A wide variety of systems and methods exist for performing biochemical analysis, for example for medical testing. A common technique is to load analytes and reagents into a microfluidic “chip” that has fluid flow channels and other structures formed in it using photolithography techniques. Such a chip may include pumps, reservoirs, valves, mixing structures, and other features useful in the performance of a certain tests.
Typically, such a chip is controlled by an external controller, through application and release of fluid pressure at key points in the chip. For example, a valve may be formed by crossing a fluid flow channel in a soft medium with a dead-end cross channel. By pressurizing the cross channel, the fluid flow channel can be pinched off, and by releasing the pressure in the cross channel, the fluid flow channel is allowed to re-open. A peristaltic pump may be formed by placing three or more such valves close together crossing a fluid flow channel in a soft medium. By sequentially pressurizing and depressurizing the valves channels to pinch off and re-open adjacent locations in the fluid flow channel, fluid can be caused to flow in the fluid flow channel.
Because of the need for complex external pressure control, such microfluidic chips are not convenient for use in routine medical testing, especially in remote locations.
According to one aspect, a cartridge for fluid manipulation comprises a cartridge body defining a holding compartment and first and second fractioning compartments formed within the cartridge body. The cartridge body also defines a number of flow channels formed within the cartridge body. The compartments and flow channels are arranged such that a predetermined quantity of fluid can be held in the holding compartment when the cartridge body is held in a first orientation, and can be poured from the holding compartment to the first fractioning compartment by rotating the cartridge body about a predefined rotation axis to a second orientation, to spill the fluid from the holding compartment to the first fractioning compartment through a first one of the flow channels. The first fractioning compartment is of a shape, size, and position such that when the cartridge body is in the second orientation, not all of the fluid can be contained in the first fractioning compartment. The first fractioning compartment is connected to the second fractioning compartment by a second one of the flow channels, such that any of the fluid that overflows the first fractioning compartment when the cartridge body is in the second orientation flows through the second flow channel to the second fractioning compartment.
In some embodiments, the first and second fractioning compartments are shaped, sized, and positioned such that the predetermined quantity of fluid can be held in substantially equal quantities in the first and second fractioning compartments when the cartridge body is held in the second orientation. In some embodiments, the cartridge body defines a third fractioning compartment; the first and second fractioning compartments are shaped, sized, and positioned such that the predetermined quantity of fluid cannot be contained within the first and second fractioning compartments when the cartridge body is in the second orientation; and the second fractioning compartment is connected by a third one of the flow channels to the third fractioning compartment, such that any of the fluid that overflows the second fractioning compartment when the cartridge body in the second orientation flows through the third flow channel to the third fractioning compartment. In some embodiments, the first, second, and third fractioning compartments are shaped, sized, and positioned such that the predetermined quantity of fluid can be held in substantially equal quantities in the first, second, and third fractioning compartments when the cartridge body is held in the second orientation. In some embodiments, the cartridge further comprises two analysis areas, one analysis area respectively for each fractioning compartment, wherein the analysis areas are connected directly or indirectly to the respective fractioning compartments by respective ones of the flow channels, and the analysis areas are positioned such that fluid held in the fractioning compartments when the cartridge body is in the second orientation can be delivered to the respective analysis areas by one or more subsequent rotations of the cartridge body about the rotation axis, to spill fluid from the fractioning compartments and into the respective connections to the analysis areas. In some embodiments, the cartridge body further defines two mixing compartments, one mixing compartment respectively for each fractioning compartment, and wherein the fluid spilled from the fractioning compartments passes through the respective mixing compartments before reaching the respective analysis areas. In some embodiments, each of the mixing compartments stores a quantity of a reagent positioned to mix with the fluid spilled from the respective fractioning compartment before the fluid flows to the respective analysis area. In some embodiments, the fluid is a first fluid; the cartridge body further defines a second set of compartments and flow channels for manipulating a second fluid in sequence through the second set of compartments in reaction to the rotations of the cartridge about the rotation axis; the cartridge body further defines a second set of outlet channels respectively connecting the last of the second set of compartments with the analysis areas; and the second set of compartments and flow channels and the outlet channels are shaped, sized, and positioned such that the second fluid reaches the analysis areas later than the first fluid when the cartridge is rotated in such a way as to deliver the first fluid to the analysis areas. In some embodiments, the lengths of the outlet channels are selected to ensure that the second fluid will reach the analysis areas later than the first fluid.
According to another aspect, a cartridge for fluid manipulation comprises a cartridge body. The cartridge body defines a first set of compartments and flow channels for manipulating a first fluid. The compartments and channels in the first set are sized, shaped, and positioned such that a sequence of rotations of the cartridge body about a predefined rotation axis will cause a quantity of the first fluid to sequentially pass through all of the compartments in the first set via the first set of flow channels to reach an outlet of the first set of compartments and flow channels. The cartridge body defines a second set of compartments and flow channels for manipulating a second fluid. The compartments and channels in the second set are sized, shaped, and positioned such that the same sequence of rotations of the cartridge body about the predefined rotation axis will cause a quantity of the second fluid to sequentially pass through all of the compartments in the second set via the second set of flow channels to reach an outlet of the second set of compartments and flow channels. In some embodiments, the outlets of the first and second sets of compartments and flow channels are joined at a junction, and the first and second sets of compartments and flow channels are shaped, sized, and positioned such that the second fluid reaches the junction at a different time than the first fluid in response to the sequence of rotations. In some embodiments, the cartridge further comprises: a reservoir holding a sample fluid and a washing buffer fluid in separate compartments of the reservoir, the reservoir including two openings sealed by puncturable sealing covers; two hollow piercing elements positioned on the cartridge body such that the two piercing elements pierce the puncturable sealing covers of the reservoir when the reservoir is joined to the cartridge body, enabling the sample fluid and the washing buffer fluid to pass through the two hollow piercing elements and to pass respectively to the first set of compartments and flow channels and the second set of compartments and flow channels, the sample fluid being the first fluid and the washing buffer fluid being the second fluid; and an analysis area at the junction; wherein at least some of the compartments in the first set of compartments store quantities of reagents for mixing with the sample fluid as the sample fluid traverses the first set of compartments and flow channels, the reagents usable to conduct an assay of the sample fluid; and wherein the analysis area enables reading of a result of the assay. In some embodiments, the analysis area comprises an absorbent medium through which the sample fluid and the washing buffer fluid can sequentially transport by capillary action.
According to another aspect, a testing system comprises a cartridge for fluid manipulation as in claim 1, a motorized mechanism for producing a rotary motion of cartridge about a rotational axis, and a controller having a processor and memory. The controller is coupled to the motorized mechanism and programmed to cause the motorized mechanism to produce a predetermined series of rotations of cartridge in accordance with a predetermined assay.
According to another aspect, a method of conducting an assay comprises providing a cartridge having a cartridge body defining a holding compartment and first and second fractioning compartments formed within the cartridge body. The cartridge body also defines a number of flow channels formed within the cartridge body. The method further comprises placing a quantity of fluid in the holding compartment and holding the cartridge body in a first orientation, and rotating the cartridge about a predefined rotation axis to a second orientation to pour at least some of the fluid from the holding compartment through a first one of the flow channels to the first fractioning compartment. The first fractioning compartment is of a shape, size, and position such that when the cartridge body is in the second orientation, not all of the fluid can be contained in the first fractioning compartment. The first fractioning compartment is connected to the second fractioning compartment by a second one of the flow channels, such that any of the fluid that overflows the first fractioning compartment when the cartridge body is in the second orientation flows through the second flow channel to the second fractioning compartment. In some embodiments, the method further comprises rotating the cartridge about the rotation axis to one or more subsequent orientations, causing the fluid to spill from the two fractioning compartments to reach respective analysis areas in the cartridge. In some embodiments, the method further comprises pausing between successive rotations of the cartridge to allow an analyte in the fluid to react with a reagent previously stored in one of the compartments. In some embodiments, the rotation axis is a first rotation axis, the method further comprising rotating the cartridge about a second rotation axis different from the first. The method may further comprise depositing an analyte in the quantity of fluid. In some embodiments, depositing the analyte in the quantity of fluid comprises injecting the analyte through a puncturable seal.
Cartridge 100 further includes a reservoir 105 having multiple isolated compartments 106. Compartments 106 may be used to hold fluids to be manipulated in cartridge 100. For example, one compartment may be loaded with a sample fluid for carrying an analyte, and another of compartments 106 may be loaded with a washing buffer fluid. Puncturable seals 107a, 107b may be placed over openings in reservoir 105, to retain the pre-loaded fluids. For example, cover 109 and puncturable seal 107a may be placed on reservoir 105, and the fluids loaded through the remaining openings in reservoir 105 (shown at the bottom of reservoir 105 in
A specimen containing an analyte may be introduced to the sample fluid using a sample injector 110. For example, sample injector may include a sharp hollow needle or similar structure 111 for puncturing puncturable seal 107a and carrying the analyte to reservoir 105. In some embodiments, the specimen may be a human blood sample and cartridge 100 is configured to perform an assay for glycated hemoglobin (HbA1c), useful in diagnosing and monitoring diabetes and capable of detecting the presence of variant forms of hemoglobin that are relevant to HbA1c measurements. It will be recognized that the invention may be embodied in many other ways as well. Example cartridge 100 also includes analysis areas 112, as will be explained in more detail below. Cover 108 may include viewing windows 114 for viewing analysis areas 112 from outside cartridge 100. In other embodiments, cover 108 may be made of a transparent material such as glass or a transparent polymer, to allow viewing of analysis areas 112.
Example cartridge body 101 also includes two hollow piercing elements 113 positioned to pierce puncturable seals 107b when reservoir 105 is mated to cartridge body 101, and to carry the respective fluids from reservoir compartments 106 to cartridge body compartments 102.
In
In
As is shown in
While reservoir 105 is shown as being joined to cartridge body 101 by a simple linear motion, it will be recognize that many other joining motions and techniques may be used. For example, reservoir 105 may undergo a rotational or sliding motion to connect with cartridge body 101 and to reach hollow piercing elements 113.
In some embodiments, one or more compartments may include structures that can aid in mixing of fluids and reagents. For example, as shown in
In
In
In
In
In
In the meantime, washing buffer fluid 202 has spilled from compartment 802 and into channels 902, to be carried by capillary action toward analysis areas 112 as well. However, because channels 902 are longer than channels 901a and 901b, washing buffer fluid 202 arrives at analysis areas 112 later than does sample fluid 201. By the time washing buffer fluid 202 arrives at analysis areas 112, sample fluid 201 may have already substantially soaked into the absorbent medium of analysis areas 112, and washing buffer fluid 202 may carry sample fluid 201 further across analysis areas 112. Washing buffer fluid 202 may serve to carry away antibodies not bound to any of the proteins present in analysis areas 112, removing stray antibodies that could otherwise interfere with interpretation of the test result. Washing buffer fluid 202 and other fluid components it carries may be exhausted into a collection area (not shown) within cartridge 100.
It will be recognized that many, many variations from this example are possible within the scope of the appended claims. The number, size, and arrangement of compartments present in a particular cartridge may be varied according to the intended use of the cartridge. Only one set of compartments and channels may be provided, or more than two sets of compartments and channels may be provided, for manipulating more than two fluids. Different kinds of analysis areas may be provided, according to the intended use of the cartridge. And while only two fractioning compartments are shown in the above example, it will be recognized that three or more fractioning compartments may be provided, so that a fluid sample can be divided in to any workable number of smaller quantities for performing different tests or for other purposes.
In some embodiments, and additional axis of rotation of cartridge 100 may be provided. For example, the rotation mechanism that provides rotations of cartridge 100 about axis 104 may also include a second rotational degree of freedom as shown in
An assay cartridge such as cartridge 100 may be particularly useful in a point-of-care or field hospital environment, because the motions required for completing an assay are simple and easily accomplished. For example, especially when cover 108 is transparent, the rotational motions and test sequence described in conjunction with
In cartridge 1200, a sample fluid 1201 may be pre-loaded in a compartment 1202 of cartridge 1200 itself, rather than in a separate reservoir. Compartment 1202 may be lined with puncturable seals 1203. An analyte 1204 may be introduced directly into compartment 1205, for example using a sample injector or needle 1206.
As shown in
In the claims appended hereto, the term “a” or “an” is intended to mean “one or more.” The term “comprise” and variations thereof such as “comprises” and “comprising,” when preceding the recitation of a step or an element, are intended to mean that the addition of further steps or elements is optional and not excluded.
It is to be understood that any workable combination of the elements and features disclosed herein is also considered to be disclosed.
The invention has now been described in detail for the purposes of clarity and understanding. However, those skilled in the art will appreciate that certain changes and modifications may be practiced within the scope of the appended claims.
This application claims the benefit of U.S. Provisional Patent Application No. 62/132,984, filed Mar. 13, 2015, and titled “Assay Cartridge”, the entire disclosure of which is hereby incorporated by reference herein for all purposes.
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