WO2022125074A1 - Dispositifs fluidiques avec fluide de colmatage non newtonien - Google Patents

Dispositifs fluidiques avec fluide de colmatage non newtonien Download PDF

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Publication number
WO2022125074A1
WO2022125074A1 PCT/US2020/063756 US2020063756W WO2022125074A1 WO 2022125074 A1 WO2022125074 A1 WO 2022125074A1 US 2020063756 W US2020063756 W US 2020063756W WO 2022125074 A1 WO2022125074 A1 WO 2022125074A1
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Prior art keywords
fluid
newtonian
plugging
volume
fluids
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PCT/US2020/063756
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English (en)
Inventor
John Michael LAHMANN
Kenneth James Faase
Paul Mark Haines
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Hp Health Solutions Inc.
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Priority to PCT/US2020/063756 priority Critical patent/WO2022125074A1/fr
Publication of WO2022125074A1 publication Critical patent/WO2022125074A1/fr

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502769—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/50273—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the means or forces applied to move the fluids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/52—Containers specially adapted for storing or dispensing a reagent
    • B01L3/527—Containers specially adapted for storing or dispensing a reagent for a plurality of reagents
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
    • B01L2200/027—Fluid interfacing between devices or objects, e.g. connectors, inlet details for microfluidic devices
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/04—Closures and closing means
    • B01L2300/041—Connecting closures to device or container
    • B01L2300/044—Connecting closures to device or container pierceable, e.g. films, membranes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0481—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure squeezing of channels or chambers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/06—Valves, specific forms thereof
    • B01L2400/0677—Valves, specific forms thereof phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
    • B01L2400/0683—Valves, specific forms thereof phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers mechanically breaking a wall or membrane within a channel or chamber

Definitions

  • isolating a component of interest from a sample fluid can be useful. Such separations can permit analysis or amplification of a component of interest. As the quantity of available assays for components increases, so does the demand for the ability to isolate components of interest from sample fluids. Fluidic devices can be used for these applications, among others. In some examples, microfluidic devices can be used to prepare and process samples with small volumes.
  • FIGS. 1 A and 1 B illustrate a cross-sectional view of an example fluidic device in accordance with the present disclosure
  • FIG. 2 illustrates a cross-sectional view of an example fluid processing system in accordance with the present disclosure
  • FIGS. 3A-3C illustrate a cross-sectional view of another example fluid processing system in accordance with the present disclosure
  • FIGS. 4A-4H illustrate the use of an example fluid processing system for processing fluids in accordance with the present disclosure.
  • FIG. 5 is a flowchart illustrating an example method of processing fluids in accordance with examples of the present disclosure.
  • the present disclosure describes fluidic devices, fluid processing systems, and methods of processing fluids.
  • the devices, systems, and methods can include the use of a non-newtonian plugging fluid to form a plug in a fluidic channel to partition fluids on either side of the plug.
  • This plug of non-newtonian fluid can act as a shutoff valve because the plug can block the fluidic channel so that fluids do not flow through the channel. Forming valves in small fluidic channels can often be difficult and expensive.
  • the use of non-newtonian plugging fluid can provide a simple and inexpensive solution for stopping flow through small fluidic channels.
  • the non-Newtonian fluid can separate fluids on either side of the plug.
  • fluidic devices and systems described herein can be a sample preparation device that is used to isolate a biological component from a biological sample.
  • a particular biological component can be separated from a biological sample, and the non-newtonian plugging fluid can be used to partition the biological component from the remainder of the biological sample.
  • the biological component can be purified and/or concentrated and this purified or concentrated biological component can be partitioned from the remainder of the biological sample using the non-newtonian plugging fluid.
  • the fluidic devices, fluid processing systems, and methods that can be used in a process of preparing samples for a PCR (polymerase chain reaction) assay. PCR assays are processes that can rapidly copy millions to billions of copies of a very small DNA or RNA sample.
  • PCR can be used for many different application, included sequencing genes, diagnosing viruses, identifying cancers, and others.
  • a small sample of DNA or RNA is combined with reactants that can form copies of the DNA or RNA.
  • the fluidic device can isolate a nucleic acid such as DNA or RNA from a biological sample fluid.
  • the nucleic acid can be concentrated and/or purified by the fluidic device, and the non-newtonian plugging fluid can be used to partition this concentrated or purified nucleic acid from the remainder of the biological sample fluid. Because the volumes of samples fluid and reactant involved in this process are very small, it can be beneficial to use small fluidic devices and systems such as those described herein.
  • the non-newtonian plugging fluids described herein can provide a useful alternative to mechanical valves in these small fluidic devices and systems.
  • a fluidic device includes interconnected volumes with a bulk fluid volume fluidical ly connected in series with a capillary volume to receive a density gradient column.
  • a reservoir of a non-newtonian plugging fluid is positioned outside the interconnected volumes.
  • a plugging fluid injection opening is also positioned at a location along a length of the capillary volume to inject the non-newtonian plugging fluid into the capillary volume.
  • the non-newtonian plugging fluid has a sufficient viscosity to partition fluid upstream of the non-newtonian plugging fluid from fluid downstream of the non-newtonian plugging fluid.
  • a sufficient viscosity can vary depending on the conditions of fluids in a particular fluidic device.
  • the fluidic device can include a fluid positioned along the density gradient column that is above (upstream) the plug of non-newtonian plugging fluid.
  • the fluid above the plug can exert a pressure on the plug due to the force of gravity on the fluid (i.e., the pressure head of the fluid).
  • the non-newtonian plugging fluid can have a sufficient viscosity to hold the plug in place, against the pressure head of the fluid above the plug.
  • the level of viscosity that is sufficient can also be affected by the diameter of capillary volume, since a smaller diameter capillary can allow a less viscous plugging fluid to support a given pressure head.
  • the capillary volume can have a column diameter from 0.2 mm to 3 mm.
  • the fluid above the non-newtonian fluid plug can exert a pressure of up to 8 inches of water, or up to 12 inches of water, or up to 16 inches of water.
  • the level of viscosity that is sufficient to partition the fluid upstream of the plug from fluid downstream of the plug can be 5,000 centipoise or greater in some examples.
  • the sufficient viscosity can be 10,000 centipoise or greater, or 15,000 centipoise or greater, or 20,000 centipoise or greater.
  • Viscosity is often referred to more specifically as dynamic viscosity, and can be measured using a viscometer such as viscometers available from AMETEK, Inc.
  • non-newtonian fluids can act as if they have an infinite viscosity when the amount of shear stress applied to the fluid is below a certain threshold.
  • the non-newtonian fluid plug can effectively have an infinite viscosity when the non-newtonian fluid is at rest in the capillary volume.
  • the non-newtonian plugging fluid can be a Bingham plastic, a viscoplastic, a shear thinning fluid, or a curable fluid.
  • the non-newtonian plugging fluid can include a mineral oil-based grease, a vegetable oil-based grease, a petroleum oil-based grease, a synthetic oil-based grease, a semi-synthetic oil-based grease, a silicone oil-based grease, or a combination thereof.
  • NLGI National Lubricating Grease Institute
  • the NLGI consistency number can be one of nine grades, included grade 000, grade 00, grade 0, grade 1 , grade 2, grade 3, grade 4, grade 5, and grade 6. The grades progress from softer consistency to harder consistency.
  • the non-newtonian plugging fluids described herein can have an NLGI consistency number from 0 to 6. In further examples, the NLGI consistency number can be from 1 to 5, from 2 to 5, from 1 to 4, or from 2 to 4.
  • the plugging fluid injection opening can be positioned to inject the plugging fluid into the capillary volume.
  • the non-newtonian plugging fluid can have a holding pressure from 1 ,000 Pa to 5,000 Pa when the non-newtonian plugging fluid is injected into the capillary volume.
  • the interconnected volumes can be within a solid device body.
  • the plugging fluid injection opening can be connected to the flexible fluid-filled blister by an injection channel formed in the solid device body.
  • the non-newtonian plugging fluid can be injectable by puncturing the sealing layer and applying pressure to the blister.
  • the density gradient column can be formed in a solid device body such as vessel that defines the interconnected volumes, e.g., unitary vessel or modular vessel, and the plugging fluid injection opening can be connected to the flexible fluid-filled blister by an injection channel formed in the solid device body.
  • the solid device body can include a sharp point adjacent to the injection channel to puncture the sealing layer when pressure is applied to the blister.
  • the device can include a second fluid injection opening formed in the solid device body and positioned along the length of the capillary volume below the plugging fluid injection opening. Any features of fluidic devices described herein can also be included in fluid processing systems and methods, in various examples.
  • a fluid processing system includes interconnected volumes having a bulk fluid volume connected in series with capillary volume, and a reservoir of a first fluid positioned outside the interconnected volumes.
  • the system also includes a first fluid injection opening to inject the first fluid into the interconnected volumes.
  • the first fluid injection opening is connected to the reservoir of the first fluid.
  • a reservoir of a non-newtonian plugging fluid in this example is positioned outside the interconnected volumes, and a plugging fluid injection opening is positioned at a location along a length of the interconnected volumes to inject the non-newtonian plugging fluid therein.
  • the non-newtonian plugging fluid has a sufficient viscosity to partition fluid upstream of the non-newtonian plugging fluid from fluid downstream of the non-newtonian plugging fluid.
  • the interconnected volumes can contain a density gradient column.
  • a reservoir of a second fluid cam be positioned outside the interconnected volumes, and a second fluid injection opening positioned along the length of the capillary volume below the plugging fluid injection opening.
  • the first fluid in this instance can have a greater density than the second fluid.
  • the interconnected volumes that carry the fluid column can be a density gradient column.
  • the system can also include a reservoir of a second fluid positioned outside the interconnected volumes and a second fluid injection opening positioned along the length of the capillary volume below the plugging fluid injection opening, wherein the wash buffer has a greater density than the second fluid.
  • the second fluid can be gas, e.g., air, or an aqueous PCR master mix solution, wherein the non-newtonian plugging fluid prevents mixing of the washing buffer and the second fluid when the non-newtonian plugging fluid is injected into the capillary volume.
  • the system can include a sample fluid, where the sample fluid includes magnetizing particles having a biological component bound thereto dispersed therein, and the sample fluid can have a lower density than the wash buffer and can be loaded or loadable above the wash buffer.
  • the sample fluid includes magnetizing particles having a biological component bound thereto dispersed therein, and the sample fluid can have a lower density than the wash buffer and can be loaded or loadable above the wash buffer.
  • a method of processing fluid incudes injecting a first fluid into interconnected volumes having a bulk fluid volume fluidically connected in series with a capillary volume to receive a density gradient column, wherein the first fluid occupies the capillary volume (which is inclusive of also allowing for occupying the bulk fluid volume in part above the capillary volume).
  • the method further includes injecting a non-newtonian plugging fluid at a location along the interconnected volumes.
  • the non-newtonian plugging fluid partitions the first fluid or portion thereof from fluid upstream and above the non-newtonian plugging fluid.
  • the method further includes injecting a second fluid into the capillary volume downstream from the non-newtonian plugging fluid.
  • the first fluid in this example has a higher density than the second fluid.
  • the non-newtonian plugging fluid does not allow the second fluid to enter fluid positioned upstream and above the non-newtonian plugging fluid.
  • the first fluid, the second fluid, the non-newtonian plugging fluid, or a combination thereof can be individually injected from respective flexible fluid-filled blister.
  • the first fluid can be a wash buffer and the method can further include loading a sample fluid including magnetizing particles having a biological component bound thereto over the wash buffer forming a density gradient column.
  • the second fluid is an aqueous PCR master mix solution.
  • the present disclosure describes fluidic devices that incorporate a non-newtonian plugging fluid that can be used to partition fluids in a fluid channel.
  • the non-newtonian plugging fluid can effectively act as a shut-off valve to stop fluid flow through a channel.
  • Small-scale fluidic devices such as microfluidic devices, can often involve fluids flowing through channels having small diameters, such as less than one millimeter or on the order of a few millimeters. Valving in such small-scale systems can be difficult for a variety of reasons. Mechanical valves that are often used with larger pipes and tubing can be complex and expensive to miniaturize in some cases.
  • microfluidic applications can call for disposable and consumable microfluidic devices, such as one-time-use devices that are used to process biological samples.
  • Using expensive miniaturized mechanical valves can add greatly to the cost of such disposable devices.
  • valves in microfluidic devices can encounter different forces than in larger devices. For example, forces due to surface tension can have a much larger effect on fluid flow in small-scale devices.
  • the fluid devices described herein can include a non-newtonian plugging fluid that can be injected into a capillary channel to stop fluid flow through the capillary channel.
  • the non-newtonian plugging fluid can form a plug in the capillary.
  • the plug of non-newtonian fluid can have a sufficiently high viscosity that the plug partitions fluid on either side of the plug so that the fluids do not contact one another or mix one with another.
  • the plug of non-newtonian fluid can have a sufficiently high viscosity that the plug can prevent fluid from flowing even under pressure, up to a certain threshold pressure.
  • the sufficiently high viscosity can be 5,000 centipoise or greater, 10,000 centipoise or greater, 15,000 centipoise or greater, or 20,000 centipoise or greater.
  • the capillary can be part of a vertically oriented fluid column of interconnected volumes, and the non-newtonian plugging fluid can have a sufficient viscosity to hold up fluid along the fluid column that is above the non-newtonian plugging fluid.
  • non-newtonian refers to a characteristic of a fluid, which is that the fluid can have a different viscosity when different magnitudes of stress are applied to the fluid.
  • the fluid can have a relatively high viscosity when no stress or a small stress is applied to the fluid.
  • Some such fluids can have a sufficiently high viscosity that the fluids do not flow under the force of gravity.
  • the fluid can be made to flow and viscosity can drop to a lower value, allowing the fluid to flow relatively easily until the stress is removed.
  • the non-newtonian plugging fluid can be capable of flowing into a capillary channel when the fluid is injected, such as by applying a sufficient pressure to the fluid.
  • the non-newtonian plugging fluid can be injected from a flexible blister, by applying a force to the blister to squeeze the non-newtonian plugging fluid out into the capillary channel.
  • the force applied to the blister can be from 10 Newtons (kg ⁇ m ⁇ s -2 ) to 40 Newtons (kg ⁇ m ⁇ s -2 ), or from 10 Newtons (kg ⁇ m ⁇ s -2 ) to 20 Newtons (kg ⁇ m ⁇ s -2 ), or from 20 Newtons (kg ⁇ m ⁇ s -2 ) to 40 Newtons (kg ⁇ m ⁇ s -2 ).
  • the viscosity of the fluid can increase so that the fluid remains as a plug in the capillary channel without flowing out.
  • fluidic devices can include devices for biological sample preparation and processing.
  • the fluidic device can be used to isolate a biological component from a biological sample.
  • the biological component can be a nucleic acid such as DNA or RNA and the biological sample can be fluid that includes the nucleic acid mixed with other materials.
  • the fluidic device can be used to isolate a nucleic acid such as DNA or RNA from a sample including lysate and lysed cells or viruses, where the nucleic acid is to be further processed in a polymerase chain reaction (PCR) nucleic acid amplification process.
  • PCR polymerase chain reaction
  • non-newtonian fluid can be injected into a capillary volume of a fluidic device, where the capillary volume has a diameter from 0.2 mm to 3 mm.
  • the non-newtonian plugging fluid can be used in larger devices, such as devices with flow channels of 1 cm or more in width.
  • non-newtonian plugging fluid can be used in smaller microfluidic devices, with microfluidic channels of 5 ⁇ m to 200 ⁇ m in width. Using a non-newtonian plugging fluid in this way can provide a cheaper and simpler alternative to miniaturized mechanical valves. This can be particularly useful in disposable and consumable fluidic devices.
  • the non-newtonian plugging fluid can also be capable of filling in any imperfections or gaps in fluid channels, such as rough fluid channel walls that may be present in 3D printed fluidic devices.
  • the non-newtonian plugging fluid can form a good seal even when the fluid channel walls are rough or have imperfections. This makes the non-newtonian plugging fluid versatile for use in forming valves in a wide variety of fluidic devices.
  • the non-newtonian plugging fluid can be used in specific fluid processing systems for processing biological samples that are described in more detail below.
  • the present disclosure includes several figures illustrating specific examples of the technologies described herein. These figures show fluidic devices and fluid processing systems that include a variety of components arranged is specific ways depending on the purpose and function of the particular examples depicted. Although the figures illustrate examples that implement the technologies described herein, these examples also include many features that are optional, which may be changed or removed depending on the particular example. Accordingly, it is understood that the technologies described herein are not limited by the examples shown in the figures.
  • FIG. 1A shows one example fluidic device 100 in accordance with the present disclosure.
  • the fluidic device includes a solid device body 102.
  • the fluidic device also includes interconnected volumes 110 to receive a fluid column with a capillary volume 112 and a bulk fluid volume 114.
  • a reservoir 120 of a non-newtonian plugging fluid 122 is positioned outside the interconnected volumes.
  • a plugging fluid injection opening 124 is positioned at a location along a length of the capillary volume. The plugging fluid injection opening can be used to inject the non-newtonian plugging fluid into the capillary volume.
  • the reservoir of non-newtonian plugging fluid is a flexible fluid-filled blister.
  • a sealing layer 116 separates the fluid in the blister from the plugging fluid injection opening.
  • a sharp point 126 formed in the solid device body is positioned under the blister. When pressure is applied to the blister, the sharp point can puncture the sealing layer, allowing the non-newtonian plugging fluid to flow out of the blister. The non-newtonian plugging fluid can then flowthrough an injection channel 128 to the plugging fluid injection opening.
  • FIG. 1B shows the same example fluidic device 100 in use.
  • the non-newtonian plugging fluid 122 has been injected from the reservoir 120 into the capillary volume 112.
  • the non-newtonian plugging fluid forms a plug in the reservoir.
  • This figure also shows a first fluid 132 upstream of the plug and a second fluid 142 downstream of the plug.
  • the plug can have a sufficient viscosity so that the plug can remain in place and separate the first fluid from the second fluid.
  • FIGS. 1 A-1 B above depict various portions of example devices and FIGS. 2-4H below depict various portions of example systems.
  • the devices and systems shown and described herein can be interchangeable with respect to structural components shown in the various examples.
  • the devices and systems can include other structures not shown that may be present upstream and/or downstream from the illustrated structures.
  • these devices and systems can be part of a sample preparation cartridge module that includes a biological sample input 170 and output 180.
  • the sample preparation cartridge module may include interconnected volumes arranged in series between the input and output in a linear direction.
  • the various volumes may include, for example, the bulk fluid volume 114 and the capillary volume 112.
  • the bulk fluid volume may include a mixing chamber (not shown) connected to the biological sample input to contain and mix a composition comprising a biological sample and a particulate substrate.
  • the mixing chamber may reside as part of the bulk fluid volume separated by a displaceable membrane, e.g., rupturable, piercable, puncturable, removable, etc., or other barrier or valve.
  • the mixing chamber may reside as part of the entire bulk fluid volume.
  • the capillary volume may include a fluidic isolation chamber connected to the mixing chamber downstream of the mixing chamber to separate particulate substrate and a biological component from the biological sample.
  • the separation may be by the introduction of a non-newtonian fluid along the fluid column, or in other examples, the introduction of a gas, e.g., gas bubble in the capillary volume to separate the mixing chamber from the fluidic isolation chamber, as described in greater detail hereinafter.
  • upstream and downstream are used merely to describe fluid on two different sides of the plug of non-newtonian plugging fluid. These terms do not imply that fluid is actively flowing in a particular direction through the fluid column.
  • the fluid or fluids on either side of the plug of non-newtonian plugging fluid can be described as “upstream” and “downstream” when fluid has previously flowed through the capillary volume, or when fluid will flow through the capillary volume in the future, or when no flow has occurred or will occur, or when fluid has flowed or will flow in multiple directions through the capillary volume, including opposing directions.
  • the fluid column can be a density gradient column, and the density gradient volume can be defined in part or fully by the interconnected volumes with the bulk fluid volume fluidically connected in series with the capillary volume, the terms “density gradient” can be used in various contexts herein but can refer to the ability of multiple fluids to remain separated in layers due to their density difference (with denser fluids being positioned vertically lower along the column).
  • a density gradient column can contain multiple fluids of different density that are in contact at a density-differential interface.
  • density gradient column can include a capillary volume.
  • the capillary volume can be a portion having a narrowed diameter in which capillary forces can be significant.
  • capillary force can allow a fluid having a lower density to occupy a position below a fluid having a higher density.
  • a fluid with a lower density can be present in the capillary volume, and capillary forces can maintain the lower density fluid in the capillary volume even when a higher density fluid is present above the capillary volume.
  • capillary force or “capillary force-supported gradient” can refer to fluid interfaces that are not maintained by density difference, but rather, the fluids of immediately adjacent layers can have different densities, but less dense fluids can be positioned below denser fluids. Less dense fluids can be constrained within the capillary volume due to the surface tension of the fluids at the fluid interface and the interaction of the fluids with walls of the capillary volume. The interface between such fluids can be a “capillary force-supported interface.”
  • the fluids may be referred to as a “first,” “second,” “third,” etc., fluid so that the fluids can be described relative to one another and for clarity in describing for understanding the disclosure.
  • first fluid and “second fluid” and so on can be interchangeable as is convenient for describing a particular example.
  • first, “second,” and so on do not imply a particular order, position, or hierarchy of the fluids.
  • adjacent fluids can have a density difference that is calculated as the difference between the density of the denser fluid and the density of the lighter fluid.
  • Example density differences between fluids in the density gradient column can be from 50 mg/mL to 3 g/mL, from 100 mg/mL to 3 g/mL, from 500 mg/mL to 3 g/mL or from 1 g/mL to 3 g/mL.
  • the “fluid density” can be measured by calibrating a scale to zero with the container thereon and then obtaining the mass of the fluid, e.g., liquid, in grams.
  • the volume of the measure mass can then be determined using a graduated cylinder.
  • the density is then calculated by dividing the mass by the volume to provide the fluid density (g/mL).
  • the column can also be referred to as a “multi-fluid density gradient” column.
  • the fluids may or may not be positioned 90 degrees from horizontal relative to one another, e.g., they may or may not be stacked or layered directly on top of one another but may be in a vessel angled at less than 90 degrees from horizontal, but the interface between the fluids can be horizontal.
  • the term “vertically layered” refers to fluids that are on top of one another relative to a force such as gravity or centripetal force in a centrifuge with a horizontal interface extending there between, even if they are not fully directly on top of one another.
  • a multi-fluid density gradient column does not include columns where all of the fluid layers include an additional structures substance may be used to separate one fluid layer from another.
  • Fluid layers of the multi-fluid density gradient portion can be phase separated from one another based on fluidic properties of the various fluids, including density of the respective fluids along the column.
  • the first fluid layer can have a first density and can form a first fluid layer of the multi-fluid density gradient portion.
  • the second fluid layer can have a second density that can be greater than a density of the first fluid layer and can form a second fluid layer of the multi-fluid density gradient portion beneath the first fluid layer.
  • An additional fluid layer(s), e.g., third, fourth, etc., can have a third, fourth, etc., density that can be greater than a density of the previous fluid layer and can form a third, fourth, etc., fluid layer of the multi-fluid density gradient portion beneath the second fluid layer.
  • this is not the case for the “capillary force-supported interface.”
  • the surface tension of the fluid relative to the size and material of the vessel provides the ability to put less dense fluids beneath fluids of greater density.
  • a density of a fluid in a fluid layer can be altered using a densifier.
  • Example densifiers can include sucrose, polysaccharides such as FICOLLTM (commercially available from Millipore Sigma (USA)), C 19 H 26 I 3 N 3 O 9 such as NYCODENZ® (commercially available from Progen Biotechnik GmbH (Germany)) or HISTODENZTM, iodixanols such as OPTIPREPTM (both commercially available from Millipore Sigma (USA)), or combinations thereof.
  • example additives that can be included in the fluid layers can include sucrose, C1-C4 alcohol, e.g., isopropyl alcohol, ethanol, etc., which can be included to adjust density, and/or to provide a function with respect to biological components or materials to pass through the column.
  • the fluid devices described herein can include interconnected volumes that includes a bulk fluid volume and a capillary volume.
  • the bulk fluid volume can be upstream of the capillary volume.
  • the density gradient column can be oriented vertically and the bulk fluid volume can be above the capillary volume.
  • the bulk fluid volume can be wider and can have a larger cross-section than a cross-section of the capillary volume.
  • the bulk fluid volume can include a conical chamber, a cylindrical chamber, or a combination thereof.
  • a cross-section of the chamber can be round, square, triangle, rectangle, or other polygonal in shape.
  • the bulk fluid volume can have a diameter at the widest cross-section of from 5 mm to 5 cm, 7 mm to 4 cm, 8 mm to 3 cm, or 8 mm to 2 cm.
  • the bulk fluid volume can be where a majority of the fluid in the density gradient column resides (by fluid volume).
  • the bulk fluid volume can connect to the capillary volume at a capillary junction.
  • the capillary volume can have a smaller cross-section than a cross-section of the bulk fluid volume.
  • the capillary volume can be an elongated tubular region and can have a round, square, triangle, rectangle, or other polygonal cross-section.
  • the capillary volume at the widest cross-section can have an interior opening diameter of from 0.1 mm to 4 mm, 0.2 mm to 3 mm, 0.5 mm to 4 mm, or 1 mm to 3 mm.
  • the capillary volume may be tapered.
  • the capillary can be tapered and can have an interior channel diameter of 4 mm at one end to an interior channel diameter at the opposite end of 1 mm.
  • the capillary can be tapered from an interior channel diameter of 3 mm at one end to an interior channel diameter at the opposite end of 1 mm, or from 2 mm at one end to an interior channel diameter at the opposite end of 1.5 mm, or from 2 mm at one end to an interior channel diameter at the opposite end of 1 mm.
  • the density gradient column can be present in the interconnected volumes, which may be defined by a solid device body, e.g., a unitary or modular vessel.
  • the solid device body can be made of various polymers (e.g. Polypropylene, TYGON, PTFE, COC, others), glass (e.g. borosilicate), metal (e.g. stainless steel), or a combination of materials.
  • the capillary volume can also be formed in the same solid device bod, or the capillary volume can be made from a different material.
  • the capillary volume can be formed from materials used in various microfluidic devices, such as silicon, glass, SU-8, PDMS, a glass slide, a molded fluidic channel(s), 3-D printed material, and/or cut/etched or otherwise formed features.
  • the solid device body, or vessel can be monolithic or may be a combination of components fitted together, thus indicating that interconnected volumes may be defined by a unitary device with multiple regions or may be defined by a modular device where vessel components are joined together to form the interconnected volumes.
  • the interconnected volumes defined by the solid device body can be operable to receive fluids, such as a sample fluid, a lysis buffer, a wash buffer, a gas, e.g., air, a reconstituted reagent, and the like.
  • Fluids can be arranged along the density gradient column in layers and individual layers can be phase separated from one another at fluid interfaces. In some examples, the phase separation can be based on fluidic properties of the various fluids, including density of the respective fluids along the column. Fluid layers can be in fluid communication with adjoining fluid layers.
  • the density gradient column can include a sample fluid positioned on top of a wash buffer, where the wash buffer has a greater density than the sample fluid.
  • the sample fluid can be a biological sample fluid where a biological component is eluted into a buffer, such as from a swab or other biological sample collection device, e.g., DNA, RNA, protein, peptides, amino acids, etc.
  • a biological component is eluted into a buffer, such as from a swab or other biological sample collection device, e.g., DNA, RNA, protein, peptides, amino acids, etc.
  • a surface tension of the fluid relative to the size and material of the vessel can provide the ability to position less dense fluids beneath fluids of greater density.
  • a separation gas bubble e.g., separation air bubble
  • the separation gas bubble can become trapped in the capillary volume due to the surface tension in the capillary volume.
  • a fluid having a density that is less than the density of fluids above the separation gas bubble can be located below the separation gas bubble.
  • the fluid that is above the gas bubble can include densifiers, as described above, and the fluid below the gas bubble can be free of densifiers so that the fluid above the gas bubble has a higher density.
  • the density difference between the fluid above the gas bubble and the fluid below the gas bubble can be from 50 mg/mL to 3 g/mL, from 100 mg/mL to 3 g/mL, from 500 mg/mL to 3 g/mL or from 1 g/mL to 3 g/mL.
  • the separation gas bubble can prevent intermixing despite the density difference.
  • the fluidic device can be a fluid processing device for mixing a biological sample with reagents.
  • the fluid in the capillary volume can include the biological sample and reagents, and the fluid in the bulk fluid volume can include a wash buffer.
  • the wash buffer may be separated from the sample and reagents by a gas bubble.
  • This particular device can also include a cap covering the bottom end of the capillary volume. The cap can be unsealed and the sample and reagents can be ejected out the bottom of the capillary volume.
  • the uncapping and ejecting process can generate back pressure in the capillary volume, which can often push the gas bubble out of the capillary volume, which can break the separation between the wash buffer and the sample/reagent mixture.
  • the non-newtonian plugging fluids described herein can provide a much more secure and robust separation compared to a gas bubble.
  • a combination of a gas bubble and a plug of non-newtonian plugging fluid can be used to separate the fluid in the bulk fluid volume from fluid in the capillary volume.
  • the non-newtonian plugging fluid can be injected into the capillary volume from a reservoir.
  • fluidic devices can also include reservoirs of other fluids that are used in the fluidic device. Reservoirs can be positioned outside of the interconnected volumes that carry the density gradient column. In various examples, the reservoirs can be fluidically connected to the density gradient column via openings, microchannels, inlets, etc., into the interconnected volumes, which can be operable to permit dispensing of fluid from the reservoir into the density gradient column. [0038] Reservoirs can vary in type.
  • a reservoir can be a chamber, a channel, a flexible blister pack, a syringe, a bag, a balloon, or a combination thereof.
  • a reservoir can be a flexible blister pack that when pushed, can open and force contents out of the reservoir and into the density gradient column.
  • the reservoir can include a sealing layer that can maintain separation of contents in the reservoir from the density gradient column until the sealing layer is broken, removed, pierced, etc. Breaking the sealing layer may allow contents of the reservoir to be released therefrom.
  • the fluidic device can include a sharp point located near the sealing layer so that the sharp point can puncture the sealing layer when pressure is applied to the blister.
  • the blister pack can be designed to release fluid from the blister in other ways.
  • the sealing layer can be easy to rupture so that the sealing layer can rupture without a sharp point to puncture the sealing layer.
  • a sharp point can be formed inside the blister, such as on the exterior flexible wall of the blister, so that the sharp point can puncture the sealing layer from the inside of the blister when pressure is applied to the blister.
  • Reservoirs can be sized and shaped to contain a fluid, a reagent, or a combination thereof.
  • Types of reservoirs can include the non-newtonian plugging fluid reservoir, a wash buffer reservoir, a gas reservoir, a dry reagent reservoir, a reconstitution buffer reservoir, or a combination thereof.
  • a non-newtonian plugging fluid reservoir can be sized and shaped to contain a non-newtonian plugging fluid.
  • the non-newtonian plugging fluid reservoir can include the non-newtonian plugging fluid sealed inside the reservoir.
  • the reservoir can be connected to a plugging fluid injection by an injection channel that can be sized appropriately to deliver the non-newtonian plugging fluid to the plugging fluid injection opening.
  • non-newtonian plugging fluids can be used as the non-newtonian plugging fluid.
  • the non-newtonian plugging fluid can have a sufficient viscosity, after the plugging fluid has been injected into the capillary volume of the interconnected volumes, to partition fluid upstream of the plugging fluid from fluid downstream of the plugging fluid.
  • Some non-newtonian plugging fluids can include Bingham plastics, viscoplastics, shear thinning fluids, or curable fluids.
  • Bingham plastics can include materials that behave as rigid bodies at low stress but which flow as a viscous fluid at high stress. The transition between the rigid body behavior and the viscous fluid behavior can occur at various different stress levels, depending on the particular Bingham plastic material. Bingham plastics can include greases, slurries, suspensions of pigments, and others.
  • Viscoplastics are a broader category of materials that can include Bingham plastics. Viscoplastic materials can experience irreversible plastic deformation when stress over a certain level is applied. When stress under this level is applied, the viscoplastic material can behave as a rigid body, as is the case with Bingham plastics, or the viscoplastic material can undergo reversible elastic deformation.
  • Shear thinning fluids are materials that behave as a fluid with a high viscosity when low stress is applied, but the viscosity of the fluid decreases when the stress is increased.
  • shear thinning fluids can include polymer solutions, molten polymers, suspensions, colloids, and others.
  • Curable fluids can include fluids that can undergo a curing process to increase the viscosity of the fluid.
  • the curing process can include thermal curing, chemical curing, ultraviolet radiation curing, or other curing methods.
  • curable fluids can include monomers that can polymerize to form polymers and/or polymers that can become crosslinked during the curing process.
  • examples of curable fluids can include two-part epoxy resins, two-part polyurethane resins, ultraviolet curing epoxies, ultraviolet curing acrylates, ultraviolet curing urethanes, ultraviolet curing thiols, and others.
  • the non-newtonian plugging fluid can prevent flow of fluids through the capillary volume in some examples.
  • the pressures of the fluids involved are relatively low.
  • the non-newtonian plugging fluid can have a sufficiently high viscosity so that the plug of non-newtonian plugging fluid will remain stationary and block fluid flow as long as the fluid pressure is below a particular holding pressure.
  • the holding pressure of the plug of non-newtonian plugging fluid can be from 1 ,000 Pa to 5,000 Pa, or from 1,000 Pa to 4,000 Pa, or from 2,000 Pa to 4,000 Pa.
  • the holding pressure can be sufficient to hold up fluids that are above the plug in a vertically oriented fluid column.
  • the fluids above the plug can have a pressure head that is less than the holding pressure of the plug.
  • the plug can hold fluids up to a pressure head of 8 inches of water, or up to a pressure head of 12 inches of water, or up to a pressure head of 16 inches of water, before the holding pressure of the plug is overcome and the plug is washed out of the capillary volume.
  • the viscosity of the non-newtonian plugging fluid can be sufficient to separate fluids above the plug of non-newtonian plugging fluid from fluids below the plug of non-newtonian. This can include holding a pressure head of the fluids above the non-newtonian fluid when the fluid column is oriented vertically. In some examples, the viscosity of the non-newtonian plugging fluid can be effectively infinite up to a threshold stress. In these examples, the non-newtonian plugging fluid can act as a rigid body when the stress on the fluid is below the threshold.
  • the non-newtonian plugging fluid can have a viscosity that is sufficient to support the fluids above the plug for an amount of time that can allow fluid below the plug to be ejected from the device without mixing the fluid above the plug.
  • the non-newtonian fluid plug can have a viscosity of greater than 5,000 centipoise, or greater than 10,000 centipoise, or greater than 15,000 centipoise, or greater than 20,000 centipoise.
  • the non-newtonian plugging fluid can be grease-based.
  • greye can refer to a dispersion of a thickening agent in a liquid lubricant.
  • Some greases can include a soap emulsified with a base oil, such as a mineral oil, vegetable oil, or petroleum oil.
  • Greases often have a high initial viscosity when at rest, and the viscosity can drop upon application of shear stress.
  • greases are often used on bearings to give the effect of an oil-lubricated bearing when the bearing is in motion, where the grease has approximately the viscosity of the base oil when the bearing is in motion.
  • the non-newtonian plugging fluid can include a mineral oil-based grease, a vegetable oil-based grease, a petroleum oil-based grease, a synthetic oil-based grease, a semi-synthetic oil-based grease, a silicone oil-based grease, or a combination thereof.
  • examples of greases that can be used can include greases available under the trade names ANTI-SEIZE TECHNOLOGYTM (A.S.T.
  • the consistency of the grease can be characterized using a NLGI consistency number.
  • the grease can have an NLGI consistency number from 0 to 6.
  • the NLGI consistency number can be from 1 to 5, from 2 to 5, from 1 to 4, or from 2 to 4.
  • Grease-based plugging fluids can also include additives such as PTFE particles, polyurea, calcium stearate, sodium stearate, lithium stearate, day, graphite, silica, molybdenum disulfide, aluminum, copper, zinc, and others.
  • additives such as PTFE particles, polyurea, calcium stearate, sodium stearate, lithium stearate, day, graphite, silica, molybdenum disulfide, aluminum, copper, zinc, and others.
  • Curable fluids can also be used as the non-newtonian plugging fluid.
  • Curable fluids can include fluids that can undergo a curing process, through which the viscosity of the fluid increases.
  • Some curing fluids can include monomers that can polymerize during curing to form a material having a higher viscosity than the original viscosity of the fluid.
  • Curing fluids can also include gels, cross-linkable polymers, and other materials.
  • a variety of curing processes can be used, such as chemical curing, thermal curing, ultraviolet radiation curing, and so on.
  • a curable adhesive such as ultraviolet curable adhesives can be used.
  • Some example ultraviolet curable adhesives can include MASTERBOND® UV adhesives from Master Bond Inc. (USA).
  • a two-part curable resin such as a two-part epoxy resin can be used. In these examples, two reservoirs can hold the two parts of the resin and the two parts can be mixed together when the two parts are injected into the capillary volume.
  • the non-newtonian plugging fluid can be insoluble in the fluids that will contact the non-newtonian plugging fluid after being injected into the capillary volume of the interconnected volumes.
  • the fluids in the fluid column can be aqueous fluids
  • the non-newtonian plugging fluid can be a non-aqueous fluid such as a grease.
  • the fluids in the fluid column can be non-aqueous fluids and the non-newtonian plugging fluid can be an aqueous fluid such as a hydrogel.
  • the present disclosure also describes fluid processing systems.
  • Fluid processing systems can include fluidic devices as described above together with additional components.
  • fluid processing systems can include a reservoir of wash buffer and an opening for injecting or introducing the wash buffer into the interconnected volumes.
  • the systems can also include a reservoir of non-newtonian plugging fluid as described above.
  • a fluid processing system can include interconnected volumes having a capillary volume fluidically connected in series with a bulk fluid volume thereabove.
  • a reservoir of a wash buffer can be positioned outside the interconnected volumes.
  • a first fluid injection opening can be into the interconnected volumes.
  • the first fluid injection opening can be connected to the reservoir of the wash buffer so that the wash buffer can be injected into the interconnected volumes through the opening.
  • a reservoir of a non-newtonian plugging fluid can also be positioned outside the interconnected volumes.
  • a plugging fluid injection opening can be positioned at a location along a length of the capillary volume to inject the non-newtonian plugging fluid into the capillary volume.
  • the non-newtonian plugging fluid can have a sufficient viscosity to partition fluid upstream of the non-newtonian plugging fluid from fluid downstream of the non-newtonian plugging fluid.
  • the fluid column held in the interconnected volumes can be a density gradient column as described above.
  • the fluid processing systems can also include additional fluid reservoirs.
  • a fluid processing system can include a reservoir of a first fluid, a reservoir of a second fluid, and a reservoir of a non-newtonian plugging fluid.
  • the first and second fluids can be a variety of fluids that can be useful depending on the function of the fluid processing system.
  • the first fluid can be a wash buffer.
  • the wash buffer can be an aqueous solution.
  • a wash buffer can include water, alcohol (such as ethanol), a binding agent, a salt, a surfactant, a stabilizing agent, buffering agents to maintain pH, or a combination thereof.
  • the wash buffer can include a densifier. Any fragments and other materials from the biological sample that may be adhere to the magnetizing particles at locations other than the interactive surface group or the ligand on the exterior surface thereof can be washed off by the wash buffer.
  • the wash buffer can be a liquid that can wash off these materials while also being safe for the biological component.
  • the second fluid can be a reconstitution buffer.
  • the reconstitution buffer can be used to reconstitute a dried reactant and then the reconstitution buffer and the reconstituted reactant can be injected into the capillary volume of the interconnected volumes.
  • the reactant can include PCR (polymerase chain reaction) master mix reactants. This type of reactant can be useful to mix with a sample containing nucleic acids in order to perform nucleic acid amplification or similar processes.
  • PCR master mix reactants can include a mixture of multiple compounds that are used in a PCR assay.
  • the reactant can be a lyophilized PCR master mix.
  • examples of commercially available PCR master mixes include TITANIUM TAQ ECODRYTM premix, ADVANTAGE 2 ECODRYTM premix (available from Takara Bio, Inc. Japan); Lyophilized Ready-to-Use and Load PCR Master Mix (available from Kerafast, Inc., USA); MAXIMOTM Dry-Master Mix (available from GenEon Technologies, USA), and others.
  • the reactant can be a dried reactant that includes all ingredients for the process other than water.
  • the reconstitution buffer can simply be water.
  • the reconstitution buffer can include additional ingredients, such as salts, surfactants, buffering agents to maintain pH, and others.
  • the second fluid can be gas, e.g., air.
  • gas can be injected into the capillary volume of the interconnected volumes to form a gas bubble to separate fluid in the bulk fluid volume from fluid in the capillary volume, as described above.
  • the gas bubble can likewise separate some of the fluid upstream but still within the capillary volume from the fluid that may be downstream from the gas bubble.
  • FIG. 2 shows one example fluid processing system 200.
  • This system includes a solid device body 102 in which interconnected volumes 110 is formed.
  • the interconnected volumes include a capillary volume 112 and a bulk fluid volume 114.
  • a reservoir 120 of non-newtonian plugging fluid 122 is positioned outside the interconnected volumes.
  • a plugging fluid injection opening 124 is positioned at a location along a length of the capillary volume.
  • the system also includes a reservoir 230 of a first fluid 232.
  • the first fluid reservoir is connected to a first fluid injection opening 234 in the interconnected volumes to inject the first fluid into the interconnected volumes.
  • the first fluid reservoir connects to the first fluid injection opening through a long fluid channel that is formed in the solid device body, but which is not shown in this cross-section.
  • the reservoirs are in the form of flexible fluid-filled blisters.
  • a sealing layer 116 keeps the fluids contained in the reservoirs. The fluids can be injected by applying sufficient pressure to the blisters to rupture the sealing
  • FIG. 3A shows another example fluid processing system 200.
  • This system includes a solid device body 102 in which the interconnected volumes 110 is formed.
  • the interconnected volumes include a capillary volume 112 and a bulk fluid volume 114.
  • a reservoir 120 of non-newtonian plugging fluid 122 is positioned outside the interconnected volumes.
  • a plugging fluid injection opening 124 is positioned at a location along a length of the capillary volume.
  • the system also includes a reservoir 230 of a first fluid 232.
  • the first fluid reservoir is connected to a first fluid injection opening 234 in the interconnected volumes to inject the first fluid into the interconnected volumes.
  • the system also includes a reservoir 240 of a second fluid 242.
  • the second fluid reservoir is connected to a second fluid injection opening 244 in the interconnected volumes.
  • the second fluid injection opening is located in the capillary volume below the plugging fluid injection opening.
  • a reactant chamber 246 and a second fluid inlet chamber 248 are located between the second fluid reservoir and the second fluid injection opening.
  • a reactant 260 is held in the reactant chamber, so that the reactant can be mixed with the second fluid.
  • the reservoirs are in the form of flexible fluid-filled blisters.
  • a sealing layer 116 keeps the fluids contained in the reservoirs. The fluids can be injected by applying sufficient pressure to the blisters to rupture the sealing layer.
  • FIG. 3B shows the system of FIG. 3A after the first fluid 232 has been injected into the interconnected volumes 110.
  • the first fluid injection opening 234 is located near the bottom of the capillary volume 112, so the first fluid fills the capillary volume and the bulk fluid volume 114 of the interconnected volumes from the bottom up.
  • pressure is applied to the reservoir 240 of second fluid 242.
  • the second fluid fills the second fluid inlet chamber 248 and the reactant chamber 246. This displaces the gas, e.g., air, that was previously in the second fluid inlet chamber and the reactant chamber.
  • the gas is injected into the capillary volume, which forms a gas gap 250 in the capillary volume. At this point, the gas gap separates the first fluid in the bulk fluid volume from the second fluid. As shown in the figure, a small amount of the first fluid is also left in the capillary volume under the second fluid injection opening.
  • FIG. 3C shows the system after the non-newtonian plugging fluid 122 has been injected into the capillary volume 112 to form a plug.
  • the plug of non-newtonian plugging fluid separates the first fluid in the bulk fluid volume 114 from the gas in the capillary volume. This can provide a much more robust way to keep the fluids separate compared to the gas gap alone.
  • the second fluid can be mixed with the small amount of first fluid at the bottom of the capillary volume, and the mixture can be ejected from the system without fear of the fluids mixing the first fluid above the non-newtonian plugging fluid.
  • the fluid referred to as the second fluid can be a reconstitution buffer as described above.
  • a blister filled with reconstitution buffer can be the reservoir of the second fluid.
  • the reconstitution buffer can flow out of the blister into a reconstitution buffer inlet chamber and reactant chamber. After this, the reconstitution buffer is injected into the capillary volume.
  • the non-newtonian plugging fluid keeps the reconstitution buffer separate from the first fluid in the bulk fluid volume throughout this process.
  • the first fluid can be a wash buffer as described above.
  • the gas that is initially present in the second fluid inlet chamber and the reactant chamber can also be considered to be the “second fluid.”
  • the second fluid inlet chamber and the reactant chamber can be considered to be the reservoir of the second fluid if the gas is considered to be the second fluid.
  • the gas can be displaced and injected into the capillary volume.
  • the non-newtonian plugging fluid can partition the first fluid in the bulk fluid volume from the gas in the capillary volume. Therefore, either the reconstitution buffer or the gas can be considered to be a second fluid in this example.
  • FIG. 4A shows a cross-sectional view of an example fluid processing system 200.
  • the system includes a fluidic device body formed of a solid device body 102.
  • the interconnected volumes 110 can be formed in the solid device body.
  • the fluid column is a density gradient column.
  • the upper part of the interconnected volumes include a bulk fluid volume 114, and the lower part includes a capillary volume 112.
  • the capillary volume includes the narrower section at the bottom of the column, where capillary forces become more significant.
  • the system also includes a wash buffer reservoir
  • the wash buffer reservoir is a flexible blister located on an exterior surface of the solid device body.
  • the wash buffer can be injected from the wash buffer reservoir into the capillary volume through a first fluid injection opening 234.
  • a non-newtonian plugging fluid reservoir 120 is also located on the surface of the solid device body. This reservoir is filled with a non-newtonian plugging fluid 122 that can be injected into the capillary volume through a plugging fluid injection opening 124.
  • a reconstitution buffer reservoir 240 contains a reconstitution buffer 242.
  • the reconstitution buffer reservoir is connected to a reconstitution buffer inlet chamber 248, which is connected to a reactant chamber 246, which is in turn connected to a second fluid injection opening 244.
  • a dried reactant 260 is held inside the reactant chamber.
  • the reservoirs of the various fluids are kept sealed with a sealing layer 116.
  • the sealing film can rupture to allow the fluids to flow into the fluid column carried by the interconnected volumes.
  • This particular system also includes a spring loaded cap 270 that holds a flexible septum 272.
  • the flexible septum can seal the bottom opening of the capillary volume. If it is desired to eject fluid out of the bottom of the capillary volume, then the spring loaded cap can be pushed upward and the bottom end of the capillary volume can push through the septum so that the capillary volume is unsealed and fluid can eject out the bottom opening.
  • FIG. 4B a front view is shown in FIG. 4B.
  • This view shows the surface of the solid device body where the sealing layer is applied.
  • the sealing layer is not shown in FIG. 4B, so that the various chambers and fluid channels are visible.
  • These chambers and fluid channels can be formed as recessed areas in the surface of the solid device body.
  • the sealing layer can be placed over this surface to enclose these chambers and fluid channels.
  • a wash buffer channel 235 is formed so that the wash buffer can flow from the wash buffer reservoir down to the first fluid injection opening 234, which is the lowest of the fluid injection openings in this example.
  • the plugging fluid injection opening 124 is adjacent to a sharp point 126 formed on the solid device body.
  • the sharp point can puncture the sealing layer to allow non-newtonian plugging fluid to flow into the capillary volume through the plugging fluid injection opening.
  • the reconstitution buffer reservoir ruptures, the reconstitution buffer can flow into the reconstitution buffer inlet chamber 248. Then, the reconstitution buffer can flow to the reactant chamber 246 through a reconstitution buffer channel 247.
  • the volume of reconstitution buffer in the reconstitution buffer reservoir can be such that squeezing the blister reservoir will cause reconstitution buffer to fill the reconstitution buffer inlet chamber and the reactant chamber, but little or no reconstitution buffer will flow into the capillary volume at this time.
  • the reconstitution buffer that is in the reactant chamber can then be pushed into the capillary volume by injecting gas, e.g., air, through a gas channel 249.
  • FIG. 4A and FIG. 4B show the fluid processing system before beginning the example process.
  • the process can begin as shown in FIG. 40, by pressing on the wash buffer reservoir blister 230 to inject wash buffer 232 into the interconnected volumes 110.
  • the wash buffer is injected in a lower part of the capillary volume 112. From there, the wash buffer fills up the capillary volume and then partially fills the bulk fluid volume 114.
  • FIG. 4D shows that after introducing the wash buffer 232 into the interconnected volumes 110, a sample fluid 204 is loaded into the interconnected volumes from the top, above the wash buffer.
  • the sample fluid can have a lower density than the wash buffer, so that the sample fluid remains in a layer on top of the wash buffer.
  • the fluids form a density gradient as explained above.
  • the sample fluid can include a biological component such as nucleic acids (such as DNA or RNA), or others. Other materials can also be present, such as lysate and components of lysed cells or viruses.
  • the preparation of the sample fluid can include lysing viruses or cells to extract nucleic acids (such as DNA or RNA) therefrom.
  • the sample fluid in this particular example can include magnetizing particles.
  • the magnetizing particles can be configured to bind or adhere to the biological component.
  • magnetizing particles having biological components bound thereto can be dispersed in the sample fluid.
  • the fluid processing device in this example can include a magnet or system of magnets that can be used to move the magnetizing particles downward through the fluid column. Accordingly, the magnet or magnets can be used to draw the magnetizing particles across the interface from the sample fluid into the wash buffer. Then, the magnets can continue to draw the magnetizing particles down through the capillary volume until the magnetizing particles are concentrated at the bottom of the capillary volume.
  • the biological component can remain bound to the magnetizing particles.
  • the reconstitution buffer reservoir 240 is pressed. This causes the reconstitution buffer 242 to flow into the reconstitution buffer inlet chamber 248 and the reactant chamber 246.
  • the dried reactant that was held in the reactant chamber is dissolved by the reconstitution buffer.
  • the dried reactant can include PCR master mix reactants.
  • the gas e.g., air
  • the gas that was present in these chambers is displaced into the capillary volume of the interconnected volumes 110. This forms a gas gap 250.
  • a small amount of the wash buffer 232 remains at the bottom of the capillary volume.
  • wash buffer was injected through the first fluid inlet opening, which is located below the second fluid inlet opening.
  • the gas flows in through the second fluid inlet opening to form the gas gap.
  • magnetizing particles with biological components bound thereon are concentrated in the wash buffer at the bottom of the capillary volume. Therefore, the biological components remain in the small volume of wash buffer at the bottom of the capillary volume, below the gas gap.
  • the non-newtonian plugging fluid reservoir 120 is pressed so that the non-newtonian plugging fluid 122 is injected into the capillary volume of the interconnected volumes 110.
  • the non-newtonian fluid can have a sufficient viscosity after being injected into the capillary volume that the non-newtonian fluid can prevent the fluid above the plug from flowing down into the capillary volume below the plug.
  • This figure shows the non-newtonian plugging fluid being injected into a region of the capillary volume that contains wash buffer, so that a small amount of wash buffer is beneath the plug between the plug and the gas gap 250.
  • the non-newtonian plugging fluid may be injected directly into the gas gap.
  • the plug can be in direct contact with gas on the bottom of the plug and with wash buffer on the top of the plug.
  • a small amount of gas from the gas gap can remain on the top on the top of plug.
  • FIG. 4G shows that after the plug of non-newtonian plugging fluid 122 has been formed, the reconstitution buffer 242 and the small volume of wash buffer 232 at the bottom of the capillary volume can be ejected or dispensed out of the device or system. This can be accomplished by uncapping the bottom end of the capillary volume.
  • the spring loaded cap 270 can be pushed upward and the bottom end of the capillary volume can penetrate through the flexible septum 272.
  • a reservoir 280 of gas can be pressed to use gas to force the reconstitution buffer and the wash buffer out of the bottom opening of the capillary volume.
  • the view of the fluid processing system shown in FIG. 4G is extended at the top to show the gas reservoir, which was not shown in the previous figures.
  • FIG. 4H shows a front view of the surface of the solid device body.
  • the gas flows from the gas reservoir, through a gas channel 249 that connects to the fluid channel 247 between the reconstitution buffer inlet chamber and the reactant chamber.
  • the reactant chamber and the fluid channel were filled with the reconstitution buffer and the dissolved reactants.
  • the gas forces the reconstitution buffer and dissolved reactants out into the capillary volume, and then out of the bottom opening of the capillary volume.
  • some of the reconstitution buffer remains in the reconstitution buffer inlet chamber, as this chamber is bypassed by the gas from the gas reservoir.
  • FIGS. 4A-4H can be used to prepare mixtures of biological components with reactants such as PCR master mix reactants. This mixture can then be further analyzed and processed using appropriate equipment. After the mixture of biological components and PCR master mix reactants is ejected from the bottom of the capillary volume as shown in FIG. 4G, the capillary volume can be re-capped and the fluidic device can be discarded.
  • reactants such as PCR master mix reactants.
  • the fluid processing systems according to the present disclosure can be used to perform a variety of other processes, and the fluid processing systems can include other components besides those described above.
  • various fluids can be added to the interconnected volumes to form a density gradient column in the fluid processing system.
  • the density gradient column can include a sample fluid and a wash buffer.
  • the density gradient column can include a sample fluid, a wash buffer, gas, and a reconstituted reagent.
  • the density gradient column can include a sample fluid, a lysate solution, a wash buffer, gas, and a reconstituted reagent, as described above.
  • the fluid layers in the density gradient column can be formulated to interact with the magnetizing particles that can be present in the sample fluid.
  • the sample fluid layer and other individual fluid layers can have different functions.
  • a fluid layer can include a lysis buffer to lyse cells.
  • a fluid layer can be a surface binding fluid layer to bind the biological component to the magnetizing particles
  • a wash fluid layer can trap contaminant from a sample fluid and/or remove contaminant from an exterior surface of the magnetizing particles
  • a surfactant fluid layer can coat the magnetizing particles
  • an elution fluid layer can remove the biological component from the magnetizing particles following extraction from the biological sample
  • a labeling fluid layer can bind labels to the biological component such as a fluorescent label (either attached to the magnetizing particles or unbound thereto), and so on.
  • individual fluid layers can provide sequential processing of a biological sample.
  • individual fluid layers can carry out individual functions, and in many cases, the functions can be coordinated to achieve a specific result.
  • sequential fluid layers from top to bottom of a density gradient column can act on the biological sample to lyse cells in a fluid layer, bind a biological component from the lysed biological material to magnetizing partides, wash the magnetizing particles with the biological material bound thereto in a fluid layer, combine biological material with a reagent, and/or elute (or separate) the biological material from the magnetizing particles.
  • a vertical height of individual fluid layers in the density gradient column can vary. Adjusting a vertical height of an individual fluid layer can affect a residence time of the paramagnetic microparticles in that fluid layer. The taller the fluid layer, the longer the residence time of the magnetizing particles in the fluid layer. In some examples, all of the fluid layers in the density gradient column can be the same vertical height. In other examples, a vertical height of individual fluid layers in a multi-fluid density gradient column can vary from one fluid layer to the next. In one example, a vertical height of the individual fluid layers can individually range from 10 ⁇ m to 50 mm. In another example, a vertical height of the individual fluid layers can individually range from 10 ⁇ m to 30 mm, from 25 ⁇ m to 1 mm, from 200 ⁇ m to 800 ⁇ m, or from 1 mm to 50 mm.
  • the interconnected volumes defined by the solid device body may further include openings, inputs, outputs, and/or ports.
  • a fluid injection opening can permit loading of a sample fluid, a wash buffer, or the like into the bulk fluid volume of the density gradient column.
  • the density gradient column can include an input or port to permit loading of fluids and reagents in the density gradient column.
  • the solid device body may also include outputs.
  • the capillary volume may include a fluidic output that can permit dispensing of a biological component, a biological sample, a fluid, magnetizing particles, or a combination thereof from the density gradient column.
  • the solid device body where the interconnected volumes reside may include an output for venting gas to relieve pressure in the density gradient column.
  • the wash buffer reservoir can be a flexible fluid-filled blister.
  • the wash buffer can be an aqueous solution.
  • a wash buffer can include alcohol (such as ethanol), a binding agent, a salt, a surfactant, a stabilizing agent, or a combination thereof.
  • the amount of wash buffer in the wash buffer reservoir can be sufficient to fill or partially fill the capillary volume and the bulk fluid volume.
  • a gas reservoir can be sized and shaped to contain gas in an amount capable of pushing out the reconstitution buffer and the volume of wash buffer that includes the concentration magnetizing particles at the end of the process described above.
  • the gas reservoir may be located to allow dispensing of gas into the capillary volume.
  • the gas reservoir can be connected by a gas channel to the reactant chamber.
  • the reactant chamber can be sized and shaped to contain a dry reagent.
  • the dry reagent reservoir can include the dry reagent.
  • a dry reagent can include PCR master mix reagents, nucleic acid primers, secondary antibodies, polymerases, magnesium salt, bovine serum albumin (BSA), or combinations thereof.
  • the reactant chamber can be connected to a reservoir of reconstitution buffer so that the reconstitution buffer can reconstitute the dry reactant.
  • the reactant chamber can also be connected to the capillary volume so that the reconstitution buffer and reconstituted reactant can be injected into the capillary volume.
  • a reconstitution buffer reservoir can be sized and shaped to contain a reconstitution buffer.
  • the reconstitution buffer reservoir can include the reconstitution buffer.
  • the reconstitution buffer can be any aqueous solvent.
  • the reconstitution buffer can be water.
  • the reconstitution buffer reservoir as previously discussed, can be arranged to allow dispensing of the reconstitution buffer into a reactant chamber to reconstitute a dry reactant.
  • Reservoirs may be arranged to allow a fluid or a reagent therein to be individually dispensed into the density gradient column, and/or can be arranged in series to allow a fluid, a reagent, or a combination thereof to be dispensed sequentially or at the same time into the density gradient column.
  • the magnetizing particles in the methods and systems described herein can be in the form of paramagnetic microparticles, superparamagnetic microparticles, diamagnetic microparticles, or a combination thereof, for example.
  • the term “magnetizing particles” is defined herein to include microparticles that may not be magnetic in nature unless and until a magnetic field is introduced at a strength and proximity to cause them to become magnetic. Their magnetic strength can be dependent on the magnetic field applied and may become stronger as the magnetic field is increased, or as the magnetizing particles move closer to the magnetic source that is applying the magnetic field.
  • paramagnetic microparticles have these properties, in that they have the ability to increase in magnetism when a magnetic field is present; however, paramagnetic microparticles are not magnetic when a magnetic field is not present. In some examples, the paramagnetic microparticles can exhibit no residual magnetism once the magnetic field is removed. A strength of magnetism of the paramagnetic microparticles can depend on the strength of the magnetic field, the distance between a source of the magnetic field and the paramagnetic microparticles, and a size of the paramagnetic microparticles.
  • “Superparamagnetic microparticles” can act similar to paramagnetic microparticles, however, they can exhibit magnetic susceptibility to a greater extent than paramagnetic microparticles in that the time it takes for them to become magnetized appears to be near zero seconds. “Diamagnetic microparticles,” on the other hand, can display magnetism due to a change in the orbital motion of electrons in the presence of a magnetic field.
  • the magnetizing particles can be surface-activated to selectively bind with a biological component or can be bound to a biological component from a biological sample.
  • An exterior of the magnetizing particles can be surface-activated with interactive surface groups that can interact with a biological component of a biological sample or may include a covalently attached ligand.
  • the ligand can include proteins, antibodies, antigens, nucleic acid primers, nucleic acid probes, amino groups, carboxyl groups, epoxy groups, tosyl groups, sulphydryl groups, or the like.
  • the ligand can be a nucleic acid probe. The ligand can be selected to correspond with and to bind with the biological component.
  • the ligand may vary based on the type of biological component targeted for isolation from the biological sample.
  • the ligand can include a nucleic acid probe when isolating a biological component that includes a nucleic acid sequence.
  • the ligand can include an antibody when isolating a biological component that includes antigen.
  • magnetizing particles that are surface-activated include those sold under the trade name DYNABEADS®, available from ThermoFischer Scientific (USA).
  • the magnetizing particles can have an average particle size that can range from 10 nm to 50,000 nm. In yet other examples, the magnetizing particles can have an average particle size that can range from 500 nm to 25,000 nm, from 10 nm to 1 ,000 nm, from 25,000 nm to 50,000 nm, or from 10 nm to 5,000 nm.
  • the term “average particle size” describes a diameter or average diameter, which may vary, depending upon the morphology of the individual particle.
  • a shape of the magnetizing particles can be spherical, irregular spherical, rounded, semi-rounded, discoidal, angular, sub-angular, cubic, cylindrical, or any combination thereof.
  • the particles can include spherical particles, irregular spherical particles, or rounded particles.
  • the shape of the magnetizing particles can be spherical and uniform, which can be defined herein as spherical or near-spherical, e.g., having a sphericity of >0.84.
  • any individual particles having a sphericity of ⁇ 0.84 are considered non-spherical (irregularly shaped).
  • the particle size of the substantially spherical particle may be provided by its diameter, and the particle size of a non-spherical particle may be provided by its average diameter (e.g., the average of multiple dimensions across the particle) or by an effective diameter, e.g., the diameter of a sphere with the same mass and density as the non-spherical particle.
  • average diameter e.g., the average of multiple dimensions across the particle
  • an effective diameter e.g., the diameter of a sphere with the same mass and density as the non-spherical particle.
  • the magnetizing particles can be unbound to a biological component when added to the density gradient column or to the sample fluid in preparation for adding to the density gradient column. Binding between the magnetizing particles and the biological component of the biological sample can occur within the density gradient column. In yet another example, the magnetizing particles and a biological sample including a biological component can be combined before the sample fluid is added to the density gradient column.
  • the fluid processing systems can also include a magnet capable of generating a magnetic field.
  • the magnetic field may be turned on and off by introducing electrical current/voltage to the magnet.
  • the magnet can be permanently placed, can be movable along the density gradient column, or can be movable in position and/or out of position to effect movement of the magnetizing particles in and through the density gradient column.
  • the magnetizing particles can be magnetized by the magnetic field generated by the magnet.
  • the magnet can also create a force capable of pulling the magnetizing particles through the density gradient column, holding the magnetizing particles at a location along the density gradient column, or a combination thereof.
  • the magnetizing particles can reside in a fluid layer until gravity pulls the magnetizing particles through fluid layers of the density gradient column, or they may remain suspended in the fluid layer in which they may reside until the magnetic field is applied thereto.
  • the rate at which gravity pulls the magnetizing particles through fluid layers can be based on a mass of the magnetizing particles, a quantity of the magnetizing particles, and a surface tension at the fluid interface between fluid layers.
  • the magnet can cause the magnetizing particles to move from one fluid layer to another or can increase a rate at which the magnetizing particles pass from one fluid layer into another.
  • Strength of the magnetic field and the location of the magnet in relation to the magnetizing particles can also affect a rate at which the magnetizing particles move through the density gradient column. The further away the magnet and the lower the strength of the magnetic field, the slower the magnetizing particles will move.
  • the magnet can be moveable in position, out of position, or at variable positions to effect downward movement, rate of movement, or to promote little to no movement of the magnetizing particles.
  • the magnet can be positioned adjacent to a side of the multi-fluid density gradient column and can move vertically to cause the magnetizing particles to move therewith.
  • the magnet can be a ring magnet that can be placed around a circumference of the interconnected volumes that carry the density gradient column.
  • a movable magnet(s) can likewise be positioned adjacent to a side of the interconnected volumes that is not a ring shape, but can be any shape effective for moving magnetizing particles along the interconnected volumes.
  • the magnet can be moved along a side and/or along a bottom of the interconnected volumes to pull the magnetizing particles in one direction or another through the density gradient column. In one example, the magnet can be used to pull the magnetizing particles downwardly through fluid layers of the density gradient column.
  • a magnet can be used to concentrate and hold the magnetizing particles near a side wall defining the interconnected volumes that carry the density gradient column.
  • the magnet can concentrate the magnetizing particles near a side wall and heat can be applied to decouple and separate an isolated biological component from the magnetizing particles.
  • the magnet can continue to hold the magnetizing particles while a biological sample outlet associated with the capillary volume can be opened thereby allowing dispensing of the isolated biological component from the density gradient column where the biological component is separated from the magnetizing particles.
  • FIG. 5 is a flowchart illustration of one example method 300 of processing fluids.
  • the method can include injecting 310 a first fluid into interconnected volumes having a bulk fluid volume fluidically connected in series with a capillary volume to receive a density gradient column, wherein the first fluid occupies the capillary volume (which is inclusive of also allowing for occupying the bulk fluid volume in part above the capillary volume).
  • the method further can further include injecting 320 a non-newtonian plugging fluid at a location along the interconnected volumes.
  • the non-newtonian plugging fluid partitions the first fluid or portion thereof from fluid upstream and above the non-newtonian plugging fluid.
  • the method may also include injecting 330 a second fluid into the capillary volume downstream from the non-newtonian plugging fluid.
  • the first fluid in this example has a higher density than the second fluid.
  • the non-newtonian plugging fluid does not allow the second fluid to enter fluid positioned upstream and above the non-newtonian plugging fluid.
  • the first fluid, the second fluid, the non-newtonian plugging fluid, or a combination thereof can be individually injected from respective flexible fluid-filled blister.
  • the second fluid can be an aqueous PCR master mix solution.
  • the first fluid can be a wash buffer and the method can further include loading a sample fluid, e.g., biological sample fluid, including magnetizing particles having a biological component bound thereto over the wash buffer forming a density gradient column.
  • the sample fluid e.g., biological sample fluid
  • the sample fluid can be prepared and/or loaded in any of a number manners.
  • the sample fluid may be prepared by combining multiple components within the bulk fluid volume, e.g., combining carrier fluid or buffer with magnetizing particles and the biological component within the bulk fluid volume (which may be or include a portion thereof that acts as a mixing chamber).
  • the biological component may become associated with the magnetizing particles in the bulk fluid volume, or the biological component may already be associated with the magnetizing particles where they are combined with the fluid carrier or buffer within the bulk fluid volume.
  • the sample fluid may first be prepared in a vial or other vessel outside of the bulk fluid volume, and then the sample fluid can be added into the bulk fluid volume over the wash buffer either before, after, or at the same time that the wash buffer is loaded, e.g., from the bottom up through the capillary volume or also loaded from the top prior to loading the sample fluid.
  • a biological sample or specimen may be collected using a swab or other biological sample collection instrument.
  • the biological sample may include the biological component of interest.
  • the released or eluted biological component can then be placed in the carrier fluid or buffer where the biological component is eluted into the carrier fluid or buffer.
  • the biological component can become associated with magnetizing particles in the vial or vessel (or even thereafter in the bulk fluid volume, in some examples). Then, the biological sample fluid that includes the eluted biological component can be loaded into the bulk fluid volume or a mixing chamber fluidically connected to or integrated as part of the bulk fluid volume. Thus, elution may occur outside of the cartridge module before sample input, or within the cartridge module where the carrier fluid or buffer is also loaded.
  • the operations in this method can be performed in any order.
  • the flowchart does not imply a particular order of performing these operations.
  • the non-newtonian plugging fluid can be injected before the second fluid in some cases, while in other cases the second fluid can be injected before the non-newtonian plugging fluid.
  • the methods of processing fluids can include any processes described above.
  • a method of processing fluids can include the process depicted in FIGS. 4A-4H. Any of the devices, materials, and components described above can be used in the methods of processing fluids.
  • the term “about” is used to provide flexibility to a numerical range endpoint by providing that a given value may be “a little above” or “a little below” the endpoint.
  • the degree of flexibility of this term can be dictated by the particular variable and determined based on experience and the associated description herein.
  • Bingham plastic refers to a class of materials that behave as rigid bodies at low stress but which flow as a viscous fluid at high stress. The transition between the rigid body behavior and the viscous fluid behavior can occur at various different stress levels, depending on the particular Bingham plastic material. Bingham plastics can include greases, slurries, suspensions of pigments, and others.
  • viscoplastic refers to a broader category of materials that can include Bingham plastics. Viscoplastic materials can experience irreversible plastic deformation when stress over a certain level is applied. When stress under this level is applied, the viscoplastic material can behave as a rigid body, as is the case with Bingham plastics, or the viscoplastic material can undergo reversible elastic deformation.
  • shear thinning fluid refers to materials that behave as a fluid with a high viscosity when low stress is applied, but the viscosity of the fluid decreases when the stress is increased.
  • shear thinning fluids can include polymer solutions, molten polymers, suspensions, colloids, and others.
  • curable fluids refers to fluids that can undergo a curing process to increase the viscosity of the fluid.
  • the curing process can include thermal curing, chemical curing, ultraviolet radiation curing, or other curing methods.
  • curable fluids can include monomers that can polymerize to form polymers and/or polymers that can become crosslinked during the curing process.
  • examples of curable fluids can include two-part epoxy resins, two-part polyurethane resins, ultraviolet curing epoxies, ultraviolet curing acrylates, ultraviolet curing urethanes, ultraviolet curing thiols, and others.
  • the term “interact” or “interaction” as it relates to a surface of the particulate substrates, such as the magnetizing particles indicates that a chemical, physical, or electrical interaction occurs where a particulate substrate surface property is modified in some manner that is different than may have been present prior to entering the fluid layer, but does not include modification of magnetic properties magnetizing particles as they are influenced by the magnetic field introduced by the magnet.
  • a fluid layer can include a lysis buffer to lyse cells, and cellular components can become bound to or otherwise associated with a surface of the magnetizing particles.
  • Lysing cells in a fluid can modify the sample fluid and thus modify or interact with a surface of magnetizing particles, e.g., the cellular component binds or becomes otherwise associated with a surface of the magnetizing particles.
  • the association between the biological component and the magnetizing particles (or other particulate substrate) can alternatively include surface adsorption, electrostatic attraction, or some other attraction between the biological component and the surface of the particulate substrate.
  • a fluid layer that would be considered to interact with the magnetizing particles could be a wash fluid layer to trap contaminates from a sample fluid and/or remove contaminates from an exterior surface of the magnetizing particles, a surfactant fluid layer to coat the magnetizing particles, a dye fluid layer to introduce visible or other markers to the fluid or surface, an elution fluid layer to remove the biological component from the magnetizing particles following extraction from the biological sample, a labeling fluid layerfor binding labels to the biological component such as a fluorescent label (either attached to the magnetizing particles or unbound thereto), a reagent fluid layer to prep a biological component for further analysis such as a master mix fluid layer to prep a biological component for PCR, and so on.
  • a wash fluid layer to trap contaminates from a sample fluid and/or remove contaminates from an exterior surface of the magnetizing particles
  • a surfactant fluid layer to coat the magnetizing particles
  • a dye fluid layer to introduce visible or other markers to the fluid or surface
  • an elution fluid layer to remove the
  • Concentrations, amounts, and other numerical data may be expressed or presented herein in a range format.
  • a range format is used merely for convenience and brevity and thus should be interpreted flexibly to include the numerical values explicitly recited as the limits of the range, and also to include individual numerical values or sub-ranges encompassed within that range as if numerical values and sub-ranges are explicitly recited.
  • a numerical range of “about 1 wt% to about 5 wt%” should be interpreted to include the explicitly recited values of about 1 wt% to about 5 wt%, and also to include individual values and sub-ranges within the indicated range.
  • a fluidic device was constructed having a design as shown in FIG. 4A.
  • the interconnected volumes included a capillary volume with an inner diameter of 1.5 mm.
  • a blister was filled with a grease.
  • the blister was connected to the capillary volume in the device through a plugging fluid inlet opening.
  • Another blister was filled with a wash buffer.
  • the wash buffer was first injected from below through the capillary volume upward into the bulk density column by rupturing the wash buffer blister and forcing the wash buffer fluid into the interconnected volumes.
  • the grease blister was then ruptured by applying force with a mechanical piston. The grease from the blister flowed into the capillary volume, forming a fluid-tight seal with the walls of the capillary volume.
  • the holding pressure of the grease plug was then tested by applying pressure to the top of the bulk fluid volume down into the fluid column using a syringe and measuring the pressure. This was repeated several times with plugs of grease having different lengths in the capillary volume.
  • the length of the grease plugs, the pressure at slip, and the maximum holding pressure are listed in Table 1.
  • the pressure at slip is the measured pressure when the grease plug first shows movement.
  • the maximum holding pressure is the measured pressure when the grease plug was flushed out the bottom of the column.

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Dispersion Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Medicinal Chemistry (AREA)
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Abstract

L'invention concerne un dispositif fluidique qui peut inclure des volumes interreliés comprenant un volume de fluide en vrac relié par voie fluidique en série avec un volume capillaire pour recevoir une colonne à gradient de densité, un réservoir d'un fluide de colmatage non newtonien positionné à l'extérieur des volumes interreliés, et une ouverture d'injection de fluide de colmatage positionnée à un emplacement sur une longueur des volumes interreliés pour y injecter le fluide de colmatage non newtonien. Le fluide de colmatage non newtonien peut présenter une viscosité suffisante pour séparer le fluide en amont du fluide de colmatage non newtonien du fluide en aval du fluide de colmatage non newtonien.
PCT/US2020/063756 2020-12-08 2020-12-08 Dispositifs fluidiques avec fluide de colmatage non newtonien WO2022125074A1 (fr)

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Cited By (1)

* Cited by examiner, † Cited by third party
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CN115235151A (zh) * 2022-07-13 2022-10-25 广州特域机电有限公司 一种防堵塞的毛细管

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US20060283486A1 (en) * 2005-06-15 2006-12-21 Lam Research Corporation Method and apparatus for cleaning a substrate using non-newtonian fluids
EP2019320B1 (fr) * 2007-07-27 2010-09-15 Hewlett-Packard Development Company, L.P. Systèmes et procédés de mesure d'hémoglobine glyquée
EP1587626B1 (fr) * 2003-01-31 2012-05-09 Hewlett-Packard Development Company, L.P. Dispositif microfluidique a dispositifs electroniques en films minces
WO2013159116A1 (fr) * 2012-04-20 2013-10-24 University Of Chicago Dispositifs fluidiques utilisés pour la conservation d'échantillons biologiques

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Publication number Priority date Publication date Assignee Title
EP1587626B1 (fr) * 2003-01-31 2012-05-09 Hewlett-Packard Development Company, L.P. Dispositif microfluidique a dispositifs electroniques en films minces
US20060283486A1 (en) * 2005-06-15 2006-12-21 Lam Research Corporation Method and apparatus for cleaning a substrate using non-newtonian fluids
EP2019320B1 (fr) * 2007-07-27 2010-09-15 Hewlett-Packard Development Company, L.P. Systèmes et procédés de mesure d'hémoglobine glyquée
WO2013159116A1 (fr) * 2012-04-20 2013-10-24 University Of Chicago Dispositifs fluidiques utilisés pour la conservation d'échantillons biologiques

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115235151A (zh) * 2022-07-13 2022-10-25 广州特域机电有限公司 一种防堵塞的毛细管
CN115235151B (zh) * 2022-07-13 2023-09-26 广州特域机电有限公司 一种防堵塞的毛细管

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