Fluid dispenser volume calibration device

Abstract

The technique relates, in part, to a device and related methods that can be used to accurately and precisely measure the volume of fluid dispensed from a fluid dispenser device, and in certain applications, to accurately and precisely measure the volume dispensed from a multi-channel fluid dispenser device. In an alternative embodiment, a fluid dispenser volume calibration device is provided, comprising: a front surface, a rear surface, a side surface, a first chamber, a second chamber, a hydrophobic membrane, and a channel, where: the first chamber comprises a front opening and a rear end, the front opening being provided at the front surface of the device; the second chamber comprises a rear end, a side wall and a front end; the hydrophobic membrane is in fluid connection with the second chamber; and the channel includes a proximal end in fluid connection with the first chamber and a distal end in fluid connection with the second chamber.

Description

[0001] Related Applications

[0002] This PCT international application claims priority to U.S. Provisional Patent Application Serial No. 63 / 342,889, filed May 17, 2022, under 35 U.S.C. § 119(e). The entire content of the above application is hereby expressly incorporated by reference herein for all purposes. All published texts, patents, and patent applications cited herein are hereby expressly incorporated by reference herein for all purposes. Technical Field

[0003] The present technology relates in part to an apparatus and related methods that can be used to accurately and precisely measure the volume of fluid dispensed from a fluid dispenser device, and in certain applications, the volume dispensed from a multi-channel fluid dispenser device. Such volume measurements can be used to calibrate fluid dispensing devices. Background Art

[0004] Fluid dispensers are commonly used in laboratory environments. Fluid dispensers are sometimes single-channel devices that deliver a volume of liquid to a single target location in one operation, and sometimes multi-channel devices that deliver a volume of liquid to each of multiple target locations in one operation. Summary of the Invention

[0005] Operators typically rely on fluid dispenser devices that accurately and precisely deliver volumes of fluid. In alternative embodiments, the volume calibration apparatus and methods provided herein can evaluate the accuracy and precision of the dispensed volume via low-error volume measurements. Without being limited by theory, in alternative embodiments, low-error volume measurements are provided by the effective transfer of sample fluid through the fluid path of the device and by other features.

[0006] In certain aspects, a fluid dispenser volume calibration apparatus is provided, comprising: a front surface, a rear surface, side surfaces, a first chamber, a second chamber, a hydrophobic membrane, and a channel, wherein: the first chamber includes a front opening and a rear end, the front opening being disposed at the front surface of the device; the second chamber includes a rear end, side walls, and a front end; the hydrophobic membrane is in fluid communication with the second chamber; and the channel includes a proximal end in fluid communication with the first chamber and a distal end in fluid communication with the second chamber.

[0007] In some aspects, a method for manufacturing the devices described herein is provided, which includes: providing a rear portion and a front portion; and joining the rear portion to the front portion. In some embodiments, the front portion includes a first chamber, and the rear portion includes a second chamber and a hydrophobic membrane. In some cases, the rear portion includes at least a part of a channel. In some aspects, a method for manufacturing the devices described herein is provided, including: providing a substrate, a front member, and a rear member; and joining the front surface of the rear member to at least a part of the rear surface of the substrate, and joining the rear surface of the front member to at least a part of the front surface of the substrate.

[0008] In some aspects, a method for determining the volume of fluid dispensed by a fluid dispenser device is provided, which includes: dispensing fluid from the fluid dispensing device into a first chamber of a volume calibration device that does not contain fluid, whereby the first chamber of the calibration device includes the dispensed fluid from the fluid dispenser device; determining the position of the fluid endpoint in the channel; and determining the volume of the dispensed fluid in the calibration device based on the position of the fluid endpoint. The dispensed fluid can flow from the first chamber into the channel, which includes a proximal end associated with the first chamber. The dispensed fluid can flow in the channel to a second chamber associated with the distal end of the channel. The dispensed fluid can flow in the channel under a pressure difference between the first chamber and the second chamber. The dispensed fluid can fill the second chamber and can fill a part of the channel in a continuous fluid path between the second chamber and the fluid endpoint until the fluid endpoint between the proximal end and the distal end of the channel.

[0009] Certain embodiments are further described in the following specification, claims, and drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] The drawings illustrate certain embodiments of the present technology, but are not limiting. For clarity and ease of illustration, the drawings are not drawn to scale, and in some cases, various aspects may be shown exaggerated or enlarged for ease of understanding a particular embodiment.

[0011] Figure 1 is a top perspective view of the volume calibration device 100. Figure 2A and Figure 2B each in is a top view of the device 100, Figure 3A and Figure 3B each in is a side view thereof, Figure 4A and Figure 4B each in is a bottom view thereof. Figure 5 is an exploded view of the device 100.

[0012] Figures 6 to 8D Shows the calibration device 100 for determining the volume of dispensed fluid. Figure 6A top perspective view of the device 100 associated with the pipette tip 200 containing the fluid 290. Figure 7 The fluid 290 dispensed from the pipette tip 200 into the volume calibration device 300 is shown. The device 300 has the same features as the device 100, except that the device 300 contains the dispensed fluid 290 and the device 100 does not contain the dispensed fluid. Figure 8A A top perspective view of the device 300 containing the dispensed fluid 290 after the pipette tip has been separated and before introducing a pressure difference into the device 300. Figure 8B A top perspective view of the device 300 showing the dispensed fluid 290 that has flowed in the fluid path in the device 300 after applying a pressure difference. Figure 8C Is Figure 8B A top perspective view and a partial exploded view of the device 300 shown in , showing the rear portion 338, which contains a part of the fluid path containing the dispensed fluid 290 and a part of the fluid path not containing the dispensed fluid. Figure 8D Is Figure 8C A top view of the rear portion 338 shown in .

[0013] Figure 9A A top view of the volume calibration device 400 containing an array of calibration units 460, Figure 9B A side view thereof, Figure 9C A bottom view thereof, Figure 9D A perspective view thereof, Figure 9E An exploded and enlarged view thereof.

[0014] Figure 10A A top view of the calibration device 500 containing an array of calibration units 501, Figure 10B A side view thereof, Figure 10C A bottom view thereof, Figure 10D A top perspective view thereof, and Figure 10E An upward perspective view thereof. Figure 11A A top exploded perspective view of the device 500, and Figure 11B Is Figure 11A An enlarged view of the portion defined by the dashed circle in . Figure 12A An exploded upward perspective view of the device 500, and Figure 12B Is Figure 12A An enlarged view of the portion defined by the dashed circle in . Figure 13A A top view of the device 500 showing the cutting plane A-A, and Figure 13B A cross-sectional view through the cutting plane A-A. Figure 13C , Figure 13D And Figure 13E Each is Figure 13B An enlarged view of a part defined by the respective dashed circle in . Figure 14Ais a top view of the device 500 showing the cutting plane J-J, Figure 14B is a cross-sectional view through the cutting plane J-J, and Figure 14C is Figure 14B an enlarged view of the portion defined by the dashed circle in Figure 15A is an enlarged top view of the device 500 showing the cutting plane L-L, Figure 15B is a cross-sectional view through the cutting plane L-L, and Figure 15C is Figure 15B an enlarged view of the portion defined by the dashed circle in

[0015] Figure 16A is a top view of the substrate 502 of the device 500 including an array of calibration units 501, Figure 16B is a side view thereof, Figure 16C is a bottom view thereof, Figure 16D is Figure 16A an enlarged view of the portion defined by the dashed circle in, and Figure 16E is Figure 16C an enlarged view of the portion defined by the dashed circle in Figure 17A is a side view of the front member 540 of the device 500, Figure 17B is a top view thereof, and Figure 17C is a top perspective view thereof. Figure 18A is a side view of the rear member 550 of the device 500, Figure 18B is a bottom view thereof, and Figure 18C is a bottom perspective view thereof.

[0016] Figure 19A is a top perspective view of the assembly 600 including the device 500 and the vacuum manifold 650, and Figure 19B is an exploded view thereof.

[0017] The following table describes exemplary elements of the devices shown in the drawings.

[0018]

[0019]

[0020]

[0021]

[0022]

[0023]

[0024]

[0025]

[0026] Like reference numerals in the various figures indicate like elements. DETAILED DESCRIPTION

[0027] In alternative embodiments, the devices and methods provided herein solve the problem of evaluating the liquid transfer performance from a liquid dispensing device. The devices and methods provided herein specifically solve the problem of evaluating the liquid transfer performance across multiple channels of a multi-channel fluid dispenser, typically for a single multi-channel dispensing event.

[0028] The devices and methods provided herein can evaluate the accuracy of the dispensed fluid volume (i.e., how closely the dispensed volume approaches the set volume of the liquid dispenser) and the precision of the dispensed fluid volume (i.e., how repeatable the dispensed volume is with respect to the set volume of the liquid dispenser). These parameters can be evaluated for multiple transfers from a single pipetting channel or for single or multiple transfers from a set of channels of a multi-channel liquid dispenser. Accuracy is typically reported as the percentage error of the average transfer volume relative to the set volume. Precision is typically reported as the coefficient of variation (CV), which is calculated by determining the standard deviation of multiple dispensed volumes (across a single channel or multiple channels) and dividing the standard deviation by the average dispensed volume, where the CV value is typically reported as a percentage. A user can use data from multiple dispensed volumes from the channel to evaluate the performance within the channel, and / or evaluate the performance within the channel for a single dispensed volume or two or more different dispensed volumes across multiple channels of a multi-channel liquid processor.

[0029] The devices and methods provided herein can be used to effectively evaluate the precision and accuracy of dispensed volumes with low error. The devices and methods provided herein can facilitate low-error measurements through the efficient transfer of sample fluid through the fluid path of the device and through other features.

[0030] The devices and methods provided herein can be used to calibrate fluid dispensing devices. For example, as part of the calibration process, a dispensing device that does not meet the precision and / or accuracy requirements of the dispensed volume can be adjusted and re-evaluated to determine whether the adjusted dispenser meets the required fluid dispensing requirements. Thus, the devices described herein are referred to as "calibration devices" because they can be used to perform calibration evaluations.

[0031] Volume Calibration Device

[0032] In certain embodiments, a fluid dispenser volume calibration device is provided, comprising: a substrate including a front surface, a rear surface, and side surfaces; a first chamber disposed in the substrate, the first chamber including a front opening disposed at the front surface of the substrate and a rear end (generally within the substrate); a second chamber disposed in the substrate, the second chamber including a rear opening disposed at the rear surface of the substrate and a front end (generally within the substrate); a hydrophobic membrane disposed at the rear surface of the substrate at the opening of the second chamber; and a channel disposed in the substrate. The channel generally includes a proximal end fluidly connected to the first chamber (e.g., fluidly connected to the rear end of the first chamber) and a distal end fluidly connected to the second chamber (e.g., fluidly connected to the front end of the second chamber; fluidly connected to the rear end of the second chamber). The proximal end of the channel may be connected to one side of the first chamber. The proximal end of the channel is sometimes connected to one side of a well, where the well includes a front opening disposed at a rear position of the first chamber. The proximal end of the channel is sometimes connected to the side of a hole and at the front end of the well. The well is sometimes disposed concentrically with respect to the front end of the first chamber. The well is sometimes integral with the first chamber, and the front end of the well may be connected to the rear end of the first chamber. The distal end of the channel is sometimes connected to one side of the second chamber, sometimes connected at one side and the front end of the second chamber, and / or sometimes connected at one side and the rear end of the second chamber.

[0033] The hydrophobic membrane is sometimes disposed outside the rear surface of the device. The hydrophobic membrane is sometimes disposed on the front surface of the substrate of the device and may be covered by another component of the device (e.g., a front component). The hydrophobic membrane and the opening of the second chamber are generally not disposed at the external front surface or external side surface of the device. The front end of the second chamber is sometimes disposed inside the device and sometimes not disposed at the external front surface of the device. The hydrophobic membrane is generally not disposed at the distal end of the channel. The hydrophobic membrane is sometimes disposed at the rear end of the second chamber, or sometimes disposed at the front end of the second chamber. The rear end of the second chamber sometimes extends together with the rear opening of the second chamber at a position on the rear surface of the device. In some cases, the rear surface of the device includes a recess, the recess including a recessed surface, and the rear opening of the second chamber is disposed at the recessed surface. The hydrophobic membrane may be disposed on the rear surface of the device surrounding the recess or may be disposed within the recess. The hydrophobic membrane is sometimes disposed adjacent to or at the end of the second chamber (e.g., the rear end of the second chamber and / or the rear end opening of the second chamber). The front end of the second chamber sometimes extends together with the front opening of the second chamber disposed at a front position of the substrate and / or at the rear surface of the front component. In some cases, the front surface of the substrate includes a recess, the recess including a recessed surface, and the front opening of the second chamber is sometimes disposed at or near the recessed surface. The hydrophobic membrane may be disposed in the recess, and the surface of the hydrophobic membrane (e.g., the rear surface of the hydrophobic membrane) is sometimes connected to the recessed surface within the recess.

[0034] In some embodiments, the device includes an optional vent connected to the end and / or opening of the second chamber, and a hydrophobic membrane is disposed at the end of the vent. For example, the substrate of the device may include an optional vent disposed between the end of the second chamber (e.g., the front end of the second chamber) and the front surface of the substrate. The front end of the optional vent may be disposed at the front surface of the substrate, or sometimes the front end of the optional vent is disposed at the recessed surface of a recess disposed at the front surface of the substrate. The proximal end of the optional vent is sometimes disposed at the front end and / or front opening of the second chamber. The optional vent is generally a void within the device, sometimes in the substrate, and has any suitable shape for fluid transmission, such as a cylinder or a frustum (e.g., a conical frustum). The surface of the hydrophobic membrane (e.g., the proximal surface of the hydrophobic membrane) is sometimes disposed at the front end of the optional vent (e.g., the hydrophobic membrane may be disposed in a recess within the substrate).

[0035] Channels generally include a length and a cross-sectional width, and generally include at least a portion that is much longer than the width. In at least a portion of the channel (e.g., at the expected location of the dispensing fluid end point (described herein)), the length of the channel is generally the primary dimension, and the width of the channel is generally the secondary dimension. In at least a portion of the channel (e.g., at the expected location of the dispensing fluid end point), the length of the channel is generally much larger than the width because a longer channel can provide an accurate determination of the dispensing volume. Without being limited by theory, relatively small differences in the dispensing volume can result in relatively large differences in the position of the dispensing fluid end point in a longer channel. The channel or a portion thereof is sometimes a microfluidic channel, and sometimes the channel or a portion thereof is a capillary or not a capillary.

[0036] The channel can be disposed in the substrate in any configuration suitable for identifying the fluid end point within the channel and suitable for achieving continuous fluid slugs (e.g., minimizing or preventing foam (e.g., bubbles) in the dispensed fluid) of the dispensed fluid in a calibration device. The channel is generally disposed in a configuration that minimizes the volume or surface area containing the channel, which is particularly applicable to devices containing an array of calibration units (described herein). In some embodiments, a single calibration unit 501 includes a single first chamber, a single axial duct, a single connected junction 570, and a single connected single axial duct 535, as shown in the lower right corner of Figure 11B and as shown in the lower right corner of Figure 16D and includes a single second chamber 516 and a single attached channel 524, as shown in the middle of Figure 12B , in the middle of Figure 13D and in the middle of Figure 13E and in the middle of Figure 16E and in the lower right corner of

[0037] The channels are sometimes helically disposed in the substrate, where the channels wind around a virtual center point from a proximal end to a distal end in a continuous and gradually widening curve. Sometimes, the virtual center point of the helically disposed channels is concentric with the virtual center point of the first chamber (e.g., the center point at the rear end of the first chamber) and / or the virtual center point of a well (described herein) disposed at a front position relative to the rear end of the first chamber to which the proximal end of the channel is connected. The helically disposed channels sometimes wind around a virtual center point on a virtual plane that coincides with the channel surface, which can be considered a two-dimensional channel orientation. The channels generally include a front surface, and sometimes the helically disposed channels wind around a virtual center point on a virtual plane that coincides with the front surface of the channel. In some cases, the helically disposed channels wind around a virtual axis perpendicular to the front surface on a virtual frustum or virtual cone that coincides with the surface of the channel (e.g., the front surface of the channel), which can be considered a non-planar three-dimensional orientation.

[0038] The channels can be configured in a non-helical configuration, such as a configuration including linear spans or substantially linear spans (also referred to herein as "linear members" or "substantially linear members", referred to as "spans" and "lateral members") each joined by a transition member, at least a portion of the transition member being sometimes curved and sometimes defined by a radius (e.g., also referred to herein as a "curved portion"). The lateral members are sometimes arranged in a folded-back orientation. The lateral members can be disposed in a virtual plane (two-dimensional orientation). The lateral members can be disposed in a non-planar three-dimensional orientation (e.g., one or more lateral members are disposed at an angle in a rear-to-front direction relative to another lateral member). Some or all of the lateral members are sometimes parallel or substantially parallel to each other in a calibration unit. The channels of the calibration unit can include any suitable number of lateral members, sometimes including 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more lateral members. The lateral members in the channels of the calibration unit sometimes have the same length or one or more different lengths. The channels of the calibration unit can include transition members between each lateral member, can include a transition member to a second chamber at the distal end of the channel, and can include a transition member from a first chamber or well at the proximal end of the channel.

[0039] In some embodiments, the channel includes a primary length and a cross-section perpendicular to the primary length. The cross-section and orientation of the channel in the substrate can be selected to maintain a continuous fluid slug of the dispensed fluid that has flowed within the device (e.g., to minimize or prevent the formation of foam (e.g., air bubbles) in the dispensed fluid). In some cases, the cross-sectional geometry and cross-sectional surface area of the channel are each independently substantially uniform or non-uniform along the length of the channel. In some embodiments, the cross-sectional surface area of the channel is non-uniform along the length of the channel. In some embodiments, the channel can include a first region (e.g., a recess) having a larger cross-sectional area adjacent to a second region (e.g., a detection region) having a smaller cross-sectional area, where the endpoint of the dispensed fluid is identified in the second region. For example, in the case of an expected dispensed fluid volume of 50 microliters, a portion of the channel can include a recess that holds 48 microliters, which transitions to an adjacent distally disposed detection region that can hold 4 microliters, which allows for precise measurement of the dispensed volume in the range of 48 to 52 microliters in the detection region while keeping most of the sample in the recess region. In some cases, the cross-sectional geometry of the channel is quadrilateral (e.g., rectangular, square) or oval (e.g., elliptical, circular) and / or can have tapered sides and / or curved sides (e.g., U-shaped, V-shaped). The channel cross-section can have a width (i.e., perpendicular to the front-to-back direction) and a depth (i.e., parallel to the front-to-back direction) that is the same as or different from the width, independently being from about 0.2 millimeters to about 1 millimeter (e.g., about 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, or 0.9 millimeters).

[0040] The first chamber has a suitable shape for facilitating the transfer of fluid into the channel and can be or include a cylinder, an ellipsoid, a cube, or a rectangular prism in some embodiments. The first chamber generally includes an inner wall that helps to dispense the fluid into the calibration device. The inner wall of the first chamber is generally smooth or substantially smooth and sometimes includes a hydrophobic surface. In some embodiments, the first chamber includes or is a frustum (e.g., a right frustum, a frustum of a cone, a frustum of a pyramid). In some cases, the first chamber is or includes a frustum (e.g., a frustum of a cone) and includes an inner wall that tapers in the front-to-back direction. The first chamber is sometimes a triangular prism or a triangular frustum (also referred to as a triangular first chamber), where the triangular portion is sometimes disposed at or near the front end and / or the rear end of the first chamber. The triangular first chamber sometimes includes a rear surface (also referred to as a bottom plate) that sometimes slopes towards the rear end of the first chamber in the front-to-back direction. One or more sides of the triangular first chamber sometimes taper in the front-to-back direction.

[0041] The first chamber is generally a void in the device, sometimes defined by one or more holes. The front opening of the first chamber is typically disposed at the front surface of the device. The front opening of the first chamber is sometimes disposed at the front surface of the front portion of the device, and sometimes the first chamber is defined by holes in the front portion (e.g., Figure 5 the device 100 shown in A). The front opening of the first chamber is sometimes disposed at the front surface of the front member of the device, and sometimes the first chamber is partially defined by holes in the front member and coextensive holes in the mating substrate of the device (e.g., Figure 13C the device 500 shown in).

[0042] The first chamber generally includes a rear opening disposed at the rear end of the first chamber. The rear opening disposed at the rear end of the first chamber is generally substantially smaller than the front opening of the first chamber. An optional well is sometimes disposed at the rear end of the first chamber. The well is sometimes disposed in the rearward direction relative to the rear end of the first chamber, where the front end of the well is sometimes disposed at the rear end opening of the first chamber. The well may include sides and a rear end, and the proximal end of the channel may be disposed at the side and / or rear end of the well. The well is generally a void, sometimes in the substrate and / or rear portion of the device, sometimes is or includes a cylinder, an ellipsoid, a cube, or a rectangular parallelepiped, sometimes is or includes a frustum (e.g., a right frustum, a frustum of a cone, a frustum of a pyramid), optionally having one or more sides that taper in the front-to-back direction. The proximal end of the channel may be disposed directly at the rear opening of the first chamber, the opening of the first chamber may be disposed at the rear end and / or side of the first chamber, and the device may not include a well adjacent to the first chamber.

[0043] The second chamber has a suitable shape to facilitate fluid transfer from the channel and may be a cylinder, an ellipsoid, a cube, or a rectangular parallelepiped in some embodiments, and may include or be a frustum (e.g., a right frustum, a frustum of a cone, a frustum of a pyramid) in some embodiments. The second chamber includes side walls, which are generally smooth or substantially smooth and permit fluid flow. In some cases, the second chamber is a frustum (e.g., a frustum of a cone) and includes an inner side wall that tapers in the rear-to-front direction (e.g., Figure 3A the second chamber in the device 100 shown in). The second chamber is sometimes a triangular prism or a triangular frustum (also referred to as a triangular second chamber), where the triangular portion is sometimes disposed at or near the front end and / or rear end of the second chamber (e.g., Figure 14A and Figure 13C the second chamber shown in). The second chamber (e.g., the triangular second chamber) sometimes includes a front surface (also referred to as a top plate), which sometimes faces the second chamber in the rear-to-front direction (e.g., Figure 13CThe front end of the second chamber (as shown in []) is inclined. One or more sides of the second chamber (e.g., triangular second chamber) sometimes taper in the rear-to-front direction (e.g., Figure 13C the second chamber of the device 500 shown in [].

[0044] The second chamber is generally a void in the device and is sometimes defined by one or more holes. The second chamber holes are sometimes provided in the rear part of the device and include openings at the front and rear surfaces of the rear part (e.g., Figure 3A and Figure 5 the device 100 shown in []. The second chamber holes are sometimes provided in the substrate of the device, sometimes include a hole opening at the rear surface of the substrate, and sometimes terminate within the substrate at a certain distance from the front substrate surface (e.g., Figure 13C the device 500 shown in []. The second chamber holes provided in the substrate of the device may include a hole opening at the rear surface of the substrate and may include an opposite opening at or near the front substrate surface or at the rear ventilation opening.

[0045] The rear opening of the second chamber is sometimes provided on the rear surface of the device and is sometimes not provided on the side of the second chamber (e.g., Figure 3A the device 100 shown in []. The rear opening of the second chamber is sometimes provided at the rear surface of the rear part of the device, and sometimes the second chamber is defined by a hole in the rear part (e.g., Figure 5 the device 100 shown in A in []. The rear opening of the second chamber is sometimes provided at the rear surface of the substrate of the device, and sometimes the rear end of the second chamber is partially defined by a hole in the substrate (which has an opening at the rear surface of the substrate) and a solid coextensive part of the connecting rear member of the device (e.g., Figure 13C the device 500 shown in [].

[0046] The second chamber may include an opening provided at or adjacent to the front end of the second chamber. In some embodiments, the opening is provided at the side and the rear end of the second chamber and at the distal end of the channel (e.g., Figure 5 the device 100 shown in A in []. In some embodiments, the opening is provided at the front end of the second chamber, sometimes not provided at the side of the second chamber, and sometimes is in fluid communication with the hydrophobic membrane (e.g., Figure 13E the device 500 shown in []. The surface of the hydrophobic membrane (e.g., the rear surface of the hydrophobic membrane) is sometimes provided at the release surface in the joint in the device. The front end opening of the second chamber can be directly provided at the release surface of the joint. In some embodiments, the front end opening of the second chamber is provided at the ventilation port, which is sometimes arranged in the forward direction relative to the front end of the second chamber (e.g., Figure 13CThe device 500 shown in []. The front end opening of the second chamber is sometimes provided at the rear end of the vent, and the front end of the vent is sometimes provided at the release surface of the joint or the front surface of the device or the substrate. The vent is usually a void, sometimes in the substrate, sometimes is or includes a cylinder, an ellipsoid, a cube or a cuboid, and sometimes is or includes a frustum (e.g., a regular frustum, a frustum of a cone, a frustum of a pyramid), optionally having one or more sides that taper in the direction from back to front.

[0047] The second chamber generally includes an opening provided at the rear end of the second chamber and / or adjacent to the rear end of the second chamber. The rear end opening of the second chamber is sometimes provided at the rear surface of the device, and sometimes provided at the rear surface of the second part of the device (e.g., Figure 3A the device 100 shown in []. The rear end opening of the second chamber is sometimes provided at the hydrophobic membrane (e.g., Figure 3A the device 100 shown in [].

[0048] The second chamber sometimes includes an opening provided at the rear end and the side wall of the second chamber, and is provided at the distal end of the channel (e.g., Figure 13C the device 500 shown in []. The rear end of the second chamber is sometimes defined by the hole opening at the rear surface of the substrate and the solid coextensive part of the joined rear member (e.g., Figure 13C the device 500 shown in []. In some embodiments, the second chamber hole opening and the channel cavity are provided at the rear surface of the substrate, and the rear end of the second chamber and the rear surface of the channel are defined by the solid coextensive part of the front surface of the rear member (e.g., Figure 12B and Figure 13C the device 500 shown in [].

[0049] The calibration device generally includes fluid paths from the first chamber to the channel and from the channel to the second chamber. In some embodiments, the second chamber includes sides, and the distal end of the channel is provided at the sides and the front end of the second chamber (e.g., Figure 5 the device 100 shown in []. In some cases, the front end of the second chamber coincides with the front surface of the distal end of the channel or is disposed in a rear position relative to the front surface of the distal end of the channel. In some embodiments, the second chamber includes sides, and the distal end of the channel is provided at the sides and the rear end of the second chamber (e.g., Figure 12Bthe device 500 shown in). In some cases, the rear end of the second chamber coincides with the rear surface of the distal end of the channel or is disposed in a rearward position relative to the rear surface of the distal end of the channel. This orientation of the distal end of the channel and the second chamber can help the second chamber to be completely filled with the dispensed fluid when the calibration device is in use. Each of the channel, the first chamber, the optional well, and the second chamber is generally located at a fixed position within the device and is non-movable within the device. The proximal and distal ends of the channel are generally in fixed (i.e., non-movable) positions relative to the first chamber, the optional well, and the second chamber. In some cases, each of the channel, the first chamber, the optional well, and the second chamber is disposed at a fixed position within the overall device.

[0050] In some cases, the calibration device includes a well fluidly connected to the first chamber. In some embodiments, the calibration device includes fluid paths from the first chamber to the well, from the well to the channel, and from the channel to the second chamber. In some cases, the well includes a side surface, and the proximal end of the channel is disposed at the side surface of the well. In some embodiments, the well includes a front opening, the front opening of the well is disposed in a rearward position relative to the rear end of the first chamber, and sometimes the front opening of the well is concentrically aligned with the rear end of the first chamber (e.g., Figure 3A the device 100 shown in). In some embodiments, the well includes a front opening, the front opening of the well is disposed in a rearward position relative to the rear end and the rear opening of the first chamber, and sometimes the front opening of the well is not concentrically aligned with the rear end of the first chamber (e.g., Figure 13D the device 500 shown in). The well is sometimes integral with the first chamber, wherein the rear end of the first chamber engages the front end of the well, and sometimes the side surface of the well engages the side surface of the first chamber. The rear end of the well sometimes coincides with the rear end of the channel at the proximal end of the channel (at the connection of the channel and the well) or is disposed in a forward position relative to the rear end of the channel, which can facilitate the emptying of the dispensed fluid from the first chamber and the well after the fluid flows through the calibration device during use. In embodiments where the device does not include a well, the rear end of the first chamber sometimes coincides with the rear end of the channel at the proximal end of the channel (at the connection of the channel and the first chamber) or is disposed in a forward position relative to the rear end of the channel, which can facilitate the emptying of the dispensed fluid from the first chamber after the fluid flows through the calibration device during use.

[0051] A hydrophobic membrane is sometimes disposed at or near the rear surface or the front surface of the device. For embodiments in which fluid is pushed through the device by a pressure difference, a vacuum device is sometimes used to provide the pressure difference. The vacuum device is sometimes disposed in the assembly having the calibration device at or near the hydrophobic membrane. In some embodiments, the hydrophobic membrane is sometimes disposed at the rear surface of the calibration device, and the vacuum device is disposed in the assembly having the calibration device at or near the hydrophobic membrane (e.g., Figure 3Athe apparatus 100) shown in. In such an embodiment, due to the proximity of the vacuum device to the hydrophobic membrane, the calibration device generally does not include ducts associated with the hydrophobic membrane and the other surface of the device. In certain embodiments, the hydrophobic membrane is disposed at or near the front surface of the calibration device, and the vacuum device is disposed at or near the rear surface of the calibration device in a component having the calibration device (e.g., Figure 13E the apparatus 500 shown in and Figure 19A the component 600 shown in). In such an embodiment, since the vacuum device is separated from the hydrophobic membrane, the calibration device generally includes one or more ducts associated with the hydrophobic membrane and the rear surface of the device. The ducts are typically voids in the device (e.g., voids in the substrate of the device) and have any suitable geometry that helps to apply a vacuum and drive fluid through the device. In certain embodiments, the ducts are sometimes or include cylinders, elliptical cylinders, cubes, or rectangular parallelepipeds, and may include or be frustum cones (e.g., right frustum cones, frustum cones, frustum pyramids). The walls of the ducts may taper in a back-to-front direction or in a front-to-back direction. The ducts sometimes include one or more curved portions. The ducts are sometimes defined by voids in the substrate and the solid surfaces of fused rear and / or front members. The ducts sometimes contact the hydrophobic membrane directly, and / or sometimes are indirectly associated with the hydrophobic membrane by a void volume. Indirect association by a void volume means that the ducts are indirectly connected to the hydrophobic membrane via another void in the device.

[0052] The calibration device may include lateral ducts (perpendicular or substantially perpendicular to the front-to-back direction), or axial ducts (parallel or substantially parallel to the front-to-back direction), or a combination of lateral and axial ducts, which are directly or by void volume associated with the hydrophobic membrane. In some cases, the device includes ducts directly connected to the hydrophobic membrane, and sometimes the ducts are connected to the side or rear surface of the hydrophobic membrane. In some cases, the device includes ducts connected to a junction that is directly connected to the ducts and directly or indirectly connected to the hydrophobic membrane. In some cases, the device includes a junction containing the hydrophobic membrane that connects the front end opening of the second chamber or an optional vent opening to a duct opening, such as a lateral duct opening. In some cases, the device includes a junction connecting a first duct opening to a second duct opening, where the first and second ducts are aligned in different orientations in the device. In certain embodiments, the lateral ducts are associated with the axial ducts. The junctions are sometimes defined by holes and / or recesses in the substrate of the device and the solid coextensive portions of the front or rear members adjacent and / or covering the holes and / or recesses (e.g., Figure 11B and Figure 13C the junction 570 of the apparatus 500 shown in). The ducts are sometimes defined by holes and / or recesses in the substrate of the device and the solid coextensive portions of the front or rear members adjacent and / or covering the holes and / or recesses (e.g., Figure 11Band Figure 13C the lateral conduit 546 in the apparatus 500 shown in, and Figure 14C the axial conduit 535) shown in. The conduits sometimes include openings in a substrate of the apparatus, and the openings are arranged adjacent to, coextensive with, and / or coaxial with holes in a front member or a rear member (e.g., the rear end 538 of the axial conduit is coaxially aligned with the rear member hole 556 in the apparatus 500, as Figure 14C shown).

[0053] In some embodiments, the substrate includes a front portion and a rear portion fused to the front portion (e.g., Figure 5 the apparatus 100 shown). The front portion sometimes includes a first chamber, and the rear portion sometimes includes a second chamber and a hydrophobic membrane. The rear portion sometimes includes at least a portion of a channel. In some cases, the channel includes a front surface, a rear surface, and side surfaces, where the rear surface and the side surfaces of the channel are disposed in the rear portion, and where the front surface of the channel is defined by the rear surface of the front portion joined to the rear portion. In some embodiments, a well is disposed in the rear portion of the calibration apparatus.

[0054] In some embodiments, the apparatus includes a substrate, a front member fused to all or a portion of the front surface of the substrate, and a rear member fused to all or a portion of the rear surface of the substrate (e.g., Figure 13CThe device 500 shown in [figure reference]. The substrate sometimes includes a first channel hole having a front opening disposed at the front substrate surface, an optional well associated with the first chamber, a channel cavity disposed at the rear substrate surface and associated with the first chamber or the optional well, a second channel hole having a rear opening disposed at the rear substrate surface and associated with the channel, an optional vent associated with the second channel hole, an optional lateral duct cavity, an optional axial duct, and an optional junction between the second chamber or the optional vent and the optional lateral duct. The front member may include a hole that coextends with the front first chamber hole opening, may include a solid portion adjacent to and / or covering the hydrophobic membrane, an optional junction between the second chamber or the optional vent and the optional lateral duct, may include a solid portion adjacent to and / or covering the optional lateral duct cavity, and / or may include a solid portion adjacent to and / or covering the front opening of the optional axial duct. The rear member may include a hole adjacent to and / or coextending with the rear end of the optional axial duct, may include a solid portion adjacent to and / or covering the rear hole opening of the second chamber of the substrate, may include a solid portion adjacent to and / or covering the channel cavity and / or recess of the substrate, and may include a solid portion adjacent to and / or covering the rear opening of the rear first chamber or the rear opening of the optional well. The fluid flow path in the calibration device is typically from the first chamber to the optional well, to the channel, and to the second chamber. Each of the first chamber, the optional well, the channel, and the second chamber is typically a void within the calibration device. The fluid flow path sometimes includes an optional vent, which is typically a void in the device between the second chamber and the hydrophobic membrane. The fluid flow path typically reaches the hydrophobic membrane in the device and typically terminates at the hydrophobic membrane in the device. The fluid flow path (e.g., from the first chamber to the optional well, to the channel, to the second chamber, to the optional vent, and to the hydrophobic membrane) is typically substantially smooth and typically contains smooth transitions that facilitate fluid flow and negligible retention of fluid in the portion of the fluid flow path near the fluid terminus (described herein). The inner surfaces in the fluid flow path typically contain smooth transitions, such as curved surfaces, tapered surfaces, flared surfaces, etc. The inner surfaces in the fluid flow path typically do not contain cavities, voids, or sharp transitions that may retain fluid in the region of the flow path near the fluid terminus disposed between the proximal end and the distal end of the channel. In some embodiments, the channel includes an inner surface, and at least a portion of the inner surface of the channel is hydrophobic (e.g., a portion or all of the inner surface of the channel is hydrophobic). In some cases, each of the first chamber and / or the second chamber includes an inner surface, and at least a portion of the inner surface of the first chamber and / or the second chamber is hydrophobic (e.g., a portion or all of the inner surface of the first chamber and / or the second chamber is hydrophobic). In some cases, the optional well includes an inner surface, and at least a portion of the inner surface of the well is hydrophobic (e.g., a portion or all of the inner surface of the well is hydrophobic).In some cases, the optional vent includes an inner surface, and at least a portion of the inner surface of the vent is hydrophobic (e.g., a portion or all of the inner surface of the vent is hydrophobic). In some cases, the inner surfaces of the chamber (first chamber, second chamber), the channel, the optional well, and / or the optional vent are not coated and are sometimes the surfaces of cavities within a substrate containing a hydrophobic material. In some cases, the substrate includes a hydrophobic material; the channel, the first chamber, the optional well, the second chamber, and the optional vent are defined by voids within the substrate; and the sidewalls of the channel, the first chamber, the optional well, the second chamber, and the optional vent include a hydrophobic material. In some cases, a portion of the channel and / or the second chamber is partially defined by a rear member, and at least a portion of the rear member that defines the portion of the channel and / or the second chamber includes a hydrophobic material. In some cases, at least a portion of the inner surface of the channel, the first chamber, the optional well, the second chamber, and / or the optional vent includes a coating containing a hydrophobic material. In some cases, a portion of the channel and / or the second chamber is partially defined by a rear member, and at least a portion of the rear member that defines the portion of the channel and / or the second chamber includes a coating containing a hydrophobic material. Any hydrophobic material can be selected that is suitable for fluid transport through the channel, the first chamber, the optional well, the second chamber, and / or the optional vent, and / or for forming a continuous fluid volume (i.e., a fluid slug) away from the fluid distribution end point and near the hydrophobic membrane. Such a hydrophobic material can facilitate the formation of a continuous fluid volume by excluding or substantially excluding the dispensed fluid from the fluid flow path near the fluid distribution end point, and / or by excluding or substantially excluding detectable foam (e.g., air bubbles) from the dispensed fluid volume away from the fluid distribution end point and near the hydrophobic membrane. Non-limiting examples of hydrophobic materials are moldable plastics such as polypropylene or polystyrene. In some embodiments, the surface of one or more of the channel, the first chamber, the optional well, the second chamber, and / or the optional vent is hydrophilic, and optionally, the surface of one or more of the channel, the first chamber, the optional well, the second chamber, and / or the optional vent is hydrophobic.

[0055] The hydrophobic membrane can include any hydrophobic material suitable for one or more of the following: (i) fluid flow through the channel into the second chamber; (ii) filling the second chamber and an optional vent without allowing the fluid to escape from the second chamber and the optional vent through the hydrophobic membrane; (iii) applying a pressure difference in the device (e.g., applying a pressure difference between the pressure in the device and the pressure outside the device); (iv) forming a continuous fluid volume (i.e., a fluid slug) away from the end point of the dispensed fluid and close to the hydrophobic membrane; and (v) excluding or substantially excluding detectable foam (e.g., air bubbles) in the dispensed fluid volume away from the end point of the dispensed fluid and close to the hydrophobic membrane. The hydrophobic membrane is typically porous, which can facilitate applying a pressure difference in the calibration device. In some cases, the hydrophobic membrane includes polytetrafluoroethylene (PTFE) and has an average pore size of from about 0.05 microns to about 0.5 microns (e.g., about 0.25 microns). In some embodiments, the hydrophobic membrane is a disk.

[0056] The hydrophobic membrane sometimes includes a width (e.g., a minor width) that is greater than the width of the opening in the device in which the membrane is disposed (e.g., a major width). In some cases, the second chamber includes a rear opening that is smaller than the width of the hydrophobic membrane in contact with the opening (e.g., Figure 3B the device 100 shown in ). The perimeter of the front surface of the hydrophobic membrane in contact with the rear surface of the substrate is sometimes joined to the rear surface of the substrate. The hydrophobic membrane is sometimes concentric with the rear opening of the second chamber of the calibration device. In some cases, the second chamber includes a front end opening and / or an optional vent front opening that is smaller than the width of the hydrophobic membrane in contact with the opening (e.g., Figure 13E the device 500 shown in ). In some embodiments, the hydrophobic membrane is disposed in a recess that is disposed at or near the surface of the device. The hydrophobic membrane is sometimes disposed in a recess that is disposed in the front surface of the substrate, and the rear surface of the membrane sometimes contacts the surface of the recess (also referred to as the recessed surface) and / or the front end opening of the second chamber or the optional vent is sometimes disposed at the recessed surface (e.g., Figure 13E the device 500 shown in ).

[0057] In some embodiments, the fluid path in the calibration device includes a first chamber, a second chamber, and a channel that includes a proximal end fluidly connected to the first chamber and a distal end fluidly connected to the second chamber. In some cases, the fluid path in the calibration device includes a first chamber, a second chamber, a well fluidly connected to the back end of the first chamber, and a channel that includes a proximal end fluidly connected to the well and a distal end fluidly connected to the second chamber. In some embodiments, the fluid path in the calibration device includes a first chamber, a second chamber, a well disposed at the back end of the first chamber and connected to the first chamber, and a channel that includes a proximal end connected to the sidewall of the well and a distal end connected to the sidewall of the second chamber. The fluid path typically terminates at a hydrophobic membrane in the calibration device. In some cases, the fluid path in the calibration device includes an optional vent hole between the second chamber and the hydrophobic membrane. In some embodiments, (i) the first chamber and / or the well is connected to a single channel and not to multiple channels, (ii) the second chamber is connected to a single channel and not to multiple channels, (iii) the first chamber and / or the well and the second chamber are connected to one channel and the same channel, (iv) the channel does not branch, and (v) a combination of two, three, or all of (i), (ii), (iii), and (iv).

[0058] In most cases, the fluid path extends from the first chamber and terminates at the hydrophobic membrane. In most cases, after the dispensed fluid has flowed through the calibration device, the continuous fluid volume extending from the fluid end point in the channel (i.e., the end point disposed between the proximal and distal ends of the channel) to the distal end of the channel, to the second chamber, to the optional vent, and ending at the hydrophobic membrane is determined based on the position of the fluid end point in the channel (as described herein). In this embodiment, the dispensed fluid volume is defined from the end point at the hydrophobic membrane back to the fluid end point within the channel.

[0059] The fluid path is generally within a cavity or void in the calibration device, and in some cases, the entire inner surface of the cavity is hydrophobic, for example, being disposed in a hydrophobic substrate and / or coated with a hydrophobic coating. The entire cavity in the calibration device sometimes does not include a hydrophilic surface and sometimes does not include capillary channels (e.g., channels through which fluid flows by capillary action). In some embodiments, the fluid is urged to flow through the calibration device by an externally applied force. The propulsion of fluid flow in the calibration device is sometimes provided by a pressure difference in the calibration device including the dispensed fluid. The pressure difference is sometimes between the pressure in the fluid flow path cavity in the calibration device and the pressure outside the device, and sometimes between the pressure in the first chamber and the pressure at the hydrophobic membrane. The pressure difference can be generated by exposing the calibration device to a vacuum (as described herein). In some embodiments, the calibration device includes a vacuum seal, which is sometimes (i), a protrusion or rib disposed at a side in the rear region of the calibration device and sometimes surrounding the perimeter of the calibration device, (ii), a protrusion or rib disposed at or near the hydrophobic membrane on the rear surface of the calibration device (e.g., an annular protrusion, a rib grid), or (iii), a combination of (i) and (ii). In some cases, the fluid is urged to flow through the calibration device by exposing the calibration device to a centrifugal force (e.g., generated by placing the calibration device in a centrifuge). In some cases, the fluid is not urged to flow through the calibration device by capillary action.

[0060] In certain embodiments, an external force (e.g., differential pressure, vacuum) is applied to the calibration device to push fluid through the fluid path of the device, and the fluid delivered to the device is retained within the device. In certain embodiments, a measurable amount of fluid does not leave the device. In certain embodiments, a measurable amount of fluid does not leave the device through a hydrophobic membrane disposed at an end of the fluid path. Fluid typically flows within the device in a first chamber and an optional well and from the first chamber and the optional well in a front-to-back direction, and typically flows in a lateral direction (i.e., perpendicular to the front-to-back direction) within a channel. Fluid sometimes flows in a front-to-back direction within a second chamber to an inner surface (e.g., front surface) of the hydrophobic membrane. In certain cases, fluid flows in a back-to-front direction within the second chamber and optionally in a back-to-front direction within an optional vent to the surface of the hydrophobic membrane (e.g., back surface). After the fluid flows within the device and the flow of the fluid stops, the fluid is typically disposed within the device as a continuous (i.e., non-interrupted) fluid volume having a proximal end and a distal end, where the proximal end is disposed within the channel as a fluid terminus, and the distal end is disposed at the inner surface (e.g., front surface, back surface) of the hydrophobic membrane. After the fluid flow stops, the fluid terminus is typically not disposed within the first chamber or the optional well, and the fluid terminus is typically disposed within a portion of the channel between the distal end and the proximal end of the channel. After the fluid flow stops, the fluid typically fills a portion of the channel in a distal orientation up to the channel fluid terminus, fills the second chamber, and fills the optional vent (if present) up to the surface of the hydrophobic membrane. The void volume from the proximal end of the channel to the inner surface of the hydrophobic membrane at the end of the fluid flow path (e.g., the total void volume of the void volumes of the channel, the second chamber, and the optional vent (if present)) is typically greater than the volume of fluid delivered to the calibration device.

[0061] In a specific embodiment, Figures 1 to 5 Device 100 in shows the features of a volume calibration device. Device 100 does not include dispensed fluid from a fluid dispenser. Device 100 includes a substrate 102, a substrate front surface 104, a substrate back surface 106, and a substrate side surface 108. In the front portion, device 100 includes a first chamber 110, and first chamber 110 includes a front opening 112, a back end 114, and an inner wall. First chamber 110 is a frustum (i.e., a right frustum, a frustum of a cone), having an inner wall that tapers from the front opening 112 to the back end 114.

[0062] At a rear position relative to the back end 114 of the first chamber 110, disposed within device 100 is a well 130 that includes a side 132 and a front opening 134. Well 130 is a cylinder, the back end of the well having the same surface area as the front opening 134, and side 132 of well 130 is perpendicular to the front surface 104.

[0063] In the rear portion, the device 100 includes a second chamber 116, which includes an opening 118, a front end 120, and an inner wall. The second chamber 116 is a frustum (i.e., a right frustum, a frustum of a cone), having an inner wall that flares from the rear opening 118 towards the front end 120. At the rear surface, and disposed at the rear opening 118, the device 100 includes a bonded hydrophobic membrane 122. The device 100 includes a front portion 136 that is joined to the rear portion 138. The front portion 136 includes a rear surface 140, and the rear portion 138 includes a front surface 142. The device 100 includes a joint 150 between the rear surface 140 and the front surface 142, where the surface 140 is joined to the surface 142 at the joint 150.

[0064] Within the body of the substrate 102, the device 100 includes a channel 124, which includes a proximal end 126 disposed at a side 132 of the well 130. The channel 124 includes a distal end 128, which is disposed at the rear end 120 and at the side of the second chamber 116. The channel 124 is helically disposed within the substrate 102, where the channel 124 winds around a virtual center point 190. The virtual center point 190 is concentric with the center point of the rear end 114 of the first chamber 110, and is concentric with the center point of the front opening 134 of the well 130. The channel 124 includes a channel front surface 144, which is disposed at the rear surface 140 of the front portion 136 and is defined by the rear surface 140 of the front portion 136. The channel 124 further includes a channel rear surface 146 and a channel side surface 148, which are disposed within the rear portion 138. The device 300 includes the same features as the device 100 (e.g., the channel 124 in the device 100 corresponds to the channel 324 of the device 300, the first chamber 110 of the device 100 corresponds to the first chamber 310 in the device 300, etc.), except that the device 300 includes a dispensed fluid 290 while the device 100 does not include a dispensed fluid.

[0065] In certain embodiments, the volume calibration device includes at least one calibration unit, where the calibration unit includes a first chamber, a second chamber, and a channel, and optionally includes a hydrophobic membrane. In some cases, the calibration device includes two or more calibration units. In the calibration device, multiple calibration units can be arranged in series (e.g., a single row of calibration units, for use with a manually operated multi-channel dispenser) and / or in an array arrangement (e.g., for use with an automated multi-channel dispenser). The device sometimes contains an array of about 8 calibration units to about 1536 calibration units (e.g., 8, 12, 96, 384, 1536 calibration units). The calibration unit array is typically a two-dimensional array, which can include any suitable number of calibration units arranged in rows and columns (e.g., an array containing 8 rows and 12 columns of units, an array containing 32 rows and 48 columns of units, an array containing 128 rows and 192 columns of units).

[0066] In an apparatus including a series of calibration units and / or an array of calibration units, the center of each calibration unit is typically aligned with a corresponding dispenser element that dispenses fluid into the apparatus. A calibration device having standardized dimensions can be registered relative to a fluid dispensing device (i.e., a liquid processor device) that has a seat configured to receive the calibration device with precise alignment. The perimeter of a calibration device having standardized dimensions sometimes conforms to the footprint of a Standardized Experimental Chamberware (i.e., 5.03 inches by 3.365 inches) of the Society for Biomolecular Screening (SBS). In some embodiments, the centers of each calibration unit are spaced a uniform distance from the centers of adjacent calibration units in the series of calibration units and / or the array of calibration units. In some embodiments, the centers of each calibration unit are spaced 9 millimeters or 4.5 millimeters from the centers of adjacent calibration units.

[0067] In a particular embodiment, the features of an apparatus 400 including an array 460 of calibration units are shown in Figures 9A to 9E FIG. 6. The apparatus 400 includes a substrate 402 that includes a front surface 404, a rear surface 406, and side surfaces 408. The apparatus 400 includes a plurality of calibration units 460 arranged in an 8×12 array (i.e., 96 calibration units 460). Each calibration unit 460 includes a first chamber 410 that includes a front opening 412 and a rear end 414. Rearward of the first chamber 410 in each calibration unit 460 is a well 430 that includes a well side surface 432 and a well front opening 434. Each calibration unit 460 also includes a second chamber 416 that includes a rear opening 418 and a front end 420. Each calibration unit 460 includes a hydrophobic membrane 422 that is joined to the rear surface 406 at the rear opening 418 of the second chamber 416. The apparatus 400 includes a front portion 436 that is joined to a rear portion 438 at a junction 450. The front portion 436 includes a rear surface 440, and the rear portion 438 includes a front surface 442, where surface 440 is joined to surface 442. The perimeter dimensions (i.e., length and width) of the rear portion 438 define the SBS footprint, where the centers of each calibration unit 460 are spaced 9 millimeters from the centers of adjacent calibration units 460. In each calibration unit 460, the well 430 is connected to a channel 424 that includes a channel proximal end 426 (connected to the well 430), a channel distal end 428 (connected to the second chamber 416), a channel front surface 444 (defined by the rear surface 440), and a channel rear surface 446 and channel side surfaces 448 (disposed in the rear portion 428). The channel 424 is disposed helically around a virtual center point 490 in each calibration unit 460.

[0068] In a particular embodiment, the features of a volume calibration device 500 are shown as Figures 10A to 18C FIG. 7. Figures 10A to 18CThe volume calibration device 500 shown does not contain fluid. The device 500 includes an array of calibration units, where each calibration unit 501 is capable of receiving a separate dispensed fluid volume to determine the delivered fluid volume. The device 500 includes a substrate 502. The front surface 504 of the substrate is joined to the rear surface 542 of the front member 540. The rear surface 506 of the substrate is joined to the front surface 552 of the rear member 550. The substrate 502 includes outer wall outer surfaces 508 and 509.

[0069] The substrate 502 includes a first chamber aperture 503 of the substrate, and the first chamber aperture 503 includes an aperture opening 505 disposed at the front surface 504 of the substrate. Each first chamber 510 (also referred to as a "sample port") of the device 500 is defined in part by the aperture 503 in the substrate 502 and a coextensive mating aperture 545 of the joined front member 540. Each aperture opening 512 in the substrate 502 and each coextensive mating aperture 545 of the joined front member 540 are triangular. Each first chamber 510 includes a front opening 512, a rear surface 513 (also referred to as a "bottom plate"), a rear end 514, and a rear opening 511 (also referred to as a port) disposed at the end 514. The rear surface 513 is triangular and slopes from one side of the first chamber 510 towards the opposite side of the first chamber 510 and slopes to the rear end 514 in the front-to-rear direction 599, facilitating fluid transfer and evacuation from the first chamber 510. At least one sidewall 515 of the first chamber 510 tapers in the front-to-rear direction 599, which can also facilitate fluid transfer and evacuation from the first chamber 510.

[0070] The rear portion of the first chamber 510 is joined to the front portion of an optional well 530 at the opening 511. The well 530 is a void or aperture in the substrate 502 and is a frustum of a cone including sidewalls 532 that taper from a front end 534 to a rear end 533 in the front-to-rear direction 599. The rear opening 511 of the first chamber 510 is disposed at the front end 534 of the well. The well 530 includes a rear port 533' disposed at the rear end 533 of the well, at the well sidewalls 532, and at the proximal end 526 of the channel. The solid portion of the front surface 552 of the rear member 550 disposed below the well 530 forms the inner rear bottom surface of the well at the rear end 533, as Figure 13D shown.

[0071] The well rear port 533' is disposed at the proximal end 526 of the channel 524. The channel 524 (also referred to as a "sample channel") includes a distal end 528 and an inner wall 529. The channel 524 is a void or groove disposed in the substrate 502 in a folded-back orientation, which includes a lateral member 525 and a transition member 527. Figure 12BThe channel 524 shown in [Fig.] includes three lateral members 525 having different lengths, each lateral member being shorter than the previous adjacent lateral member in the proximal-to-distal direction of the channel 524. The lateral members 525 each have a length different from that of another lateral member 525 in a single calibration unit 501, and have successively smaller lengths from the proximal end 526 of the channel to the distal end 528 of the channel. Each of the two lateral members 525 is connected by a curved transition member 527, and the channel 524 in the device 500 includes a rectangular cross-section. The channel 524 includes a front surface 560, a rear surface 562, and side walls 564 as inner surfaces. The solid portion of the front surface 552 of the rear member 550 disposed below the channel 524 forms the inner rear bottom surface 562 of the channel, as Figure 13D shown.

[0072] The distal end 528 of the channel 524 is disposed at the second chamber 516. The second chamber 516 is partially defined by a hole 517 including a rear opening 519. The second chamber 516 includes a rear end 518, a front surface 521 (also referred to as the "top plate"), a front end 520, and side walls 523. The second chamber 516 includes two openings: a rear opening 531 disposed at the side wall and the rear end 518 of the second chamber 516, and a front opening 520' disposed at the front end 520 and the front surface 521. The rear opening 519 is triangular. The front surface 521 is triangular and slopes and inclines from one side of the second chamber 516 toward the opposite side of the second chamber 516 in the rear-to-front direction 598 to the front end 520, thereby facilitating fluid transfer to the front opening 520'. At least one side wall 523 of the second chamber 516 tapers in the rear-to-front direction 598, as Figure 14C and Figure 15C shown (since the side walls taper from the cutting plane in the rear-to-front direction, the rear portion of the side wall 523 of the second chamber 516 is shown). The solid portion of the front surface 552 of the rear member 550 disposed below the second chamber 516 forms the inner rear bottom surface at the rear end 518 of the second chamber 516 and at the rear opening 519 of the substrate hole, as Figure 13D shown.

[0073] In each calibration unit 501 of the device 500, the first chamber 510 and the second chamber 516 have a triangular cross-section, where each triangular cross-section is a right triangle or substantially a right triangle. The hypotenuse of the triangular portion of the first chamber 510 faces the hypotenuse of the triangular portion of the second chamber 516, thereby forming a compact calibration unit structure. Non-limiting examples of this orientation of the first chamber and the second chamber are shown in Figure 16A and Figure 16Cis shown, where the first chamber 510 is disposed at a front position relative to the channel 524. The opposing sidewalls of the first chamber 510 and the second chamber 516 are tapered, where the opposing sidewalls of the first chamber 510 taper in a front-to-distal direction 599, and the opposing sidewalls of the second chamber taper in a rear-to-front direction 598. The opposing sidewalls of the first chamber and the second chamber may follow or substantially follow the same or a similar draft angle. These features of the first chamber and the second chamber contribute to a compact calibration unit structure.

[0074] The apparatus 500 includes a hydrophobic membrane 522, also referred to as a "porous membrane", associated with the interstitial volume of the front opening 520' of the second chamber. The hydrophobic membrane 522 is indirectly associated with the interstitial volume of the front opening 520' of the second chamber through a vent 575. The vent 575 is disposed between the hydrophobic membrane 522 and the front opening 520' of the second chamber and is disposed at a front position relative to the opening 520'. The rear end 577 of the vent is disposed at the opening 520' of the second chamber 516 and includes an inner sidewall 578. The vent 575 is a void in the substrate 502 and is cylindrical. The vent 575 is integral with the second chamber 516 and the junction 570 and is disposed at a rear position of the junction 570 in the apparatus 500. The junction 570 is a hole in the substrate 502 at the front surface 504 and includes a release surface 573 and sidewalls 571. The sidewalls 571 include an opening 572. The sidewalls 571 are perpendicular to the front surface 504 of the substrate 502, and the hole of the junction 570 is cylindrical. The hole of the junction 570 may be another geometry, such as a frustum (e.g., a conical frustum), and the sidewalls may taper in a front-to-rear direction. The junction 570 contains the hydrophobic membrane 522, and the depth of the sidewalls 571 is generally the same as or substantially the same as the depth of the hydrophobic membrane 522. The release surface 573 of the junction 570 includes an opening 574 concentric with the release surface 573 and the vent 575. The front end 576 of the vent is disposed at the opening 574 in the release surface 572. The solid portion of the rear surface 542 of the front member 540 disposed above the junction 570 forms the inner front roof surface of the junction 570 and captures the hydrophobic membrane 522, as Figure 13E shown. The hydrophobic membrane 522 may be adhered to the release surface 573 by an adhesive, heat melt, or other joining method. In an alternative embodiment, the hydrophobic membrane 522 may be disposed in a counterbore that is disposed at a rear position relative to the junction 570 and extends in a front-to-rear direction from the release surface 573. The counterbore may include a counterbore release surface disposed at the rear end of the counterbore. The counterbore may be cylindrical or other geometry, such as a frustum, e.g., having sidewalls that taper in a front-to-rear direction. The counterbore may hold the hydrophobic membrane by a press fit.

[0075] The hydrophobic membrane 522 disposed at the front of the device 500 is indirectly associated with the interstitial volume of the opening at the rear surface of the device through a piping system. The device 500 includes a lateral pipe 546 disposed at the joint 570. The lateral pipe 546 is a groove formed in the substrate 502 at the front surface 504 and includes side walls 547, a proximal end 548, and a distal end 549, as Figure 11B , Figure 15C and Figure 16D shown. The lateral pipe 546 has a rectangular cross-section. The lateral pipe 546 includes a curved portion closer to the joint 570 in the proximal part and a linear or substantially linear portion closer to the axial pipe 535 in the distal part. The proximal end 548 of the lateral pipe 546 is disposed at an opening 572 in the side wall 571 of the joint 570. As Figure 14C shown, the solid portion of the rear surface 542 of the front member 540 disposed above the lateral pipe 546 forms the internal front surface of the lateral pipe 546.

[0076] The lateral pipe 546 is connected to another pipe, the axial pipe 535. The axial pipe 535 is a void in the substrate 502 and is partially defined by a hole extending from the front surface 504 of the substrate to the rear surface 506 of the substrate. The axial pipe 535 includes side walls 536, a front end 537 disposed at the front surface 504 of the substrate, and a rear end 538 disposed at the rear surface 554 of the rear member. The axial pipe 535 is a frustum of a cone with side walls 536 that taper in the front-to-rear direction 599. The axial pipe 535 includes a front opening 536' disposed in the side wall 536 at the front end 573, and the distal end 549 of the lateral pipe 546 is disposed at the front opening 536' of the axial pipe 535. The solid portion of the rear surface 542 of the front member 540 disposed above the front opening of the axial pipe 535 in the front surface 504 of the substrate forms the internal front top surface of the axial pipe 535 at the front end 537, as Figure 14C shown. The hole 556 of the rear member 550 is disposed at the rear end 538 of the axial pipe in the rear surface 506 of the substrate. The hole 556 is larger than the opening at the rear end 538 of the axial pipe in the rear surface 506 of the substrate, and the portion 539 of the rear surface 506 of the substrate that surrounds the opening at the rear end 538 of the axial pipe and is bounded by the perimeter of the hole 566 is exposed, as Figure 14C shown.

[0077] In device 500, a portion of the front surface 504 of the substrate is joined to the rear surface 542 (also referred to as the "joining surface") of the front member 540, forming a joining portion 580 (also referred to as the "joining layer") between the front member 540 and the substrate 502. The front member 540 includes a front surface 544 and a hole 545 that partially defines the front opening 512 of the first chamber. Most or all of the rear surface 506 of the substrate is joined to the front surface 552 (also referred to as the "joining surface") of the rear member 550, forming a joining portion 582 (also referred to as the "joining layer") between the rear member 550 and the substrate 502. The rear member 550 includes a rear surface 554 and a hole 556 adjacent to the rear end 538 of the axial duct.

[0078] In device 500, the substrate 502 includes outer sidewalls having adjacent outer sidewalls 508 or 509 and inner surfaces 587 and 588 on the adjacent outer sidewalls. The substrate 502 also includes inner sidewalls, each inner sidewall being opposite to an adjacent outer sidewall and having surfaces 584 and 584 on the adjacent inner sidewalls. The inner sidewalls are disposed at a rear position relative to the front surface 504 of the substrate, each inner sidewall being offset from the outer sidewall by an insertion surface 586, and each inner sidewall being offset from the inner surface of the outer sidewall by a space or void. The inner wall surface partially defines the perimeter of the rear surface 506 of the substrate, and due to the offset distance between the outer wall and the inner wall, the surface area of the rear surface 506 of the substrate is smaller than the surface area of the front surface 504 of the substrate.

[0079] In a calibration unit 501, the front member hole 545 that partially defines the front opening 512 of the first chamber has an offset distance from the outer sidewall of the substrate that is different from the offset distance from the adjacent outer sidewall of the substrate, and the array of front member holes may not be centered on the front surface 504 of the substrate or the front surface 544 of the front member. The offset distance 593 between the outer surface 508 of the substrate and the front member hole 545 and the offset distance 594 between the outer surface 509 of the substrate and the front member hole 545 are different, and are shown in Figure 10A A rear member hole 556 in a calibration unit 501 may have an offset distance from the inner sidewall of the substrate that is different from the offset distance from the adjacent inner sidewall of the substrate, and the array of rear member holes may not be centered on the rear surface 506 of the substrate or the rear surface 554 of the rear member. The offset distance 591 between the inner surface 584 of the substrate and the rear member hole 556 and the offset distance 592 between the inner surface 585 of the rear substrate and the rear member hole 556 are different, and are shown in Figure 10C are shown.

[0080] The calibration device 500 can be connected to a device that applies a force that pushes the fluid being conveyed through a cavity in the calibration device, such as the vacuum manifold 650 in the assembly 600. The vacuum manifold 650 includes a front surface 652, a release surface 660, an inner cavity 670, and an inner surface 662. The rear portion of the calibration device 500 (which generally includes a continuous perimeter around the rear of the outer sides 508 and 509) can be fitted onto the release surface 660 of the vacuum manifold 650, whereby the inner surface 662 contacts the continuous perimeter around the rear of the outer sides 508 and 509 of the calibration device 500. A gasket is sometimes provided at the release surface 660 and / or the inner surface 662 to facilitate the seal between the calibration manifold 650 and the calibration device 500. The vacuum manifold 650 can be connected to a vacuum (e.g., applied by a vacuum generator) and includes a vacuum connector 682, a vacuum connector orifice 684, an optional pressure gauge 686, and a vacuum connector link 688 that connects the vacuum connector 682 to the sidewall of the manifold 650.

[0081] Method of manufacturing a volume calibration device

[0082] The calibration device is sometimes manufactured by a process that includes preparing the rear portion by an injection molding process, an embossing process (e.g., a hot embossing process), and / or a die cutting process. In certain embodiments, the calibration device is manufactured by a process that includes preparing the front portion by an injection molding process, an embossing process (e.g., a hot embossing process), and / or a die cutting process. In certain cases, the calibration device is manufactured by a process that includes providing a rear portion and a front portion and joining the rear portion to the front portion. In certain embodiments, the calibration device is manufactured by a process that includes joining a hydrophobic membrane to the rear surface of the calibration device. The hydrophobic membrane can be joined to the rear surface of the calibration device after the front portion is joined to the rear portion, or before the front portion is joined to the rear portion. Any suitable joining process can be employed, including the joining processes described herein.

[0083] In certain embodiments, a front portion, a middle portion, and a rear portion are manufactured and then joined to each other to form the device, with the middle portion sandwiched between the front portion and the rear portion. The front portion can include a first chamber and a well (e.g., the well and the rear end of the first chamber are concentric) optionally disposed at a front position relative to the rear end of the first chamber. The middle portion can be a membrane that has been cut (e.g., by a die or a laser or other means) to form a channel and an optional well. The rear portion, which generally includes a second chamber and a hydrophobic membrane, can be joined to the middle portion before or after the middle portion and the front portion are joined. These portions are generally aligned with each other such that the first chamber and / or the well of the front portion is aligned with the proximal end of the channel, and the second chamber of the rear portion is aligned with the distal end of the channel. Any suitable joining method as described herein can be used to join the portions.

[0084] In some embodiments, a substrate, a front member, and a rear member are provided and then joined to form a device including the substrate disposed between the front member and the rear member. The front surface of the rear member is typically joined to all or sometimes a portion of the rear surface of the substrate. The rear surface of the front member is typically joined to a portion or sometimes all of the front surface of the substrate. The substrate is sometimes injection molded. The substrate can include holes that partially define a first chamber, a second chamber, optional wells, optional vents, optional axial ducts, and / or optional junctions between the second chamber and / or optional vents and optional lateral ducts. The substrate sometimes includes one or more recesses and / or grooves that partially define channels and optional lateral ducts. The front member typically includes a hole that is aligned with and generally coextensive with a first chamber hole opening disposed at the front surface of the substrate. The solid portion of the front member where no hole is provided can define the front surface of a lateral duct, can define the front surface of a junction, and / or can capture a hydrophobic membrane disposed in the junction. One or more hydrophobic membrane units can be joined to the substrate (e.g., within a recessed surface disposed on the front surface of the substrate (e.g., at the junction)) and optionally joined (or e.g., not joined) to the substrate before the front member is joined to the substrate. The rear member typically includes a hole that is aligned (e.g., coaxially aligned) with an axial vent rear opening. The solid portion of the rear member where no hole is provided can define the rear end of an optional well, can define the rear end of the first chamber, and / or can define the rear surface of a channel. The front member and / or the rear member are sometimes polymer films in which holes are introduced by a suitable process (e.g., a stamping process).

[0085] Parts of the device, the substrate, and / or the members (e.g., the rear member and / or the front member) can be made of a polymer. Non-limiting examples of polymers include polypropylene (PP), polyethylene (PE, high-density PE, low-density PE), polyethylene terephthalate (PET), polystyrene (PS), etc.

[0086] The joining process can be a solvent joining process, a laser joining process (e.g., interfacial laser joining), an adhesive joining process (e.g., pressure-sensitive adhesive joining), an ultrasonic welding process, and / or a plasma joining process. In the solvent joining process, a solvent that dissolves the polymer used to make the part is applied to one or both of the surfaces to be joined. When the surfaces are wetted with the solvent, the parts are pressed together so that the wet surfaces contact. Pressure is applied to keep the surfaces in close proximity and ensure contact of the entire surface to be joined. The solvent evaporates and / or is absorbed into the body of the material. The polymer at the interface that is mildly dissolved by the solvent re-hardens and causes the adjacent surfaces to join. This solvent joining is typically strong and waterproof. The solvent strength and polymer type are selected to ensure sufficient joining between the layers.

[0087] In the laser welding process, a laser is used to heat and melt the polymer at the interface between two parts. The material of the first part is chosen to be transparent to the light wavelength of the laser, so it transmits the laser energy without absorbing it. The polymer of the second part is chosen such that it will absorb the laser and heat until it is above its melting point. Through this process, the two materials can be joined at the interface surface with substantially no melting and distortion of the surrounding materials. Additionally, the laser can be focused to a fine spot and programmed to selectively join the two surfaces in a precisely controlled area. In this way, channels can be joined on either side of the channel geometry, forming a liquid-tight seal at the edges of the channel.

[0088] In the pressure-sensitive adhesive joining process, the parts of the device are joined using a pressure-sensitive adhesive that is designed to firmly join the parts while also providing a liquid-tight seal at the edges of the channel. The pressure-sensitive adhesive can be cut (die-cut or laser-cut) to precisely control the area of the adhesive.

[0089] Ultrasonic welding is a method of introducing high-frequency vibrations into two (or more) polymer components to be joined. The vibrations cause relative movement between the components, especially at the interface where the components are in contact. The friction generated by the relative movement produces heat, which melts the areas of the contacting components. Once the ultrasonic energy is removed, the polymer re-solidifies and joins together. Features such as beads and energy directors can be used to localize the heat and precisely control the location where the joining occurs. The energy director can be used to create a liquid-tight seal along the edges of the channels of the device.

[0090] In the plasma joining process, a plasma (e.g., a plasma of oxygen) is used to modify the surface of one or both components to be joined. The plasma treatment makes the surfaces reactive so that when they are pressed closely together, the surfaces react with each other, join tightly, and form a liquid-tight joint.

[0091] Any suitable joining process can be used to join the hydrophobic membrane to the calibration device. For example, a suitable joining process can be used to join the hydrophobic membrane at the rear surface area of the calibration device above the rear opening of the second chamber. One method is to apply a pressure-sensitive adhesive to the membrane of the polymer that adheres to the calibration device. Another method is to apply heat and pressure through a properly shaped heating element that melts the membrane and the calibration device surface, which joins the membrane to the calibration device. Another joining method is to apply ultrasonic energy to generate frictional heat at the interface between the membrane and the calibration device part, which melts the materials of the membrane and the calibration device together.

[0092] Distributor volume calibration method

[0093] In some embodiments, a method for determining the volume of fluid dispensed by a fluid dispensing device is provided. The method includes dispensing fluid from the fluid dispensing device into a first chamber of a volume calibration device that does not contain fluid, whereby the first chamber of the calibration device includes the fluid dispensed from the fluid dispensing device. Thereafter, the dispensed fluid generally flows in the calibration device under the influence of a fluid driving force (e.g., an applied fluid driving force), such as a pressure differential (e.g., an applied vacuum). The fluid generally flows from the first chamber into a channel (e.g., via an optional intermediate well) in a fluid path in the calibration device, where the channel generally includes a proximal end fluidly associated with the first chamber (e.g., the proximal end of the channel is sometimes disposed at the rear end and / or side of the first chamber or the optional intermediate well). In the fluid path, the fluid generally flows in the channel into a second chamber fluidly associated with the distal end of the channel. The fluid generally fills the second chamber, and when the proximal end of the fluid contacts the surface of the hydrophobic membrane, the proximal end of the fluid generally no longer transmits. The second chamber is generally fluidly associated with the surface of the hydrophobic membrane (e.g., via an optional intermediate vent). In some cases, the rear opening or the front opening of the second chamber contacts the surface of the hydrophobic membrane. In some embodiments, the rear opening or the front opening of the second chamber contacts the vent end (e.g., the rear vent opening), and the opposite vent end (e.g., the front vent opening) is associated with or contacts the surface of the hydrophobic membrane.

[0094] The fluid can be dispensed into the first chamber of the calibration device in any suitable manner by the fluid dispenser. The fluid dispensing device is generally coupled to a pipette tip or a probe or a plurality of pipette tips or probes, and the fluid is aspirated into the pipette tip or the probe, and the aspirated fluid is dispensed from the pipette tip or the probe into the calibration device. The distal portion (e.g., the distal end) of the pipette tip or the probe is sometimes positioned near the first chamber and / or sometimes positioned to contact the inner surface (e.g., the sidewall) of the first chamber as part of the process of dispensing the fluid into the first chamber. The front opening of the first chamber can be slightly offset from the center of perfect concentric alignment with the pipette tip or the probe such that the pipette tip or the probe contacts the tapered inner sidewall surface of the first chamber. This orientation allows for touching and allows the distal end of the pipette tip to be very close to or in contact with the inner surface of the first chamber, increasing the likelihood that the dispensed fluid is completely transferred to the first chamber by minimizing or preventing droplets from being retained by the pipette tip or the probe.

[0095] A pipette tip or probe containing fluid to be dispensed into the first chamber is typically lowered (or the calibration device is raised) until the pipette tip or probe is near or in contact with the inner sidewall of the first chamber. The liquid handler can be commanded to dispense the fluid at a rate that expels the fluid from the pipette tip or probe into the first chamber. After the fluid dispensing event, a blow-out may occur, where air is expelled from the pipette tip or probe to increase the likelihood of complete expulsion of the fluid from the pipette tip or probe. Then, the liquid handler can retract the pipette tip or probe away from the first chamber.

[0096] Any fluid can be dispensed into the calibration device, which is adapted to accurately and precisely determine the volume of fluid in the calibration device. After the fluid has stopped flowing in the calibration device, fluid that does not dwell or substantially does not dwell in the first chamber or a portion of the channel near the end point of fluid dispensing can be selected. The dispensed fluid typically has a high surface tension and is generally hydrophilic, such as water or other aqueous solutions (e.g., purified water, distilled water). The high surface tension of the dispensed fluid helps prevent the dispensed fluid from penetrating and / or passing through the hydrophobic membrane disposed at the rear opening of the second chamber. The dispensed fluid can contain one or more components (e.g., dyes or other optically detectable components) suitable for determining the position of the end point of fluid dispensing in the channels of the device. For embodiments in which hydrophilic fluids and / or fluids with high surface tension are dispensed, the walls of one or more voids and / or cavities (e.g., the first chamber, the second chamber, the channels, optional wells, optional vents) of the calibration device are sometimes hydrophobic (e.g., contain a hydrophobic coating and / or are made of a hydrophobic material).

[0097] The dispensed fluid typically flows in the channel under a driving force, which is typically an applied driving force (e.g., applied by the user of the calibration device). The driving force is sometimes a pressure difference between the first chamber and the second chamber. The pressure difference between the first chamber and the second chamber can be applied in any manner suitable for driving the fluid dispensed in the first chamber through the channel and into the second chamber. In some cases, the pressure difference is established by applying a vacuum to the second chamber. In some embodiments, the vacuum is applied outside (i.e., at the rear side) of the hydrophobic membrane. The hydrophobic membrane can hold or substantially hold the fluid in the device and can block or substantially block the fluid from leaving the device.

[0098] A vacuum can be applied to the hydrophobic membrane by connecting a calibration device to a vacuum device (e.g., a vacuum manifold) designed to sealably connect to the calibration device. The calibration device sometimes includes an SBS standard footprint and can be connected to a vacuum manifold designed for an SBS standard footprint microplate (e.g., a commercially available manifold for the bottom of a filter microplate). The manifold sometimes includes a gasket designed to seal the perimeter of the rear of the calibration device after the vacuum is applied. The calibration device sometimes includes a sealing member, such as a continuous rib around the perimeter of the rear of the calibration device, which contributes to the sealing integrity. Each calibration unit of the calibration device is generally within the perimeter of the rear surface of the calibration device and is thus exposed to the vacuum of the vacuum device. A first chamber typically provided in the front of the calibration device is typically exposed to atmospheric pressure. After applying a vacuum to the rear of the calibration device by the vacuum device, a pressure differential can be created between a fluid path proximal starting position (e.g., the first chamber) and a fluid path distal end (e.g., the hydrophobic membrane surface), which drives the dispensed fluid within the calibration device to the distal fluid end point (e.g., the hydrophobic membrane surface). A pressure differential is typically created between the front (e.g., the first chamber) and the rear (e.g., the hydrophobic membrane on the rear surface of the device, the rear opening of the conduit at or near the rear surface of the device, and / or the rear member aperture) of the calibration device.

[0099] As an alternative to applying a vacuum after dispensing fluid into the calibration device, a calibration device can be provided in which the fluid path within the device is under a negative pressure (e.g., a pressure below atmospheric pressure). Such a device can include seals (e.g., frangible seals) provided at each void opening that maintain the negative pressure within the internal voids of the device. The seal sometimes covers the front opening of the first chamber and can include a seal covering the rear opening of the device. The hydrophobic membrane of the device can serve as a seal. In use, a fluid dispenser member (e.g., a pipette tip, a probe) can rupture the seal at the front opening of the first chamber, and the ruptured seal can seal around the fluid dispenser member. Fluid can then be dispensed into the first chamber, and the pressure differential between the fluid dispenser member (e.g., sometimes at atmospheric pressure) and the internal fluid path within the calibration device (e.g., sometimes at a pressure below atmospheric pressure) drives the dispensed fluid within the fluid path of the calibration device.

[0100] The pressure differential is sometimes about 10% to about 90% of atmospheric pressure (about 14.7 pounds per square inch (psi)) and can be about 20%, 30%, 40%, 50%, 60%, 70%, or 80% of atmospheric pressure. The pressure differential can partially determine the rate at which the dispensed fluid flows through the fluid path of the calibration device (e.g., from the first chamber to an optional well to a channel to a second chamber to an optional vent and the hydrophobic membrane), and different calibration devices can operate at different optimal pressure differentials.

[0101] The dispensing fluid flowing in the fluid path of the device typically fills the second chamber. After the dispensing fluid stops flowing in the calibration device, the fluid in the device typically has filled the entire volume or substantially the entire volume of the second chamber, and typically fills a portion of the channel up to the fluid terminus between the proximal end and the distal end of the channel. For devices that include an optional vent hole between the second chamber and the hydrophobic membrane, the entire volume or substantially the entire volume of the vent hole is typically also filled with the dispensing fluid. After the dispensing fluid stops flowing in the device, the dispensing fluid is typically a continuous fluid volume (e.g., a slug) that extends from the surface of the hydrophobic membrane through the distal end of the channel and reaches the fluid terminus at a location in the channel between the distal end and the proximal end of the channel. The continuous fluid volume (e.g., a slug) typically contains no or substantially no detectable foam (e.g., air bubbles). After the dispensing fluid stops flowing in the device, the portion of the channel near the fluid terminus, as well as the first chamber and the optional well, typically do not include or substantially do not include detectable sample fluid.

[0102] The volume determination method typically includes determining the location (i.e., position) of the fluid terminus in the channel. The proximal fluid terminus is typically in the channel between the proximal channel end and the distal channel end, and the distal fluid terminus is typically at the surface of the hydrophobic membrane. The location of the proximal fluid terminus in the channel is typically determined after the fluid stops flowing in the device and the proximal fluid terminus in the channel is in a fixed position. The user can determine when the fluid stops flowing in the device by any suitable technique, including but not limited to: (i) waiting a predetermined amount of time between dispensing the fluid from the dispenser into the first chamber and determining the position of the fluid terminus in the channel, and / or (ii) determining the fluid terminus at several time points and determining that the fluid terminus has not changed in at least two determinations.

[0103] The position of the fluid terminus in the channel can be determined using an optical detector. In some embodiments, an image of the channel containing the fluid sample is obtained and computer software is used to determine the position of the fluid terminus in the channel. Such software can analyze the pixels of the image and identify the difference between the fluid-filled region of the channel (the region of the channel remote from the dispensed fluid terminus) and the region not containing fluid (the region of the channel proximal to the dispensed fluid terminus). There are several ways to enhance the contrast between the fluid-filled and non-fluid-filled regions of the channel, such as (i) including a detectable component (e.g., a dye) in the dispensed fluid, (ii) providing a clear foreground layer and a translucent or opaque background layer (e.g., a white or other light-colored background), (iii) backlighting of the translucent or transparent layer, and / or (iv) including molded features in the channel. The molded features in the channel can include (i) a matte surface, for which a portion of the channel containing fluid is optically distinguishable from a portion of the channel not containing fluid, and / or (ii) registration marks, which can appear or disappear depending on the presence or absence of fluid. The registration marks can be included on the outer or inner surface of a calibration device (e.g., on the channel surface).

[0104] The volume of the dispensed fluid in the calibration device can be determined based on the position of the dispensed fluid terminus in the channel. After the dispensed fluid stops flowing in the device, the dispensed fluid generally occupies the entire volume of the second chamber and optionally the vent, or substantially occupies the entire volume of the second chamber and optionally the vent. The volume of the dispensed fluid from the distal fluid terminus (e.g., at the hydrophobic membrane surface) to the distal end of the channel is known and / or can be determined as known in the art. After the fluid stops flowing in the device, the volume of the dispensed fluid contained in the portion of the channel between the distal end of the channel and the fluid terminus between the distal and proximal ends of the channel can be determined by methods known in the art. The volume of the dispensed fluid can be determined based on the channel geometry, the second chamber geometry, the optional vent geometry, and the geometry of any other applicable components (e.g., the rear member geometry) as well as the precise position of the dispensed fluid terminus in the channel.

[0105] In some embodiments, the length of the channel containing the fluid is determined (i.e., from the distal end of the channel to the endpoint of the dispensed fluid in the channel), and the sample volume in the channel containing the fluid can be calculated based on the known cross-sectional area of the channel. The cross-section does not need to be constant over the entire length of the channel, and the sample fluid volume can be calculated in part based on the known cross-sectional area of the measured channel length. In some cases, a computer-aided design (CAD) model of the channel and other applicable structures (e.g., a second chamber, an optional vent, and any other structure that will contain the fluid volume after the fluid has flowed through the device) are used to calculate the fluid volume. In some embodiments, the dispensed fluid volume is determined (calibrated) empirically by filling the fluid path cavity (i.e., the second chamber and part of the channel) with a known volume of liquid and measuring the position of the endpoint of the dispensed fluid over the length of the channel or part thereof. A hydrophobic membrane can be used as a known starting point in the cavity volume, simplifying the measurement process to one desired dimension. The hydrophobic membrane typically positions the dispensed fluid in a known region of the cavity.

[0106] In certain embodiments, the fluid volume determination method can be implemented as Figures 6 to 8D shown. The distal portion 202 of the pipette tip 200 connected to the fluid dispensing device (also referred to as a "liquid handling device") can be inserted through the opening 112 into the first chamber 110 of the calibration device 100, e.g., as Figure 6 shown. Although the pipette tip can dispense fluid into the device 100, the dispensing device can dispense fluid into the device 100 via a probe, needle, or other dispensing structure. At this point in the method, the calibration device 100 does not contain the fluid 290 in the pipette tip 200.

[0107] Then, the fluid dispenser dispenses the fluid 290 (also referred to as a "liquid sample") into the device 100. The device 300 includes the same structural features as the device 100, except that the device 300 includes the dispensed fluid 290 and the device 100 does not include the dispensed fluid. As Figure 7 shown, the fluid 290 dispensed from the pipette tip 200 is in the first chamber 310 of the device 300. The dispensed fluid 290 typically fills a portion of the first chamber 310 and sometimes fills the well 330 at this point in the method, as Figure 8A shown.

[0108] The pressure difference between the first chamber 310 and the second chamber 316 pushes the dispensed fluid from the first chamber 310 through the well 330 into the channel 324 and into the second chamber 316. As Figure 8AAs shown, a pressure differential can be applied to the device 300 by contacting the rear surface 306 at the rear opening 318 of the second chamber and at the hydrophobic membrane 322 with a device that provides a vacuum or otherwise creates a pressure differential between the first chamber 310 and the second chamber 316. The hydrophobic membrane 322 retains the fluid within the second chamber 316 by not allowing the fluid to flow out of the rear end 318 of the device 300. Thus, the first chamber 310 is typically emptied of the dispensed fluid 290, the second chamber 316 is filled with a portion 294 of the dispensed fluid 290, and a portion 296 of the dispensed fluid 290 forms a continuous fluid path from the distal end 328 of the channel 324 at the second chamber 316 to the dispensed fluid terminus 292 in the channel 324, as Figure 8B shown. The portion 296 of the dispensed fluid 290 in the channel 324 fills the distal portion 325 of the channel 324 between the distal end 328 of the channel 324 and the dispensed fluid terminus 292, resulting in the proximal portion 327 of the channel 324 being free of or substantially free of the dispensed fluid 290, as Figure 8C and Figure 8D shown.

[0109] The volume of the portion 294 of the dispensed fluid 290 that fills or substantially fills the second chamber 316 is equal to or substantially equal to the volume of the second chamber 316. The volume of the portion 296 of the dispensed fluid 290 from the distal end 328 of the channel 324 to the fluid terminus 292 in the channel 324 is determined by the position of the terminus 292 in the channel 324. The total volume of the dispensed fluid 290 in the device 300 is equal to or substantially equal to the volumes of the portion 294 and the portion 296.

[0110] In some embodiments, the fluid dispensing device is a multi-channel dispensing device that includes a plurality of dispenser units arranged in series and / or in an array (e.g., Figures 9A to 9E the device 400 shown). The calibration device that receives the dispensed fluid from the dispensing device typically includes calibration units arranged in a paired series and / or in an array, where each calibration unit in the calibration device receives a volume of the dispensed fluid from one of the dispenser units in one dispensing event. In some embodiments, the calibration device includes the same, more, or fewer number of calibration units as the dispenser units of the dispensing device. In some cases, the dispensed volume from each dispenser unit is determined for one dispensing event. In some cases, the dispensed volume from each dispenser unit is determined in each of a plurality of dispensing events.

[0111] In certain embodiments, the fluid volume determination method can be implemented by utilizing a calibration device 500. For example, the distal portion 202 of the pipette tip 200 connected to the fluid dispensing device can be inserted into the first chamber 510 of the calibration device 500 through the first chamber front opening 512. Although the pipette tip can dispense fluid into the device 500, the dispensing device can dispense fluid into the device 500 via a probe, a needle, or other dispensing structures. In some embodiments, the fluid dispensing device is a multi-channel dispensing device including a plurality of dispenser units arranged in series and / or in an array, where each dispenser unit is paired with a corresponding first chamber 510 in the calibration unit 510 of the device 500. Then, the fluid dispenser can dispense fluid 290 into the device 500. The fluid 290 dispensed from the pipette tip 200 is in the first chamber 510 of the device 500. The dispensed fluid 290 generally fills a portion of the first chamber 510 and sometimes fills the well 530.

[0112] Then a driving force can be applied to the device 500, which pushes the fluid from the first chamber 510 through the void to the hydrophobic membrane 522 in each calibration unit 501. The pressure difference between the first chamber 510 and the hydrophobic membrane 522 can be applied by introducing a vacuum into the device 500. The pressure difference can be applied by introducing a vacuum introduced by a vacuum manifold 650 into the device 500. The device 500 can be connected to the vacuum manifold 650 before or after the fluid is dispensed into the first chamber 510. Applying the pressure difference can push the dispensed fluid to flow from the first chamber 510 through the well 530, through the channel 524, through the second chamber 516 and into the vent 575. The hydrophobic membrane 522 holds the fluid in the vent 575 by acting as a barrier to fluid flow. Thus, the proximal portions of the first chamber 510, the well 530, and the channel 524 are generally free of the dispensed fluid, and the dispensed fluid generally fills and is disposed in a portion of the channel 524, the second chamber 516, and the vent 575 in a continuous fluid path from the proximal fluid end point in the channel 524 to the distal fluid end point at the hydrophobic membrane 522. Then the volume of the dispensed fluid in the continuous fluid path can be determined from the proximal fluid end point as described herein.

[0113] Exemplary embodiments

[0114] The following are non-limiting examples of certain exemplary embodiments of the technology provided herein.

[0115] A0.1. A fluid dispenser volume calibration device, comprising:

[0116] A front surface, a rear surface, side surfaces, a first chamber, a second chamber, a hydrophobic membrane, and a channel, wherein:

[0117] The first chamber includes a front opening and a rear end, the front opening being provided at the front surface of the device;

[0118] The second chamber includes a rear end, side walls, and a front end;

[0119] The hydrophobic membrane is in fluid communication with the second chamber; and

[0120] The channel includes a proximal end in fluid communication with the first chamber and a distal end in fluid communication with the second chamber.

[0121] A0.2. The device according to embodiment A0.1, comprising a well, wherein:

[0122] The first chamber includes a rear port provided at the rear end of the first chamber;

[0123] The well includes a well front end, well side walls, and a well rear end;

[0124] The well front end is provided at the rear port of the first chamber;

[0125] The well includes a well rear port, the well rear port being provided at the well rear end and the well side walls; and

[0126] The proximal end of the channel is provided at the well rear port.

[0127] A0.3. The device according to embodiment A0.1 or A0.2, wherein:

[0128] The second chamber includes a front port provided at the front end of the second chamber and a rear port provided at the rear end and the side walls of the second chamber; and

[0129] The distal end of the channel is provided at the rear port of the second chamber.

[0130] A0.4. The device according to embodiment A0.3, comprising a vent, wherein:

[0131] The vent includes a vent front end, vent side walls, and a vent rear end; and

[0132] The vent rear end is provided at the front port of the second chamber.

[0133] A0.5. The device according to any one of embodiments A0.1 - A0.4, comprising a substrate, wherein:

[0134] The substrate includes a front surface and a rear surface;

[0135] The substrate optionally includes a recess;

[0136] The recess is optionally provided at the front surface of the substrate;

[0137] The optional recess includes a recessed surface and a recess side wall; and

[0138] The recessed surface optionally includes a recessed surface port.

[0139] A0.6. The device according to embodiment A0.5, wherein the front end of the vent is disposed at the recessed surface port.

[0140] A0.7. The device according to embodiment A0.5 or A0.6, wherein the recess includes a recess side wall port disposed in the recess side wall.

[0141] A0.8. The device according to any one of embodiments A0.5 - A0.7, wherein the hydrophobic membrane is disposed in the recess.

[0142] A0.9. The device according to any one of embodiments A0.5 - A0.7, comprising a pipe, wherein the pipe includes a pipe end associated with the void volume of the hydrophobic membrane.

[0143] A0.9.1. The device according to embodiment A0.9, wherein the pipe end is disposed at the hydrophobic membrane, or at the recess, or at the recess side wall port.

[0144] A0.10. The device according to embodiment A0.9, comprising a lateral pipe, wherein:

[0145] The lateral pipe includes a lateral pipe proximal end, a lateral pipe distal end, and a lateral pipe side wall; and

[0146] The lateral pipe proximal end is disposed at the recess side wall port.

[0147] A0.11. The device according to embodiment A0.10, comprising an axial pipe, wherein:

[0148] The axial pipe includes an axial pipe front end, an axial pipe rear end, an axial pipe side wall, and an axial pipe front port disposed in the axial pipe side wall at the axial pipe front end; and

[0149] The axial pipe rear end is disposed at the rear surface of the substrate.

[0150] A0.12. The device according to any one of embodiments A0.1 - A0.12, comprising a front member, wherein:

[0151] The front member includes a front surface, a rear surface, and a front member hole;

[0152] The rear surface of the front member is joined to at least a portion of the front surface of the substrate; and

[0153] The substrate includes a first chamber hole that includes a hole front opening disposed at the front surface of the substrate;

[0154] The front member hole is aligned with and co - extends with the first chamber front hole of the substrate and is the first chamber front opening of the device.

[0155] A0.13. The device according to embodiment A0.12, wherein the front member includes a solid portion covering the recess and a hydrophobic membrane disposed in the recess.

[0156] A0.14. The device according to embodiment A0.12 or A0.13, wherein the front member includes a solid portion covering the lateral duct, and the rear surface of the front member at that portion is the lateral duct top plate surface.

[0157] A0.15. The device according to any one of embodiments A0.12 - A0.14, wherein the front member includes a solid portion covering the front end of the axial duct, and the rear surface of the front member at that portion is the axial duct top plate surface.

[0158] A0.16. The device according to any one of embodiments A0.12 - A0.15, comprising a rear member, wherein:

[0159] The rear member includes a front surface, a rear surface, and a rear member hole;

[0160] The front surface of the rear member is joined to at least a portion of the rear surface of the substrate; and

[0161] The rear member hole is coaxially and / or concentrically disposed at the rear end of the axial duct.

[0162] A0.17. The device according to embodiment A0.16, wherein the rear member includes a solid portion disposed at the channel, and the front surface of the rear member at that portion is the channel bottom plate.

[0163] A0.18. The device according to embodiment A0.16 or A0.17, wherein the rear member includes a solid portion disposed at the rear end of the second chamber, and the front surface of the rear member at that portion is the second chamber bottom plate.

[0164] A0.19. The device according to any one of embodiments A0.16 - A0.18, wherein the rear member includes a solid portion provided at the rear end of the well, and the front surface of the rear member at that portion is the bottom plate of the well.

[0165] A0.20. The device according to any one of embodiments A0.1 - A0.19, including at least one calibration unit

[0166] A0.21. The device according to embodiment A0.20, wherein each calibration unit includes the first chamber, the second chamber, the channel, and the hydrophobic membrane.

[0167] A0.22. The device according to embodiment A0.21, wherein each calibration unit includes the well, the vent, the recess, the lateral duct, and the axial duct.

[0168] A0.23. The device according to any one of embodiments A0.20 - A0.22, including two or more units arranged in an array.

[0169] A0.24. The device according to any one of embodiments A0.1 - A0.23, wherein the channel includes an inner surface, and at least a portion of the inner surface of the channel is hydrophobic.

[0170] A0.25. The device according to embodiment A0.24, wherein at least a portion of the inner surface of the channel includes a coating, and the coating includes a hydrophobic material.

[0171] A0.26. The device according to any one of embodiments A0.1 - A0.25, wherein:

[0172] the channel includes two or more substantially linear members or linear members and a transition member connecting two substantially linear members or linear members; and

[0173] the substantially linear members or linear members are parallel or substantially parallel to each other.

[0174] A0.27. The device according to any one of embodiments A0.1 - A0.26, wherein the first chamber and the second chamber are each independently a triangular prism or a triangular frustum.

[0175] A0.28. The device according to embodiment A0.27, wherein:

[0176] the first chamber and the second chamber have a triangular cross - section;

[0177] the triangular cross - section is a right - angled triangular cross - section including the hypotenuse; and

[0178] The hypotenuse of the right-angled triangle cross-section of the first chamber faces and is parallel or substantially parallel to the hypotenuse of the right-angled triangle cross-section of the second chamber.

[0179] A1. A fluid dispenser volume calibration device, comprising:

[0180] A substrate including a front surface, a rear surface, and side surfaces;

[0181] A first chamber disposed in the substrate, the first chamber including a front opening and a rear end disposed at the front surface of the substrate;

[0182] A second chamber disposed in the substrate, the second chamber including a rear opening and a front end disposed at the rear surface of the substrate;

[0183] A hydrophobic membrane disposed at the rear surface of the substrate at the opening of the second chamber; and

[0184] A channel disposed in the substrate, the channel including a proximal end fluidly connected to the first chamber and a distal end fluidly connected to the second chamber.

[0185] A1.1. The device according to embodiment A1, wherein the channel winds in a continuous and gradually widening curve from the proximal end to the distal end.

[0186] A2. The device according to A1.1, wherein the channel winds around a virtual center point on a virtual plane coinciding with the surface of the channel.

[0187] A3. The device according to embodiment A2, wherein the channel includes a front surface, and the channel winds around a virtual center point on a virtual flat plane coinciding with the front surface of the channel.

[0188] A4. The device according to any one of embodiments A1 - A3, including at least one calibration unit.

[0189] A5. The device according to embodiment A4, wherein each calibration unit includes the first chamber, the second chamber, and the channel.

[0190] A6. The device according to embodiment A5, wherein each calibration unit includes a hydrophobic membrane.

[0191] A7. The device according to any one of embodiments A4 - A6, including two or more units arranged in an array.

[0192] A8. The device according to any one of embodiments A1 - A7, wherein the channel includes an inner surface, and at least a portion of the inner surface of the channel is hydrophobic.

[0193] A9. The device according to embodiment A8, wherein at least a portion of the inner surface of the channel includes a coating, and the coating includes a hydrophobic material.

[0194] A10. The device according to any one of embodiments A1 - A9, wherein:

[0195] the channel includes a major length and a cross - section perpendicular to the major length, and

[0196] the cross - sectional geometry and the cross - sectional surface area of the channel are each substantially uniform along the length of the channel.

[0197] A11. The device according to embodiment A10, wherein the cross - sectional geometry is quadrilateral.

[0198] A12. The device according to any one of embodiments A1 - A11, wherein the first chamber includes a frustum - of - a - cone geometry.

[0199] A13. The device according to embodiment A12, wherein the frustum of the cone includes sides that taper from the front opening of the first chamber to the rear end.

[0200] A14. The device according to any one of embodiments A1 - A13, wherein the second chamber includes a frustum - of - a - cone geometry.

[0201] A15. The device according to embodiment A14, wherein the frustum of the cone includes sides that flare from the rear opening of the second chamber towards the front end.

[0202] A16. The device according to any one of embodiments A1 - A15, wherein the hydrophobic membrane is a disk.

[0203] A17. The device according to any one of embodiments A1 - A16, wherein the hydrophobic membrane includes a perimeter joined to the front surface of the substrate.

[0204] A18. The device according to any one of embodiments A1 - A17, wherein the second chamber includes sides, and the distal end of the channel is disposed at the sides and the front end of the second chamber.

[0205] A19. The device according to any one of embodiments A1 - A18, wherein the substrate includes a well, the well includes sides, and the proximal end of the channel is disposed at the sides of the well.

[0206] A20. The device according to embodiment A19, wherein the well includes a front opening, the front opening of the well is disposed at a rear position relative to the rear end of the first chamber, and the front opening of the well is concentrically aligned with the rear end of the first chamber.

[0207] A21. The device according to any one of embodiments A1 - A20, wherein the substrate includes a front portion and a rear portion fused to the front portion.

[0208] A22. The device according to embodiment A21, wherein the front portion includes the first chamber.

[0209] A23. The device according to embodiment A21 or A22, wherein the rear portion includes the second chamber and the hydrophobic membrane.

[0210] A24. The device according to any one of embodiments A21 - A23, wherein:

[0211] The channel includes a front surface, a rear surface, and side surfaces;

[0212] The rear surface and the side surfaces of the channel are disposed in the rear portion; and

[0213] The front surface of the channel is defined by the rear surface of the front portion fused to the rear portion.

[0214] A25. The device according to any one of embodiments A21 - A24, wherein the well is disposed in the rear portion.

[0215] B0.1. A method of manufacturing the device according to any one of embodiments A0.1 - A0.28, including preparing a substrate by an injection molding process, an embossing process, and / or a die-cutting process.

[0216] B0.2. A method of manufacturing the device according to any one of embodiments A0.1 - A0.28, including preparing a front member and a rear member by a stamping process.

[0217] B0.3. A method of manufacturing the device according to any one of embodiments A0.1 - A0.28, including:

[0218] Providing a substrate, a front member, and a rear member; and

[0219] Bonding the front surface of the rear member to at least a portion of the rear surface of the substrate, and bonding the rear surface of the front member to at least a portion of the front surface of the substrate.

[0220] B0.4. The method according to embodiment B0.3, wherein the joining includes solvent joining, laser joining, adhesive joining, ultrasonic welding, and / or plasma joining.

[0221] B1. A method for manufacturing the device according to any one of embodiments A1 - A25, comprising preparing the rear part according to any one of embodiments A21 - A25 by an injection molding process, an embossing process, and / or a die-cutting process.

[0222] B2. A method for manufacturing the device according to any one of embodiments A1 - A25, comprising preparing the front part according to any one of embodiments A21 - A25 by an injection molding process, an embossing process, and / or a die-cutting process.

[0223] B3. A method for manufacturing the device according to any one of embodiments A1 - A25, comprising: providing a rear part and a front part according to any one of embodiments A21 - A25; and

[0224] joining the rear part to the front part.

[0225] B4. The method according to embodiment B3, wherein the joining includes solvent joining, laser joining, adhesive joining, ultrasonic welding, and / or plasma joining.

[0226] C1. A method for determining the volume of fluid dispensed by a fluid dispenser device, comprising:

[0227] dispensing fluid from the fluid dispensing device into a first chamber of a volume calibration device that does not contain fluid, whereby the first chamber of the calibration device includes the dispensed fluid from the fluid dispenser device; wherein:

[0228] the dispensed fluid flows from the first chamber into a channel, the channel including a proximal end associated with the first chamber;

[0229] the dispensed fluid flows in the channel to a second chamber associated with the distal end of the channel;

[0230] the dispensed fluid flows in the channel under a pressure difference between the first chamber and the second chamber; and

[0231] the dispensed fluid fills the second chamber and a portion of the channel in a continuous fluid path between the second chamber and a proximal fluid termination point until the proximal fluid termination point between the proximal end and the distal end of the channel.

[0232] determining the position of the proximal fluid termination point in the channel; and

[0233] Determine the volume of the dispensed fluid in the calibration device based on the position of the proximal fluid end point.

[0234] C2. The method according to embodiment C1, including applying a vacuum to the volume calibration device.

[0235] C3. The method according to embodiment C1 or C2, wherein:

[0236] The fluid dispensing device is a multi-channel dispensing device including a plurality of dispenser units;

[0237] The calibration device includes an array of calibration units; and

[0238] Each calibration unit in the calibration device receives the volume of the dispensed fluid from one of the dispenser units.

[0239] C4. The method according to any one of embodiments C1 - C3, wherein the volume calibration device includes a hydrophobic membrane associated with the second chamber, and the distal fluid end point of the dispensed fluid is disposed at the hydrophobic membrane.

[0240] C5. The method according to any one of embodiments C1 - C4, wherein the volume calibration device is the device according to any one of embodiments A1 - A25.

[0241] C6. The method according to any one of embodiments C1 - C5, wherein the volume calibration device is the device according to any one of embodiments A0.1 - A0.28.

[0242] The entire content of each patent, patent application, published text, and literature cited herein is incorporated herein by reference. The citation of patents, patent applications, published texts, and literature is not an admission that any of the foregoing is relevant prior art, nor does it constitute any admission as to the content or date of these published texts or literature. Their citation does not represent a search of the relevant disclosures. All statements regarding the date or content of the documents are based on available information and do not represent an admission of their accuracy or correctness.

[0243] The technology has been described with reference to specific embodiments. The terms and expressions used herein to describe the technology are descriptive and not necessarily restrictive. Certain modifications to the disclosed embodiments may be considered within the scope of the technology. Certain aspects of the disclosed embodiments may be practiced appropriately in the presence or absence of certain elements not specifically disclosed herein.

[0244] Each of the terms "comprising", "consisting essentially of", and "consisting of" may be replaced by any one of the other two terms. The term "a" or "an" may refer to one or more of the elements it modifies (e.g., "a reagent" may mean one or more reagents), unless the context clearly dictates one or more than one of the elements. As used herein, the term "about" refers to a value within 10% of the underlying parameter (i.e., plus or minus 10%; e.g., a weight of "about 100 grams" may include weights between 90 grams and 110 grams). When the term "about" modifies each value at the beginning of a list of values (e.g., "about 1, 2, and 3"), it refers to "about 1, about 2, and about 3". When describing a list of values, the list includes all intermediate values and all fractional values thereof (e.g., a list of values "80%, 85%, or 90%" includes the intermediate value 86% and the fractional value 86.4%). When the term "or more" follows a list of values, the term "or more" applies to each of the values listed (e.g., a list of "80%, 90%, 95%, or more" or "80%, 90%, 95%, or more" or "80%, 90%, or 95%, or more" means "80% or more, 90% or more, or 95% or more"). When describing a list of values, the list includes all ranges between any two of the values listed (e.g., a list of "80%, 90%, or 95%" includes the ranges "80% to 90%", "80% to 95%", and "90% to 95%").

[0245] Exemplary embodiments of the present technology are set forth in the following claims. Multiple embodiments of the present invention have been described. However, it is understood that various modifications may be made without departing from the spirit and scope of the present invention. Accordingly, other embodiments are within the scope of the following claims.

Claims

1. A fluid dispenser volume calibration device, Comprising: A front surface, a rear surface, side surfaces, a first chamber, a second chamber, a hydrophobic membrane, and a channel, wherein: The first chamber includes a front opening and a rear end, and the front opening is disposed at the front surface of the device; The second chamber includes a rear end, side walls, and a front end; The hydrophobic membrane is in fluid connection with the second chamber; and The channel includes a proximal end in fluid connection with the first chamber and a distal end in fluid connection with the second chamber.

2. The device according to claim 1, Wherein, The second chamber includes a front port disposed at the front end of the second chamber and a rear port disposed at the rear end and the side walls of the second chamber, and the distal end of the channel is disposed at the rear port of the second chamber.

3. The device according to claim 1 or claim 2, Comprising: (i), a well, wherein: The well includes a well front end, well side walls, and a well rear end; The first chamber includes a rear port disposed at the rear end of the first chamber; The well front end is disposed at the rear port of the first chamber; The well includes a well rear port disposed at the well rear end and the well side walls; and The proximal end of the channel is disposed at the well rear port; or (ii), including a vent, wherein: The vent includes a vent front end, vent side walls, and a vent rear end; and The vent rear end is disposed at the front port of the second chamber; or (iii), a combination of (i) and (ii).

4. The device according to any one of claims 1-3, including a substrate, Wherein: The substrate includes a front surface and a rear surface; The substrate includes a recess; The recess is optionally disposed at the front surface of the substrate; The recess includes a recessed surface and recess side walls; and The recessed surface includes a recessed surface port; and Optionally: (i), the vent front end is disposed at the recessed surface port; or (ii), the recess includes a side wall port disposed in the recess side walls; Or (iii), the hydrophobic membrane is disposed in the recess; or (iv), a combination of (i) and (ii), a combination of (ii) and (iii), a combination of (i) and (iii), or a combination of (i), (ii), and (iii).

5. The device according to any one of claims 1-4, including a pipe, wherein the pipe includes a pipe end associated with the void volume of the hydrophobic membrane; and Optionally, wherein the pipe end is disposed at the hydrophobic membrane, or at the recess, or at the recess side wall port.

6. The device according to claim 5, including a lateral pipe, Wherein: The lateral pipe includes a lateral pipe proximal end, a lateral pipe distal end, and lateral pipe side walls; and The lateral pipe proximal end is disposed at the recess side wall port.

7. The device according to claim 5 or 6, including an axial pipe, Wherein: The axial pipe includes an axial pipe front end, an axial pipe rear end, axial pipe side walls, and an axial pipe front port, and the axial pipe front port is disposed in the axial pipe side walls at the axial pipe front end; and The rear end of the axial pipe is disposed at the rear surface of the substrate.

8. The device according to any one of claims 1-7, comprising a front member, wherein: The front member includes a front surface, a rear surface, and a front member hole; The rear surface of the front member is joined to at least a portion of the front surface of the substrate; and The substrate includes a first chamber hole, the first chamber hole including a hole front opening disposed at the front surface of the substrate; and The front member hole is aligned with and extends coextensively with the first chamber front hole in the substrate, and is the first chamber front opening of the device; and Optionally: (i), the front member includes a solid portion covering the recess and a hydrophobic membrane disposed in the recess; or (ii), the front member includes a solid portion covering the lateral pipe, and the rear surface of the front member at that portion is the lateral pipe top plate surface; or (iii), the front member includes a solid portion covering the front end of the axial pipe, and the rear surface of the front member at that portion is the axial pipe top plate surface; or (iv), a combination of (i) and (ii), a combination of (ii) and (iii), a combination of (i) and (iii), or a combination of (i), (ii), and (iii).

9. The device according to any one of claims 1-8, comprising a rear member, wherein: The rear member includes a front surface, a rear surface, and a rear member hole; The front surface of the rear member is joined to at least a portion of the rear surface of the substrate; and The rear member hole is coaxially and / or concentrically disposed at the rear end of the axial pipe; and Optionally: (i), the rear member includes a solid portion disposed at the channel, and the front surface of the rear member at that portion is the channel bottom plate; or (ii), the rear member includes a solid portion disposed at the rear end of the second chamber, and the front surface of the rear member at that portion is the second chamber bottom plate; or (iii), the rear member includes a solid portion disposed at the rear end of the well, and the front surface of the rear member at that portion is the well bottom plate; or (iv), a combination of (i) and (ii), a combination of (ii) and (iii), a combination of (i) and (iii), or a combination of (i), (ii), and (iii).

10. The device according to any one of claims 1-9, comprising at least one calibration unit, and optionally, two or more calibration units in an array.

11. The device according to claim 10, wherein, Each calibration unit includes the first chamber, the second chamber, the channel, and the hydrophobic membrane, and optionally, each calibration unit includes the well, the vent, the recess, the lateral pipe, and the axial pipe.

12. The device according to any one of claims 1-11, wherein, The channel includes an inner surface, and at least a portion of the inner surface of the channel is hydrophobic, and optionally a portion of the inner surface of the channel includes a coating comprising a hydrophobic material.

13. The device according to any one of claims 1-12, wherein: the channel includes two or more substantially linear members or linear members, and a transition member connecting the two substantially linear members or linear members; and the substantially linear members or linear members are parallel or substantially parallel to each other.

14. The device according to any one of claims 1-13, wherein, the first chamber and the second chamber are each independently a triangular prism or a triangular frustum.

15. The device according to claim 14, wherein: the first chamber and the second chamber have a triangular cross-section; the triangular cross-section is a right-angled triangular cross-section including a hypotenuse; and the hypotenuse of the right-angled triangular cross-section of the first chamber is opposite to and parallel or substantially parallel to the hypotenuse of the right-angled triangular cross-section of the second chamber.

16. A method for manufacturing the device according to any one of claims 9-15, comprising: providing the substrate, the front member and the rear member; and joining the front surface of the rear member to at least a part of the rear surface of the substrate, and joining the rear surface of the front member to at least a part of the front surface of the substrate.

17. A method for determining the volume of fluid dispensed by a fluid dispenser device, comprising: dispensing fluid from the fluid dispenser device into a first chamber of a volume calibration device that does not contain fluid, whereby the first chamber of the calibration device includes the dispensed fluid from the fluid dispenser device; wherein: the dispensed fluid flows from the first chamber into a channel, the channel including a proximal end associated with the first chamber; the dispensed fluid flows in the channel to a second chamber associated with the distal end of the channel; the dispensed fluid flows in the channel under a pressure difference between the first chamber and the second chamber; and the dispensed fluid fills the second chamber, and fills a part of the channel in a continuous fluid path between the second chamber and the fluid end point until the fluid end point between the proximal end and the distal end of the channel. determining the position of the fluid end point in the channel; and determining the volume of the dispensed fluid in the calibration device based on the position of the fluid end point.

18. The method according to claim 17, including applying a vacuum to the volume calibration device.

19. The method according to claim 17 or claim 18, wherein: the fluid dispenser device is a multi-channel dispenser device including a plurality of dispenser units; the calibration device includes an array of calibration units; and each calibration unit in the calibration device receives a volume of dispensed fluid from one of the dispenser units.

20. The method according to any one of claims 1-3, wherein, the volume calibration device includes a hydrophobic membrane associated with the second chamber, and the distal fluid end point of the dispensed fluid is provided at the hydrophobic membrane.

21. The method according to any one of claims 17-20, wherein, the volume calibration device is the device according to any one of claims 1-15.

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