .

[0203] According to examples described herein, a liquid handling device (e.g., microfluidic cartridge 100) includes a first reservoir container (e.g., a first reservoir container of a reservoir capsule 120) and a second reservoir container (e.g., a second reservoir container of a reservoir capsule 120), wherein each of the first reservoir container and the second reservoir container is configured to be opened by creating an inlet and an outlet in the reservoir container; and a fluidic layer (e.g., fluidic layer 300) including: a network of channels (e.g., channels 304); and a pneumatic port (e.g., pneumatic port 312b) configured to receive a positive pressure; wherein the network of channels includes: a first reservoir container inlet channel (e.g., a first reservoir capsule inlet channel of reservoir capsule inlet channels 360, 362, 364 described below) configured to provide a fluidic connection between the pneumatic port and the inlet in the first reservoir container when the first reservoir container is opened; and a second reservoir container inlet channel (e.g., a second reservoir capsule inlet channel of reservoir capsule inlet channels 360, 362, 364 described below) configured to provide a fluidic connection between the pneumatic port and the inlet in the second reservoir container when the second reservoir container is opened. This arrangement is not limited to liquid handling devices including reservoir capsules 120 having the particular structure of those described herein (i.e., having chambers 122, 124, 126 and restrictions 123, 125) and can be implemented with any reservoir container configured to be opened by creating an inlet and an outlet in the reservoir container.

[0204] FIG. 26 Channels 304 in the first surface 308 of the fluidic layer 300 are shown. Channels 304 in the first surface of the fluidic layer include a first reservoir container inlet channel 360 (also referred to herein as a first reservoir capsule inlet channel 360), a second reservoir container inlet channel 362 (also referred to herein as a second reservoir capsule inlet channel 362), and a third reservoir container inlet channel 364 (also referred to herein as a third reservoir capsule inlet channel 364). The first reservoir capsule inlet channel 360 provides a fluidic connection between an inlet chamber 122a (see FIG. 8 ) of a first reservoir capsule 120a and the second pneumatic port 312b. The second reservoir capsule inlet channel 362 provides a fluidic connection between an inlet chamber 122b of a second reservoir capsule 120b and the second pneumatic port 312b. The third reservoir capsule inlet channel 364 provides a fluidic connection between an inlet chamber 122c of a third reservoir capsule 120c and the second pneumatic port 312b.

[0205] The cartridge 100 also includes a well 602 that is partially defined by the opening 324 in the fluid layer 300. In the illustrated example, the well 602 is defined by the opening 324 and a second slot 514b (also referred to as a “cavity”) in the second portion 500. Referring back to FIG. 4, it can be seen that the second slot 514b includes one of the absorbent waste pads 160, which means that the well 602 includes absorbent material. As described above, the second pneumatic port 312b is in fluid communication with the second slot 514b via a connection channel 518b in the second pneumatic port support region 516b, which means that the second pneumatic port 312b is in fluid communication with the well 602. Each of the reagent capsule inlet channels 360, 362, 364 is connected to the opening 324, which means that each of the reagent capsule inlet channels 360, 362, 364 is in fluid communication with the well 602. Thus, each of the reagent capsule inlet channels 360, 362, 364 is in fluid communication with the second pneumatic port 312b via the well 602. FIG. 26 FIG. 27 In the illustrated example, the opening 324 extends through the entire thickness of the fluid layer 300 (as shown in the side view of FIG. 6). In this example, the well 602 is defined by the opening 324 and the second slot 514b in the second portion 500. Referring back to FIG. 4, it can be seen that the second slot 514b includes one of the absorbent waste pads 160, which means that the well 602 includes absorbent material. As described above, the second pneumatic port 312b is in fluid communication with the second slot 514b via a connection channel 518b in the second pneumatic port support region 516b, which means that the second pneumatic port 312b is in fluid communication with the well 602. Each of the reagent capsule inlet channels 360, 362, 364 is connected to the opening 324, which means that each of the reagent capsule inlet channels 360, 362, 364 is in fluid communication with the well 602. Thus, each of the reagent capsule inlet channels 360, 362, 364 is in fluid communication with the second pneumatic port 312b via the well 602. FIG. 4

[0206] Each of the reagent capsule inlet channels 360, 362, 364 includes a plurality of regions in the form of one or more first channel regions 366a and one or more second channel regions 366b. Each first channel region 366a is disposed in the first surface 308 of the fluid layer 300 and has a depth that is a portion of the thickness of the fluid layer 300. Each second channel region 366b is disposed in the first surface 308 of the fluid layer 300 and has a depth that is greater than the depth of the first channel regions 366a and a width that is greater than the width of the first channel regions 366a. In the illustrated example, the reagent capsule inlet channels 360, 362, 364 each include two first channel regions 366a and two second channel regions 366b. In addition, in this example, the second channel regions 366b have a depth that is equal to the thickness of the fluid layer 300 and are disposed in the form of circular holes that extend through the fluid layer 300 (as shown in FIG. 6). Thus, the second channel regions 366b are sealed using the first and second sealing layers 400, 410. FIGS. 25-27 FIG. 27

[0207] ​​​​During opening of the reagent reservoir 120, some liquid from the reagent reservoir 120 can flow through the respective reagent reservoir inlet channel 360, 362, 364 in the direction of the second pneumatic port 312b. The first and second channel regions 366 and the well 602 of the reagent reservoir inlet channel 360, 362, 364 prevent liquid from flowing to the second pneumatic port 312b, and thus from reaching the pressure applying analyzer via the second pneumatic port 312b. The depth of the second channel region 366b and the well 602 means that liquid collects in the second channel region 366b and the well 602, and the absorbent material in the well 602 absorbs any liquid in the well 602. The lower depth of the first channel region 366a compared to the second channel region 366b provides a barrier for liquid to move from the second channel region 366b to the first channel region 366a. Thus, the arrangement of the reagent reservoir inlet channel 360, 362, 364 and the well 602 reduces the risk of liquid reaching the pressure applying analyzer via the second pneumatic port 312b.

[0208] FIG. 27 The bottom view of the fluidic layer 300 shown illustrates the channels 304 in the second surface 310 of the fluidic layer 300, which are referenced FIG. 28 in more detail. FIG. 27 Also shown is the opening 350 over which the reagent reservoir 120 is positioned, the opening 316 of the pneumatic port 312, the second channel region 366b, and the opening 324 that partially defines the well 602, each as described above.

[0209] FIG. 28 is a schematic illustration of the fluidic network defined by the channels 304 in the fluidic layer 300. In FIG. 28 the channels 304 in the first surface 308 are represented by a plurality of lines (as in FIG. 26 the channels 304 in the second surface 310 are represented by a single line. The dashed lines represent fluidic connections through the second portion 500 (i.e., the connecting channel 518). FIG. 29 is a schematic illustration of the fluidic network shown in FIG. 30

[0210] The fluidic network includes a first pneumatic port 312a (P1), a second pneumatic port 312b (P2), and a third pneumatic port 312c (P3). The fluidic network also includes a first chamber 332a (M1), a second chamber 332b (M2), and a third chamber 332c (M3). One or more of the first chamber 332a (M1), the second chamber 332b (M2), and the third chamber 332c (M3) can contain a dry reagent, such as a lyophilized reagent provided in the form of a reagent pellet.

[0211] ​Having a third chamber 332c available to store reagents allows the cartridge 100 to run more complex assay methods that require additional reagents that are not mixed with the sample in the first reaction (e.g., the method 1500 and the method 1800 described below). Moreover, having chambers 332 configured to store dry reagents is advantageous because: (i) dry reagents are generally more stable for long-term storage, and (ii) different dry reagents can be stored in different chambers and re-suspended with the same liquid buffer, allowing different liquid reagents to be formed during operation of the cartridge 100.

[0212] One or more of the first chamber 332a (Ml), the second chamber 332b (M2), and the third chamber 332c (M3) can be used as a mixing chamber for mixing a sample and a liquid (e.g., a reagent, a diluent), or for mixing a dry reagent with a liquid for re-suspending the dry reagent.

[0213] In addition to the three measurement chambers 610 in the form of the first flow unit 610a (Fl), the second flow unit 610b (F2), and the sample flow unit 610c (F3), the fluidic network also includes the liquid reagent capsule inlet channels 360, 362, 364 described above. In addition, the fluidic network includes a first waste chamber 508a and a second waste chamber 508b. As described above, the first waste chamber 508a is vented using the first pneumatic port 312a. The first waste chamber 508a (also referred to as the sample flow unit waste chamber) is accessed via a first waste chamber port 358a (Wl) in the fluid layer 300, and the second waste chamber 508b (also referred to as the flow unit waste chamber) is accessed via a second waste chamber port 358b (W2) and a third waste chamber port 358c (W3) in the fluid layer 300. As described above, the second waste chamber 508b is permanently vented. Permanently venting the second waste chamber 508b frees up a pneumatic port 312 for other operations. In other words, fewer pneumatic ports 312 can be needed.

[0214] The sample inlet channel 367 provides fluidic connection to the permanently vented sample inlet chamber 236 (SI). The sample inlet valve region 302a (V2) is used to control flow through the sample inlet channel 367. The sample inlet channel 367 provides fluidic connection to the first junction 306a. Two other channels are connected to the first junction 306a, as described below.

[0215] The chamber inlet channel 368 provides fluidic connection between the first junction 306a and the second chamber 332b (M2), also referred to herein as the "mixing chamber". Flow through the chamber inlet channel 368 is controlled using the chamber inlet valve 302b (V6). The second chamber 332b (M2) is in permanent fluidic communication with the third pneumatic port 312c (P3), meaning there is no valve between the second chamber 332b (M2) and the third pneumatic port 312c (P3).

[0216] The sample flow channel 370 provides fluidic connection between the first junction 306a and the second junction 306b. In addition, the sample flow cell inlet channel 372 provides fluidic connection between the second junction 306b and the sample flow cell 610c (F3), while the sample flow cell outlet channel 374 provides fluidic connection between the sample flow cell 610c (F3) and the first waste chamber 508a. Flow through the sample flow cell inlet channel 372, the sample flow cell 610c (F3), and the sample flow cell outlet channel 374 is controlled using the sample flow cell valve 302c (V9).

[0217] Implementing a sample flow cell (F3) that can be filled with sample allows performing additional measurements directly on the sample (without dilution). This allows measuring additional biomarkers (e.g. hematocrit) on the same cartridge.

[0218] The second chamber 332b (M2) is fluidically connected to the first chamber 332a (M1) using the mixing channel 376. In addition, the chamber outlet channel 378 provides fluidic connection between the first chamber 332a (M1) and the third junction 306c. Flow through the chamber outlet channel 378 is controlled using the chamber outlet valve 302d (V12). The first chamber 332a (M1) is in permanent fluidic communication with the first pneumatic port 312a (P1), meaning there is no valve between the first chamber 332a (M1) and the first pneumatic port 312a (P1).

[0219] Having two chambers 332a, 332b (M1, M2) in permanent fluidic communication with different pneumatic ports 312a, 312c (P1, P3), respectively, allows mixing by moving the liquid back and forth between the two chambers 332a, 332b (M1, M2). This can be achieved, for example, by venting one of the pneumatic ports 312a, 312c (P1, P3) in permanent fluidic communication with one of the chambers 332a, 332b (M1, M2) and applying a cycle of positive and negative pressure through the other pneumatic port 312a, 312c (P1, P3) in permanent fluidic communication with the other chamber 332a, 332b (M1, M2).

[0220] The first flow cell inlet channel 380 provides fluidic connection between the third junction point 306c and the first flow cell 610a (Fl). Thus, the first flow cell 610a (Fl) is in fluid communication with the first chamber 332a (Ml) via the chamber outlet channel 378 and the first flow cell inlet channel 380. Having the first flow cell 610a (Fl) connected to the first chamber 332a (Ml) in this way (i.e., by providing a direct connection between the first flow cell 610a (Fl) and the first chamber 332a (Ml)) allows the length of the fluidic path of the dilution sample moving to the first flow cell 610a (Fl) to be minimized, thereby reducing the loss of analyte or reagent along the fluidic path. In contrast, some existing liquid handling devices have a coating on the channel surface to avoid loss of analyte or reagent by adsorption to the surface of the channel wall. This approach requires a complex production process, thus increasing the cost of such liquid handling devices.

[0221] Furthermore, the first flow cell 610a (Fl) is connected to the second flow cell 610b (F2) using a flow cell connection channel 382. A bypass channel 384 fluidically connects the fourth junction point 306d on the flow cell connection channel 382 to the second waste chamber 508b via the third waste chamber port 358c (W3). The bypass channel 384 allows liquid to bypass the second flow cell 610b (F2) after flowing through the first flow cell 610a (Fl). The flow through the bypass channel 384 is controlled using a bypass valve 302e (Vll). Thus, the bypass valve 302e (Vll) controls the flow of liquid from the first flow cell 610a (Fl) into the second waste chamber 508b.

[0222] The bypass channel 384 provides the option of which liquids to pass through only the first flow cell 610a (Fl) and which liquids to pass through the first flow cell 610a (Fl) and the second flow cell 610b (F2). Another advantage relates to the use of magnetic beads in an analysis performed using the cartridge 100. The configuration of the bypass channel 384 allows the first flow cell 610a (Fl) to be used as a sensorless washing cell and, once the magnetic beads have reacted with the detection solution, allows the magnetic beads to be transferred only to the measurement flow cell (e.g., the second flow cell 610b (F2)).

[0223] The second flow cell outlet passage 386 fluidically connects the second flow cell 610b (F2) to the second waste chamber 508b via the second waste chamber port 358b (W2). Flow through the second flow cell outlet passage 386 is controlled using the second flow cell outlet valve 302f (V10). Thus, the flow circuit comprising the second flow cell 610b (F2) can be connected to a permanently vented waste chamber (i.e. the second waste chamber 508b) using a valve (i.e. the second flow cell outlet valve 302f (V10)) and does not require the use of a pneumatic port 312.

[0224] The second pneumatic port 312b (P2) is in permanent fluid communication with the first, second and third reservoir capsules 120a (B1), 120b (B2) and 120c (B3), meaning there is no valve between the second pneumatic port 312b and each reservoir capsule 120. The second pneumatic port 312b is in permanent fluid communication with the first, second and third reservoir capsules 120a (B1), 120b (B2) and 120c (B3) via the first, second and third reservoir capsule inlet passages 360, 362 and 364 respectively.

[0225] When the first and third pneumatic ports 312a (P1) and 312c (P3) are in permanent fluid communication with the first and second chambers 332a (M1) and 332b (M2) respectively, it allows the reservoir capsules 120 to be controlled by the second pneumatic port 312b (P2), allowing liquid to move from the reservoir capsules 120 to the first and / or second chambers 332a, 332b, whilst still being able to mix between the two chambers 332a, 332b.

[0226] The permanent fluid connections between the ports 312 and the features of the fluidic network (chambers 332, reservoir capsules 120) reduces the number of valves required to control other functions (i.e. fewer valves are required).

[0227] The first reservoir capsule outlet passage 388 (also referred to herein as a first reservoir container outlet passage) provides an outlet from the first reservoir capsule 120a (B1). Flow through the first reservoir capsule outlet passage 388 is controlled using the first reservoir capsule valve 302g (V4) (also referred to herein as a first reservoir container valve). Thus, the first reservoir capsule 120a (B1) can be set to be in fluid communication with the first flow cell 610a (F1) via the first flow cell inlet passage 380 using the first reservoir capsule valve 302g (V4).

[0228] The second reservoir capsule 120b (B2) is in fluid communication with a fifth junction 306e on the second flow cell outlet channel 386 via a second reservoir capsule outlet channel 390 (also referred to herein as a second reservoir container outlet channel). Flow through the second reservoir capsule outlet channel 390 is controlled using a second reservoir capsule valve 302h (V5) (also referred to herein as a second reservoir container valve). Thus, the second reservoir capsule 120b (B2) can be placed in fluid communication with the second flow cell 610b via the second flow cell outlet channel 386 using the second reservoir capsule valve 302h (V5).

[0229] A third reservoir capsule outlet channel 392 (also referred to herein as a third reservoir container outlet channel) provides an outlet from the third reservoir capsule 120c (B3). Flow through the third reservoir capsule outlet channel 392 is controlled using a third reservoir capsule valve 302i (V7) (also referred to herein as a third reservoir container valve).

[0230] The third reservoir capsule outlet channel 392 and the first reservoir capsule outlet channel 388 are each in fluid communication with a third chamber inlet channel 394, which provides a fluid connection to the third chamber 332c (M3). Flow through the third chamber inlet channel 394 is controlled using a third chamber inlet valve 302j (V8) to control the flow into the third chamber 332c (M3). The third chamber 332c (M3) is in permanent fluid communication with the third pneumatic port 312c (P3), meaning there is no valve between the third pneumatic port 312c and the third chamber 332c (M3).

[0231] The third reservoir capsule outlet channel 392 and the first reservoir capsule outlet channel 388 are also each in fluid communication with a priming channel 396. The priming channel 396 provides a fluid connection between a second junction 306b and a sixth junction 306f on an air / wash supply channel 398. Flow through the priming channel 396 (V1) is controlled by a priming channel valve 302k. This means that the third reservoir capsule 120c (B3) and the first reservoir capsule 120a (B1) can be brought into fluid communication with the second chamber 332b (M2) by opening the first reservoir capsule valve 302g (V4) or the third reservoir capsule valve 302i (V7), the priming channel valve 302k (V1), and the chamber inlet valve 302b (V6). The air / wash supply channel 398 provides a fluid connection between the second pneumatic port 312b and the third junction 306c. Air flow through the air / wash supply channel 398 is controlled using an air supply valve 302L (V3), and liquid (e.g. wash liquid) flow through the air / wash supply channel 398 is controlled using the first reservoir capsule valve 302g (V4).

[0232] Connecting the air / wash supply channel 398 to the fluidic circuit allows any remaining liquid to be drained from the channels (purged) between operations, thereby avoiding mixing or contamination between different liquids.

[0233] Each channel 304 in the fluidic layer 300 described above is sealed using the first sealing layer 400 and / or the second sealing layer 410. Once sealed, a conduit is formed. For example, the sample inlet channel 367 is sealed by the first sealing layer 400 to form a sample inlet conduit. Accordingly, reference to a "conduit" of the fluidic network refers to a channel 304 of the fluidic network that has been sealed using the first sealing layer 400 and / or the second sealing layer 410.

[0234] FIG. 30 is a side view of the fluidic layer 300 showing the protrusions 330 extending from the first surface 308 of the fluidic layer 300. The protrusions 330 can also be seen in FIG. 31 The protrusions 314 of the pneumatic port 312 can be seen in the side view shown. FIG. 31 is a cross-section through the fluidic layer 300 showing a cross-section through one of the chambers 332. As FIG. 33 shown, the chamber 332 includes a plurality of protrusions 338 extending from the inner wall 340 of the chamber 332. When the chamber 332 is used to house a reagent ball 344 (e.g., as FIG. 34 and FIG. 32 shown), the protrusions 338 prevent the reagent ball 344 from contacting the sealing layer (i.e., the sealing band 130) used to seal the chamber 332.

[0235] The protrusions 330 defining the chamber 332 extend from the first surface 308 of the fluidic layer 300 and are thus integrally formed with the fluidic layer 300. Likewise, the protrusions 338 extending from the inner wall 340 of the chamber 332 are thus integrally formed with the protrusions 330. As a result, the protrusions 338 are elastically deformable because the material of the fluidic layer 300 is an elastomer (e.g., a thermoplastic elastomer). This allows the reagent ball 344 to be inserted into the chamber 332 through the protrusions 338 when a force is applied to the reagent ball 344. For example, the reagent ball 344 can be inserted through the protrusions 338 prior to sealing the chamber 332 using the sealing band 130.

[0236] FIG. 33 is a top view of a fluidic layer assembly including the fluidic layer 300 with the sealing layer 410 attached, while FIG. 32 is a cross-section through the fluidic layer assembly shown in FIG. 34 along the line Z-Z. FIG. 33 is a close-up view of the region T in FIG. 33

[0237] As FIG. 34 and FIG. 33 ​As shown, each chamber 332 includes an opening 342 at its upper end 343 (i.e., adjacent to the seal band 130 when the cartridge 100 is assembled). The opening 342 provides an entry point into the hole defined by the protrusion 330, which means that the opening 342 allows for the insertion of a reagent ball 344 into the chamber 332. The seal band 130 is configured to seal the opening 342 of the chamber 332. Given the low humidity conditions (typically less than 10% humidity) required to handle such reagents, handling dry reagents is challenging. As excessive exposure to low humidity conditions is harmful to human health, the use of strictly controlled humidity results in limited time for an operator to handle dry reagents. Reagent handling can be automated, but still requires strictly controlled humidity, which increases the cost and complexity of the reagent handling process. The use of the elastically deformable protrusion 338 to allow for the insertion of a reagent ball 344 into the chamber 332 and to hold the reagent ball 344 in place during the application of the seal band 130 reduces the time required to handle the reagent ball 344. As such, these features also reduce the cost and complexity of the manufacturing process, as well as reducing the time for human exposure to low humidity conditions.

[0238] Each protrusion 338 extends from the inner wall 340 at the upper end 343 of the chamber 332. In other words, each protrusion 338 extends from the inner wall 340 adjacent to the opening 342. As shown, the protrusion 338 is elastically deformable, which means that the protrusion 338 can be deformed to allow for the insertion of a reagent ball 344 into the chamber 332. FIG. 34 and FIG. 33 As shown, in effect, the opening 342 is defined by the protrusion 338, as the protrusion 338 defines the perimeter of the opening 342. As such, when the seal band 130 is attached to the protrusion 330 to seal the chamber 332, the seal band 130 contacts the protrusion 338.

[0239] Each protrusion 338 includes a first protrusion portion 339a extending from the inner wall 340 of the chamber 332 and a second protrusion portion 339b extending from a distal end of the first protrusion portion 339a. The second protrusion portion 339b extends away from the upper end 343 of the chamber 332, such that the protrusion 338 has a “claw” shape. The use of the second protrusion portion 339b minimizes the amount of material in contact with the reagent ball 344. This means that the reagent ball 344 can be resuspended without wetting the protrusion 338 (as approximately half to two-thirds of the reagent ball 344 needs to be wetted to resuspend the reagent).

[0240] The inner wall 340 of each chamber 332 also includes at least one liquid flow aperture 346 that allows liquid to flow into and out of the chamber 332. For the first chamber 332a, one liquid flow aperture 346 allows liquid to flow from the first chamber 332a into the chamber outlet channel 378 and from the chamber outlet channel 378 into the first chamber 332a, while the other liquid flow aperture 346 allows liquid to flow from the first chamber 332a into the mixing channel 376 and from the mixing channel 376 into the first chamber 332a. For the second chamber 332b, one liquid flow aperture 346 allows liquid to flow from the second chamber 332b into the chamber inlet channel 368 and from the chamber inlet channel 368 into the second chamber 332b, while the other liquid flow aperture 346 allows liquid to flow from the second chamber 332b into the mixing channel 376 and from the mixing channel 376 into the second chamber 332b. For the third chamber 332c, the liquid flow aperture 346 allows liquid to flow from the third chamber 332c into the third chamber inlet channel 394 and from the third chamber inlet channel 394 into the third chamber 332c.

[0241] FIG. 34 and FIG. 33 One liquid flow aperture 346 is shown for each of the first chamber 332a and the second chamber 332b, in a form that allows liquid to flow from the chamber 332a, 332b into the mixing channel 376 and from the mixing channel 376 into the chamber 332a, 332b. Since each channel 304 connected to the chamber 332 is in the same plane, all of the liquid flow apertures 346 are disposed at the same height (relative to the height of the chamber 332) as the liquid flow apertures 346 shown. FIG. 34 and FIG. 33 As shown in Figs. 33 and 34, each liquid flow aperture 346 is located below the protrusion 338, such that the distance between the upper end 343 and the liquid flow aperture 346 is greater than the distance between the upper end 343 and the distal end of the second protrusion 339b. FIG. 34 and FIGS. 26-28 As shown in Figs. 33 and 34, each liquid flow aperture 346 is located below the protrusion 338, such that the distance between the upper end 343 and the liquid flow aperture 346 is greater than the distance between the upper end 343 and the distal end of the second protrusion 339b.

[0242] Each chamber 332 also includes an air aperture 348, which is also located in the inner wall 340. The air aperture 348 provides a fluid connection to a pneumatic connection conduit 349, which extends perpendicularly between the air aperture 348 and the second surface 310 of the fluid layer 300 (as shown in Fig. 33) and provides a fluid connection to a pneumatic port 312 to which the chamber 332 is connected (i.e., via a corresponding slot 514 in the second portion 500). As shown in Figs. 33 and 34, the air aperture 348 is located below the protrusion 338, such that the distance between the upper end 343 and the air aperture 348 is greater than the distance between the upper end 343 and the distal end of the second protrusion 339b. FIG. 33 FIG. 34 and FIG. 35 ​As shown, a first maximum distance (measured perpendicular to the first surface 308) between the air orifice 348 and the upper end 343 of the chamber 332 is less than or equal to a second maximum distance (measured perpendicular to the first surface 308) between the protrusion 338 (specifically, the second protruding portion 339b) and the upper end 343. In other words, the protrusion 338 extends away from the upper end 343 such that a distal end of the second protrusion 339b is located horizontally below the air orifice 348. Thus, during re-suspension of the reagent spheres 344, liquid does not flow into the air orifice 348.

[0243] FIG. 36 is a flowchart of a method 1000 of controlling liquid flow in a liquid handling device, such as the microfluidic cartridge 100 described above. At 1002, an inlet and an outlet are formed in a first reservoir (e.g., a reservoir capsule 120) of the liquid handling device to form a fluid connection between a pneumatic port (e.g., 312b) of the liquid handling device and the inlet in the first reservoir.

[0244] At 1004, an inlet and an outlet are formed in a second reservoir (e.g., a reservoir capsule 120) of the liquid handling device to form a fluid connection between the pneumatic port 312b of the liquid handling device and the inlet in the second reservoir.

[0245] Optionally, at 1006, an inlet and an outlet are formed in a third reservoir (e.g., a reservoir capsule 120) of the liquid handling device to form a fluid connection between the pneumatic port 312b of the liquid handling device and the inlet in the third reservoir.

[0246] At 1008, liquid from the outlet in the first reservoir is dispensed by applying positive pressure via the pneumatic port 312b. Dispensing the liquid from the outlet in the first reservoir can include opening a first reservoir valve configured to control flow of the liquid from the outlet in the first reservoir. Dispensing the liquid from the outlet in the first reservoir can include opening one or more valves of the liquid handling device to establish a fluid connection between the outlet in the first reservoir and a second pneumatic port (e.g., 312a) of the liquid handling device, and venting the second pneumatic port during the application of positive pressure via the (first) pneumatic port 312b.

[0247] At 1010, liquid from the outlet in the second reservoir is dispensed by applying positive pressure via pneumatic port 312b. Dispensing liquid from the outlet in the second reservoir can include opening a second reservoir valve configured to control flow of liquid from the outlet in the second reservoir. Dispensing liquid from the outlet in the second reservoir can include opening one or more valves of the liquid handling device to establish a fluid connection between the outlet in the second reservoir and a second pneumatic port (e.g., 312a) of the liquid handling device, and venting the second pneumatic port during the application of positive pressure via the (first) pneumatic port 312b.

[0248] Optionally, at 1012 (if 1006 has been performed), liquid from the outlet in the third reservoir can be dispensed by applying positive pressure via pneumatic port 312b. Dispensing liquid from the outlet in the third reservoir can include opening a third reservoir valve configured to control flow of liquid from the outlet in the third reservoir. Dispensing liquid from the outlet in the third reservoir can include opening one or more valves of the liquid handling device to establish a fluid connection between the outlet in the third reservoir and a third pneumatic port (e.g., 312c) of the liquid handling device, and venting the third pneumatic port during the application of positive pressure via the (first) pneumatic port 312b.

[0249] FIG. 28 is a flowchart of a method 1100 of metering liquid in a channel. At 1102, a first pressure is applied to move a first liquid from a first chamber through a first fluid path comprising two junctions. The first pressure can be applied to aspirate the first liquid through the first fluid path from the chamber (e.g., a vented sample chamber) (i.e., using negative pressure). Alternatively, the first pressure can be applied to dispense (i.e., using positive pressure) the first liquid from the chamber, such as a liquid reagent capsule.

[0250] At 1104, a second pressure is applied to move a fluid through a second fluid path comprising a portion of the first fluid path between the two junctions, such that the fluid displaces a volume of the first liquid between the two junctions. The fluid can be a second liquid, which can be dispensed from a second chamber (e.g., a liquid reagent capsule). Alternatively, the fluid can be air, which can be supplied via an air supply conduit in fluid communication with a pneumatic port.

[0251] The fluid can be displaced by a volume of the first liquid by moving the volume of the first liquid to the chamber. Displacing the volume of the first liquid using the second fluid means that an accurately metered volume of liquid can be moved to the chamber. This accurate volume is FIG. 37The volume of liquid between the two junctions of the fluidic network (e.g., junctions 306a and 306b) is shown. Steps 1102 and 1104 can be repeated in order to measure a particular volume of the first liquid that is a multiple of the volume of the first fluidic path between the two junctions. For example, if the volume of the first fluidic path between the two junctions is 5 pL, then steps 1102 and 1104 can be performed in order to move 20 pL of the first liquid volume into the chamber.

[0252] FIG. 38 is a flowchart of a method 1200 of priming a fluidic circuit. At 1202, a first pressure is applied to move liquid from a chamber into a first channel. At 1204, the first pressure is continued to be applied to move the liquid until the liquid fills the first channel past a junction on the first channel, where the junction connects the first channel to a second channel. At 1206, a second pressure is applied to move the liquid from the chamber into the second channel via the first channel and the junction.

[0253] FIG. 39 is a flowchart of a method 1300 of diluting a sample. At 1302, a negative pressure is applied to aspirate a first liquid volume into a first flow cell. Optionally, at 1304, a detection signal is measured in the first flow cell. At 1306, a negative pressure is applied to aspirate a volume of a first liquid (e.g., a metered volume) into a first chamber. At 1308, a positive pressure is applied to dispense a volume of a second liquid (e.g., a metered volume) into the first chamber. At 1310, a positive pressure is applied to dispense the first liquid and the second liquid from the first chamber to a second chamber. At 1312, a negative pressure is applied to aspirate the first liquid and the second liquid from the second chamber to the first chamber. At 1314, steps 1310 and 1312 are repeated one or more times. At 1316, a positive pressure is applied to move the mixture from the first chamber or the second chamber to a second flow cell. Optionally, at 1318, a detection signal is measured in the second flow cell.

[0254] FIGS. 26-28 is a flowchart of a method 1400 of resuspending dried reagents. At 1402, a first pressure is applied to transfer a metered volume of a first liquid to a first chamber to resuspend one or more first dried reagents, where resuspension of the one or more first dried reagents in the first liquid forms a first reagent. At 1404, a second pressure is applied to transfer a metered volume of the first liquid to a second chamber to resuspend one or more second dried reagents, where resuspension of the one or more second dried reagents in the first liquid forms a second reagent. At 1406, a third pressure is applied to move the first reagent through at least one flow cell. At 1408, a fourth pressure is applied to move the second reagent through the at least one flow cell. At 1410, a detection signal is measured in the at least one flow cell.

[0255] The cartridge 100 having the fluidic circuit described above can be used to perform the assay described above. FIGS. 35-39 The cartridge 100 having the fluidic circuit described above can be used to perform the assay described above. FIGS. 40-43 The cartridge 100 having the fluidic circuit described above can be used to perform the assay described above. The assay described below can include one or more of the methods 1000, 1100, 1200, 1300, 1400 described above. Thus, the methods 1000, 1100, 1200, 1300, 1400 described above can also include one or more steps of the exemplary assay described above. FIGS. 26-28 The cartridge 100 having the fluidic circuit described above can be used to perform the assay described above. FIGS. 40-43 The cartridge 100 having the fluidic circuit described above can be used to perform the assay described above. The assay described below can include one or more of the methods 1000, 1100, 1200, 1300, 1400 described above. Thus, the methods 1000, 1100, 1200, 1300, 1400 described above can also include one or more steps of the exemplary assay described above. FIG. 40 The cartridge 100 having the fluidic circuit described above can be used to perform the assay described above. The assay described below can include one or more of the methods 1000, 1100, 1200, 1300, 1400 described above. Thus, the methods 1000, 1100, 1200, 1300, 1400 described above can also include one or more steps of the exemplary assay described above.

[0256] FIG. 41 is a flowchart of a method 1500 to perform a first assay. At 1502, a negative pressure is applied to the first pneumatic port 312a (P1) to flow a sample from the permanently vented sample inlet chamber 236 (S1) to the first waste chamber port 358a (W1) and to fill the sample flow unit 610c (F3), with valves 302a (V2) and 302c (V9) open. At 1504, a detection signal is measured from the sample present in the sample flow unit 610c (F3).

[0257] At 1506, a positive pressure is applied to the second pneumatic port 312b (P2) to flow a diluent from the outlet of the third reagent reservoir capsule 120c (B3B) to the first waste chamber port 358a (W1), with valves 302k (V1), 302i (V7), and 302c (V9) open and to vent the first pneumatic port 312a (P1). Since the vented state of the first waste chamber 508a is controlled through the first pneumatic port 312a (P1), venting of the first pneumatic port 312a (P1) is required to allow flow to the first waste chamber port 358a (W1). At 1508, a positive pressure is applied to the second pneumatic port 312b (P2) to transfer a metered volume of diluent from the outlet of the third reagent reservoir capsule 120c (B3B) to the second chamber 332b (M2), with valves 302k (V1), 302b (V6), and 302i (V7) open and to vent the third pneumatic port 312c (P3). In this example, the second chamber 332b (M2) contains two dry reagents in the form of beads: one is a magnetic bead coated with a capture agent and the other has a sample processing reagent. In this step, both beads are hydrated.

[0258] At 1510, positive pressure is applied to the second pneumatic port 312b (P2) to transfer a metered volume of diluent from the outlet of the third reagent reservoir capsule 120c (B3B) to the third chamber 332c (M3), where valves 302i (V7) and 302j (V8) are open and the third pneumatic port 312c (P3) is vented. In this embodiment, the third chamber 332c (M3) contains a dry reagent in the form of a bead having a secondary antibody and an enzyme label. In this step, the bead is hydrated.

[0259] At 1512, negative pressure is applied to the third pneumatic port 312c (P3) to flow a metered volume of sample from the permanently vented sample inlet chamber 236 (SI) to the second chamber 332b (M2), where valves 302a (V2) and 302b (V6) are open. At 1514, positive and negative pressure is cycled to the third pneumatic port 312c (P3), where the first pneumatic port 312a (PI) is vented to mix the sample with the diluent and the dry reagent stored in the first chamber 332a (Ml) and / or the second chamber 332b (M2) by moving the liquid back and forth between the first chamber 332a (Ml) and the second chamber 332b (M2). Step 1514 can also be performed by cycling positive and negative pressure to the first pneumatic port 312a (PI) and venting the third pneumatic port 312c (P3).

[0260] At 1516, positive pressure is applied to the third pneumatic port 312c (P3) to flow the mixture from the first chamber 332a (Ml) or the second chamber 332b (M2) through the first flow cell 610a (Fl) to the third waste chamber port 358c (W3), where valves 302e (Vl l) and 302d (V12) are open and a magnet on the first flow cell 610a (Fl) is activated. The magnet can be actuated by the analyzer device that receives the cartridge 100 and performs the method 1500 of the first assay using the cartridge 100. The magnet can be actuated by the analyzer device to apply a localized magnetic field to the first flow cell 610a (Fl). The magnet can be a moveable permanent magnet or a fixed electromagnet. At 1518, positive pressure is applied to the second pneumatic port 312b (P2) to flow a wash liquid from the outlet of the first reagent reservoir capsule 120a (BlB) to the first waste chamber port 358a (Wl), where valves 302k (Vl), 302g (V4), and 302c (V9) are open and the first pneumatic port 312a (PI) is vented.

[0261] At 1520, positive pressure is applied to the second pneumatic port 312b (P2) to flow the wash solution from the outlet of the first reservoir bladder 120a (B1B) through the first flow cell 610a (F1) to the third waste chamber port 358c (W3), with valves 302g (V4) and 302e (V11) open and the magnet on the first flow cell 610a (F1) activated. At 1522, positive pressure is applied to the second pneumatic port 312b (P2) to flow air through the first flow cell 610a (F1) to the third waste chamber port 358c (W3), with valves 302L (V3) and 302e (V11) open and the magnet on the first flow cell 610a (F1) activated.

[0262] At 1524, positive pressure is applied to the third pneumatic port 312c (P3) to flow the detection solution from the third chamber 332c (M3) through the first flow cell 610a (F1) to the third waste chamber port 358c (W3), with valves 302j (V8) and 302e (V11) open and the magnet on the first flow cell 610a (F1) activated. In this example, the detection solution is prepared from a diluent solution and detection beads in the third chamber 332c (M3) in step 1510. At 1526, positive pressure is applied to the second pneumatic port 312b (P2) to flow the wash solution from the outlet of the first reservoir bladder 120a (B1B) through the first flow cell 610a (F1) to the third waste chamber port 358c (W3), with valves 302g (V4) and 302e (V11) open and the magnet on the first flow cell 610a (F1) activated. At 1528, positive pressure is applied to the second pneumatic port 312b (P2) to flow air through the first flow cell 610a (F1) to the third waste chamber port 358c (W3), with valves 302L (V3) and 302e (V11) open and the magnet on the first flow cell 610a (F1) activated.

[0263] At 1530, positive pressure is applied to the second pneumatic port 312b (P2) to flow wash solution from the outlet of the first reservoir bladder 120a (B1B) through the first flow cell 610a (F1) and the second flow cell 610b (F2) to the second waste chamber port 358b (W2), with valves 302g (V4) and 302f (V10) open and the magnet on the second flow cell 610b (F2) activated. This step transfers the magnetic beads from the first flow cell 610a (F1) to the second flow cell 610b (F2). At 1532, positive pressure is applied to the second pneumatic port 312b (P2) to flow air through the first flow cell 610a (F1) and the second flow cell 610b (F2) to the second waste chamber port 358b (W2), with valves 302L (V3) and 302f (V10) open and the magnet on the second flow cell 610b (F2) activated.

[0264] At 1534, positive pressure is applied to the second pneumatic port 312b (P2) to flow signal solution from the outlet of the second reservoir bladder 120b (B2B) to the second waste chamber port 358b (W2), with valve 302h (V5) and valve 302f (V10) open. At 1536, positive pressure is applied to the second pneumatic port 312b (P2) to flow signal solution from the outlet of the second reservoir bladder 120b (B2B) through the first flow cell 610a (F1) and the second flow cell 610b (F2) to the first waste chamber port 358a (W1), with valves 302k (V1), 302h (V5), and 302c (V9) open, the first pneumatic port 312a (P1) vented, and the magnet on the second flow cell 610b (F2) activated. At 1538, a detection signal is measured in the flow cell 610a (F1) and / or 610b (F2).

[0265] FIG. 42 is a flowchart of a method 1600 of performing a second assay. At 1602, negative pressure is applied to the first pneumatic port 312a (P1) to flow sample from the permanently vented sample inlet chamber 236 (S1) to the first waste chamber port 358a (W1) and to fill the sample flow cell 610c (F3), with valves 302a (V2) and 302c (V9) open. At 1604, a detection signal is measured from the sample present in the sample flow cell 610c (F3).

[0266] At 1606, positive pressure is applied to the second pneumatic port 312b (P2) to flow diluent from the outlet of the third reservoir capsule 120c (B3B) to the first waste chamber port 358a (Wl), with valves 302k (Vl), 302i (V7), and 302c (V9) open and the first pneumatic port 312a (Pl) vented. At 1608, positive pressure is applied to the second pneumatic port 312b (P2) to transfer the metered diluent from the outlet of the third reservoir capsule 120c (B3B) to the second chamber 332b (M2), with valves 302k (Vl), 302b (V6), and 302i (V7) open and the third pneumatic port 312c (P3) vented. The diluent will move the portion of the sample metered and held in the U-shaped channel between valves 302k (Vl), 302a (V2), and 302b (V6).

[0267] At 1610, positive and negative pressure is cycled to the third pneumatic port 312c (P3), with the first pneumatic port 312a (Pl) vented, to mix the sample with the diluent and resuspended reagents stored in the first chamber 332a (Ml) and / or the second chamber 332b (M2) by moving the liquid back and forth between the first chamber 332a (Ml) and the second chamber 332b (M2). Step 1610 can also be performed by cycling positive and negative pressure to the first pneumatic port 312a (Pl) and venting the third pneumatic port 312c (P3).

[0268] At 1612, positive pressure is applied to the third pneumatic port 312c (P3) to flow the mixture from the first chamber 332a (Ml) or the second chamber 332b (M2) through the first flow cell 610a (Fl) to the third waste chamber port 358c (W3), with valves 302e (Vl l) and 302d (V12) open and the magnet on the first flow cell 610a (Fl) activated. At 1614, positive pressure is applied to the second pneumatic port 312b (P2) to flow wash solution from the outlet of the first reservoir capsule 120a (BlB) to the first waste chamber port 358a (Wl), with valves 302k (Vl), 302g (V4), and 302c (V9) open and the first pneumatic port 312a (Pl) vented. After this step, the wash buffer can now be pre-charged.

[0269] At 1616, positive pressure is applied to the second pneumatic port 312b (P2) to flow the wash solution from the outlet of the first reservoir capsule 120a (B1B) through the first flow cell 610a (F1) to the third waste chamber port 358c (W3), with valves 302g (V4) and 302e (V11) open and the magnet on the first flow cell 610a (F1) activated. At 1618, positive pressure is applied to the second pneumatic port 312b (P2) to flow air through the first flow cell 610a (F1) to the third waste chamber port 358c (W3), with valves 302L (V3) and 302e (V11) open and the magnet on the first flow cell 610a (F1) activated.

[0270] At 1620, positive pressure is applied to the second pneumatic port 312b (P2) to flow the wash solution from the outlet of the first reservoir capsule 120a (B1B) through the first flow cell 610a (F1) and the second flow cell 610b (F2) to the second waste chamber port 358b (W2), with valves 302g (V4) and 302f (V10) open and the magnet on the second flow cell 610b (F2) activated. This step transfers the magnetic beads from the first flow cell 610a (F1) to the second flow cell 610b (F2). At 1622, positive pressure is applied to the second pneumatic port 312b (P2) to flow air through the first flow cell 610a (F1) and the second flow cell 610b (F2) to the second waste chamber port 358b (W2), with valves 302L (V3) and 302f (V10) open and the magnet on the second flow cell 610b (F2) activated.

[0271] At 1624, positive pressure is applied to the second pneumatic port 312b (P2) to flow the signal solution from the outlet of the second reservoir capsule 120b (B2B) to the second waste chamber port 358b (W2), with valves 302h (V5) and 302f (V10) open. At 1626, positive pressure is applied to the second pneumatic port 312b (P2) to flow the signal solution from the outlet of the second reservoir capsule 120b (B2B) through the first flow cell 610a (F1) and the second flow cell 610b (F2) to the third chamber 332c (M3), with valves 302h (V5) and valve 302j (V8) open, venting the third pneumatic port 312c (P3), and the magnet on the second flow cell 610b (F2) activated. At 1628, the detection signal is measured in the first flow cell 610a (F1) and the second flow cell 610b (F2).

[0272] FIG. 43is a flowchart of a method 1700 to perform a third assay. At 1702, a negative pressure is applied to the first pneumatic port 312a (P1) to flow sample from the permanently vented sample inlet chamber 236 (S1) to the first waste chamber port 358a (W1) and fill the sample flow cell 610c (F3), with valves 302a (V2) and 302c (V9) open. At 1704, a detection signal is measured from the sample present in the flow cell 610c (F3).

[0273] At 1706, a positive pressure is applied to the second pneumatic port 312b (P2) to flow diluent from the outlet of the third reservoir bladder 120c (B3B) to the first waste chamber port 358a (W1), with valves 302k (V1), 302i (V7), and 302c (V9) open and the first pneumatic port 312a (P1) vented. At 1708, a positive pressure is applied to the second pneumatic port 312b (P2) to transfer a metered amount of diluent from the outlet of the third reservoir bladder 120c (B3B) to the second chamber 332b (M2), with valves 302k (V1), 302b (V6), and 302i (V7) open and the third pneumatic port 312c (P3) vented. At 1710, a positive pressure and a negative pressure are applied to the third pneumatic port 312c (P3) in a cycle, with the first pneumatic port 312a (P1) vented, to mix the sample with the diluent and resuspend the dry reagent stored in the first chamber 332a (M1) and / or the second chamber 332b (M2) by moving the liquid back and forth between the first chamber 332a (M1) and the second chamber 332b (M2).

[0274] At 1712, a positive pressure is applied to the third pneumatic port 312c (P3) to flow the mixture from the first chamber 332a (Ml) or the second chamber 332b (M2) through the first flow cell 610a (Fl) and the second flow cell 610b (F2) to the second waste chamber port 358b (W2), with valves 302f (V10) and 302e (Vll) open. At 1714, a positive pressure is applied to the second pneumatic port 312b (P2) to flow the wash solution from the first reservoir bladder 120a (BlB) to the first waste chamber port 358a (Wl), with valves 302k (Vl), 302g (V4), and 302c (V9) open, and the first pneumatic port 312a (Pl) vented. At 1716, a positive pressure is applied to the second pneumatic port 312b (P2) to flow the wash solution from the first reservoir bladder 120a (BlB) through the first flow cell 610a (Fl) and the second flow cell 610b (F2) to the second waste chamber port 358b (W2), with valves 302g (V4) and 302f (V10) open. At 1718, a positive pressure is applied to the second pneumatic port 312b (P2) to flow air through the first flow cell 610a (Fl) and the second flow cell 610b (F2) to the second waste chamber port 358b (W2), with valves 302L (V3) and 302f (V10) open.

[0275] At 1720, a positive pressure is applied to the second pneumatic port 312b (P2) to flow the signal solution from the outlet of the second reservoir bladder 120b (B2B) to the second waste chamber port 358b (W2), with valves 302h (V5) and 302f (V10) open. At 1722, a positive pressure is applied to the second pneumatic port 312b (P2) to flow the solution from the outlet of the second reservoir bladder 120b (B2B) through the first flow cell 610a (Fl) and the second flow cell 610b (F2) to the third chamber 332c (M3), with valves 302h (V5) and 302j (V8) open, and the third pneumatic port 312c (P3) vented. At 1724, a detection signal is measured from the flow cells of the first flow cell 610a (Fl) and the second flow cell 610b (F2).

[0276] FIG. 3 is a flowchart of a method 1800 to perform a fourth assay. At 1802, a negative pressure is applied to the first pneumatic port 312a (Pl) to flow the sample from the permanently vented sample inlet chamber 236 (Sl) to the first waste chamber port 358a (Wl) and to fill the sample flow cell 610c (F3), with valves 302a (V2) and 302c (V9) open. At 1804, a detection signal is measured from the sample present in the sample flow cell 610c (F3).

[0277] At 1806, positive pressure is applied to the second pneumatic port 312b (P2) to flow diluent from the outlet of the third reservoir bladder 120c (B3B) to the first waste chamber port 358a (Wl), with valves 302k (Vl), 302i (V7), and 302c (V9) open and the first pneumatic port 312a (Pl) vented. At 1808, positive pressure is applied to the second pneumatic port 312b (P2) to transfer the metered volume of diluent from the outlet of the third reservoir bladder 120c (B3B) to the second chamber 332b (M2), with valves 302k (Vl), 302b (V6), and 302i (V7) open and the third pneumatic port 312c (P3) vented. At 1810, positive pressure is applied to the second pneumatic port 312b (P2) to transfer the metered volume of diluent from the outlet of the third reservoir bladder 120c (B3B) to the third chamber 332c (M3), with valves 302i (V7) and 302j (V8) open and the third pneumatic port 312c (P3) vented.

[0278] At 1812, negative pressure is applied to the third pneumatic port 312c (P3) to flow a metered volume of sample from the permanently vented sample inlet chamber 236 (Sl) to the second chamber 332b (M2), with valve 302a (V2) and valve 302b (V6) open. At 1814, positive and negative pressure is applied to the third pneumatic port 312c (P3) in a cyclical fashion, with the first pneumatic port 312a (Pl) vented, to mix the sample with the diluent and resuspend dry reagents stored in the first chamber 332a (Ml) and / or the second chamber 332b (M2) by moving the liquid back and forth between the first chamber 332a (Ml) and the second chamber 332b (M2).

[0279] At 1816, positive pressure is applied to the third pneumatic port 312c (P3) to flow the mixture from the first chamber 332a (Ml) or the second chamber 332b (M2) through the first flow cell 610a (Fl) and the second flow cell 610b (F2) to the second waste chamber port 358b (W2), with valves 302f (V10) and 302e (Vl l) open. At 1818, positive pressure is applied to the second pneumatic port 312b (P2) to flow wash solution from the outlet of the first reservoir bladder 120a (BlB) to the first waste chamber port 358a (Wl), with valves 302k (Vl), 302g (V4), and 302c (V9) open and the first pneumatic port 312a (Pl) vented.

[0280] At 1820, positive pressure is applied to the second pneumatic port 312b (P2) to flow the wash solution from the outlet of the first reservoir capsule 120a (B1B) through the first flow cell 610a (F1) and the second flow cell 610b (F2) to the second waste chamber port 358b (W2) with valves 302g (V4) and 302f (V10) open. At 1822, positive pressure is applied to the second pneumatic port 312b (P2) to flow air through the first flow cell 610a (F1) and the second flow cell 610b (F2) to the second waste chamber port 358b (W2) with valves 302L (V3) and 302f (V10) open.

[0281] At 1824, positive pressure is applied to the third pneumatic port 312c (P3) to flow the detection solution from the third chamber 332c (M3) through the first flow cell 610a (F1) and the second flow cell 610b (F2) to the second waste chamber port 358b (W2) with valves 302j (V8) and 302f (V10) open. At 1826, positive pressure is applied to the second pneumatic port 312b (P2) to flow the wash solution from the outlet of the first reservoir capsule 120a (B1B) through the first flow cell 610a (F1) and the second flow cell 610b (F2) to the second waste chamber port 358b (W2) with valves 302g (V4) and 302f (V10) open. At 1828, positive pressure is applied to the second pneumatic port 312b (P2) to flow air through the first flow cell 610a (F1) and the second flow cell 610b (F2) to the second waste chamber port 358b (W2) with valves 302L (V3) and valve 302f (V10) open.

[0282] At 1830, positive pressure is applied to the second pneumatic port 312b (P2) to flow the signal solution from the outlet of the second reservoir capsule 120b (B2B) to the second waste chamber port 358b (W2) with valves 302h (V5) and valve 302f (V10) open. At 1832, positive pressure is applied to the second pneumatic port 312b (P2) to flow the solution from the outlet of the second reservoir capsule 120b (B2B) through the first flow cell 610a (F1) and the second flow cell 610b (F2) to the first waste chamber port 358a (W1) with valves 302k (V1), 302h (V5), and 302c (V9) open and the first pneumatic port 312a (P1) vented. At 1834, the detection signal is measured in the first flow cell 610a (F1) and the second flow cell 610b (F2).

[0283] Each of methods 1500, 1600, 1700, and 1800 can be implemented using the fluid loop of cartridge 100 described above. Therefore, cartridge 100 is capable of performing many different analytical methods. Depending on the sensor configuration, the same analytical method can be used for single detection (one biomarker) or for multiplex detection (two or more biomarkers). Methods 1500 and 1600 are designed to enable a liquid-phase reaction of the sample with a trapping agent (e.g., antibody or antigen) coated on magnetic beads. Methods 1700 and 1800 are designed to enable a solid-phase reaction of the sample with a trapping agent (e.g., antibody or antigen) coated on the sensor surface. The sensor and flow unit can be configured for different types of detection, such as electrochemical detection (e.g., ...). FIGS. 35-43 (as shown) or optical detection (e.g., photometry or surface plasmon resonance). It should be understood that the steps in methods 1500, 1600, 1700, and 1800 may be performed in a different order than described above. That is, the specific order of the method steps described above is not restrictive but optional.

[0284] The embodiments described herein also include an analyzer device configured to house cartridge 100. The analyzer device may be configured to operate cartridge 100 to perform reference operations. FIGS. 35-43 One or more methods described. Therefore, the analyzer device may include a computer-readable medium storing computer-executable instructions that, when executed by the processor of the analyzer device, cause the analyzer device to perform a reference... ​ One or more methods are described. The analyzer device may include one or more of the following hardware components: one or more valve actuators configured to deform a valve region 302 of cartridge 100 to close a valve of cartridge 100; one or more capsule actuators configured to actuate an actuable portion 240 of a first portion 200 of cartridge 100 to create an inlet and an outlet in each of one or more reservoir capsules 120; one or more pneumatic actuators configured to supply positive or negative pressure to each pneumatic port 312 of cartridge 100 to ventilate and / or close the pneumatic port 312 of cartridge 100; one or more magnets (e.g., movable permanent magnets or electromagnets) actuated to apply a local magnetic field to one or more of the flow units 610 of cartridge 100 (e.g., retaining a magnetic bead within one or more flow units 610); and a detection module for measuring detection signals in one or more flow units 610 of cartridge 100. The specific hardware requirements of the analyzer will depend on which of the above methods the analyzer is used to implement, and those skilled in the art will understand this from the steps of the above methods.

[0285] The described methods can be implemented using computer-executable instructions. A computer program product or computer-readable medium can include or store computer-executable instructions. The computer program product or computer-readable medium can include a hard disk drive, flash memory, read-only memory (ROM), CD, DVD, cache, random access memory (RAM), and / or any other storage medium that stores information for any duration (e.g., for extended periods of time, for short periods of time, for

[0286] The singular terms "a," "an," and "the" should not be construed as "only one" unless otherwise indicated. Rather, they should be read to mean "at least one" or "one or more." The term "includes" and its derivatives mean that the claimed feature is included, but not that other features are excluded.

[0287] As used herein, the term "channel" refers to a recess disposed in a surface having an open cross-section (i.e., the cross-section is not sealed). As used herein, the term "conduit" refers to: (i) a channel that has been sealed (e.g., by a sealing layer), thereby providing a closed cross-section; or (ii) a hole or tunnel that extends at least partially through a body.

[0288] The above implementations have been described by way of example only, and are not intended to be limiting. It will be appreciated that changes can be made to the described implementations without departing from the scope of the present disclosure. It will also be appreciated that there are many alternative ways of implementing the application which will be apparent to those skilled in the art without departing from the scope of the application. It will be appreciated that the scope of the application is not limited to the specific implementations described above.

[0289] The present disclosure also includes the following numbered clauses, which describe combinations of features that are helpful in understanding the present disclosure:

[0290] A1. A liquid treatment device, comprising:

[0291] a fluid layer comprising:

[0292] a plurality of channels; and

[0293] an opening in a surface of the fluid layer, wherein the opening extends through at least a portion of a thickness of the fluid layer to one of the plurality of channels;

[0294] a rigid layer comprising an actuatable portion; and

[0295] a liquid reservoir capsule positioned between the fluid layer and the actuatable portion, wherein the liquid reservoir capsule comprises:

[0296] a body defining a volume to store a liquid, the body comprising a first deformable portion, a main liquid storage portion, and a restriction portion connecting the first deformable portion to the main liquid storage portion; and

[0297] a sealing layer configured to seal the volume;

[0298] wherein the liquid storage capsule is positioned above the opening such that a portion of the sealing layer covers the opening;

[0299] wherein the actuatable portion comprises a protrusion extending towards the liquid storage capsule, wherein the actuatable portion is actuatable from a first position in which the protrusion does not deform the first deformable portion to a second position in which the protrusion deforms the first deformable portion into contact with the portion of the sealing layer to cause the portion of the sealing layer to rupture, wherein the protrusion does not deform the restriction portion when the actuatable portion is in the second position.

[0300] A2. The liquid handling device of clause Al, wherein the protrusion is unable to pass through the opening.

[0301] A3. The liquid handling device of clause Al or clause A2, wherein the liquid storage capsule is positioned above the opening such that the first deformable portion and the restriction portion are at least partially positioned above the opening.

[0302] A4. The liquid handling device of any one of clauses Al to A3, wherein the opening has an oblong shape.

[0303] A5. The liquid handling device of any one of clauses Al to A4, wherein the first deformable portion comprises a first region and a second region, wherein a first distance between the first region and the sealing layer is greater than a second distance between the second region and the sealing layer.

[0304] A6. The liquid handling device of clause A5, wherein the first region and the second region are concentric such that the first region surrounds the second region.

[0305] A7. The liquid handling device of any one of clauses Al to A6, wherein a width of the opening is less than a width of the protrusion.

[0306] A8. The liquid handling device of any one of clauses Al to A7, wherein the protrusion comprises an end portion that engages the first deformable portion of the liquid storage capsule.

[0307] A9. The liquid handling device of clause A6, wherein the end portion of the protrusion is flat.

[0308] A10. The liquid treatment device of clause A8 or clause A9, wherein the end of the protrusion has a cross-section that is a main scallop shape.

[0309] A11. The liquid treatment device of any one of clauses A8 to A10, wherein the protrusion comprises a groove that defines the cross-section of the end of the protrusion.

[0310] A12. The liquid treatment device of clause A11, wherein the groove is aligned with the restriction of the reservoir capsule.

[0311] A13. The liquid treatment device of clause A11 or clause A12 when dependent on clause A10, wherein the groove defines the main scallop shape of the cross-section of the end of the protrusion.

[0312] A14. The liquid treatment device of any one of clauses A8 to A13, wherein the cross-section of the end of the protrusion comprises one or more outwardly extending protruding ribs.

[0313] A15. The liquid treatment device of clause A14 when dependent on clause A10, wherein the one or more outwardly extending protruding ribs extend from a curved portion of the main scallop shape.

[0314] A16. The liquid treatment device of any one of clauses A1 to A15, wherein:

[0315] the protrusion is a first protrusion and the actuatable portion comprises a second protrusion extending towards the reservoir capsule;

[0316] the body of the reservoir capsule comprises a second deformable portion;

[0317] the opening is a first opening and the fluid layer comprises a second opening extending through at least a portion of a thickness of the fluid layer to one of the plurality of channels;

[0318] the portion of the seal layer is a first portion of the seal layer and the reservoir capsule is positioned over the second opening such that a second portion of the seal layer covers the second opening;

[0319] wherein, in the first position, the second protrusion does not deform the second deformable portion;

[0320] wherein, in the second position, the second protrusion deforms the second deformable portion to contact the second portion of the seal layer, thereby causing the second portion of the seal layer to rupture.

[0321] A17. The liquid treatment device of any of clauses A1-A16, wherein:

[0322] the reservoir capsule is a first reservoir capsule, the liquid treatment device comprising a second reservoir capsule, the second reservoir capsule comprising:

[0323] a body defining a volume to store liquid, the body comprising a first deformable portion and a second deformable portion; and

[0324] a sealing layer configured to seal the volume;

[0325] the actuable portion comprising:

[0326] a third protrusion extending toward the first deformable portion of the second reservoir capsule; and

[0327] a fourth protrusion extending toward the second deformable portion of the second reservoir capsule;

[0328] the fluid layer comprising:

[0329] a third opening extending through at least a portion of a thickness of the fluid layer to one of the plurality of channels; and

[0330] a fourth opening extending through at least a portion of a thickness of the fluid layer to one of the plurality of channels;

[0331] wherein the second reservoir capsule is positioned over the third opening and the fourth opening such that a first portion of a sealing layer of the second reservoir capsule covers the third opening and a second portion of the sealing layer of the second reservoir capsule covers the fourth opening;

[0332] wherein, in the first position, the third protrusion does not deform the first deformable portion of the body of the second reservoir capsule and the fourth protrusion does not deform the second deformable portion of the body of the second reservoir capsule;

[0333] wherein, in the second position, the third protrusion deforms the first deformable portion of the body of the second reservoir capsule to be in contact with the first portion of the sealing layer of the second reservoir capsule, thereby causing the first portion of the sealing layer of the second reservoir capsule to rupture; and

[0334] wherein, in the second position, the fourth protrusion deforms the second deformable portion of the body of the second reservoir capsule to be in contact with the second portion of the sealing layer of the second reservoir capsule, thereby causing the second portion of the sealing layer of the second reservoir capsule to rupture.

[0335] B1. A liquid treatment device comprising:

[0336] a fluid layer comprising:

[0337] a plurality of channels; and

[0338] an opening in a surface of the fluid layer, wherein the opening extends through at least a portion of a thickness of the fluid layer to one of the plurality of channels;

[0339] a rigid layer comprising an actuatable portion; and

[0340] a liquid reservoir capsule positioned between the fluid layer and the actuatable portion, wherein the liquid reservoir capsule comprises:

[0341] a body defining a volume to store a liquid; and

[0342] a sealing layer configured to seal the volume;

[0343] wherein the liquid reservoir capsule is positioned over the opening such that a portion of the sealing layer covers the opening;

[0344] wherein the actuatable portion comprises a protrusion extending toward the liquid reservoir capsule, wherein the actuatable portion is actuatable from a first position in which the protrusion does not deform the body to a second position in which the protrusion deforms the body into contact with the portion of the sealing layer to cause the portion of the sealing layer to rupture; and

[0345] wherein the protrusion is unable to pass into the opening.

[0346] B2. The liquid treatment device of clause B1, wherein the opening has an oblong shape.

[0347] B3. The liquid treatment device of clause B1 or clause B2, wherein the body of the liquid reservoir capsule comprises a first deformable portion configured to be deformed by the protrusion when the actuatable portion is in the second position.

[0348] B4. The liquid treatment device of clause B3, wherein the body further comprises a main liquid reservoir portion and a restriction portion connecting the first deformable portion to the main liquid reservoir portion, wherein the liquid reservoir capsule is positioned over the opening such that the first deformable portion and the restriction portion are positioned at least partially over the opening.

[0349] B5. The liquid handling device of clause B3 or clause B4, wherein the first deformable portion comprises a first region and a second region, wherein a first distance between the first region and the sealing layer is greater than a second distance between the second region and the sealing layer.

[0350] B6. The liquid handling device of clause B5, wherein the first region and the second region are concentric, such that the first region encloses the second region.

[0351] B7. The liquid handling device of any one of clauses B1 to B6, wherein a width of the opening is less than a width of the protrusion.

[0352] B8. The liquid handling device of any one of clauses B1 to B7, wherein the protrusion comprises an end portion that engages the body of the liquid storage capsule.

[0353] B9. The liquid handling device of clause B8, wherein the end portion of the protrusion is flat.

[0354] B10. The liquid handling device of clause B8 or clause B9, wherein the end portion of the protrusion has a cross-section that is a main sector shape.

[0355] B11. The liquid handling device of any one of clauses B8 to B10, wherein the protrusion comprises a groove that defines the cross-section of the end portion of the protrusion.

[0356] B12. The liquid handling device of clause B11, wherein the groove is aligned with the restriction of the liquid storage capsule.

[0357] B13. The liquid handling device of clause B11 or clause B12 when dependent on clause B10, wherein the groove defines a main sector shape of the cross-section of the end portion of the protrusion.

[0358] B14. The liquid handling device of any one of clauses B8 to B13, wherein the cross-section of the end portion of the protrusion comprises one or more outwardly extending protruding ribs.

[0359] B15. The liquid handling device of clause B14 when dependent on clause B10, wherein the one or more outwardly extending protruding ribs extend from a curved portion of the main sector shape.

[0360] B16. The liquid handling device of any one of clauses B1 to B15, wherein:

[0361] the protrusion is a first protrusion, and the actuable portion comprises a second protrusion that extends towards the liquid storage capsule;

[0362] the opening is a first opening, and the fluid layer includes a second opening extending through at least a portion of a thickness of the fluid layer to one of the plurality of channels;

[0363] the portion of the sealing layer is a first portion of the sealing layer, and the reservoir capsule is positioned over the second opening such that a second portion of the sealing layer covers the second opening;

[0364] wherein, in the first position, the second protrusion does not deform a body of the reservoir capsule;

[0365] wherein, in the second position, the second protrusion deforms the body to contact the second portion of the sealing layer, thereby causing the second portion of the sealing layer to rupture.

[0366] B17. The liquid treatment device of any of clauses B1-B16, wherein:

[0367] the reservoir capsule is a first reservoir capsule, the liquid treatment device comprising a second reservoir capsule, the second reservoir capsule comprising:

[0368] a body defining a volume to store liquid; and

[0369] a sealing layer configured to seal the volume;

[0370] the actuatable portion comprises:

[0371] a third protrusion extending toward the second reservoir capsule; and

[0372] a fourth protrusion extending toward the second reservoir capsule;

[0373] the fluid layer comprises:

[0374] a third opening extending through at least a portion of a thickness of the fluid layer to one of the plurality of channels; and

[0375] a fourth opening extending through at least a portion of a thickness of the fluid layer to one of the plurality of channels;

[0376] wherein the second reservoir capsule is positioned over the third opening and the fourth opening such that a first portion of a sealing layer of the second reservoir capsule covers the third opening and a second portion of the sealing layer of the second reservoir capsule covers the fourth opening;

[0377] wherein, in the first position, the third protrusion does not deform the body of the second reservoir capsule, and the fourth protrusion does not deform the body of the second reservoir capsule;

[0378] wherein, in the second position, the third protrusion deforms the body of the second reservoir capsule to contact the first portion of the seal layer of the second reservoir capsule to cause the first portion of the seal layer of the second reservoir capsule to rupture; and

[0379] wherein, in the second position, the fourth protrusion deforms the body of the second reservoir capsule to contact the second portion of the seal layer of the second reservoir capsule to cause the second portion of the seal layer of the second reservoir capsule to rupture.

[0380] C1. A liquid handling device, comprising:

[0381] a fluid layer comprising a chamber, wherein the chamber comprises:

[0382] an opening at an upper end of the chamber; and

[0383] a seal layer configured to seal the opening of the chamber;

[0384] wherein the chamber comprises one or more protrusions, wherein each of the one or more protrusions extends from an inner wall of the chamber;

[0385] wherein, when a reagent sphere is contained in the chamber, the one or more protrusions prevent the reagent sphere from contacting the seal layer.

[0386] C2. The liquid handling device of clause C1, wherein the opening is configured to allow the reagent sphere to be inserted into the chamber.

[0387] C3. The liquid handling device of clause C1 or clause C2, wherein the one or more protrusions are elastically deformable.

[0388] C4. The liquid handling device of any one of clauses C1 to C3, wherein the fluid layer is formed from a thermoplastic elastomer.

[0389] C5. The liquid handling device of any one of clauses C1 to C4, wherein each of the one or more protrusions extends from an inner wall of the chamber at an upper end of the chamber.

[0390] C6. The liquid handling device of clause C5, wherein the opening is partially defined by each of the one or more protrusions.

[0391] C7. The liquid treatment device of any of clauses CI to C6, wherein the fluid layer comprises a protrusion extending from an upper surface of the fluid layer, wherein the protrusion partially defines the chamber.

[0392] C8. The liquid treatment device of any of clauses CI to C7, wherein the chamber further comprises:

[0393] a fluid inlet positioned below the one or more protrusions; and

[0394] an air outlet positioned in the inner wall;

[0395] wherein a first maximum distance between the air outlet and the upper end base is less than or equal to a second maximum distance between the one or more protrusions and the upper end.

[0396] C9. The liquid treatment device of any of clauses CI to C8, wherein each of the one or more protrusions comprises:

[0397] a first protrusion portion extending from an inner wall of the chamber; and

[0398] a second protrusion portion extending from a distal end of the first protrusion portion, wherein the second protrusion portion extends away from an upper end of the chamber.

[0399] D1. A liquid treatment device, comprising:

[0400] a first reservoir and a second reservoir, wherein each of the first reservoir and the second reservoir is configured to be opened by forming an inlet and an outlet in the reservoir; and

[0401] a fluid layer comprising:

[0402] a network of channels; and

[0403] a pneumatic port configured to receive positive pressure;

[0404] wherein the network of channels comprises:

[0405] a first reservoir inlet channel configured to provide a fluid connection between the pneumatic port and an inlet in the first reservoir when the first reservoir is opened; and

[0406] a second reservoir inlet channel configured to provide a fluid connection between the pneumatic port and an inlet in the second reservoir when the second reservoir is opened.

[0407] D2. The liquid treatment device of clause Dl, further comprising:

[0408] a first reservoir valve configured to control flow of liquid from an outlet of the first reservoir; and

[0409] a second reservoir valve configured to control flow of liquid from an outlet of the second reservoir.

[0410] D3. The liquid handling device of clause D2, wherein the network of channels further comprises:

[0411] a first reservoir outlet channel configured to be in fluid communication with the outlet in the first reservoir when the first reservoir is open, wherein the first reservoir valve is on the first reservoir outlet channel.

[0412] D4. The liquid handling device of clause D2 or clause D3, wherein the network of channels further comprises:

[0413] a second reservoir outlet channel configured to be in fluid communication with the outlet in the second reservoir when the second reservoir is open, wherein the second reservoir valve is on the second reservoir outlet channel.

[0414] D5. The liquid handling device of any one of clauses D1-D4, wherein the liquid handling device further comprises a well, wherein the well is at least partially defined by an opening extending through at least a portion of a thickness of the fluid layer, wherein the well is in fluid communication with the pneumatic port, the first reservoir inlet channel, and the second reservoir inlet channel.

[0415] D6. The liquid handling device of clause D5, wherein the opening, the first reservoir inlet channel, and the second reservoir inlet channel are each disposed in an upper surface of the fluid layer, and wherein a depth of the opening is greater than a depth of the first reservoir inlet channel and the second reservoir inlet channel.

[0416] D7. The liquid handling device of clause D5 or clause D6, wherein:

[0417] the liquid handling device further comprises a lower portion comprising a slot in an upper surface of the lower portion;

[0418] the fluid layer is positioned on top of the lower portion; and

[0419] the well is at least partially defined by the opening in the fluid layer and the slot in the lower portion.

[0420] D8. The liquid treatment device of clause D7, wherein the opening in the fluid layer extends through a thickness of the fluid layer.

[0421] D9. The liquid treatment device of clause D7 or clause D8, wherein the pneumatic port extends through a thickness of the fluid layer, wherein the pneumatic port is in fluid communication with the slot via a connection channel in the lower portion.

[0422] D10. The liquid treatment device of any one of clauses D5-D9, wherein the well portion comprises an absorbent material.

[0423] D11. The liquid treatment device of any one of clauses D1-D10, wherein each of the first reservoir inlet channel and the second reservoir inlet channel comprises:

[0424] one or more first channel regions; and

[0425] a second channel region, wherein a first depth of the one or more first channel regions is less than a second depth of the second channel region.

[0426] D12. The liquid treatment device of clause D11, wherein the second depth is equal to a thickness of the fluid layer.

[0427] D13. The liquid treatment device of clause D11 or clause D12, wherein a first width of the one or more first channel regions is less than a second width of the second channel region.

[0428] D14. The liquid treatment device of any one of clauses D1-D13, wherein the liquid treatment device further comprises a third reservoir, wherein the third reservoir is configured to be opened by creating an inlet and an outlet in the reservoir, and wherein the channel network further comprises a third reservoir inlet channel configured to provide a fluid connection between the pneumatic port and the inlet in the third reservoir when the third reservoir is opened.

[0429] D15. The liquid treatment device of any one of clauses D1-D14, wherein the pneumatic port is a first pneumatic port, and wherein the liquid treatment device further comprises a second pneumatic port configured to be in fluid communication with the outlet in the first reservoir.

[0430] D16. The liquid treatment device of clause D15, wherein the second pneumatic port is configured to be in fluid communication with the outlet in the second reservoir.

[0431] D17. The liquid handling device of clause D15 or clause D16, wherein the liquid handling device further comprises a third pneumatic port configured to be in fluid communication with the outlet in the second reservoir.

[0432] E1. A method of controlling liquid flow in a liquid handling device, the method comprising:

[0433] creating an inlet and an outlet in a first reservoir of the liquid handling device to create a fluid connection between a pneumatic port of the liquid handling device and the inlet in the first reservoir;

[0434] creating an inlet and an outlet in a second reservoir of the liquid handling device to create a fluid connection between the pneumatic port and the inlet in the second reservoir;

[0435] dispensing liquid from the outlet in the first reservoir by applying positive pressure via the pneumatic port; and

[0436] dispensing liquid from the outlet in the second reservoir by applying positive pressure via the pneumatic port.

[0437] E2. The method of clause E1, wherein dispensing liquid from the outlet in the first reservoir comprises opening a first reservoir valve configured to control flow of liquid from the outlet in the first reservoir.

[0438] E3. The method of clause E1 or clause E2, wherein dispensing liquid from the outlet in the second reservoir comprises opening a second reservoir valve configured to control flow of liquid from the outlet in the second reservoir.

[0439] E4. The method of any one of clauses E1-E3, further comprising:

[0440] creating an inlet and an outlet in a third reservoir of the liquid handling device to create a fluid connection between the pneumatic port and the inlet of the third reservoir; and

[0441] dispensing liquid from the outlet in the third reservoir by applying positive pressure via the pneumatic port.

[0442] E5. The method of any one of clauses E1-E4, wherein the pneumatic port is a first pneumatic port, wherein the liquid handling device comprises a second pneumatic port, and wherein dispensing liquid from the outlet in the first reservoir comprises:

[0443] opening one or more valves of the liquid handling device to establish a fluid connection between the outlet in the first reservoir and the second pneumatic port; and

[0444] venting the second pneumatic port during application of positive pressure via the first pneumatic port.

[0445] E6. The method of clause E5, wherein dispensing liquid from an outlet in a second reservoir comprises:

[0446] opening one or more valves of the liquid handling device to establish a fluid connection between the outlet in the second reservoir and the second pneumatic port; and

[0447] venting the second pneumatic port during application of positive pressure via the first pneumatic port.

[0448] E7. The method of clause E5 or E6, wherein the liquid handling device comprises a third pneumatic port, and wherein dispensing liquid from an outlet in the second reservoir comprises:

[0449] opening one or more valves of the liquid handling device to establish a fluid connection between the outlet in the second reservoir and the third pneumatic port; and

[0450] venting the third pneumatic port during application of positive pressure via the first pneumatic port.

[0451] F1. A liquid handling device comprising:

[0452] a first chamber in fluid communication with a second chamber;

[0453] a first pneumatic port in fluid communication with one of the first chamber and the second chamber;

[0454] a second pneumatic port in fluid communication with the other of the first chamber and the second chamber;

[0455] a chamber inlet conduit configured to allow liquid to flow into the first chamber;

[0456] a chamber outlet conduit configured to allow liquid to flow out of the second chamber; and

[0457] a third pneumatic port in fluid communication with a first liquid reagent capsule, wherein the first liquid reagent capsule is in selective fluid communication with the first chamber or the second chamber when the first liquid reagent capsule is open.

[0458] F2. The liquid handling device of clause Fl, wherein liquid flow through the chamber inlet conduit is controlled using a chamber inlet valve.

[0459] F3. The liquid handling device of clause Fl or clause F2, wherein liquid flow through the chamber outlet conduit is controlled using a chamber outlet valve.

[0460] F4. The liquid handling device of any one of clauses Fl to F3, wherein liquid flow from the first liquid reagent capsule is controlled using a first liquid reagent capsule valve.

[0461] F5. The liquid handling device of any one of clauses Fl to F4, wherein the first pneumatic port is in permanent fluid communication with the first chamber.

[0462] F6. The liquid handling device of any one of clauses Fl to F5, wherein the second pneumatic port is in permanent fluid communication with the second chamber.

[0463] F7. The liquid handling device of any one of clauses Fl to F6, further comprising:

[0464] a first flow cell in fluid communication with the second chamber via the chamber outlet conduit;

[0465] a second flow cell in fluid communication with the first flow cell; and

[0466] a bypass conduit configured to allow liquid to bypass the second flow cell after flowing through the first flow cell.

[0467] F8. The liquid handling device of clause F7, wherein liquid flow through the bypass conduit is controlled using a bypass conduit valve.

[0468] F9. The liquid handling device of clause F7 or clause F8, wherein the first flow cell is in fluid communication with a waste liquid chamber via a first flow cell outlet conduit.

[0469] F10. The liquid handling device of clause F9 when dependent on clause F8, wherein liquid flowing into the waste liquid chamber via the first flow cell outlet conduit is controlled using the bypass conduit valve.

[0470] F11. The liquid handling device of clause F9 or clause F10, wherein the waste liquid chamber is permanently vented.

[0471] F12. The liquid handling device of any one of clauses F7-F11, wherein the second flow cell is in fluid communication with the waste liquid chamber via a second flow cell outlet conduit.

[0472] F13. The liquid handling device of clause F12, wherein liquid flow into the waste liquid chamber via the second flow cell outlet conduit is controlled using a second flow cell outlet valve.

[0473] F14. The liquid handling device of any one of clauses F1-F13, wherein the third pneumatic port is in fluid communication with a second liquid reagent capsule.

[0474] F15. The liquid handling device of clause F14, wherein liquid flow from the second liquid reagent capsule is controlled using a second liquid reagent capsule valve.

[0475] F16. The liquid handling device of clause F14 or clause F15 when dependent on clause F7, wherein the second liquid reagent capsule is in selective fluid communication with the second flow cell when the second liquid reagent capsule is open.

[0476] F17. The liquid handling device of clause F16 when dependent on clause F12, wherein the second liquid reagent capsule is in fluid communication with the second flow cell via the second flow cell outlet conduit when the second liquid reagent capsule is open.

[0477] F18. The liquid handling device of any one of clauses F14-F17, wherein the third pneumatic port is in fluid communication with a third liquid reagent capsule.

[0478] F19. The liquid handling device of clause F18, wherein liquid flow from the third liquid reagent capsule is controlled using a third liquid reagent capsule valve.

[0479] F20. The liquid handling device of clause F18 or clause F19, wherein the third liquid reagent capsule is in selective fluid communication with the first chamber when the third liquid reagent capsule is open.

[0480] F21. The liquid handling device of any one of clauses F18-F20, wherein the third liquid reagent capsule is in selective fluid communication with the first flow cell when the third liquid reagent capsule is open.

[0481] F22. The liquid handling device of any one of clauses F1-F21, further comprising a sample inlet chamber in fluid communication with the chamber inlet conduit.

[0482] F23. The liquid handling apparatus according to Clause F22, wherein liquid flow from the sample inlet chamber is controlled using a sample inlet valve.

[0483] F24. The liquid handling apparatus according to Clause F22 or Clause F23, wherein the sample inlet chamber is permanently vented.

[0484] F25. The liquid handling apparatus according to any one of Clauses F22 to F24, further comprising a sample flow unit in fluid communication with the sample inlet chamber via a sample flow unit inlet conduit.

[0485] F26. The liquid handling apparatus according to Clause F25, wherein liquid flow into the sample flow unit is controlled using a sample flow unit valve.

[0486] F27. The liquid handling apparatus according to Clause F25 or Clause F26, wherein the sample flow unit is in fluid communication with a sample flow unit waste chamber via a sample flow unit outlet conduit.

[0487] F28. The liquid handling apparatus according to Clause F27 when dependent on Clause F26, wherein liquid flow into the sample flow unit waste chamber is controlled using a sample flow unit valve.

[0488] F29. The liquid handling apparatus according to Clause F27 or Clause F28, wherein the sample flow unit waste chamber is in fluid communication with the second pneumatic port.

[0489] F30. The liquid handling apparatus according to any one of Clauses F1 to F29, further comprising a third chamber, wherein the third chamber is in fluid communication with the first pneumatic port.

[0490] F31. The liquid handling apparatus according to Clause F30, wherein the first pneumatic port is in permanent fluid communication with the third chamber.

[0491] F32. The liquid handling apparatus according to Clause F30 or Clause F31, wherein liquid flow into the third chamber is controlled using a third chamber valve.

[0492] F33. The liquid handling apparatus according to any one of Clauses F30 to F32, wherein the third chamber is in fluid communication with the first liquid reagent capsule.

[0493] F34. The liquid handling apparatus according to any one of Clauses F30 to F33, wherein the third chamber contains a dried reagent.

[0494] F35. The liquid handling apparatus of any of clauses F1 to F34, wherein the first liquid reagent capsule is in fluid communication with the first chamber via a prime conduit.

[0495] F36. The liquid handling apparatus of clause F35, wherein liquid flow through the prime conduit is controlled using a prime conduit valve.

[0496] F37. The liquid handling apparatus of clause F35 or clause F36 when dependent on clause F25, wherein the prime conduit is in fluid communication with the sample flow unit inlet conduit via the prime conduit valve.

[0497] F38. The liquid handling apparatus of any of clauses F1 to F37, wherein the third pneumatic port is in fluid communication with an air / wash solution supply conduit.

[0498] F39. The liquid handling apparatus of clause F38, wherein air flow through the air / wash solution supply conduit is controlled using an air supply valve.

[0499] F40. The liquid handling apparatus of clause F38 or clause F39, wherein the air / wash solution supply conduit is in fluid communication with the chamber outlet conduit.

[0500] F41. The liquid handling apparatus of any of clauses F1 to F40, wherein one or more of the first chamber and the second chamber contain a dry reagent.

[0501] G1. A method of metering a liquid in a channel of a liquid handling apparatus, the method comprising:

[0502] applying a first pressure to move a first liquid from a first chamber through a first fluid path comprising two junctions;

[0503] applying a second pressure to move fluid through a second fluid path comprising a portion of the first fluid path between the two junctions, such that the fluid displaces a volume of the first liquid between the two junctions.

[0504] G2. The method of clause G1, wherein the first pressure is applied using a first pneumatic port of the liquid handling apparatus, wherein the first pneumatic port is in selective fluid communication with the first chamber.

[0505] G3. The method of clause G1 or clause G2, wherein the second pressure is applied using a second pneumatic port of the liquid handling apparatus, wherein the second pneumatic port is in selective fluid communication with the second chamber.

[0506] G4. The method of any one of clauses G1-G3, wherein a second pressure is applied to dispense the fluid and the first liquid to a third chamber.

[0507] G5. The method of any one of clauses G1-G4, wherein the first pressure is applied to draw the first liquid from the first chamber through the first fluid path.

[0508] G6. The method of clause G5, wherein the first chamber is permanently vented.

[0509] G6. The method of any one of clauses G1-G4, wherein the first pressure is applied to dispense the first liquid from the first chamber through the first fluid path.

[0510] G7. The method of any one of clauses G1-G6, wherein the fluid is a second liquid.

[0511] G8. The method of clause G7, wherein a second pressure is applied to dispense the second liquid from a second chamber through a second fluid path.

[0512] G9. The method of any one of clauses G1-G6, wherein the fluid is air.

[0513] G10. The method of clause G9, wherein a second pressure is applied to supply air to the second fluid path through an air supply conduit.

[0514] G11. The method of any one of clauses G1-G10, further comprising metering a plurality of volumes of the first liquid by repeating one or more times:

[0515] applying the first pressure to move the first liquid from the first chamber through the first fluid path including the two junctions;

[0516] applying the second pressure to move the fluid through the second fluid path, the second fluid path including the portion of the first fluid path between the two junctions, such that the fluid displaces a volume of the first liquid between the two junctions.

[0517] H1. A method of priming a fluid circuit of a liquid treatment device, the method comprising:

[0518] applying a first pressure to move a liquid from a chamber to a first channel;

[0519] continuing to apply the first pressure to move the liquid until a meniscus of the liquid has moved past a junction on the first channel, wherein the junction connects the first channel to a second channel; and

[0520] applying a second pressure to move liquid from the chamber into the second channel via the first channel and the junction.

[0521] H2. The method of clause HI, wherein applying a first pressure comprises applying a negative pressure to move the liquid.

[0522] H3. The method of clause H2, wherein the chamber is permanently vented.

[0523] H4. The method of any of clauses HI to H3, wherein applying a second pressure comprises applying a negative pressure to move the liquid.

[0524] H5. The method of clause H4 when dependent on clause H2, wherein applying a first pressure comprises applying a negative pressure to a first pneumatic port of the liquid handling device, and wherein applying a second pressure comprises applying a negative pressure to a second pneumatic port of the liquid handling device.

[0525] H6. The method of clause HI, wherein applying a first pressure comprises applying a positive pressure to move the liquid.

[0526] H7. The method of clause H6, wherein applying a first pressure comprises applying a positive pressure to a first pneumatic port of the liquid handling device while venting a second pneumatic port of the liquid handling device.

[0527] H8. The method of clause H6 or clause H7, wherein applying a second pressure comprises applying a positive pressure to move the liquid.

[0528] H9. The method of clause H8 when dependent on clause H7, wherein applying a second pressure comprises applying a positive pressure to a first pneumatic port of the liquid handling device.

[0529] H10. The method of clause H9, wherein applying a second pressure comprises applying a positive pressure to a first pneumatic port of the liquid handling device while venting a third pneumatic port of the liquid handling device.

[0530] I1. A method of diluting a sample using a liquid handling device, the method comprising:

[0531] applying a negative pressure to draw a volume of a first liquid into a first flow cell of the liquid handling device;

[0532] applying negative pressure to draw a volume of the first liquid into a first chamber of the liquid handling device;

[0533] applying positive pressure to dispense a volume of a second liquid into the first chamber;

[0534] repeating the following steps one or more times:

[0535] applying positive pressure to dispense the first liquid and the second liquid from the first chamber to a second chamber; and

[0536] applying negative pressure to draw the first liquid and the second liquid from the second chamber to the first chamber;

[0537] applying positive pressure to move a mixture comprising the first liquid and the second liquid from the first chamber or the second chamber to the second flow cell.

[0538] I2. The method of clause II, further comprising measuring a detection signal in the first flow cell.

[0539] I3. The method of clause II or clause I2, further comprising measuring a detection signal in the second flow cell.

[0540] I4. The method of any one of clauses II to I3, wherein the volume of the first liquid drawn into the first chamber is a metered volume of the first liquid.

[0541] I5. The method of any one of clauses II to I4, wherein the volume of the second liquid dispensed into the first chamber is a metered volume of the second liquid.

[0542] J1. A method of resuspending dried reagents using a liquid handling device, the method comprising:

[0543] applying a first pressure to transfer a metered volume of a first liquid into a first chamber to resuspend one or more first dried reagents, wherein resuspending the one or more first dried reagents forms a first reagent;

[0544] applying a second pressure to transfer a metered volume of the first liquid into a second chamber to resuspend one or more second dried reagents, wherein resuspending the one or more second dried reagents forms a second reagent;

[0545] applying a third pressure to move the first reagent through at least one flow cell;

[0546] applying a fourth pressure to move the second reagent through the at least one flow cell; and

[0547] measuring a detection signal in the at least one flow cell.

[0548] J2. The method of clause J1, wherein the volume of the first liquid transferred into the first chamber is a metered volume of the first liquid.

[0549] J3. The method of clause J1 or clause J2, wherein the volume of the first liquid transferred into the second chamber is a metered volume of the first liquid.

Claims

1. A liquid handling apparatus, comprising: A fluid layer comprising: a plurality of channels, and an opening in the surface of the fluid layer, wherein the opening extends through at least a portion of the thickness of the fluid layer to one of the plurality of channels; Rigid layer, which includes an actuable part; and A reservoir capsule positioned between the fluid layer and the actuable part, wherein the reservoir capsule comprises: a body and a sealing layer; The main body defines the volume of the stored liquid, and the main body includes a first deformable portion, a main liquid storage portion, and a limiting portion connecting the first deformable portion to the main liquid storage portion; and The sealing layer is configured to seal the volume; The liquid storage capsule is positioned above the opening, such that a portion of the sealing layer covers the opening; The actuable portion includes a protrusion extending toward the reservoir capsule, wherein the actuable portion is actuated from a first position where the protrusion does not deform the first deformable portion to a second position where the protrusion deforms the first deformable portion into contact with the portion of the sealing layer, causing the portion of the sealing layer to rupture, wherein when the actuable portion is in the second position, the protrusion does not deform the restricting portion.

2. The liquid handling apparatus according to claim 1, wherein, The protrusion cannot penetrate the opening.

3. The liquid handling apparatus according to claim 1 or 2, wherein, The liquid reservoir is positioned above the opening such that the first deformable portion and the limiting portion are at least partially positioned above the opening.

4. The liquid handling apparatus according to any one of claims 1 to 3, wherein, The first deformable portion includes a first region and a second region, wherein a first distance between the first region and the sealing layer is greater than a second distance between the second region and the sealing layer.

5. The liquid handling apparatus according to claim 4, wherein, The first region and the second region are concentric, such that the first region surrounds the second region.

6. The liquid handling apparatus according to any one of claims 1 to 5, wherein, The width of the opening is smaller than the width of the protrusion.

7. The liquid handling apparatus according to any one of claims 1 to 6, wherein, The protrusion includes an end that engages with the first deformable portion of the reservoir capsule.

8. The liquid handling apparatus according to claim 7, wherein, The end of the protrusion is flat.

9. The liquid handling apparatus according to claim 7 or 8, wherein, The end of the protrusion has a main fan-shaped cross-section.

10. The liquid handling apparatus according to any one of claims 7 to 9, wherein, The protrusion includes a groove that defines the cross-section of the end of the protrusion.

11. The liquid handling apparatus according to claim 10, wherein, The groove is aligned with the limiting portion of the liquid storage capsule.

12. The liquid handling apparatus according to claim 10 or 11 when referring to claim 9, wherein, The groove defines the main fan-shaped shape of the cross-section of the end of the protrusion.

13. The liquid handling apparatus according to any one of claims 7 to 12, wherein, The cross-section of the end of the protrusion includes one or more outwardly extending protruding ribs.

14. The liquid handling apparatus according to claim 13 when referring to claim 9, wherein, The one or more outwardly extending protruding ribs extend from the curved portion of the main fan shape.

15. The liquid handling apparatus according to any one of claims 1 to 14, wherein: The protrusion is a first protrusion, and the actuable portion includes a second protrusion extending toward the reservoir capsule; The main body of the reservoir capsule includes a second deformable portion; The opening is a first opening, and the fluid layer includes a second opening that extends through at least a portion of the thickness of the fluid layer to reach one of the plurality of channels; The portion of the sealing layer is the first portion of the sealing layer, and the liquid reservoir is positioned above the second opening such that the second portion of the sealing layer covers the second opening; In the first position, the second protrusion does not deform the second deformable part; In the second position, the second protrusion deforms the second deformable portion to contact the second portion of the sealing layer, thereby causing the second portion of the sealing layer to rupture.

16. A liquid handling apparatus, comprising: A first liquid storage container and a second liquid storage container, wherein each of the first liquid storage container and the second liquid storage container is configured to be opened by forming an inlet and an outlet in the liquid storage container; and A fluid layer comprising: a channel network and pneumatic ports configured to receive positive pressure; The channel network includes: a first liquid storage container inlet channel and a second liquid storage container inlet channel; The first liquid reservoir inlet channel is configured to provide a fluid connection between the pneumatic port and the inlet in the first liquid reservoir when the first liquid reservoir is opened; and The second liquid reservoir inlet channel is configured to provide a fluid connection between the pneumatic port and the inlet in the second liquid reservoir when the second liquid reservoir is opened.

17. The liquid handling apparatus according to claim 16, further comprising: A first liquid storage container valve is configured to control the flow of liquid from the outlet of the first liquid storage container; and The second liquid storage container valve is configured to control the flow of liquid from the outlet of the second liquid storage container.

18. The liquid handling apparatus according to claim 17, wherein, The channel network also includes: A first liquid storage container outlet channel is configured to be in fluid communication with the outlet of the first liquid storage container when the first liquid storage container is opened, wherein the first liquid storage container valve is located on the first liquid storage container outlet channel.

19. The liquid handling apparatus according to claim 17 or 18, wherein, The channel network also includes: The second liquid storage container outlet channel is configured to be in fluid communication with the outlet of the second liquid storage container when the second liquid storage container is opened, wherein the second liquid storage container valve is on the second liquid storage container outlet channel.

20. The liquid handling apparatus according to any one of claims 16 to 19, wherein, The liquid handling apparatus further includes a well section, wherein the well section is at least partially defined by an opening extending through at least a portion of the thickness of the fluid layer, wherein the well section is in fluid communication with the pneumatic port, the first liquid storage container inlet channel, and the second liquid storage container inlet channel.

21. The liquid handling apparatus according to claim 20, wherein, The opening, the first liquid storage container inlet channel, and the second liquid storage container inlet channel are each disposed in the upper surface of the fluid layer, and wherein the depth of the opening is greater than the depth of the first liquid storage container inlet channel and the second liquid storage container inlet channel.

22. The liquid handling apparatus according to claim 20 or 21, wherein: The liquid handling apparatus further includes a lower portion, the lower portion comprising a groove in the upper surface of the lower portion; The fluid layer is positioned on top of the lower portion; and The well section is at least partially defined by an opening in the fluid layer and a groove in the lower section.

23. The liquid handling apparatus according to claim 22, wherein, The pneumatic port extends through the thickness of the fluid layer, wherein the pneumatic port is in fluid communication with the tank via a connecting channel in the lower portion.

24. The liquid handling apparatus according to any one of claims 16 to 23, wherein, Each of the first liquid storage container inlet channel and the second liquid storage container inlet channel includes: One or more first passage zones; and The second channel region, wherein the first depth of the one or more first channel regions is less than the second depth of the second channel region.

25. The liquid handling apparatus according to claim 24, wherein, The first width of one or more first channel regions is less than the second width of the second channel region.

26. The liquid handling apparatus according to any one of claims 16 to 25, wherein, The pneumatic port is a first pneumatic port, and the liquid handling device further includes a second pneumatic port configured to be in fluid communication with the outlet in the first liquid storage container.

27. The liquid handling apparatus according to claim 26, wherein, The second pneumatic port is configured to be in fluid communication with the outlet in the second liquid storage container.

28. The liquid handling apparatus according to claim 26 or 27, wherein, The liquid handling device further includes a third pneumatic port configured to be in fluid communication with the outlet in the second liquid storage container.

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