CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims the benefit of priority under 35 U.S.C. §119(e), to U.S. Provisional Application No. 63/424,669, as filed November 11 , 2022, the contents of which are incorporated herein by reference in its entirety.

BACKGROUND

[0002] Transferring and combining fluids in a system in an accurate and reliable manner can be difficult, and systems for doing in a reliable manner so can be complicated and expensive.

SUMMARY OF THE INVENTION

[0003] Accordingly, new systems, apparatus, and methods for microfluidic handling of fluids using valve systems are desirable.

[0004] One embodiment provides a membrane valve system, including: an etched substrate including a network of channels defining a plurality of valve connection points arranged in an arcuate pattern; a deformable membrane including a plurality of raised valve elements complementary to the plurality of valve connection points; and a rotary valve pin including a valve face including a plurality of valve controlling mechanisms, the plurality of valve controlling mechanisms including a pair of valve controlling mechanisms complementary to a pair of raised valve elements of the plurality of raised valve elements, the valve face being configured to depress each of the plurality of raised valve elements to maintain each in a closed position, and the pair of valve controlling mechanisms of the valve face being configured to permit the pair of raised valve elements of the plurality of raised valve elements to be in an open position.

[0005] Another embodiment provides a fluidic handling system, including: an etched substrate including a network of channels defining a plurality of valve connection points; a plurality of valves located at each of the respective plurality of valve connection points, each of the plurality of valves being fluidically coupled to the network of channels and comprising a shape memory alloy (SMA); and a plurality of chambers fluidically coupled to the network of channels.

[0006] Yet another embodiment provides a fluidic handing system, including: a rotary

1

SUBSTITUTE SHEET (RULE 26) valve including a valve body with a valve insert disposed therein, the valve body including a plurality of side ports, and the valve insert including at least one channel; a plurality of chambers each fluidly coupled to a respective side port of the plurality of side ports; and a motor coupled to the valve insert and configured to rotate the valve insert within the valve body to align the at least one channel of the valve insert with at least one of the plurality of side ports of the valve body to fluidly couple at least one chamber of the plurality of chambers to permit fluid to flow to or from the at least one chamber.

BRIEF DESCRIPTION OF THE DRAWINGS

[0007] Various objects, features, and advantages of the disclosed subject matter can be more fully appreciated with reference to the following detailed description of the disclosed subject matter when considered in connection with the following drawings, in which like reference numerals identify like elements.

[0008] FIG. 1 A shows a diagram of a construction of the valve face of a valve pin (an example of which is shown in perspective view, inset). FIG. 1B shows a diagram of a construction of a network of channels on an etched substrate. While the ports are shown as being arranged in a circle, in some instances the ports (green circles) may be arranged along the sides of a rectangular substrate as shown in FIG. 1C.

[0009] FIG. 2, left, shows the valve pin face of FIG. 1A and FIG. 2, right, shows an alternative construction of a network of channels on an etched substrate.

[0010] FIG. 3 shows a diagram of a membrane valve system such as those disclosed herein which can be used to combine, analyze, and deliver fluids.

[0011] FIG. 4A shows a diagram of a valve face with two notches whose radial positions can be set independently of one another; the valve face of FIG. 4A can be used with a number of channel networks including that shown in FIG. 4B.

[0012] FIG. 5 shows a diagram of a construction of a network of channels on an etched substrate which includes a single radial array of valve connection points, two metered reservoirs (50pl_ and 200pL), and a pump.

[0013] FIG. 6 shows a diagram of a construction of a network of channels on an etched substrate in which fluid can flow through the pump between different portions of the network of channels.

[0014] FIG. 7 shows a diagram of a construction of a network of channels on an etched substrate which includes two concentric radial arrays of valve connection points, two metered

2

SUBSTITUTE SHEET (RULE 26) reservoirs (50pL and 200pL), and a pump.

[0015] FIG. 8 shows a diagram of a construction of a network of channels on an etched substrate which includes two concentric radial arrays of valve connection points, two metered reservoirs (50pL and 200pL), and a pump.

[0016] FIG. 9 shows a diagram of a construction of a network of channels on an etched substrate in which fluid can flow through the pump.

[0017] FIG. 10 shows a diagram of a construction of a network of channels on an etched substrate in which fluid can flow through the pump between different portions of the network of channels, which is controlled by setting a position of the valve pin which includes two notches, e.g. whose radial positions can be set independently of one another (inset).

[0018] FIG. 11 shows a diagram of a construction of a network of channels on an etched substrate which includes two non-concentric radial arrangements of valve connection points, each of which is connected to the same pump (e.g. a peristaltic pump) as well as to a number of external ports as shown. While the reservoirs are labeled for a particular application, in practice this system can be used with a wide variety of complex systems.

[0019] FIG. 12A shows a technical drawing of a channel network for one construction of a rotary membrane valve (RMV) and FIG. 12B shows a technical drawing of a valve face of a valve pin for use with an RMV.

[0020] FIG. 13A shows a comparison between a schematic diagram of a channel network (top) and a particular implementation of the network (bottom). FIG. 13B shows a comparison between a schematic diagram of a valve face of a valve pin (top) and a particular implementation of the valve pin (bottom).

[0021] FIG. 14 shows a construction of a valve pin/pusher showing the valve face (left) with a motor engagement surface (right) on the side opposite the valve face.

[0022] FIG. 15A shows a valve pin in which the valve face includes two notches as well as a radial slot. FIG. 15B shows the valve pin of FIG. 15A rotated such that the notches are aligned with a pair of outer valve connection points to cause external ports A and B to be connected to one another; FIG. 15C shows the valve pin of FIG. 15A rotated such that the radial slot is aligned with a pair of inner and outer valve connection points to cause port C to be connected to the center port.

[0023] FIG. 16A shows the fluid path between external ports A and B when the valve pin is in the position shown in FIG. 15B; FIG. 16B shows the fluid path between external port C and the center port when the valve pin is in the position shown in FIG. 15C.

3

SUBSTITUTE SHEET (RULE 26) [0024] FIGS. 17A and 17B show exploded views at different angles of (from bottom to top) a valve pin, a positioning pin, a membrane layer, and a channels/ports layer.

[0025] FIG. 18A shows a cross-sectional side view of a construction of the membrane valve system and FIG. 18B shows a detailed view of the valve pin, membrane, and substrate with a pressure sensitive adhesive (PSA) applied.

[0026] FIG. 19A shows a cross-sectional side view of another construction of the membrane valve system and FIG. 19B shows a detailed view of the substrate, membrane, valve pin, and ports extension block with a pressure sensitive adhesive (PSA) applied.

[0027] FIG. 20A shows a perspective view of a valve pin attached to a substrate from which a series of external ports emerge; in this construction, the valve pin is between the membrane layer and the chambers. Openings in the membrane layer can then be coupled to chambers, as shown in FIG. 20B, which shows a cross-sectional side view of a construction of the membrane valve system.

[0028] FIG. 21A shows a perspective view of a valve pin/pusher attached to a substrate from which a series of external ports emerge; in this construction, the valve pin/pusher is disposed within the block containing the chambers so that a port extension block is not needed. Openings in the membrane layer can be coupled to chambers, as shown in FIG. 21 B, which shows a cross-sectional side view of a construction of the membrane valve system.

[0029] FIG. 22A shows a construction of the membrane valve system in which the valve is actuated from the bottom; FIG. 22B shows a construction of the membrane valve system in which the valve is actuated from the top.

[0030] FIG. 23 shows another construction of a membrane valve system in which the valve pin includes movable actuators which can be used instead of, or in addition to, the notches and slot design of other constructions.

[0031] FIG. 24A shows an alternative peristaltic-type pump which uses several rollers and which can be incorporated into constructions of the membrane valve system. FIG. 24B shows the pump of FIG. 24A attached to a substrate.

[0032] FIG. 25A shows a test of cell viability either as a control (left-hand bar at each cell concentration) or when the cells are passed through the membrane valve at the two different concentrations (second and third bars, respectively, at each concentration) for one pass (second bars) or two passes (third bars). FIG. 25B shows a test of cell recovery following one (left-hand bars at each cell concentration) or two (right-hand bars at each cell concentration) valve passes.

4

SUBSTITUTE SHEET (RULE 26) [0033] FIG. 26 shows a diagram of a construction of a microfluidic handling system for implementation on an etched substrate using shape memory alloy (SMA) based valves.

[0034] FIG. 27A shows a diagrammatic side view of a construction of an SMA based valve in a closed state (top) and an open state (bottom). FIG. 27B shows a photograph of two valves such as those diagrammed in FIG. 27A alongside a ball-point pen for size comparison, with one valve turned on its side to show the input and output connection points. FIG. 27C shows bottom and side view drawings of an SMA valve such as those shown in FIGS. 27A and 27B with various dimensions indicated (in mm).

[0035] FIG. 28 shows a diagram of another construction of a microfluidic handling system for implementation on an etched substrate using SMA based valves.

[0036] FIG. 29 shows a side view diagram of another construction of an SMA based valve for use with a microfluidic system.

[0037] FIG. 30 shows a perspective view (top) of a microfluidic handling system as well as a view of the underside (lower left) and an exploded view (lower right).

[0038] FIG. 31 A shows a diagram of a microfluidic system in which valves (red and while circles) are located under the chambers (larger blue circles) and FIG. 31 B shows a diagram of a microfluidic system in which valves (orange circles) are located between the chambers (larger blue circles).

[0039] FIG. 32A shows a diagram of a microfluidic system in which valves are located under (red and white circles) as well as between (orange circles) the chambers (larger blue circles). FIG. 32B shows a perspective view diagram from the underside of the placement of valves in the under-chamber or between-chamber configurations.

[0040] FIG. 33A shows a diagram of an under-chamber type valve which includes a bypass pathway (i.e. by which fluid in the microchannel can flow around the valve whether it is open or closed) and FIG. 33B shows a diagram of an under-chamber type valve which does not include a bypass pathway.

[0041] FIG. 34A shows an SMA valve that is mounted under a chamber in an open position (due to the SMA being heated, as indicated by the red line, and consequently flattened) in which fluid is flowing from the chamber into the channel through the valve seat (and also may flow through the channel around the valve seat). FIG. 34B shows the SMA valve of FIG. 34A in a closed position (due to the SMA being cooled, as indicated by the blue line, and consequently deformed) such that fluid is no longer flowing from the chamber.

[0042] FIG. 35A shows an embodiment of an SMA valve in a closed position (SMA is

5

SUBSTITUTE SHEET (RULE 26) cold and deformed, blue line), closing a chamber, and FIG. 35B shows the SMA valve in an open position (SMA is heated and relatively flat, red line), permitting the chamber to be open. [0043] FIG. 36A shows an example of a rotary valve coupled to a motor (e.g. a servo motor) and a controller (circuit board at top). FIG. 36B shows a diagram of a rotary valve system depicting the motor (center), a switching valve in cross-section shown in two different positions (top left), a distribution valve (top center), an on/off valve in cross-section (top right), and a valve shown from the back side (i.e. the side that couples to the motor) (lower left).

[0044] FIG. 37A shows a diagram of a rotary valve in which a valve insert (white T- shaped structure in the center of the valve) couples one of the side ports to the end port when the insert is rotated (indicated by the curved arrow) as driven by the motor. FIG. 37B shows a cutaway side view of an embodiment of a rotary valve system showing how the valve insert can connect two side ports to one another (and optionally to the end port) (left) to create a switch valve, or connect a side port to the end port (right) to create a distribution valve.

[0045] FIGS. 38A and 38B show embodiments of a rotary valve in which the valve insert includes channels that allow the insert to act as either a distribution valve or a switch valve, depending on the position to which the insert is turned.

[0046] FIG. 39 shows a diagram of a fluidic handling system based on the use of a rotary valve system.

[0047] FIG. 40 shows a diagram of the valve from FIG. 39 in a side view (top panel) and in a top view (bottom panel) showing the position of the valve insert and in particular how the valve insert acts as a distribution valve in this position.

[0048] FIGS. 41A-41C show how the channels leading to the side ports may be blocked at or near their outer ends so that they instead may be joined with channel segments leading to the end face of the valve body.

[0049] FIG. 42 shows a motor and controller coupled to a bracket for use with a fluidic handling system based on the use of a rotary valve system.

[0050] FIG. 43 shows the motor, controller, and bracket of FIG. 42 coupled to a chamber block (left), as an exploded view from the side (center), and as an exploded view from underneath (right).

[0051] FIG. 44 shows an exploded view of the fluidic handling system using a rotary valve from different angles.

DETAILED DESCRIPTION

6

SUBSTITUTE SHEET (RULE 26) [0052] In accordance with some embodiments of the disclosed subject matter, mechanisms (which can include systems, apparatus, and methods) for microfluidic handling of fluids using valve systems are provided.

[0053] Disclosed herein are embodiments of systems for transferring and combining fluids in a reliable, automated manner using volumes ranging from tens of microliters to hundreds of milliliters. The systems include those based on membrane valves, shape memory alloy valves, and rotary valves.

[0054] Membrane Valve Systems

[0055] In various embodiments, systems for microfluidic handling of fluids using membrane valves are provided (see FIGS. 1-35). The system may include an etched substrate having a network of channels formed thereon, where the network defines a plurality of valve connection points, generally arranged in an arcuate pattern. The valve connection points may include the ends of two adjacent channels which do not connect on the substrate. Instead, a deformable membrane including a plurality of raised valve elements that are complementary to the plurality of valve connection points is placed over substrate and pressure is selectively applied to the raised valve elements to open or close the connections at the valve connection points. Pressure may be selectively applied by a rotary valve pin which includes a valve face having a plurality of valve controlling mechanisms.

[0056] In one embodiment, the plurality of valve controlling mechanisms may include a pair of valve controlling mechanisms that are complementary to a pair of raised valve elements of the plurality of raised valve elements (FIG. 1 A). The valve face may be configured to depress each of the plurality of raised valve elements to maintain each in a closed position while the pair of valve controlling mechanisms of the valve face is configured to permit the pair of raised valve elements of the plurality of raised valve elements to be in an open position.

[0057] In some embodiments, the plurality of valve connection points may be a plurality of inner valve connection points and the etched substrate may further include a plurality of outer valve connection points. Each of the plurality of outer valve connection points is radially aligned with and fluidical ly coupled to a respective inner valve connection point of the plurality of inner valve connection points (FIG. 1 B). In certain embodiments, the plurality of raised valve elements may include a plurality of raised inner valve elements, and the deformable membrane may further include a plurality of raised outer valve elements complementary to the plurality of outer valve connection points.

[0058] In various embodiments, the pair of valve controlling mechanisms may include a

7

SUBSTITUTE SHEET (RULE 26) pair of notches in the valve face and the plurality of valve controlling mechanisms of the rotary valve pin may further include a radial slot formed therein (FIG. 1 A). The radial slot may be configured to permit a raised outer valve element of the plurality of raised outer valve elements and a radially-aligned raised inner valve element of the plurality of raised inner valve elements to each be in the open position. In some embodiments, the valve face may include a circular raised band which is wide enough to contact both the inner and outer raised valve elements at the same time, where the raised band may include notches and/or a slot (e.g. as shown in FIG. 13B, top). In other embodiments the valve face may include two separate circular raised bands, where one of the raised bands may include two notches (e.g. see the outer raised band in FIG. 13B, bottom) and/or an aligned pair of notches in the two bands that act as a "slot" (e.g. see the inner and outer bands in FIG. 13B, bottom). In general, terms such as "notch" and "slot" encompass a variety of mechanisms that permit the raised portion(s) of the membrane to move away from the substrate in order to permit a valve to transition to the open state. These mechanisms include various features in the valve pin face including notches and slots as well as various other depressions having other shapes such as circles, ovals, squares, rectangles, etc. In some embodiments, the network of channels may include a central channel, where the radial slot may be configured to fluidical ly couple the raised outer valve element and the raised inner valve element to the central channel. For example, as shown in FIG. 2 (right) the network of channels is depicted in a hub and spoke pattern where the hub represents a central channel to which all of the external ports can be connected (depending on the position of the valve pin); the central channel can then be directed to an external port as shown in FIG. 2 (right) which may be connected to the central channel via a continuous fluid path which does not include any valve elements.

[0059] In various embodiments, the pair of notches may be adjacent to one another as shown in FIG. 1A. By having the notches adjacent to one another, when the valve pin is aligned with a pair of raised valve elements then this fluidly couples two adjacent external ports to one another (FIG. 16A). However, in other embodiments the notches may not be adjacent to one another and the valve face can be set up with valve controlling mechanisms (such as notches or, as disclosed below, actuators) that are in other positions relative to one another.

[0060] In other embodiments, the radial slot may be disposed opposite the pair of notches as shown in FIG. 1A. The radial slot may be placed at such an angle relative to the notches that when the notches are engaged with a pair of adjacent raised valve elements, the radial slot is not aligned with any of the raised valve elements. Similarly, when the radial slot is

8

SUBSTITUTE SHEET (RULE 26) aligned with a pair of radially-aligned raised valve elements, the notches are not aligned with any of the raised valve elements. Although the radial slot is shown as being located opposite the pair of notches, in various embodiments the radial slot may be arranged in other orientations relative to the notches, generally sufficiently offset so that the notches and slot are not aligned with the raised valve elements at the same time.

[0061] In various embodiments, the plurality of valve controlling mechanisms may include a pair of notches in the valve face in which a first radial position of a first notch of the pair of notches of the rotary valve pin can be set independently of a second radial position of a second notch of the pair of notches (FIG. 4A). In some embodiments, the first notch may permit a raised outer valve element of the plurality of raised outer valve elements to be in the open position and the second notch may permit an inner raised valve element of the plurality of raised inner valve elements to be in the open position. In certain embodiments, the first radial position may be different from the second radial position. To permit independent adjustment, each of the valve controlling mechanisms/notches may be attached to separately movable portions of the valve pin, such that the notches or other valve controlling mechanisms can be rotatably set in different orientations.

[0062] In various embodiments, the system may include a pump fluidically coupled to the network of channels (FIG. 5). The pump may be used to draw fluid from one of the external ports and transfer the fluid to another one of the external ports, e.g. for combining fluids from two different parts of the system. In some embodiments such as the system of FIG. 5, the fluid may not come into contact with the pump and instead the fluid may be transiently stored in one or both of the metered reservoirs of various internal volumes while being transferred between external ports. In other embodiments, the fluid may move through the pump to one or more external ports coupled to the pump on its output side (e.g. FIG. 6).

[0063] In certain embodiments, the pump may include an arcuate raised portion of the deformable membrane (e.g. as shown in FIG. 1 C). To move fluid (including liquids, such as saline or reagents, or gases such as air) the system may include a pump pin which includes one or more raised portions which, when the pump pin is pressed against the membrane and rotated, pushes along the arcuate raised portion of the deformable membrane to move the fluid. The raised portion of the membrane that makes up the pump is generally arcuate so that the rotating pump pin is able to push fluid through the pump; nevertheless, in some embodiments the pump portion of the membrane can have other shapes, such as a straight line, in which case the pump pin is suitably adapted to operate with the alternate shaped pump. In one

9

SUBSTITUTE SHEET (RULE 26) embodiment, the arcuate raised portion of the deformable membrane includes a first end fluidically coupled to the network of channels and a second end fl uidically coupled to an external port. In another embodiment, the arcuate raised portion of the deformable membrane may include a first end fluidically coupled to the network of channels and a second end fluidically coupled to the network of channels. That is, one end of the raised portion of the membrane that operates as a pump may be associated with a channel that is part of the network of channels and the other end of the raised portion may be associated either with a separate channel that leads directly to an external port (e.g. as in FIG. 5) or with a channel that is part of another portion of the network of channels (e.g. as in FIG. 6).

[0064] Thus, in certain embodiments the network of channels may include a first portion fluidically coupled to the first end of the arcuate raised portion and a second portion fluidically coupled to the second end of the arcuate raised portion (FIG. 6). Accordingly, movement of the pump pin may push fluid from the first portion of the network of channels through the arcuate raised portion to the second portion of the network of channels.

[0065] As noted above, in some embodiments the etched substrate may further include at least one reservoir fluidically coupled to the network of channels. In some embodiments, the at least one reservoir may include a first reservoir and a second reservoir (e.g. see FIG. 5). In certain embodiments, the at least one reservoir may be disposed between the network of channels and the pump such that the pump is fluidically coupled to the network via the at least one reservoir (FIG. 5).

[0066] In various embodiments, the etched substrate may further include a plurality of external ports where each valve connection point of the plurality of valve connection points is fluidically coupled to a respective external port of the plurality of external ports, for example when there is a single circular arrangement of valve connection points as in FIG. 5. In certain embodiments in which there are two or more circular arrangements of valve connection points including inner and outer valve connection points, each outer valve connection point of the plurality of outer valve connection points may be fluidically coupled to a respective external port of the plurality of external ports (e.g. see FIG. 1 B). In particular embodiments in which independent fluidic channels are provided between each external port and the respective inner and outer valve connection points (e.g. as shown in FIG. 4B), each inner valve connection point of the plurality of inner valve connection points may also be fluidically coupled to a respective external port of the plurality of external ports.

[0067] In various embodiments, the system may further include a plurality of chambers

10

SUBSTITUTE SHEET (RULE 26) (e.g. see FIG. 18A). Each of the plurality of chambers may be fluidically coupled to a respective external port of the plurality of external ports and at least one of the plurality of chambers may include a reagent disposed therein, for example any of the reagents shown in FIGS. 3-8 and 11. [0068] In various embodiments of the system, different arrangements are provided for coupling a motor to the valve pin in order to control the position of the valve pin during operation. Thus, in some embodiments the system may further include a motor coupled to the rotary valve pin, where the motor may be configured to rotate the rotary valve pin. In certain embodiments, the valve pin and the motor may be disposed on a first side of the substrate and the plurality of chambers may be disposed on a second side of the substrate opposite the first side, as shown in FIG. 18A. In other embodiments, the valve pin and the motor may be disposed on the same side of the substrate as the plurality of chambers, as shown in FIG. 19A. [0069] In some embodiments, the pump may include a peristaltic pump including a flexible tube, where the flexible tube may be fluidically coupled to the network of channels as shown in FIG. 11. In other embodiments, the pump may include a rotating pump pin which includes one or more roller on a face of the pump pin to push against the arcuate raised portion of the deformable membrane or tube (FIGS. 24A, 24B).

[0070] In some embodiments of the system, the substrate may include two or more non- concentric groupings of valve connection points, as shown in FIG. 11 , which would include separate valve pins for each grouping of valve connection points. Thus, in various embodiments the plurality of valve connection points may include a first plurality of valve connection points, where the plurality of raised valve elements complementary to the plurality of valve connection points on the deformable membrane may include a first plurality of valve elements complementary to the first plurality of valve connection points, and where the rotary valve pin may include a first rotary valve pin having a first valve face. The etched substrate may further include a second plurality of valve connection points, where the deformable membrane may further include a second plurality of raised valve elements complementary to the second plurality of valve connection points. The system may further include a second rotary valve pin including a second valve face having a plurality of valve controlling mechanisms, where the plurality of valve controlling mechanisms may be complementary to a respective pair of raised valve elements of the second plurality of raised valve elements. The valve face may be configured depress each of the second plurality of raised valve elements to maintain each in a closed position, and the plurality of valve controlling mechanisms of the valve face may be configured to permit the pair of raised valve elements of the second plurality of raised valve

11

SUBSTITUTE SHEET (RULE 26) elements to be in the open position. In various embodiments, the first plurality of valve connection points may be fluidically coupled to the second plurality of valve connection points using a pump, as shown in FIG. 11.

[0071] In certain embodiments, the valve controlling mechanisms may include movable actuators as shown in FIG. 23. Accordingly, in various embodiments the plurality of valve controlling mechanisms may include a plurality of movable actuators, where each of the plurality of movable actuators may be configured to move between an engaged position in which a movable actuator of the plurality of movable actuators presses against a raised valve element of the pair of raised valve elements to put the raised valve element into the closed position, and a disengaged position in which the movable actuator moves away from the raised valve element to permit the raised valve element to move into the open position. In some embodiments, the pair of valve controlling mechanisms may include a first pair of movable actuators, and the plurality of movable actuators may further include a second pair of movable actuators configured to actuate a raised outer valve element of the plurality of raised outer valve elements and a radially-aligned raised inner valve element of the plurality of raised inner valve elements between the open position and the closed position. In certain embodiments, the pair of valve controlling mechanisms may include a first pair of movable actuators, where radial positions of each of the pair of movable actuators can be set independently of one another.

[0072] FIGS. 1-35 show examples of various embodiments of the disclosed system. FIG. 1 A shows a diagram of an embodiment of the valve face of a valve pin (an example of which is shown in perspective view, inset) having a pair of notches (top and top right) and a radial slot (lower left); the dark portions of the circle a raised relative to the lighter portions. FIG. 1B shows a diagram of an embodiment of a network of channels on an etched substrate along with a number of valve connection points arranged in two concentric rings, with a complementary array of external ports each of which is coupled to a valve connection point. While the ports are shown as being arranged in a circle, in some instances the ports (green circles) may be arranged along the sides of a rectangular substrate as shown in FIG. 1C. FIG. 1C shows a region of a substrate which includes the external ports as well as an arcuate raised portion of the membrane which forms the pump. FIG. 2, left, shows the valve pin face of FIG. 1 A and FIG. 2, right, shows an alternative embodiment of a network of channels on an etched substrate which includes an additional channel extending from the center which leads to an additional port.

[0073] In various embodiments, the disclosed membrane valve system may be used to

12

SUBSTITUTE SHEET (RULE 26) combine reagents and deliver them to a reaction chamber as well as to perform analysis on fluids in the system, as shown in FIGS. 3-8 and 11. FIG. 3 shows a diagram of a membrane valve system such as those disclosed herein which can be used to combine, analyze, and deliver fluids. FIG. 4A shows a diagram of a valve face with two notches whose radial positions can be set independently of one another. The valve face of FIG. 4A can be used with a number of channel networks including that shown in FIG. 4B, which includes independent connections between each external port and each of the inner and outer valve connection points, in addition to a port that is connected to the center of the network of channels. While FIG. 4A shows a valve pin with two independently-controllable notches, in other embodiments the valve can include three or more independently-controllable notches.

[0074] FIG. 5 shows a diagram of an embodiment of a network of channels on an etched substrate which includes a single radial array of valve connection points, two metered reservoirs (50pL and 200pL), and a pump which can be used to move fluids from the external ports, store and/or mix the fluids in the reservoirs, and move the stored fluids to other external ports, which is controlled by setting a position of the valve pin. Although FIGS. 5, 7, and 8 show two reservoirs with volumes of 50pL and 200pL, in various embodiments the system may include other numbers of reservoirs (e.g. from 1-20) having various volumes (e.g. between 10pL and 1000pL).

[0075] FIG. 6 shows a diagram of an embodiment of a network of channels on an etched substrate in which fluid can flow through the pump between different portions of the network of channels, for example from an external port associated with the upper portion of the network of channels, through the pump, and to an external port associated with the lower portion of the network of channels, which is controlled by setting a position of the valve pin. Given that fluid can flow directly from one external port to another through the pump, there is less need in this embodiment for storage reservoirs. As a result, larger volumes of fluid can be handled with a system such as that shown in FIG. 6 in which fluid moves directly from one external port to another, such that volumes of anywhere from 10pL to 100mL or more can be handled using embodiments of the disclosed system.

[0076] FIG. 7 shows a diagram of an embodiment of a network of channels on an etched substrate which includes two concentric radial arrays of valve connection points, two metered reservoirs (50pL and 200pL), and a pump which can be used to move fluids from the external ports, store the fluids in the reservoirs, and move the stored fluids to other external ports (including the center port, which is in line with the reservoirs), which may be controlled by

13

SUBSTITUTE SHEET (RULE 26) setting a position of the valve pin (inset) as well as by opening or closing a valve associated with the pump (represented by a red dot).

[0077] FIG. 8 shows a diagram of an embodiment of a network of channels on an etched substrate which includes two concentric radial arrays of valve connection points, two metered reservoirs (50pL and 200pL), and a pump which can be used to move fluids from the external ports, store the fluids in the reservoirs, and move the stored fluids to other external ports (including the center port, which is in a separate branched channel from the reservoirs), which is controlled by setting a position of the valve pin (inset) as well as by opening or closing a valve associated with the pump (represented by a red dot).

[0078] For the embodiments of FIGS. 7 and 8, when a liquid needs to be transferred to the center port this can be accomplished by closing the red circle valve shown in the figures adjacent to the arcuate raised portion of the pump. To close the valve, the raised portion of the pump pin is placed onto the raised valve portion of the pump (indicated in red in FIGS. 7 and 8) to put the valve into the closed position; because the pump valve is in the same path as the arcuate raised portion of the pump, the valve can be contacted by the raised portion of the pump pin. Once the pump valve has been closed, liquid can be transferred via an external port from any chamber to the center port and chamber by creating a pressure differential (e.g. using an external pneumatic source) between the two chambers without using the on-chip pump. In various embodiments, liquids can be transferred between any two chambers in the systems of FIGS. 7 and 8 either by using the on-chip pump (which may include metered transfers) and/or an external pneumatic source can be used to apply positive or negative pressure to any of the chambers to move liquid between chambers; in the latter case, the pump valve needs to be placed in the closed position and valves associated with particular chambers need to be placed in the open position when the positive and/or negative pressure is applied to the chambers.

[0079] FIG. 9 shows a diagram of an embodiment of a network of channels on an etched substrate in which fluid can flow through the pump. In this embodiment, the external ports (larger gray circles) are disposed between respective inner and outer valve connection points. Also in this embodiment, the outer valve connection points are connected to the upper portion of the network of channels and the inner valve connection points are connected to the lower portion of the network of channels. The inner and outer valve connection points may be controlled by a valve pin which includes two notches, e.g. whose radial positions can be set independently of one another (inset).

[0080] FIG. 10 shows a diagram of an embodiment of a network of channels on an

14

SUBSTITUTE SHEET (RULE 26) etched substrate in which fluid can flow through the pump between different portions of the network of channels, for example from an external port associated with the upper portion of the network of channels, through the pump, and to an external port associated with the lower portion of the network of channels and/or to an external port associated with the center of the network, which is controlled by setting a position of the valve pin which includes two notches, e.g. whose radial positions can be set independently of one another (inset).

[0081] FIG. 11 shows a diagram of an embodiment of a network of channels on an etched substrate which includes two non-concentric radial arrangements of valve connection points, each of which is connected to the same pump (e.g. a peristaltic pump) as well as to a number of external ports as shown.

[0082] FIG. 12A shows a technical drawing of a channel network for one embodiment of a rotary membrane valve (RMV) and FIG. 12B shows a technical drawing of a valve face of a valve pin for use with an RMV.

[0083] FIG. 13A shows a comparison between a schematic diagram of a channel network (top) and a particular implementation of the network (bottom). FIG. 13B shows a comparison between a schematic diagram of a valve face of a valve pin (top) and a particular implementation of the valve pin (bottom).

[0084] FIG. 14 shows an embodiment of a pusher/valve pin showing the valve face (left) with a motor engagement surface (right) on the side opposite the valve face. The motor engagement surface may provide access to a shaped bore (e.g. a primarily hexagonal cross- sectional shape as shown) into which a shaft with a complementary shape can be inserted and serve to couple the motor to the valve pin in a manner that couples rotational movement of the motor to rotation of the valve pin.

[0085] FIG. 15A shows a valve pin in which the valve face includes two notches as well as a radial slot. FIG. 15B shows the valve pin of FIG. 15A rotated such that the notches are aligned with a pair of outer valve connection points to cause external ports A and B to be connected to one another (note that when the notches are aligned with a pair of external ports, the radial slot is not aligned with any of the valve connection points). FIG. 15C shows the valve pin of FIG. 15A rotated such that the radial slot is aligned with a pair of inner and outer valve connection points to cause port C to be connected to the center port (note that when the radial slot is aligned with a pair of inner and outer valve connection points, the pair of notches are not aligned with any of the valve connection points). FIG. 16A shows the fluid path between external ports A and B when the valve pin is in the position shown in FIG. 15B. FIG. 16B shows the fluid

15

SUBSTITUTE SHEET (RULE 26) path between external port C and the center port when the valve pin is in the position shown in FIG. 15C.

[0086] FIGS. 17A and 17B show exploded views of (from bottom to top) a valve pin, a positioning pin, a membrane layer, and a channels/ports layer. FIG. 17A shows the components at an angle which permits the valve face of the valve pin to be seen while FIG. 17B shows the components from a different angle which permits the etched channels of the substrate (channels/ports layer) to be seen. As can be discerned from the exploded views, the raised portions of the membrane coincide with discontinuities in the segments of the etched channels on the substrate such that compressing the raised portion closes the connection between the segments of the channels (putting the valve in the closed position) while permitting the raised portion to expand allows fluid to flow between the segments of the channels (putting the valve in the open position). As disclosed herein, various mechanisms including valve pins with notches, slots, or other features that are formed by actuators and/or raised portions of the valve pin face can be used to compress and release the raised portions of the membrane to close and open particular valves.

[0087] FIG. 18A shows a cross-sectional side view of an embodiment of the membrane valve system and FIG. 18B shows a detailed view of the valve pin, membrane, and substrate with a pressure sensitive adhesive (PSA) applied. FIG. 18A shows a motor attached to the valve pin via a shaft as well as a number of chambers adjacent to the substrate (and connected to the substrate via the PSA), each of which is coupled to an external port. In this embodiment, referred to as 'actuation from the bottom,' the motor and the valve pin are on one side of the substrate while the chambers are on the other side of the substrate.

[0088] FIG. 19A shows a cross-sectional side view of another embodiment of the membrane valve system and FIG. 19B shows a detailed view of the substrate, membrane, valve pin, and ports extension block with a pressure sensitive adhesive (PSA) applied. FIG. 19A shows a motor attached to the valve pin via a shaft as well as a number of chambers adjacent to the substrate (and connected to the ports extension block via the PSA), each of which is coupled to an external port via openings in the ports extension block. In this embodiment, referred to as 'actuation from the top, ¹ the motor and the valve pin are on the same side of the substrate as the chambers, where the ports extension block acts as a spacer to facilitate placing the valve pin between the chambers and the substrate.

[0089] FIG. 20A shows a perspective view of a valve pin attached to a substrate from which a series of external ports emerge; in this embodiment, the valve pin is between the

16

SUBSTITUTE SHEET (RULE 26) membrane layer and the chambers. Openings in the membrane layer can then be coupled to chambers, as shown in FIG. 20B. FIG. 20B shows a cross-sectional side view of an embodiment of the membrane valve system in which the chambers are coupled to the ports via openings in the top of the membrane layer; a port extension block such as that shown in FIGS. 19A and 19B may be used to complete the connection from the chambers to the external ports. [0090] FIG. 21A shows a perspective view of a valve pin/pusher attached to a substrate from which a series of external ports emerge. In this embodiment, the valve pin/pusher is disposed within the block containing the chambers so that a port extension block is not needed. Openings in the membrane layer can be coupled to chambers, as shown in FIG. 21 B. FIG. 21 B shows a cross-sectional side view of an embodiment of the membrane valve system in which the chambers are coupled to the ports via openings in the top of the membrane layer.

[0091] FIG. 22A shows an embodiment of the membrane valve system in which the valve is actuated from the bottom, that is, the motor and valve pin are on opposite sides of the membrane layer from the chambers. FIG. 22B shows an embodiment of the membrane valve system in which the valve is actuated from the top, that is, the motor and valve pin are on the same side of the membrane layer as the chambers.

[0092] FIG. 23 shows another embodiment of a membrane valve system in which the valve pin includes movable actuators which may be used instead of, or in addition to, the notches and slot design of other embodiments. The movable actuators (which may be activated in various ways including electrically or magnetically) are disposed within the valve pin (rotational valve head) so that they will align with two adjacent raised valve elements of the deformable membrane that are associated with respective separate external ports (equivalent to the notches) or will align with two adjacent raised valve elements that are associated with the same external port (equivalent to the slot), although other arrangements of the actuators are also possible. In various embodiments, the actuators can be actuated through one or more of mechanical, hydraulic, electrical, electromagnetic, acoustic, pneumatic, or shape memory alloy mechanisms. Upon actuation, the particular actuator will release pressure from the associated raised membrane portion to permit the valve to open. In some embodiments, actuation may cause the actuator to move from the open position to the closed position. In some embodiments, the actuators can be pulled up and/or removed from the through holes in the valve pin, in which case the through holes would be in the open position. The various embodiments disclosed herein which employ notches and/or slots in the valve face can also be implemented using one or more actuators instead of, or in addition to, the notches and/or slots.

17

SUBSTITUTE SHEET (RULE 26) Among other advantages, use of movable actuators to control opening and closing of valves permits a greater degree of time-based control of the valve opening and closing, since the valve pin does not need to be rotated, and also permits the valves controlled by notches and/or slots to be opened or closed at different times instead of having to be actuated together.

[0093] FIG. 24A shows an alternative peristaltic-type pump which uses several rollers and which can be incorporated into embodiments of the membrane valve system. FIG. 24B shows the pump of FIG. 24A attached to a substrate.

[0094] FIG. 25A shows a test of cell viability either as a control (left-hand bar at each cell concentration) or when the cells are passed through a valve from an inlet to the outlet at the two different concentrations (second and third bars, respectively, at each concentration) for one pass (second bars) or two passes (third bars). FIG. 25B shows a test of cell recovery following one (left-hand bars at each cell concentration) or two (right-hand bars at each cell concentration) valve passes.

[0095] Based on the above evaluations and other information, various embodiments of the system are expected to have parameters that are consistent with those listed in Table 1.

Liquid Volu 50 . to >50 ml chambers

Flow rates

Pressure differential Up to 5 psi

[0096] Table 1

[0097] Shape Memory Alloy Valve Systems

[0098] Additional embodiments of systems for microfluidic handling of are shown in FIGS. 26-35. While the examples are presented using shape memory alloy valve systems, in various embodiments other types of valves could also be used with the disclosed microfluidic handling arrangements instead of, or in addition to, shape memory alloy valves, including but

18

SUBSTITUTE SHEET (RULE 26) not limited to the membrane valves disclosed above.

[0099] FIG. 26 shows a diagram of a microfluidic handling system which can be implemented on an etched substrate which includes a series of normally-closed (NC) valves (see FIGS. 27A-27C) that are based on a shape memory alloy (SMA) mechanism and which are located at valve connection points. In one embodiment, the valves may be arranged in a continuous looped circuit (in a circular shape in FIGS. 26 and 28, although other shape configurations are also possible). In another embodiment, the valves may be arranged in an end to end track as shown in FIGS. 31 A, 31 B, and 32A. The microfluidic handling system in FIG. 26 includes a network of etched channels similar to those described above which are connected to various chambers which may include reagents, a mixing chamber, an assay chamber (e.g. a cuvette), and/or a buffer and collection chamber. Some of the chambers may be coupled to external sources via Routing Connections (RC), RC1 and RC2. Unlike the membrane valve systems in which a valve pin is used to open only one or two valves at the same time, in embodiments of the present system which are based on shape memory alloy valves, it is possible to open or close any combination of the valves at the same time. Thus, it is possible to open any combination of valves NC1-NC8 shown in FIG. 26 simultaneously. When the valves or combinations of valves are opened, fluid can be moved between chambers in various ways, for example by gravity and/or by application of positive or negative pressure to one or more of the chambers. Given that the valves can be opened or closed in any pattern, in various embodiments it is possible to simultaneously perform separate tasks in different branches of the network of etched channels, for example delivering material to a sampling chamber for analysis while at the same time transferring other materials to the mixing chamber.

[0100] FIG. 27A shows a diagrammatic side view of a shape memory alloy (SMA) valve in a closed state (top) and an open state (bottom). FIG. 27B shows a photograph of two valves such as those diagrammed in FIG. 27A alongside a ball-point pen for size comparison, with one valve turned on its side to show the input and output connection points. FIG. 27C shows bottom and side view drawings of an SMA valve such as those shown in FIGS. 27A and 27B with various dimensions indicated (in mm).

[0101] In the SMA valve embodiment shown in FIGS. 27A-27C, the SMA presses against a plug (depicted by the black ball in FIG. 27A) to press the plug against a fluid channel to maintain the fluid channel in the closed state (FIG. 27A, top); in the closed state, no power is applied to the SMA, which is "cold" (i.e. at ambient temperature) and in a first configuration. In the open state (FIG. 27A, bottom), power is applied to the SMA to heat (i.e. raise above ambient

19

SUBSTITUTE SHEET (RULE 26) temperature) and thereby deform the alloy into a second configuration so that the SMA pulls away from the fluid channel, at which point a spring element coupled to the plug pulls the plug away from the fluid channel to permit the fluid channel to be in the open state. In this embodiment, ends of segments of the microfluidic channels are aligned with the input and output openings on the bottom of the SMA valve so that when the valve is in the open state fluid can move from a first etched microfluidic channel segment, through the SMA valve, and into a second etched microfluidic channel segment. A membrane is disposed between the plug and the fluid channel in order to keep the flowing fluid material away from the plug and the SMA, to avoid contamination of the fluid and to keep the valve mechanism from being contaminated or damaged; in the closed state the plug pushes against the membrane which in turn closes the end of the fluid channel and in the open state the membrane moves with the plug away from the opening of the fluid channel to permit fluid to flow. In some embodiments the spring element may bias the plug into the fluid channel in the closed state and in the open state the activated SMA may pull the plug away from the fluid channel against the bias of the spring element. In other embodiments, the spring element may bias the plug away from the fluid channel in the closed state while the deactivated SMA may maintain the plug in the fluid channel to prevent fluid flow, and upon activation and deformation the SMA may permit the spring element to pull the plug away from the fluid channel to permit fluid flow.

[0102] FIGS. 28 and 29 show another embodiment of a microfluidic handling system based on SMA valves. As with the system shown in FIG. 26, the microfluidic handling system of FIG. 28 includes a number of normally closed valves (NC1-NC8) that are integrated into a network of microfluidic channels (e.g. etched onto a substrate) which connect to a series of chambers which may include reagents, a mixing chamber, an assay chamber (e.g. a cuvette), and a buffer and collection chamber. It is worth noting that, while the valves in FIGS. 26 and 27 include inlet and outlet ports, the valves of FIGS. 28-35 are single actuation point valves (as is the case with the membrane valves disclosed above). As with the system of FIG. 26, the valves of the system of FIG. 28 can be opened and closed in any combination so as to permit various chambers to be coupled at different times (or different portions of the system to be coupled to one another simultaneously, independent of other portions) so that fluid can be moved through the system (e.g. moved by gravity or by negative or positive pressure applied to one or more of the chambers).

[0103] FIG. 29 shows an embodiment of a normally closed SMA based valve in the open position (e.g. when energy is applied to heat and thereby deform the material). In the

20

SUBSTITUTE SHEET (RULE 26) closed state, a plunger/spring element presses a plug (black ball in FIG. 29) against a membrane which in turn is pressed against a valve seat that is in the flow path, blocking flow of fluid through the valve and thereby blocking the channel. In the open state shown in FIG. 29, the heated SMA deforms and thereby presses against the plunger/spring element and moves the plug and membrane away from the valve seat, opening the valve and permitting fluid to flow through the channel. As with the SMA valve embodiment of FIG. 27, an intervening membrane maintains separation between the fluid and the valve components to prevent contamination of the fluid and contamination and possible damage of the valve. Given that the plunger/spring element and the plug are located below the fluid path in this embodiment, this embodiment of the SMA valve can be coupled to the underside of the etched substrate whereas the valve in FIG. 27 may be coupled to the top side of the etched substrate.

[0104] As shown in the embodiment of FIG. 30, the SMA valves do not need to be arranged in a circular or arcuate pattern. FIG. 30 shows a perspective view (top) of a microfluidic handling system as well as a view of the underside (lower left) and an exploded view (lower right). The embodiment of a microfluidic handling system of FIG. 30 includes a cartridge block which includes multiple chambers (7 chambers in the specific example, although it is possible to have more or fewer chambers) with a membrane layer adjacent to the underside of the cartridge block followed by a valve holder plate with a number of valves attached thereto placed adjacent to the membrane layer. In various embodiments of the microfluidic handling system of FIG. 30, valves may be placed under each chamber (FIG. 31A), between the chambers (FIG. 31 B), or a combination of under and/or between the chambers (FIG. 32A). FIG. 32B provides a diagram (viewed from under the chamber) of the placement of valves in the "under chamber" or "between chamber" configurations. In some embodiments, microfluidic channels may be etched into the underside of the cartridge block to connect the valves with the chambers and in other embodiments a separate substrate layer (e.g. between the membrane layer and the cartridge block) may be provided to facilitate fluid flow. In certain embodiments, a series of channels may be provided on the top side of the cartridge block to facilitate delivery of fluids to the individual chambers from a clustered grouping of filling holes.

[0105] The valves in this embodiment may be provided in a version having a bypass component (FIG. 33A) or with no bypass (FIG. 33B), where the selection of one valve or the other may depend on the application and the location of the valve within the system. However, even with the bypass as in FIG. 33A, the valve portion that couples to the chamber is still provided in a normally closed state, whereas the channel to which the valve is coupled includes

21

SUBSTITUTE SHEET (RULE 26) a bypass so that when the chamber valve is closed it is still possible for fluid to move through the system; for valves that are located between chambers, it may be preferable to use a valve as in FIG. 33B which does not have a bypass so as to block fluid flow when the valve is in the closed state. In the diagrams shown in FIGS. 33A and 33B, the valve is similar to that shown in FIG. 29 and includes a valve seat disposed in the fluid path. In the embodiments of FIGS. 33A and 33B, the valve seat may include a central opening which is coupled to the chamber (see FIGS. 34A and 34B).

[0106] FIG. 34A shows an SMA valve that is mounted under a chamber in an open position (due to the SMA being heated (red line) and flattened) in which fluid is flowing from the chamber into the channel through the valve seat and also may flow through the channel around the valve seat. FIG. 34B shows the SMA valve of FIG. 34A in a closed position (due to the SMA being cooled (blue line) and deformed) such that fluid is no longer flowing from the chamber; in some embodiments fluid may continue to flow around the valve seat (e.g. as in FIG. 33A) even when the valve is close, or fluid flow may stop and not flow around the valve seat (e.g. as in FIG. 33B), depending on the type of valve that has been provided.

[0107] FIG. 35A shows an embodiment of an SMA valve in a closed position (SMA is cold and deformed, blue line) and FIG. 35B shows the SMA valve in an open position (SMA is heated and relatively flat, red line). In this embodiment, the valve is coupled to a consumable (e.g. a container having one or more chambers and which can be used once and disposed) and the valve and consumable are separated from one another by a membrane. Although no valve seat is depicted in FIGS. 35A and 35B, a valve seat is needed, either under the chamber (e.g. at the end of the cone) or between chambers.

[0108] Rotary Valve Systems

[0109] In certain embodiments, systems for microfluidic handling of fluids using rotary valves are provided (FIGS. 36A, 36B). FIG. 36A shows an example of a rotary valve coupled to a motor (e.g. a servo motor) and a controller (circuit board at top). FIG. 36B shows a diagram of a rotary valve system depicting the motor (center), a switching valve in cross-section shown in two different positions (top left), a distribution valve (top center), an on/off valve in cross-section (top right), and a valve shown from the back side (i.e. the side that couples to the motor) (lower left). As seen in the back side view, the valve may include a flat, square, or rectangular portion which firmly connects to the motor to permit the motor to rotate the valve to various positions. [0110] The rotary valve generally includes a valve insert rotatably disposed within a valve body. The valve body may include one or more side ports (which can be fluidly coupled to

22

SUBSTITUTE SHEET (RULE 26) chambers as described below) and the valve insert may include one or more channel including an end channel, where the valve insert is rotated to align one or more of the channels of the valve insert with one or more of the side ports of the valve body to permit fluid to flow (typically to flow to, from, or between one or more chambers or external compartments or sources). [0111] FIG. 37A shows a diagram of a rotary valve in which a valve insert (white T- shaped structure in the center of the valve) couples one of the side ports to the end port when the insert is rotated (indicated by the curved arrow) as driven by the motor. FIG. 37B shows a cutaway side view of an embodiment of a rotary valve system showing how the valve insert can connect two side ports to one another (and optionally to the end port) (left) to create a switch valve, or connect a side port to the end port (right) to create a distribution valve, where the left and right panels of FIG. 37B use two different valve inserts to create the different fluid diversion pathways. In either version of the valve in FIG. 37B, the valve insert is rotated to determine which ports are connected to one another via the channels in the insert.

[0112] FIGS. 38A and 38B show embodiments of a rotary valve in which the valve insert includes channels that allow the insert to act as either a distribution valve (generally for connecting one chamber to potentially multiple chambers) or a switch valve (generally for connecting one chamber to another on a one-to-one basis), depending on the position to which the insert is turned. FIG. 38A shows an embodiment in which the valve insert can be actuated from the top (as shown in the top panel) and FIG. 38B shows another embodiment in which the valve insert can be actuated from the bottom (as shown in the top panel). The lower panels of FIGS. 38A and 38B also show one possible arrangement of ports in the valve, including six side ports (shown in red, yellow, light green, dark green, light blue, and dark blue) and an end port (shown in magenta, also referred to as a center port), where the end port is shown as a dotted line connected to the center of the valve insert. As shown in the lower panels of FIGS. 38A and 38B, the valve insert in this embodiment includes a side channel (curved channel on left side of valve insert) which connects two adjacent ports to create a switch valve and also a straight radial channel emanating from the center (straight line on right side of valve insert) which connects to an axial channel (represented by the magenta dashed line) which connects to the end coupler. Depending on the position of the valve insert, either two adjacent side ports will be connected to one other or the end port will be connected to one of the side ports. Based on the relative positions of the two channels in the valve insert shown in FIGS. 38A and 38B, only one of the two channels can be connected to a side port in a given position. However, in other embodiments the two channels may be aligned such that both channels may be connected to

23

SUBSTITUTE SHEET (RULE 26) side ports at the same time so that one side port may be connected to the end port and two other side ports may be connected to one another.

[0113] FIG. 39 shows a diagram of a complete fluidic handling system based on the use of a rotary valve system. In FIG. 39, the motor and valve insert are below the valve while the chambers containing reagents and other solutions are above the valve. As seen in FIG. 39, the valve insert is rotated so that it is acting as a distribution valve, connecting a side port on the left side of the drawing to the end port. The switch valve channel can be seen on the right as a small segment which in this particular position is not connected to any of the side ports (see FIG. 40). The through holes in the valve are provided to accommodate fasteners to tightly couple the valve to the chamber block (e.g. using screws coupled to threaded heat-set inserts mounted in the block as indicated in FIG. 39), to minimize the risk of leakage.

[0114] FIG. 40 shows a diagram of the valve from FIG. 39 in a side view (top panel) and in a top view (bottom panel) showing the position of the valve insert and in particular how the valve insert acts as a distribution valve in this position, connecting the right side port to the end port (bottom panel). In various embodiments, the channels may be 0.5-1 mm in diameter and the system may be capable of withstanding pressures of up to 10 PSI (0.35 bar). As with other embodiments, fluids may be moved between chambers using positive and/or negative pressure applied to the system (e.g. applied to the chambers or to connected channels). The components in various embodiments are generally made of biocompatible materials that are suitable for a single use and resistant to gamma irradiation and in certain embodiments the materials may include UHMW-PE, PEEK, or PVDF.

[0115] In the embodiments of the rotary valve disclosed above, the side ports are arranged around the perimeter of the valve. In other embodiments, however, the valve body may be reconfigured so that the side ports emerge from the valve body at the end, as shown in FIGS. 41A-41C. As shown in FIGS. 41A-41C, the channels leading to the side ports may be blocked at or near their outer ends and instead may be joined with channel segments leading to the end face of the valve body. In some embodiments, some of the channels leading to the side ports may be completely blocked (see FIG. 41 B, solid red channels). FIG. 41 C shows a fluid flow path (indicated by orange arrows) when the valve insert is rotated so that two side ports are connected to one another.

[0116] FIGS. 42-44 show an embodiment of a fluidic handling system using a rotary valve. FIG. 42 shows a motor and controller coupled to a bracket. FIG. 43 shows the motor, controller, and bracket of FIG. 42 coupled to a chamber block (FIG. 43, left). FIG. 43, center,

24

SUBSTITUTE SHEET (RULE 26) shows an exploded view of the fluidic handling system in which the relative positions of the motor/controller, the valve insert, the valve body, the bracket, a pressure-sensitive adhesive (PSA, magenta rectangle), and the chamber block are depicted. FIG. 43, right, shows the exploded view from underneath, which makes it possible to see holes in the PSA for the connections to the side ports of the valve (pattern of smaller holes in the center) and the holes for the attachment fasteners (pattern of larger holes on the outside).

[0117] FIG. 44 shows an exploded view of the fluidic handling system using a rotary valve from different angles. FIG. 44, left and right, show views from above and FIG. 44, center, shows a view from underneath.

[0118] Thus, while the invention has been described above in connection with particular embodiments and examples, the invention is not necessarily so limited, and that numerous other embodiments, examples, uses, modifications and departures from the embodiments, examples and uses are intended to be encompassed by the claims attached hereto.

25

SUBSTITUTE SHEET (RULE 26)