WO2019174400A1 - 高度并行的微流体血液分离装置 - Google Patents

高度并行的微流体血液分离装置 Download PDF

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WO2019174400A1
WO2019174400A1 PCT/CN2019/072361 CN2019072361W WO2019174400A1 WO 2019174400 A1 WO2019174400 A1 WO 2019174400A1 CN 2019072361 W CN2019072361 W CN 2019072361W WO 2019174400 A1 WO2019174400 A1 WO 2019174400A1
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channel
separation
fluid
sample
flow
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PCT/CN2019/072361
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English (en)
French (fr)
Inventor
瓦希迪·B
唐东江
叶莘
范献军
周燕玲
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珠海圣美生物诊断技术有限公司
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Priority to CN201980000198.4A priority Critical patent/CN110494756A/zh
Publication of WO2019174400A1 publication Critical patent/WO2019174400A1/zh

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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    • 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
    • B01L3/502715—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 interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
    • B—PERFORMING OPERATIONS; TRANSPORTING
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    • 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
    • B01L3/502776—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 specially adapted for focusing or laminating flows
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C1/00—Magnetic separation
    • B03C1/005—Pretreatment specially adapted for magnetic separation
    • B03C1/01—Pretreatment specially adapted for magnetic separation by addition of magnetic adjuvants
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C1/00—Magnetic separation
    • B03C1/02—Magnetic separation acting directly on the substance being separated
    • B03C1/28—Magnetic plugs and dipsticks
    • B03C1/288—Magnetic plugs and dipsticks disposed at the outer circumference of a recipient
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
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    • B03C1/00—Magnetic separation
    • B03C1/32—Magnetic separation acting on the medium containing the substance being separated, e.g. magneto-gravimetric-, magnetohydrostatic-, or magnetohydrodynamic separation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5002—Partitioning blood components
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • G01N33/54326—Magnetic particles
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    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/80—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood groups or blood types or red blood cells
    • 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
    • B01L2200/04—Exchange or ejection of cartridges, containers or reservoirs
    • 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
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
    • B01L2200/0652—Sorting or classification of particles or molecules
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0829—Multi-well plates; Microtitration plates
    • 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
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0893—Geometry, shape and general structure having a very large number of wells, microfabricated wells
    • 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
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/043—Moving fluids with specific forces or mechanical means specific forces magnetic forces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
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    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
    • 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
    • B01L3/502707—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 manufacture of the container or its components
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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    • B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C2201/00—Details of magnetic or electrostatic separation
    • B03C2201/18—Magnetic separation whereby the particles are suspended in a liquid
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C2201/00—Details of magnetic or electrostatic separation
    • B03C2201/26—Details of magnetic or electrostatic separation for use in medical applications

Definitions

  • the disclosure relates to a blood separation device, and in particular to a device that uses sheath flow technology in a highly parallel configuration.
  • Sheath flow blood separation techniques have proven effective in clinical analysis for a variety of applications. Such techniques can benefit from flux improvements in both the separation operation and the separate analyte collection.
  • a highly parallel microfluidic blood separation device for the isolation of various analytes for large scale clinical trials.
  • the device can be used to isolate a plurality of different target analytes, which can be starting materials for various diagnostic methods including NGS, PCR, FISH, and IHC. Separation is achieved via the principle of magnetic sheath flow and is highly parallel.
  • the separation device achieves efficient magnetic separation, acts during the flow of the sample within the vertical geometry, and can be fastened to standard multiwell plates for highly multiplexed sample recovery. Operates in the vertical direction, allowing multiplexing, allowing more cards/slots/spaces on the instrument, allowing real estate to be used effectively on the countertop and instrument.
  • a parallel microfluidic blood separation device comprising at least two parallel microfluidic channels including a flow region configured to direct fluid flow in a substantially vertical direction; each channel having at least one sample container a (hopper) in fluid communication with the flow region; at least one buffer fluid input port in fluid communication with the flow region; and at least one fluid output port; wherein the device is configurable to receive sample fluid at a substantially top end of each channel, Buffer fluid is received at the side of each channel, wherein a pressure is applied between the buffer input port and an output port located substantially at the bottom of each channel, creating a sheath of sample and buffer fluid between the flow region and the output port Flow, and wherein the device is configured for use with a separation device to capture an analyte of interest from a sample fluid in the device.
  • the separation device can be magnetic, configured to attract the magnetically labeled analyte to the sidewall of each channel, and to maintain the analyte in place after the flow operation is completed To remain in the device.
  • the device may be made of at least two plastic parts, one part injection molded with a hopper, an input port, an output port and a flow area structure, and the other part being a cover piece, and injection molded The components are connected to obtain a finished device.
  • the at least one flexible gasket material is injection molded to the at least one port.
  • the flexible gasket can be on the output port that is disposed substantially on the bottom of each channel and the gasket is configured to seal to the orifice container.
  • the plurality of separation devices containing the separated analytes can be nested together in a retaining fixture and sealed to the multi-row orifice container.
  • the rotating fixture can be configured to rotate a plurality of sets of separation devices sealed to the orifice plate together for highly parallel sample extraction.
  • the apparatus can be configured to engage the output port of each channel to the collection device after the flow separation is complete, wherein the separation device and the collection device can be rotated to deliver the analyte into the collection device.
  • the collection device can be an orifice plate and the separation device output port seals each channel to the rows of orifice plate containers.
  • a method of highly parallel microfluidic blood separation using a separation device comprising at least two parallel microfluidic channels including a flow region configured to be in a substantially vertical direction
  • the method comprising the steps of: filling a sample hopper of each channel in fluid communication with the flow region; adding a buffer fluid to an input port of each channel in fluid communication with the flow region; and extracting a buffer from the output port And a sample fluid; wherein the sample fluid can be received at a substantially top end of each channel, and a buffer fluid is received at a side of each channel, wherein the buffer input port and the output port located substantially at the bottom of each channel Pressure is applied to create a sheath flow of sample and buffer fluid in the flow region to the output port, and wherein the separation device is used to capture the analyte of interest from the sample fluid in the device.
  • the separation device can be magnetic, the separation device attracts the magnetically labeled analyte to the sidewall of each channel, and after the flow operation is completed, the analyte is maintained in a suitable The location is left in the device.
  • the method may further comprise the step of, after the flow separation is completed, engaging the output port of each channel to the collection device, wherein the separation device and the collection device are rotated to deliver the analyte Into the collection device.
  • the collection device can be an orifice plate and the separation device output port seals each channel to the rows of orifice plate containers.
  • the plurality of separation devices with the separated analytes can be nested together in a retaining fixture and sealed to the multi-row orifice container.
  • FIG. 1A and 1B illustrate an exemplary embodiment of a channel of a vertical, highly parallel blood separation device
  • FIGS. 2A and 2B illustrate an exemplary embodiment of a vertical, highly parallel blood separation device
  • Figure 3 shows the construction details of an exemplary embodiment of a vertical, highly parallel blood separation device
  • Figure 4 illustrates the compatibility between an exemplary embodiment of a vertical, highly parallel blood separation device and a standard orifice plate
  • Figure 5 illustrates an exemplary embodiment of a device for holding and rotating a plurality of vertically parallel height blood separation devices that can be used for unloading of separated analytes
  • Figure 6 depicts the seal between a vertical highly parallel blood separation device and an orifice plate for sample analyte unloading.
  • the present disclosure relates to a highly parallel blood separation device configured for vertical flow operation, including a plurality of parallel flow channels and compatible with standard sample analysis devices, such as orifice plates.
  • Patent 8,263,287 describes a technique for the flow of a sample fluid coated by one or more buffer fluid streams.
  • Analytes of interest such as cancer cells in the blood, are labeled in some manner, such as in combination with magnetic particles.
  • a separation mechanism such as a magnetic field
  • the analyte of interest is separated from the sample stream into one of the buffer streams and captured within the sheath flow device.
  • the pellets of the analyte remain in the device and can be removed for analysis.
  • One type of removal technique is the spin elution technique described in U.S. Provisional Application Serial No. 61/702,730, filed on Sep. This application incorporates it in its entirety by reference.
  • a smaller sample volume can be provided at a higher sample density.
  • these features allow for much faster detection.
  • the sample fluid can be end loaded and the ends depleted.
  • this arrangement does not require reorientation of sample flow, reducing manufacturing complexity and cost.
  • the collection of isolated analytes of interest is compatible with standard multi-sample analysis, such as well plates.
  • this arrangement allows the sample to be delivered directly into the high throughput analytical device.
  • sample loss missing analyte/cell of interest
  • present disclosure also introduces a direct transfer method that introduces minimal sample loss into standard container designs, such as PCR tubes and multiwell plates.
  • the present disclosure provides a highly parallel microfluidic blood separation device for isolating various analytes for large scale clinical trials.
  • the device can be used to isolate a variety of different target analytes, such as cfDNA, CTCs, RNA, and exosomes, which can be used in a variety of diagnostic methods (including NGS, PCR, FISH, IHC, and other methods). Starting material. Separation is achieved by the principle of magnetic sheath flow via the incorporated references, but enhanced to a high degree of parallelism.
  • the apparatus of the present disclosure provides enhanced magnetic separation that functions during flow of the sample within the vertical geometry and is fastened to a standard multiwell plate for highly multiplexed sample recovery.
  • the vertical orientation of the system eliminates the need for end buffers, which reduces costs and improves automatic compatibility. Operation in the vertical direction allows multiplexing, allowing more cards/slots/spaces on the instrument, allowing maximum use of real estate on the table and instrument.
  • the separation device is optimized for detection performance, low cost/detection and ease of manufacture, and provides an ingenious and efficient way to collect analyte pellets.
  • FIG. 1A, 1B, 2A, and 2B various views illustrate a cross-section of one channel of the present separation device 100, the overall configuration of an exemplary multi-channel blood separation device, and flow characteristics of the sample and buffer.
  • Each channel has 2 side inlet ports 110 and a single outlet port 150 on the bottom serves each flow area 120 of each channel.
  • the top portion of each channel 125 is in communication with the atmosphere and functions as a sample container.
  • the sample container (hopper) is an integral part of the device (minimizing sample loss and simplifying liquid handling).
  • samples containing target analytes are injected/concentrated in 2 inert fluids by fluid dynamics (via positive pressure). Between the dotted arrows).
  • the device 100 shown in the embodiment is a linearly arranged channel.
  • the center distance can be chosen to match the center hole distance for the standard orifice plate.
  • the channel-to-channel center distance at the bottom is 0.18 inches.
  • an example specification may be that the thickness of the device is -0.3 inches or less, and the total length of the 24-channel device is -4.3 inches with a height of -2.4 inches per channel.
  • Ordinary orifice sizes include an array of holes having 8, 16, or 24 holes and other dimensions along one dimension. The number of channels and channel-to-channel distances can be selected to be compatible with these standards or other collection devices.
  • an exemplary workflow for magnetic separation is employed, starting with blood/other body fluids that arrive in the laboratory in standard and separate containers.
  • the sample is then passed to a manufacturing process to label the cell/analyte of interest with a target-specific antibody modified with ferromagnetic beads.
  • the different samples are then passed through the device 100 in parallel or in any desired order.
  • the sheath flow region 120 of the device such as the region between the device side buffer input and the bottom end outlet port
  • the external magnetic field generator 130 only attracts the target cells/analyte 140 bound to the magnetic beads.
  • These magnetic beads have been previously bound to specific antibodies that are complementary to specific antigens on the cells/analytes of interest. Due to the sheath flow effect described above, non-specific absorption of non-target cells/analytes is minimized, resulting in highly purified target cells/analyte 140. Buffer and sample fluid flow from bottom outlet 125 to each channel.
  • the flow rate is applied to the analyte run cycle, and the flow rate is 3-15 mL/h, driven by the difference between the two positive pressure side flows and the suction (-pressure flow) at the outlet port, which is the flow rate of the analyte.
  • a series of rinses are performed at a higher flow rate.
  • the pressure is also small. About 0-600Pa. After the instrument is automatically rinsed, the small pellets of the selected cells/analyte 140 remain in the device 100.
  • All of the composite/complex components of device 100 are located on one side/half side of device 170, which is injection molded. These include ribs on the back side of the flow area of the device 100 as energy guiding means for good soldering and precise guidance of the device 100 to external devices such as microfluidic systems, orifice plates or analytical instruments.
  • a more flexible plastic material 180 can be injected through the mold to form a gasket around the outlet port on the base of the card or also near other ports.
  • an important element is to avoid chemical contamination. The sample should not be contaminated by debris/contaminants from the outside, and the instrument and its components should not be contaminated by the analyte.
  • the biological sample is only in contact with the gasket, so the gasket is part of the device and can be discarded after use.
  • the gasket 180 on the outlet of the device 100 can also be seamless (cells are not captured/leave) and integrated into the device 100.
  • the final step is to ultrasonically weld the plastic flat cover sheet 160 to the structural side of the device 170.
  • the material used in some embodiments of the device may be polystyrene, which is the same material used for many multiwell plates.
  • Polycarbonate can also be used because polystyrene and polycarbonate have the same shrinkage in injection molding.
  • Suitable gasket materials are KRATON and 32A SHORE, but there are many more options for suitable flexible materials.
  • the process is called overmolding/overmolding, in which PS is injected into the mold, cured, and then a second injection of a different material. It achieves a seamless connection while providing high precision/tolerance for board connections.
  • the cover plate thickness is 0.02", ie the distance between the sides of the magnet and the captured analyte cells, the sample hopper volume is -300 uL per channel, and the side ports are rectangular and connected to the fluid via a distribution manifold.
  • each treated channel will have a pellet of target analyte remaining in the flow region that needs to be unloaded.
  • unloading of the treated sample can include fastening to a standard multiwell plate via an adapter.
  • device 100 can correspond to and match a row of orifice plates 200. Every other row on the board will be occupied by the box/adapter.
  • 8 cards can be assembled to a 384-well plate (16 x 24). There are 192 samples per plate/centrifuge.
  • the process can be highly multiplexed and allows for rapid and highly parallel recovery of the cells/analytes of interest from the microfluidic device into a standard well plate.
  • the device 100 can be bundled into a carrier that is designed to separate the linear devices from each other with a center-to-center hole distance of the orifice plate 200.
  • many of the containers of the orifice plates can be immediately filled from a suitable bundle of treated devices 100.
  • a portion of the orifice container can vary, with some of the rows being used as connection areas, up to all orifice containers. However, any number of devices can be used, from one to as many as the entire orifice plate.
  • the multi-beam rotary elution fixture 300 can simultaneously elute the treated analyte into a plurality of intact well plates. Although each channel has a slow flow rate, each card can process 8/16/24 samples in less than 5 minutes. Different fluid manifolds can be used. For example, a manifold for an 8-channel device can divide the line from 1 to 8. This is because the analyte is concentrated to ⁇ 300 uL (hopper volume/channel).
  • Figure 6 shows details of the output port of device 100 that is matched to the orifice plate 280 container.
  • the internal volume/channel of ⁇ 30uL needs to meet the requirements of matching the orifice plate, and no hole overflow occurs.
  • the lip of each of the overmolded washers 180 protrudes into the inner wall of the bore and the lip of the orifice is pressed into the washer.
  • a small passage in the gasket allows air to be trapped in the hole. As the air escapes the orifice during the rotational elution of the device (e.g., with the rotary elution fixture of Figure 5), centrifugal force pushes the agglomerates of the target portion into the pores.
  • conditional language used herein (such as including “can”, “may”, “may” and “such as”, etc.) is generally intended to mean Some embodiments include certain features, elements and/or states, while other embodiments do not include these. Therefore, these conditional languages are generally not intended to imply that the features, elements, and/or states may be applied to one or more embodiments in any desired manner, or one or more embodiments necessarily include logic for the decision, whether or not There are author input or prompts, whether or not these features, elements and/or states are included, or will be performed in any particular implementation.
  • an antisense conjunction such as the phrase "at least one of X, Y, or Z" is generally understood in the context as an item, term, etc., and may be X, Y, or Z, or any combination thereof ( For example, X, Y, and/or Z).
  • antisense conjunctions are generally not intended and should not imply that certain embodiments require at least one of X, at least one of Y, or at least one of Z to exist.
  • ⁇ or “approximately” and the like are synonymous and are used to indicate that the value of the term modification has a range of understanding associated therewith, wherein the range can be ⁇ 20%, ⁇ 15%, ⁇ 10%, ⁇ 5% or ⁇ 1%.
  • the term “substantially” is used to mean that a result (eg, a measured value) is close to a target value, where proximity may mean, for example, that the result is within 80% of the value, within 90% of the value, at 95 of the value Within % or within 99% of this value.
  • a phrase such as "a device configured as” is intended to include one or more of the devices. Such one or more of the described devices may also be configured together to perform the description.
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Abstract

一种用于大规模临床试验的离析各种分析物的高度并行的微流体血液分离装置，可以用于离析多种不同的靶分析物，靶分析物可以是包括NGS、PCR、FISH和IHC等等的各种诊断方法的起始材料。分离由磁鞘流原理实现并且是高度并行的。分离装置实现有效的磁分离，在垂直几何形状的样品流中起作用，并且可以紧固到标准多孔板，以实现高度多路复用的样品回收。在垂直方向上进行操作，允许多路复用，允许仪器上更多的卡/槽/间格，允许在台面和仪器方面有效使用不动产。

Description

高度并行的微流体血液分离装置
相关申请的交叉引用
本申请要求于2018年03月15日提交美国专利商标局的申请号为15/922,322、名称为“HIGHLY PARALLEL MICROFLUIDIC BLOOD SEPARATION DEVICE(高度并行的微流体血液分离装置)”的美国专利申请的优先权，其全部内容通过引用结合在本申请中。
背景技术
该公开涉及一种血液分离装置，并且特别是一种以高度并行配置使用鞘流技术的装置。
鞘流血液分离技术已被证明在用于各种应用的临床分析中有效。这样的技术可以通过分离操作和分离的分析物收集两者的通量改进受益。
发明内容
一种用于大规模临床试验的离析各种分析物的高度并行的微流体血液分离装置。该装置可以用于离析多种不同的靶分析物，该靶分析物可以是包括NGS、PCR、FISH和IHC等的各种诊断方法的起始材料。分离经由磁鞘流原理而实现并且高度并行。分离装置实现有效的磁分离，在样品在垂直几何结构内流动期间起作用，并且可以紧固到标准多孔板，以进行高度多路复用的样品回收。在垂直方向进行操作，允许多路复用，允许仪器上更多的卡/槽/间格，允许在台面和仪器方面有效使用的不动产。
在第一方面，提供一种并行的微流体血液分离装置，包括至少两个并行的微流体通道，其包括流动区域，配置成沿基本上垂直方向引导流体流动；每个通道有至少一个样品容器(料斗)，与流动区域流体连通；至少一个缓冲液流体输入端口，与流动区域流体连通；以及至少一个流体输出端口；其中该装置可以配置为在每个通道的基本上顶端处接收样品流体，在每个通道的侧面接收缓冲液流体，其中在缓冲液输入端口与基本上位于每个通道底部处的输出端口之间施加压力，在流动区域至输出端口之间产生样品和缓冲液流体的鞘流，并且其中该装置配置为与分离装置一起使用以从装置中的样品流体中捕获感兴趣的分析物。
在第一方面的一种实施方式中，装置中可以存在至少线性布置的8个并行通道，并且其中出口端口的间隔与标准孔板容器分离规格兼容。在第一方面的另一实施方式中，装置中可以存在线性布置的16或24个并行通道中的至少一种，并且其中出口端口的间隔与标准孔板容器分离规格兼容。在第一方面的一种实施方式中，分离装置可以是磁性的，配置为将磁性标记的分析物吸引到每个通道的侧壁，并且在流动操作完成之后，将分析物保持在合适的位置以保留在装置中。
在第一方面的另一实施方式中，装置可以由至少两个塑料部件制成，一个部件注塑成 型有料斗、输入端口、输出端口和流动区域结构，并且另一部件是盖件，与注塑成型部件连接以获得完成装置。在第一方面的一种实施方式中，至少一个柔性垫圈材料注塑成型到至少一个端口。在第一方面的另一实施方式中，柔性垫圈可以在输出端口上，该输出端口基本上设置在每个通道的底部上，并且垫圈配置为密封到孔板容器。
在第一方面的另一实施方式中，装有分离的分析物的多个分离装置可以在保持固定件中组套在一起并密封到多排孔板容器。在第一方面的一种实施方式中，旋转固定件可以配置为一起旋转被密封到孔板的分离装置的多个组套，以进行高度并行的样品提取。
在第一方面的一种实施方式中，可以存在两个侧缓冲液流体输入，位于每个通道上的流动区域的相对侧上，配置为在两侧上包覆样品流。在第一方面的另一实施方式中，装置可以配置为在流动分离完成后将每个通道的输出端口接合到收集装置，其中可以旋转分离装置和收集装置以将分析物送入收集装置中。在第一方面的一种实施方式中，收集装置可以是孔板，并且分离装置输出端口将每个通道密封到成排的孔板容器。
在第二方面，可以提供一种使用分离装置进行高度并行的微流体血液分离的方法，该分离装置包括至少两个并行的微流体通道，其包括流动区域，被配置为沿基本上垂直的方向引导流体流；该方法包括以下步骤：填充与流动区域流体连通的每个通道的样品料斗；向与流动区域流体连通的每个通道的输入端口添加缓冲液流体；以及，从输出端口提取缓冲液和样品流体；其中可以在每个通道的基本上顶端处接收样品流体，在每个通道的侧面接收缓冲液流体，其中在缓冲液输入端口与基本上位于每个通道的底部处的输出端口之间施加压力，在流动区域至输出端口中产生样品和缓冲液流体的鞘流，并且其中分离装置用于从装置中的样品流体中捕获感兴趣的分析物。
在第二方面的一种实施方式中，装置中可以存在线性布置的至少8个并行通道，并且其中出口端口的间隔与标准孔板容器分离规格兼容。在第二方面的另一实施方式中，装置中可以存在线性布置的16或24个并行通道中的至少一种，并且其中出口端口的间隔与标准孔板容器分离规格兼容。
在第二方面的一种实施方式中，分离装置可以是磁性的，该分离装置将磁性标记的分析物吸引到每个通道的侧壁，并且在流动操作完成之后，将分析物保持在合适的位置以保留在装置中。在第二方面的另一实施方式中，可以存在两个侧缓冲液流体输入，位于每个通道流动区域的相对侧上，并且缓冲液流体在两侧上包覆样品流。
在第二方面的另一实施方式中，该方法还可以包括以下步骤，在流动分离完成后，将每个通道的输出端口接合到收集装置，其中旋转该分离装置和收集装置以将分析物送入收集装置中。在第二方面的另一实施方式中，收集装置可以是孔板，并且分离装置输出端口将每个通道密封到成排的孔板容器。在第二方面的一种实施方式中，带有分离的分析物的 多个分离装置可以在保持固定件中组套在一起并密封到多排孔板容器。
附图说明
参考与附图结合的以下详细说明来描述本文提供的实施方式的方面和优点。在整个附图中，附图标记可以重新使用以指示所引用元件之间的对应关系。提供附图是为了说明本文描述的实例实施方式，并不旨在限制本公开的范围。
图1A和图1B示出了垂直的高度并行的血液分离装置的一个通道的示例性实施方式；
图2A和图2B示出了垂直的高度并行的血液分离装置的示例性实施方式；
图3示出了垂直的高度并行的血液分离装置的示例性实施方式的构造细节；
图4示出了垂直的高度并行的血液分离装置的示例性实施方式与标准孔板之间的兼容性；
图5示出了用于保持和旋转大量垂直的高度并行的血液分离装置的装置的示例性实施方式，该装置可用于分离的分析物的卸载；
图6描绘了垂直的高度并行的血液分离装置和用于样品分析物卸载的孔板之间的密封。
具体实施方式
本公开涉及一种高度并行的血液分离装置，其配置为垂直流动操作，包含多个并行流动通道并且与标准样品分析装置(诸如孔板)兼容。
本公开基于2012年9月11日公布的题为“Sheath Flow Devices and Methods”的美国专利8,263,287中描述的鞘流血液分离装置。专利8,263,287通过引用其全部内容而被并入。专利8,263,287描述了由一个或多个缓冲液流体流包覆的样品流体流动的技术。感兴趣的分析物(诸如血液中的癌细胞)以一些方式被标记，诸如与磁性颗粒结合。当暴露于分离机制(诸如磁场)时，感兴趣的分析物从样品流分离到缓冲液流之一中，并在鞘流装置内被捕获。一旦流动完成，分析物的团粒保留在装置中并且可以移除用于分析。一种移除技术是于2012年9月18日提交的题为“Spin Elute Tube”的美国临时申请61/702,730中描述的旋转洗脱技术，后来公布为国际公开WO2014/046942 A1。该申请通过引用而将其整体并入。
本申请基于所并入的参考文献的基础，但进一步公开了关键的有益特征，该关键的有益特征获得了显著的性能优势。
在一些实施方式中，可以以较高的样品密度提供较小的样品体积。有利地，这些特征允许快得多的检测。
在一些实施方式中，样品流体可以末端加载且末端耗尽。有利地，该布置不需要样品流动的重定向，降低了制造的复杂性和成本。
在一些实施方式中，分离的感兴趣分析物的收集与标准多样品分析(诸如孔板)兼容。有利地，该布置允许将样品直接输送到高通量分析装置中。
在诊断中，只有当检测的经济性有利并且存在处理大量临床样品的能力时才能实现商业规模化。与当前系统相比，本公开针对选定的应用组，并且满足人口规模的检测，具有改进的性能。
对于现有系统，随着检测材料/分析物的量的减少，样品损失(丢失的感兴趣的分析物/细胞)迅速变得更加成问题。本公开还引入了实现最小样品损失引入标准容器设计(如PCR管和多孔板等)的直接转移方法。
本公开提供了一种用于大规模临床试验的离析各种分析物的高度并行的微流体血液分离装置。该装置可以用于离析多种不同的靶分析物，诸如cfDNA、CTCs、RNA和外泌体，这些靶分析物可以是用于各种诊断方法(包括NGS、PCR、FISH、IHC和其他方法)的起始材料。分离通过经由所并入的参考文献的磁鞘流原理，但增强为高度并行来实现。本公开的装置提供了增强的磁分离，在样品在垂直几何结构内流动期间起作用，并且紧固到标准多孔板，以进行高度多路复用的样品回收。系统的垂直方向消除了对于末端缓冲液的需求，从而降低了成本并提高了自动兼容性。在垂直方向进行操作允许多路复用，允许仪器上更多的卡/槽/间格，允许在台面和仪器方面最大化使用不动产。本分离装置对检测性能、低成本/检测和易制造性进行了优化，并且提供了收集分析物团粒的巧妙且有效的方式。
参考图1A、图1B、图2A和图2B，多个视图示出了本分离装置100的一个通道的截面，示例性多通道血液分离装置的整体配置以及样品和缓冲液的流动特征。每个通道有2个侧入口端口110，并且底部上的单个出口端口150服务于每个通道的每个流动区域120。每个通道125的顶部部分与大气连通并且起到样品容器的作用。样品容器(料斗)是装置的整体部分(最小化样品损失，简化液体处理)。类似于过去十年中已有重大创新/发展的FACS技术，含有靶分析物的样品(实线箭头)通过侧入口注入(经由正压力)，被流体动力学夹带/集中在2股惰性流体(虚线箭头)之间。
如图2A和图2B所示，实施方式中所示的装置100为线性布置的通道。可以选择中心距离以匹配用于标准孔板的中心孔距离。例如，对于480孔板，底部处的通道到通道中心距离为0.18英寸。对于480孔板的兼容实施方式，示例规格可以是装置的厚度为～0.3英寸或更小，并且24通道装置的总长度是～4.3英寸，其中每个通道的高度为～2.4英寸。普通孔板尺寸沿一个维度包括具有8、16或24个孔以及其他尺寸的孔阵列。可以选择通道的数量以及通道到通道距离，以与这些标准或其他收集装置兼容。
对于示例性装置实施方式，采用了磁分离的示例工作流程，以处于标准和分别的容器中到达实验室的血液/其他体液为起始。然后样品进入制备工艺，对感兴趣的细胞/分析物用铁磁性珠修饰的靶特异性抗体标记。随后，不同样品并行或者是以任何期望的顺序相继地通过装置100。随着样品通过装置的鞘流动区域120，如在装置侧面缓冲液输入与底部末端 出口端口之间的区域，外部磁场发生器130仅吸引结合到磁珠的靶细胞/分析物140。这些磁珠已预先结合到与感兴趣的细胞/分析物上的特异性抗原互补的特异性抗体。由于上述的鞘流效应，非靶细胞/分析物的非特异性吸收被最小化，可得到高度纯化的靶细胞/分析物140。缓冲液和样品流体从底部出口125流出每个通道。
流速施压于分析物运行循环，在出口端口两个正压力侧流和吸力(-压力流)差的驱动下，流速为3-15mL/h，是分析物的流速。一旦样品通过，在较高的流速下进行一系列冲洗。压力也很小。大约0-600Pa。对仪器自动冲洗后，选定的细胞/分析物140的小团粒保留在装置100中。
参照图3，将对本装置的示例性实施方式的构造细节进行讨论。采用了注塑/注塑-压缩成型、热压花和铸造技术，这些技术既可规模化又经济。
装置100的所有复合/复杂的部件诸如料斗、入口和出口端口以及流动区域，均位于装置170的一侧/半侧，该装置是注塑成型的。其中包括位于装置100流动区域背侧上的肋，作为能量引导装置，利于良好焊接以及将装置100精确引导到外部装置(诸如微流体系统、孔板或分析仪器)。可以通过模具注入更柔性的塑料材料180，在卡的基部上围绕出口端口或者也在其他端口附近形成垫圈。关于装置100的布置，一个重要要素是要避免化学污染。样品不应被来自外部的碎屑/污染物所污染，并且仪器和其组件不应被分析物污染。生物样品仅与垫圈接触，因此垫圈是装置的一部分，其在使用后可以丢弃。装置100出口上的垫圈180也可以是无缝的(细胞不会被捕获/留下)并且集成到装置100。由更柔性的材料包覆成型，其可以提供与仪器仪表和多孔板的气密密封，该仪器仪表在操作中用于样品输送和分离，该多孔板用于处理之后的样品回收。最后的步骤是将塑料平盖片160超声波焊接到装置170的结构侧面。
用于装置的一些实施方式的材料可以是聚苯乙烯，其与用于许多多孔板的材料相同。也可以使用聚碳酸酯，因为聚苯乙烯和聚碳酸酯在注塑成型中具有相同的收缩率。合适的垫圈材料是KRATON和32A SHORE，但有许多适合的柔性材料的更多选择。工艺称为二次成型/包覆成型，其中PS注入模具中，固化，然后第二次注入不同材料。它实现了无缝连接，同时为板的连接提供了高精度/容限。在一个实施方式中，盖板厚度为0.02”，即磁体侧面与捕获的分析物细胞之间的距离，样品料斗容积为～300uL/通道，并且侧面端口为矩形并经由分配集合管连接至流体。
一旦装置通道经过处理用于分离后，每个经处理的通道将具有留在流动区域中的靶分析物的团粒，需要进行卸载。在一些实施方式中，经处理样品的卸载可以包括经由适配器紧固到标准多孔板。如图4所示，装置100可以对应于孔板200的一排并与其匹配。板上每隔一排都将被盒/适配器占用。例如，对于所示的24通道装置，可以组装8个卡到384 孔板(16×24)。每板/离心桶有192个样品。
在一个实施方式中，该工艺可以高度多路复用，并且允许从微流体装置快速且高度并行地回收感兴趣的细胞/分析物到标准孔板中。如图5所示。装置100可以集束到承载件中，承载件设计成以孔板200的中心对中心孔距离，将线性装置彼此隔开。如图所示，这些孔板的容器中的许多个可以立即从经处理的装置100的合适的束填充。孔板容器的一部分可以变化，其中一些排可以用作连接区域，最多包括所有孔板容器。不过，可以使用任何数量的装置，从一个到多达覆盖整个孔板的数量。还如图5中所示出的多束旋转洗脱固定装置300，可以将经处理的分析物同时洗脱到多个完整孔板中。尽管每个通道流速缓慢，但每个卡可在5分钟之内处理8/16/24个样品。可以使用不同的流体集合管。例如，用于8通道装置的集合管可以将线路从1分为8。这是因为分析物被浓缩至～300uL(料斗体积/通道)。
图6示出了匹配到孔板280容器的装置100输出端口的细节。～30uL的内部体积/通道需满足与孔板匹配的要求，不出现孔溢流。各个二次注塑的垫圈180的唇部突出到孔的内壁中，并且孔板唇部压入垫圈。垫圈中的小通道允许空气被捕获在孔中。随着空气在(例如具有图5的旋转洗脱固定装置)装置旋转洗脱期间逸出孔外，离心力将靶部分的团粒推入孔中。
本文描述的实施方式是示例性的。可以对这些实施方式进行修改、重新排序、替换材料和替换元件等，并且它们仍然包含在本文阐述的教导中。
除非另有明确说明或者使用时在上下文中以其他方式理解，否则本文使用的条件性语言(诸如，包括“能够”、“可以”、“可能”和“例如”等在内)通常旨在表达某些实施方式包括某些特征、元件和/或状态，而其他实施方式不包括这些。因此，这些条件性语言通常不旨在意味着特征、元件和/或状态可以任何要求的方式适用于一个或多个实施方式，或者一个或多个实施方式必然包括用于决定的逻辑，不论是否有作者输入或提示，不论是否包括这些特征、元件和/或状态，或将以任何特定实施方式执行。术语“包括”、“包含”、“具有”和“涉及”等等是同义词并且以开放式方式包含地使用，并且不排除其他元件、特征、动作和操作等等。此外，术语“或”以其包含性意义(而不是以其独有性意义)使用，因此当使用时，例如，为了连接一列元件，术语“或”意味着该列中元件中的一个、一些或所有。
除非另有明确说明，否则诸如短语“X、Y或Z中的至少一个”之类的反义连接词在上下文通常可理解为项目和术语等，可以是X、Y或Z或其任何组合(例如，X、Y和/或Z)。因此，这种反义连接词通常不旨在并且不应该暗示某些实施方式需要X中的至少一个、Y中的至少一个或Z中的至少一个至每个都存在。
术语“约”或“近似”等等是同义词，并且用于表示该术语修饰的值具有与其关联的 理解范围，其中该范围可以是±20％、±15％、±10％、±5％或±1％。术语“基本上”用于表示结果(例如，测量值)接近于目标值，其中接近可以意味着，例如，结果在该值的80％内、在该值的90％内、在该值的95％内或在该值的99％内。
除非另有明确说明，否则冠词诸如“a”或“an”通常应被解释为包括一个或多个所描述的项目。因此，短语诸如“配置为...的装置”旨在包括一个或多个所述装置。这样的一个或多个所述装置也可以共同配置为进行所述的叙述。例如，“配置为进行叙述A、B和C的处理器”可以包括配置为进行叙述A的第一处理器，结合配置为进行叙述B和C的第二处理器一起工作。
虽然上面的详细描述已经示出、描述并指出了应用于说明性实施方式的新颖特征，但是应该理解，可以在不脱离本公开主旨的情况下，对所示装置和组件的形式和细节进行各种省略、替换和改变。如将认识到的，本文描述的某些实施方式可以体现在不提供本文所阐述的所有特征和益处的形式内，因为一些特征可以与其他特征分开使用或操作。在权利要求的等同的范围和含义内的所有变化都包含在其范围内。

Claims (20)

  1. 一种并行的微流体血液分离装置，包括：
    至少两个并行的微流体通道，包括流动区域，所述流动区域配置为沿基本上垂直的方向引导流体流动；
    每个通道中至少一个样品料斗，与所述流动区域流体连通；
    至少一个缓冲液流体输入端口，与所述流动区域流体连通；以及，
    至少一个流体输出端口；其中
    所述装置配置为在每个通道的基本上顶端处接收样品流体，在每个通道的侧面接收缓冲液流体，在所述缓冲液输入端口与基本上位于每个通道底部处的输出端口之间施加压力，在所述流动区域至所述输出端口之间产生所述样品和缓冲液流体的鞘流，并且其中所述装置配置为与分离装置一起使用以从所述装置中的所述样品流体捕获感兴趣的分析物。
  2. 根据权利要求1所述的装置，其中，所述装置中存在线性布置的至少8个并行通道，并且其中所述出口端口的间隔与标准孔板容器分离规格兼容。
  3. 根据权利要求2所述的装置，其中，所述装置中存在线性布置的16或24个并行通道中的至少一种，并且其中所述出口端口的间隔与标准孔板容器分离规格兼容。
  4. 根据权利要求2所述的装置，其中，所述分离装置是磁性的，所述分离装置配置为将磁性标记的分析物吸引到每个通道的侧壁，并且在流动操作完成之后，将所述分析物保持在合适的位置以保留在所述装置中。
  5. 根据权利要求1所述的装置，其中，所述装置由至少两个塑料部件制成，一个部件注塑成型有料斗、输入端口、输出端口和流动区域结构，并且另一部件是盖件，与注塑成型部件连接以获得完成装置。
  6. 根据权利要求5所述的装置，其中，至少一个柔性垫圈的材料注塑成型到至少一个端口。
  7. 根据权利要求5所述的装置，其中，所述柔性垫圈在所述输出端口上，所述输出端口基本上设置在每个通道的底部，并且所述垫圈配置为密封到孔板容器。
  8. 根据权利要求1所述的装置，其中，存在两个侧缓冲液流体输入，位于每个通道上的所述流动区域的相对侧上，配置为在两侧上包覆所述样品流。
  9. 根据权利要求1所述的装置，配置为在流动分离完成后将每个通道的输出端口接合到收集装置，其中旋转所述分离装置和所述收集装置以将所述分析物送入所述收集装置中。
  10. 根据权利要求9所述的装置，其中，所述收集装置是孔板，并且所述分离装置输出端口将每个通道密封到成排的孔板容器。
  11. 根据权利要求10所述的装置，其中，装有分离的分析物的多个分离装置在保持固定件中组套在一起并密封到多排孔板容器。
  12. 根据权利要求11所述的装置，其中，旋转固定件配置为旋转被密封到孔板的分离装置的多个组套，以进行高度并行的样品提取。
  13. 一种使用分离装置进行高度并行的微流体血液分离的方法，所述分离装置包括至少两个并行的微流体通道，包括流动区域，所述流动区域配置为沿基本上垂直方向引导流体流动，所述方法包括：
    填充与所述流动区域流体连通的每个通道的样品料斗；
    向与所述流动区域流体连通的每个通道的输入端口添加缓冲液流体；以及，
    从输出端口提取缓冲液和样品流体；其中
    样品流体在每个通道的基本上顶端处接收，缓冲液流体在每个通道的侧面接收，在缓冲液输入端口与基本上位于每个通道底部处的输出端口之间施加压力，在所述流动区域至所述输出端口之间产生所述样品和缓冲液流体的鞘流，并且其中分离装置用于从所述装置中的所述样品流体中捕获感兴趣的分析物。
  14. 根据权利要求13所述的方法，其中，所述装置中存在线性布置的至少8个并行通道，并且其中所述出口端口的间隔与标准孔板容器分离规格兼容。
  15. 根据权利要求14所述的方法，其中，所述装置中存在线性布置的16或24个并行通道中的至少一种，并且其中所述出口端口的间隔与标准孔板容器分离规格兼容。
  16. 根据权利要求13所述的方法，其中，所述分离装置是磁性的，所述分离装置将磁性标记的分析物吸引到每个通道的侧壁，并且在流动操作完成之后，将所述分析物保持在合适的位置以保留在所述装置中。
  17. 根据权利要求13所述的方法，其中，存在两侧缓冲液流体输入，位于每个通道上的所述流动区域的相对侧上，并且所述缓冲液流体在两侧上包覆所述样品流。
  18. 根据权利要求13所述的方法，还包括在流动分离完成后将每个通道的输出端口接合到收集装置，其中旋转所述分离装置和所述收集装置以将所述分析物送入所述收集装置中。
  19. 根据权利要求13所述的方法，其中，所述收集装置是孔板，并且所述分离装置输出端口将每个通道密封到成排的孔板容器。
  20. 根据权利要求19所述的方法，其中，装有分离的分析物的多个分离装置在保持固定件中组套在一起并密封到多排孔板容器。
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