WO2006105110A2 - Dispositifs de dosage et procedes - Google Patents

Dispositifs de dosage et procedes Download PDF

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Publication number
WO2006105110A2
WO2006105110A2 PCT/US2006/011322 US2006011322W WO2006105110A2 WO 2006105110 A2 WO2006105110 A2 WO 2006105110A2 US 2006011322 W US2006011322 W US 2006011322W WO 2006105110 A2 WO2006105110 A2 WO 2006105110A2
Authority
WO
WIPO (PCT)
Prior art keywords
detection reagent
analyte
reagent
particle
region
Prior art date
Application number
PCT/US2006/011322
Other languages
English (en)
Other versions
WO2006105110A3 (fr
Inventor
Victor Manneh
Simon Burnell
Khaled A. Yamout
Original Assignee
Inverness Medical Switzerland Gmbh
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Inverness Medical Switzerland Gmbh filed Critical Inverness Medical Switzerland Gmbh
Publication of WO2006105110A2 publication Critical patent/WO2006105110A2/fr
Publication of WO2006105110A3 publication Critical patent/WO2006105110A3/fr

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Classifications

    • 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
    • G01N33/80—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
    • 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
    • B01L3/502746—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 for controlling flow resistance, e.g. flow controllers, baffles
    • 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
    • B01L3/502753—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 bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation
    • 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
    • B01L2200/0621—Control of the sequence of chambers filled or emptied
    • 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
    • B01L2200/16—Reagents, handling or storing thereof
    • 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
    • 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
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0681—Filter
    • 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
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0825—Test strips
    • 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
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/087—Multiple sequential 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
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0406—Moving fluids with specific forces or mechanical means specific forces capillary forces
    • 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
    • B01L2400/0688—Valves, specific forms thereof surface tension valves, capillary stop, capillary break
    • 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
    • B01L2400/0694—Valves, specific forms thereof vents used to stop and induce flow, backpressure valves
    • 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
    • B01L2400/08—Regulating or influencing the flow resistance
    • B01L2400/084—Passive control of flow resistance
    • B01L2400/086—Passive control of flow resistance using baffles or other fixed flow obstructions

Definitions

  • This invention relates to devices, methods and systems for performing an assay.
  • An assay device can be designed to rapidly and accurately perform an assay using a small volume of sample.
  • the device can be operated with minimal training.
  • a device with a microfluidic flow region can produce accurate measurements of an analyte from a small volume of sample.
  • Other assays e.g., for blood sugar testing by diabetics have been adapted for use at home by a patient.
  • a device can be configured for rapid detection of an analyte utilizing a small volume of biological samples and a microfluidic transport structure.
  • the device can include a microfabricated cell separation device, a solid phase pre-treatment chamber to remove interferences, and a sensitive immunoassay that takes place in an efficient light capturing detection chamber. Incubation of the sample with assay components and timely movements of fluid is achieved through a mechanically time-gated structure.
  • a biological sample such as blood travels uni-directionally and passively through the various sections of the device driven by capillary force.
  • Cells or other particulates
  • the remaining plasma is treated to remove all biological interferences and transported to a resuspension chamber containing the reagents needed to perform the assay.
  • the assay could be configured to perform homogeneous, quasi-homogeneous or, heterogeneous immunoassays.
  • Reagents for all assay platforms are configured to produce a detectable change upon interaction with the analyte.
  • the assay can be qualitative or quantitative and can measure an analyte at picomolar concentrations in a single drop of whole blood.
  • the device and assay can be particularly useful for in-home or point-of-care diagnostic tests because of its small size, rapid operation, ease of use, accurate results, no specialized equipment or user training requirement and can operate with or without temperature control.
  • the detection reagent can be lyophilized or dried in a well configured with optimized geometry to ensure efficient reconstitution, assay kinetics and optical design to insure highly efficient optical capacity.
  • a device for detecting an analyte includes a sample inlet including a filter, a reconstitution region including a detection reagent capable of producing a detectable change upon interaction with the analyte, a time gate configured to restrain fluid flow for a predetermined period of time, and an analysis region configured to reveal the detectable change.
  • the sample inlet is fluidly connected to a collection region, and the fluid connection includes a capillary.
  • a method of detecting an analyte comprising depositing a sample on a device including a sample inlet including a filter, a reconstitution region including a detection reagent capable of producing a detectable change upon interaction with the analyte, a time gate configured to restrain fluid flow for a predetermined period of time, and an analysis region configured to reveal the detectable change.
  • the sample inlet is fluidly connected to a collection region, and the fluid connection includes a capillary.
  • a method of manufacturing a device for detecting an analyte includes forming a base including a sample inlet including a filter, a reconstitution region including a detection reagent capable of producing a detectable change upon interaction with the analyte, a time gate configured to restrain fluid flow for a predetermined period of time, and an analysis region configured to reveal the detectable change.
  • the sample inlet is fluidly connected to a collection region, and the fluid connection includes a capillary.
  • the device, system and methods can include one or more of the following variations.
  • the method can include depositing a detection reagent configured to produce a detectable change upon interaction with the analyte in the reconstitution region.
  • the method can include comprising forming a lid configured to cover the base.
  • the lid can be sealed the lid to the base.
  • the seal can be an airtight seal.
  • a system for detecting an analyte includes a cartridge and a cartridge reader.
  • the cartridge includes a sample inlet including a filter, a reconstitution region including a detection reagent capable of producing a detectable change upon interaction with the analyte, a time gate configured to restrain fluid flow for a predetermined period of time, and an analysis region configured to reveal the detectable change.
  • the sample inlet is fluidly connected to a collection region, and the fluid connection includes a capillary.
  • the cartridge reader includes a slot configured to accept the cartridge, an optical system configured to measure the detectable change, an output display configured to communicate a result of the measurement to a user.
  • the time gate can substantially completely restrain fluid flow for the predetermined period of time.
  • the time gate can include a vent configured to be opened after a predetermined period of time.
  • the time gate can include a side channel.
  • the time gate can be configured to restrain fluid flow in the reconstitution region.
  • the filter can include a membrane.
  • the membrane can contact a plurality of structures having a capillary dimension.
  • the filter can include a reagent for agglomerating red blood cells.
  • the reagent for agglomerating red blood cells can include a lectin, an antibody, or a carbohydrate.
  • the filter can be configured to alter the rate of fluid flow.
  • the detection reagent can include a first particle configured to release a diffusible excited species upon excitation.
  • the detection reagent can further include a second particle configured to emit light of a predetermined wavelength when contacted with the diffusible excited species.
  • the detection reagent can be configured to bring the first particle and second particle in close proximity when contacted with the analyte.
  • the detection reagent can include an antibody.
  • the detection reagent can include a metal nanoparticle, a fluorescent compound, or a colored compound.
  • a method for detecting an analyte includes contacting a sample with a device including a sample inlet including a filter, a transfer region fluidly connected to the collection region, and an analysis region including a detection reagent configured to produce a detectable change upon interaction with the analyte, and measuring the detectable change.
  • the sample inlet is fluidly connected to a collection region, and the fluid connection includes a capillary.
  • a device for detecting an analyte in another aspect, includes a sample inlet including a filter, a transfer region fluidly connected to the collection region, an analysis region including a detection reagent configured to produce a detectable change upon interaction with the analyte, and a wash path configured to deliver a wash reagent to the analysis region.
  • the sample inlet is fluidly connected to a collection region, and the fluid connection includes a capillary.
  • the transfer region includes a plurality of capillary structures.
  • a method of detecting an analyte includes depositing a sample on a device including a sample inlet including a filter, a transfer region fluidly connected to the collection region, an analysis region including a detection reagent configured to produce a detectable change upon interaction with the analyte, and a wash path configured to deliver a wash reagent to the analysis region.
  • the sample inlet is fluidly connected to a collection region, and the fluid connection includes a capillary.
  • the transfer region includes a plurality of capillary structures.
  • a method of manufacturing a device for detecting an analyte includes forming a base including a sample inlet including a filter, a transfer region fluidly connected to the collection region, an analysis region including a detection reagent configured to produce a detectable change upon interaction with the analyte, and a wash path configured to deliver a wash reagent to the analysis region.
  • the sample inlet is fluidly connected to a collection region, and the fluid connection includes a capillary.
  • the transfer region includes a plurality of capillary structures.
  • a system for detecting an analyte includes a cartridge and a cartridge reader.
  • the cartridge includes a sample inlet including a filter, a transfer region fluidly connected to the collection region, an analysis region including a detection reagent configured to produce a detectable change upon interaction with the analyte, and a wash path configured to deliver a wash reagent to the analysis region.
  • the sample inlet is fluidly connected to a collection region, and the fluid connection includes a capillary.
  • the transfer region includes a plurality of capillary structures.
  • the cartridge reader includes a slot configured to accept the cartridge, an optical system configured to measure the detectable change, and an output display configured to communicate a result of the measurement to a user.
  • the cartridge reader can include a wash reagent reservoir arranged to be fluidly connected to the wash path when the cartridge is accepted by the slot.
  • the reservoir can include the wash reagent.
  • the detection reagent can include a metal nanoparticle, a fluorescent compound, a colored compound, or a first particle configured to release a diffusible excited species upon excitation.
  • the device, system and methods can include one or more of the following variations.
  • the device can include a wash reagent reservoir fluidly connected to the wash path.
  • the reservoir includes the wash reagent.
  • the device can include a time gate configured to delay fluid flow for a predetermined period of time.
  • the filter can include a membrane.
  • the membrane can contact a plurality of structures having a capillary dimension.
  • the filter can include a reagent for agglomerating red blood cells.
  • the reagent for agglomerating red blood cells can include a lectin, an antibody, or a carbohydrate.
  • the device can include a hydrophilic layer on a surface of the device, the hydrophilic layer having a contact angle of between 5 and 20 degrees.
  • the filter can be configured to alter the rate of fluid flow.
  • the detection reagent can include a first particle configured to release a diffusible excited species upon excitation.
  • the detection reagent can further include a second particle configured to emit light of a predetermined wavelength when contacted with the diffusible excited species.
  • the detection reagent can be configured to bring the first particle and second particle in close proximity when contacted with the analyte.
  • the device further can include a time gate configured to delay fluid flow for a predetermined period of time.
  • the detection reagent can include an antibody.
  • the device can include a wash path configured to deliver a wash reagent to the analysis region.
  • the device can include a wash reagent reservoir fluidly connected to the wash path, the reservoir including the wash reagent.
  • a method of manufacturing a device for detecting an analyte includes forming a base including a sample inlet including a filter and fluidly connected to a collection region, a transfer region fluidly connected to the collection region, and an analysis region configured to accept a detection reagent configured to produce a detectable change upon interaction with the analyte.
  • the fluid connection includes a capillary
  • the transfer region includes a plurality of capillary structures.
  • the method can include depositing a detection reagent configured to produce a detectable change upon interaction with the analyte in the analysis region.
  • the detection reagent can include a metal nanoparticle, a fluorescent compound, a colored compound, or a first particle configured to release a diffusible excited species upon excitation.
  • the method of can include forming a lid configured to cover the base.
  • the lid can be sealed to the base.
  • the seal can be an airtight seal.
  • the method can include applying a hydrophilic layer on a surface of the base, the hydrophilic layer having a contact angle of between 5 and 20 degrees.
  • the device can include a wash reagent reservoir fluidly connected to the analysis region.
  • the device can include a wash reagent reservoir fluidly connected to the wash path.
  • the method can include adding a wash reagent to the wash reagent reservoir.
  • a device for detecting an analyte in another aspect, includes a sample inlet including a filter, a transfer region fluidly connected to the collection region, and an analysis region including a detection reagent configured to produce a detectable change upon interaction with the analyte.
  • the sample inlet is fluidly connected to a collection region, and the fluid connection includes a capillary.
  • the transfer region includes a plurality of capillary structures.
  • the filter is configured to restrain cells in a sample.
  • a method of detecting an analyte includes depositing a sample on a device including a sample inlet including a filter, a transfer region fluidly connected to the collection region, and an analysis region including a detection reagent configured to produce a detectable change upon interaction with the analyte.
  • the sample inlet is fluidly connected to a collection region, and the fluid connection includes a capillary.
  • the transfer region includes a plurality of capillary structures.
  • the filter is configured to restrain cells in a sample.
  • a method of manufacturing a device for detecting an analyte includes forming a base including a sample inlet including a filter, a transfer region fluidly connected to the collection region, and an analysis region including a detection reagent configured to produce a detectable change upon interaction with the analyte.
  • the sample inlet is fluidly connected to a collection region, and the fluid connection includes a capillary.
  • the transfer region includes a plurality of capillary structures.
  • the filter is configured to restrain cells in a sample.
  • the device, system and methods can include one or more of the following variations.
  • the method can include depositing a detection reagent configured to produce a detectable change upon interaction with the analyte in the analysis region.
  • the method can include forming a lid configured to cover the base.
  • the lid can be sealed to the base.
  • the seal can be an airtight seal.
  • the method can include applying a hydrophilic layer on a surface of the base, the hydrophilic layer having a contact angle of between 5 and 20 degrees.
  • a system for detecting an analyte includes a cartridge and a cartridge reader.
  • the cartridge includes a sample inlet including a filter, a transfer region fluidly connected to the collection region, and an analysis region including a detection reagent configured to produce a detectable change upon interaction with the analyte.
  • the sample inlet is fluidly connected to a collection region, and the fluid connection includes a capillary.
  • the transfer region includes a plurality of capillary structures.
  • the filter is configured to restrain cells in a sample.
  • the cartridge reader includes a slot configured to accept the cartridge, an optical system configured to measure the detectable change, and an output display configured to communicate a result of the measurement to a user.
  • the device can include a hydrophilic layer on a surface of the device, the hydrophilic layer having a contact angle of between 5 and 20 degrees.
  • the filter can include a membrane.
  • the membrane can contact a plurality of structures having a capillary dimension.
  • the filter can include a reagent for agglomerating red blood cells.
  • the reagent for agglomerating red blood cells can include a lectin, an antibody, or a carbohydrate.
  • the filter can be configured to alter the rate of fluid flow.
  • the detection reagent can include a first particle configured to release a diffusible excited species upon excitation.
  • the detection reagent can further include a second particle configured to emit light of a predetermined wavelength when contacted with the diffusible excited species.
  • the detection reagent can be configured to bring the first particle and second particle in close proximity when contacted with the analyte.
  • the detection reagent can include an antibody.
  • the detection reagent can include a metal nanoparticle, a fluorescent compound, or a colored compound.
  • An immunoassay system includes a cartridge and a cartridge reader. The cartridge includes assay reagents. The cartridge reader can be separate from the cartridge and measures the result of the assay and communicates the result to a user.
  • the immunoassay system can be used with any analyte that can be detected with an immunoassay.
  • the system can be used quickly and easily at home by a patient, by a health care professional (for example, in a doctor's office), or by a nurse who visits patients.
  • the system could also be used as a triage device by ambulance crews prior to delivering a patient to an emergency room.
  • the cartridge can adapted to assay for one or more of a variety of markers, such as, for example, a stroke marker, a cancer marker, a marker of ischemia, or a cardiac marker, depending on the immunoassay reagents included on the cartridge.
  • the system is designed so that a minimum of user actions is required.
  • the cartridge reader can provide a user interface, for example, on a full color dot matrix display. The user interface can be simple and straightforward for use at home by a patient.
  • a system for detecting an analyte includes a cartridge and a cartridge reader.
  • the cartridge includes a microstructured flow path which includes a resuspension zone and a detection zone. At least a portion of the flow path is sealed.
  • the cartridge also includes a selectively openable port configured to vent the sealed portion of the flow path when open.
  • the resuspension zone includes a detection reagent configured to produce a detectable change upon binding to the analyte.
  • the cartridge reader includes a tray adapted to hold the cartridge and an optical system configured to measure the detectable change.
  • a method of detecting an analyte includes applying a sample to a cartridge and measuring the extent of the detectable change with a cartridge reader.
  • the cartridge includes a microstructured flow path which includes a resuspension zone and a detection zone. At least a portion of the flow path is sealed.
  • the cartridge also includes a selectively openable port configured to vent the sealed portion of the flow path when opened.
  • the resuspension zone includes a detection reagent configured to produce a detectable change upon binding to the analyte.
  • the cartridge reader includes a tray adapted to hold the cartridge, and an optical system configured to measure the detectable change.
  • the device, system and methods can include one or more of the following variations.
  • the selectively openable port can include a selectively meltable covering.
  • the cartridge reader can be configured to open the selectively openable port at a predetermined time.
  • the cartridge reader can include a display.
  • the display can be configured to communicate a result to a user.
  • the cartridge further can include an orientation feature and the tray includes a complementary orientation feature.
  • a portion of the flow path can be sealed with an airtight seal.
  • the detection reagent can be selected to detect a cardiac marker.
  • the detection reagent can be selected to detect a natriuretic peptide.
  • a method of manufacturing a cartridge for use in an assay system includes forming a base including a sample inlet fluidly connected to a microstructured flow path, depositing a detection reagent on the microstructured flow path, and forming a seal around at least a portion of the microstructured flow path.
  • the seal can be an airtight seal. Forming the airtight seal can include applying a membrane to a surface of the base.
  • the method can include forming a lid adapted to fit the base. Forming the airtight seal can include sealing the base to the lid. Sealing the base to the lid can include ultrasonic welding.
  • the optical system can include a reflectance measuring structure or photomultiplier.
  • FIG. 1 is a schematic view of a microfluidic assay device.
  • FIG. 2 is a schematic view of a portion of a microfluidic assay device.
  • FIG. 3 is a schematic view of a portion of a microfluidic assay device.
  • FIG. 4 is a schematic view of a portion of a microfluidic assay device.
  • FIG. 5 is a schematic view of a portion of a microfluidic assay device.
  • FIG. 6 is a schematic view of a portion of a microfluidic assay device.
  • FIG. 7 is a schematic view of a microfluidic assay device.
  • FIGS. 8A-8B are schematic views of a microfluidic cross-channel time gate.
  • FIG. 9 is a schematic view of a microfluidic assay device including a side channel time gate.
  • FIG. 10 is a schematic view of a microfluidic assay device.
  • FIG. 11 is a schematic view of a base of a microfluidic assay device.
  • FIG. 12 is a schematic view of a portion of a microfluidic assay device.
  • FIG. 13 is a schematic view of a portion of a microfluidic assay device.
  • FIG. 14 is a schematic view of a portion of a microfluidic assay device.
  • FIG. 15 is a schematic view of the underside of a base of a microfluidic assay device.
  • FIG. 16 is a schematic view of a microfluidic assay device.
  • FIG. 17 is a schematic view of a reader for use in conjunction with a microfluidic assay device.
  • FIG. 17 is a schematic drawing of an assay system.
  • FIG. 18 is a partial cutaway view of an assay system.
  • FIG. 19 is a schematic drawing of an optical detection system.
  • FIG. 20 is a schematic drawing of a cartridge for use in an assay system.
  • FIG. 21 is a schematic drawing of abase of a cartridge for use in an assay system.
  • FIG. 22 is a schematic drawing illustrating fluid flow in a cartridge for use in an assay system.
  • FIG. 23 is a schematic drawing of abase of a cartridge for use in an assay system.
  • FIG. 24 is a schematic drawing of a base of a cartridge for use in an assay system.
  • FIG. 25 is a schematic drawing of a base of a cartridge for use in an assay system.
  • FIG. 26 is a photomicrograph showing a portion of a base of a cartridge for ⁇ se in an assay system.
  • FIG. 27 is a graph depicting optical measurements from an assay system.
  • a device for measuring an analyte in a sample can include a microstructured flow path.
  • the flow path is a predefined path along which fluid can flow.
  • the flow path can be defined, for example, by a depression or depressed region, or by elevations forming walls that can contain a liquid.
  • the flow path can be defined by an elevated region surrounded by a depressed region, for example, a moat.
  • the fluid can be held on the elevated flow path by surface tension.
  • the flow path can include a sample inlet, a sample preparation region and an analysis region. When a sample is applied to the sample inlet, fluid can flow into the microstructured flow path.
  • the microstructured flow path includes capillary structures.
  • fluid flow along the flow path can be driven and controlled by capillary forces.
  • the flow path includes an analysis zone where the presence of an analyte in the sample results in a detectable change.
  • the extent of the change can be related to the concentration of analyte in the sample.
  • the flow path can optionally include a sample preparation region between the sample inlet and analysis region.
  • the sample preparation region can filter or selectively trap components of the sample while allowing other components to flow past the sample preparation region to the analysis region. Filtered components can be mechanically filtered, chemically or physically bound to a surface, immobilized, or otherwise prevented from traveling to the analysis region.
  • the flow path can optionally include a resuspension region.
  • the resuspension region can include a reagent that is stored in the device in a dry state until a liquid sample is introduced to the device. The liquid sample dissolves and resuspends the dry reagent.
  • the reagent can travel with the liquid sample to the analysis region.
  • the resuspension region can optionally overlap or be contiguous with the analysis region.
  • the flow path can optionally include a sink configured to collect a predetermined volume of liquid. The sink can ensure that a sufficient volume of sample is introduced to the device for the device to work as intended.
  • the flow path can optionally include a time gate.
  • a time gate is a structure that operates to delay fluid flow for a predetermined period of time.
  • the time gate can operate passively, i.e., fluid flow is gated by virtue of the fluid interacting with the structure of the gate, or can be actively controlled.
  • An actively controlled gate can be opened or closed, for example, by a valve.
  • the flow path can optionally include a control region.
  • the control region can be configured provide a detectable change when a sample is introduced to the device regardless of the amount of analyte in the sample.
  • the control region can thus reveal to a user that the device operates normally, even when no detectable change is apparent in the analysis region.
  • the flow path can optionally include a wash path.
  • the wash path can be configured to deliver a wash reagent to the analysis region.
  • the device can be configured to detect a cardiac biomarker.
  • a cardiac biomarker can be an indicator of cardiac health status. For example, an elevated level of creatine kinase can be an indicator of cardiac distress, such as a myocardial infarction.
  • the marker can be creatine kinase, creatine kinase-MB, troponin I, troponin T, myoglobin, f ⁇ brinopeptide, fibrinogen, C reactive protein, serum amyloid A, interleukin- 6, intercellular adhesion molecule- 1, vascular cell adhesion molecule- 1, E-selectin, soluble P-Selectin, soluble CD40 ligand, activated platelets, monocyte-platelet aggregates, oxidized-LDL, MDA-modified LDL, ischemia-modified albumin, free fatty acid, oxygen-regulated peptide 150, natriuretic peptides, or electrocardiogram.
  • the marker can be a marker of left ventricular volume overload or myocardial stretch; of myocardial apoptosis or injury; of inflammation; of anemia; of renal function; of electrolyte balance; or of sodium retention.
  • the marker of left ventricular volume overload or myocardial stretch can include a natriuretic peptide.
  • the marker of myocardial apoptosis or injury can include a troponin, urotensin, or a urotensin-related peptide.
  • the marker of myocardial ischemia can include ischemia-modified albumin, oxygen-regulated peptide (ORP 150), free fatty acid, Nourin-1, urotensin, or a urotensin- related peptide.
  • the marker of inflammation can include E-selectin, P-selectin, intracellular adhesion molecule- 1 , vascular cell adhesion molecule- 1 , Nourin- 1 , interleukin-l ⁇ , interleukin-6, interleukin-8, interleukin-10, tumor necrosis factor-alpha, hs-CRP, neutrophils, or white blood cell count.
  • the marker of anemia can include hemoglobin or hematocrit.
  • the marker of renal function can include creatinine or Cystatin C.
  • the marker of electrolyte balance can include Na + or K + .
  • the marker of sodium retention can include uroguanylin.
  • the natriuretic peptides include A-type- (ANP), B-type- (BNP), and C-type- (CNP) natriuretic peptide and their N-terminal prohormones (N-ANP, N-BNP, and N- CNP).
  • BNP the active peptide
  • N-BNP the inactive peptide
  • Both peptides are derived from the intact precursor, proBNP, which is released from cardiac myocytes in the left ventricle.
  • a microstructured device 10 for analysis of a sample includes a base 20 and a cover 30.
  • the device includes a sample inlet 40 and predefined flow path 50.
  • Flow path 50 can include filter 55, which can be a mechanical filter, a chemical filter, or other structure designed to selectively trap a component of the sample and prevent the selected component from traveling beyond the filter in the flow path.
  • the mechanical filter can be a membrane, for example a porous membrane.
  • Sample inlet 40 is fluidly connected to collection region 60, which in turn is fluidly connected to transfer region 90.
  • Base 20 and cover 30 are optionally sealed together. The seal can be an airtight seal.
  • the seal can be formed by, for example, applying an adhesive and contacting the base and the cover, by ultrasonic welding, laser welding, pressure sensitive tape, or heat seal tape.
  • the device can prepared and operated without a lid.
  • An adhesive seal can be selectively reversed (i.e., opened), for example, by melting the adhesive.
  • membrane filter 55 separates sample inlet 40 from collection region 60.
  • collection region 60 can include a plurality of membrane supports 70.
  • Each membrane support 70 includes a central post 72 and a plurality of projections 74 extending radially from post 72. Projections 74 are positioned so as to form vertical grooves 76 that extend from the uppermost end of membrane support 70 to base 20.
  • the membrane 50 contacts and is supported by the supports 70.
  • Vertical grooves 76 can have a capillary dimension; in other words, the grooves can have a width that acts as a capillary.
  • Collection region 60 can also include a plurality of longitudinal grooves 80. Longitudinal grooves 80 can have a capillary dimension.
  • a groove has a capillary dimension if it has a width that can provide capillary forces to a fluid, such as an aqueous fluid.
  • the capillary forces can be influenced by the surface of the grooves, for example, by the hydrophilicity or hydrophobicity of the surface.
  • filter 55 is a chemical filter, for example, including a compound immobilized on a surface of the flow path.
  • the immobilized compound can be immobilized in a transfer region 90.
  • Transfer region 90 can be configured to perform a sample preparation function.
  • the transfer region can include a structure designed to prevent a component of the sample from flowing into the analysis region.
  • the transfer region can be designed to alter the rate at which a liquid flows across the transfer region.
  • a change in the rate of liquid flow can cause red blood cells to form rouleaux, cylindrical stacks of cells.
  • the aggregated red blood cells in a rouleaux formation can be too large to flow into the analysis region.
  • the transfer region can include a reagent designed to trap or immobilize a component of the sample, such as red blood cells.
  • the transfer region can include, for example, a lectin, an antibody, or a carbohydrate immobilized on a surface of the base. As the sample flows past the lectin, red blood cells can be bound by the lectin and thus prevented from flowing to the analysis region.
  • Base 20 includes a transfer region 90 adjacent to and in fluid communication with collection region 60.
  • Transfer region preferably includes a plurality of elevated posts 100. Elevated posts 100 can be disposed in a regular array. The horizontal dimensions of posts 100 and the spacing between posts in the array can selected such that the array has capillary dimension.
  • the dimensions of posts 100 and the spacing between posts can be chosen to control the capillary properties and therefore the rate of travel of a liquid front across the transfer region.
  • Transfer region 60 can also include perpendicular ridges 110.
  • Perpendicular ridges 110 can modify the rate of travel of a liquid front along the device.
  • Base 20 includes an analysis region 120 adjacent to and in fluid communication with transfer region 90.
  • Analysis region 120 preferably includes a plurality of elevated posts 130. Elevated posts 130 can be disposed in a regular array.
  • the horizontal dimensions of posts 130 and the spacing between posts in the array can be selected such that the array has capillary dimensions.
  • the capillary dimension can be chosen to control the rate of travel of a liquid front across the analysis region.
  • a sponge-like flow-promoting pad at the far end of the device can soak up liquid, helping draw liquid through the device to the analysis region.
  • liquid sample When a liquid sample is placed in sample inlet 40, capillary action draws it to filter 55. Components that are detrimental to analysis (such as any solids or large particles, for example, cells) in the liquid are retained by the filter, while liquid (e.g., plasma) can pass through the membrane to the collection region below.
  • liquid e.g., plasma
  • the filter when the liquid sample is whole blood, cells can be retained by the filter, allowing plasma to continue to flow through the remainder of the flow path. Only a small volume of the liquid sample is required, for example, 1 mL or less, 0.5 mL or less, 0.1 mL or less, 50 microliters or less, or 10 microliters or less.
  • the volume of blood obtained by a finger stick is a sufficient volume for performing the assay.
  • the sample can be applied from a pipette.
  • vertical grooves in the membrane supports draw the liquid by capillary action out of the membrane and into the collection region.
  • the liquid can collect in the longitudinal grooves of the collection region.
  • the capillary action provided by the longitudinal grooves moves the liquid along the grooves, until it reaches the transfer region.
  • the size, shape, and spacing of the posts in the transfer region is selected to provide a desired rate of movement of the liquid across the transfer region.
  • the surface of the base, of the lid, or both, can be modified with a compound that can affect the rate of movement. For example, a hydrophilic compound can promote movement of an aqueous liquid, whereas a hydrophobic compound can inhibit movement of the aqueous liquid.
  • the degree of hydrophilicity or hydrophobicity of a compound can be expressed in terms of a surface contact angle.
  • a contact angle is the angle formed at the edge of a drop of a liquid (for example, water) on a flat surface. The more hydrophilic the surface, the more the fluid will tend to spread. Conversely, fluid will tend to bead on a hydrophobic surface.
  • the compound that modifies the surface of the base can be selected to provide a contact angle of between 5 and 20 degrees, such as between 8 and 15 degrees.
  • the surface contact angle can range from the native contact angle of the material down to 1 degree.
  • the base can be modified with a compound such that the base has a contact angle of 11 degrees.
  • the motive force for the fluid in a microfluidic device is the surface energy in the meniscus, which results from the interaction between the solid surface and the fluid. Opposing the fluid movement is the skin friction created at the wall/ fluid interface.
  • the velocity of the fluid can be controlled by the addition of forces. Pressure (e.g., hydrostatic pressure or air pressure) can be either added to act with or against the fluid, which will increase or decrease the fluid velocity, respectively. To use pressure control for fluid control requires the capability to seal the lid of the microfluidic device.
  • device 200 includes a base and cover.
  • the device includes sample inlet 240 connected by capillary flow path 250 to first reaction zone 260.
  • First reaction zone 260 can include a textured region with protrusions being spaced a capillary distance apart.
  • First reaction zone can also include a removal reagent designed to capture components of the sample that can interfere with the analysis.
  • First time gate 270 is designed to stop liquid flow for a predetermined length of time, after which time gate 270 allows liquid to flow to second reaction zone 280.
  • the length of time for which the first time gate 270 stops liquid flow can be selected to insure that any reaction taking place in reaction zone 260 attains a desired degree of completion. For example, if it is desired that the reaction in reaction zone 260 is substantially complete before the liquid flows to the second reaction zone 280, time gate 270 is designed to pause liquid flow for a length of time sufficient to ensure substantial completion of the reaction in reaction zone 260.
  • Second reaction zone 280 can include an analysis zone.
  • An affinity molecule can be disposed in the analysis zone, for example, the affinity molecule can be anchored to a surface of base 220 in the analysis zone. Any analyte in the sample can be captured by the anchored affinity molecule.
  • a surface in the analysis zone can be structured to immobilize only analyte-bound label in the analysis zone. The amount of label immobilized can correlate to the concentration of the analyte in the sample.
  • the free label i.e., label not bound to an analyte
  • flows through the analysis zone into the sink. The movement of the label and the fluid is controlled by the time gates.
  • a number of zones can be provided downstream for any fluid pretreatment.
  • a second affinity molecule can be included in the analysis zone.
  • the second affinity molecule can be soluble, and form a sandwich complex.
  • the sandwich complex can be formed from the anchored affinity molecule, the analyte, and a soluble second affinity molecule.
  • a soluble second affinity molecule can be labeled, such that an immobilized sandwich produces a detectable change.
  • the amount of sandwich complex formed, and therefore the degree of detectable change is related to the amount (for example, concentration) of analyte in the sample.
  • a measurement of the degree of detectable change can be used to calculate the amount of analyte in the sample.
  • second time gate 290 will allow the liquid to flow into sink region 300.
  • Sink region 300 can have a predetermined volume. Liquid can flow until the predetermined volume of liquid is in the sink, stopping the flow. The flow of liquid into the sink can wash away any unbound affinity molecules (i.e., soluble affinity molecules that did not bind an analyte and become immobilized as part of a sandwich complex). By washing unbound affinity molecules, in particular, unbound labeled affinity molecules, the sink helps ensure that the measurement of a detectable change in analysis region 280 is an accurate measurement.
  • unbound affinity molecules i.e., soluble affinity molecules that did not bind an analyte and become immobilized as part of a sandwich complex.
  • a time gate can be constructed as a side channel to the flow path.
  • a single central horizontal channel 300 is shown, in which the fluid moves from right to left.
  • the area which surrounds the central channel is a moat 320 which creates a raised central channel, defining the flow path.
  • the fluid flows between the base and the lid, driven by the capillary force. In this configuration the base is hydrophilic and the lid hydrophobic.
  • the cross channel When the fluid reaches the cross channel it stops.
  • the fluid in the main channel overcomes the capillary stop and moves forward. This is repeated a further 2 times.
  • a small fiber can be added to the cross channel. This has the effect of acting as a bridge and the fluid will then move over the capillary stop.
  • base 20 is shown with an alternative time gate structure. Fluid is added in the sample inlet 40 and then progresses to the chamber 350. Once the chamber 350 is filled, fluid will then exit the chamber 350, where it will be held in place by the capillary stop 360. As the chamber is filling, a side channel 370 also fills. The cross-section and length of the side channel are chosen to give the required time delay. The speed at which the fluid moves up the side channel is much slower than that through the main channel. Once the fluid in the side channel 370 reaches the end, it joins with the fluid at the capillary stop 360, overcoming it and allowing the fluid to progress to the next chamber.
  • the device will be prepared in a dry state, and an analysis is initiated by placing a liquid sample in the sample inlet.
  • Capillary action transports the liquid along the flow path, from the sample inlet into the collection region, through the transfer region and to the analysis region.
  • a microfluidic assay device 500 can include a sample inlet 510 fluidly connected to a dual flow path.
  • the device can include result windows 520, where the detectable change can be detected.
  • the device is designed for use with a sample of whole blood, plasma, serum or urine.
  • the sample can be whole blood from a finger stick.
  • the dual flow path can divide fluid flowing from the sample inlet into two paths.
  • the total required blood volume for the two channels to run can be less than 100 ⁇ L, less than 75 ⁇ L, or less than 50 ⁇ L (based on a 50% hematocrit and a 50% blood separation efficiency).
  • Sample can be added to the port either as a single sample or in discrete volumes.
  • the sample inlet is designed not to entrap air.
  • the sample bifurcates to the two channels, 530 and 532. At this point the sample is brought from the center of the device to the outer edge. The primary reason for this is to ensure high efficiency blood separation using a filter membrane.
  • the added benefit of bringing the sample to the outside of the device is that the instrument that reads the device can have a built in detector, a photodiode or similar to detect whether sufficient sample has been applied.
  • the membrane is bonded to the lid of the device.
  • the outer edge of the membrane should be crushed to prevent red blood cells from escaping.
  • Microstructure in the base is in direct physical contact with the membrane, which helps to improve the efficiency of separation.
  • the microstructure includes pillars 550 having V-shaped grooves which run from their base to their top. At the base of these pillars are very small scale grooves, having a dimension on the order of 10 ⁇ m. Plasma is initially drawn through the membrane, down the V-shaped grooves in the pillars and immediately starts to wet the base of the chamber through the small grooves in the base of the chamber.
  • a connection is thus made between the base and the membrane which yields a fluid meniscus which has a high surface energy. As the fluid tries to reduce the surface energy in the meniscus, it draws more fluid through the membrane. A plasma pool is created with sufficient volume to start to run the assay.
  • the blood separation zone is full and ready to be run through the assay. Timing of fluid flow in the device is controlled, for example, by selective venting.
  • the membrane Once the membrane is applied to the device, the channel after the blood separator is a sealed volume once the plasma fills the blood separator. Fluid cannot enter the channel as the capillary force cannot overcome the pressure which builds up as the fluid tries to fill the channel, unless a vent is opened.
  • the mechanism chosen to vent the cartridge is a membrane which has fusible tracks printed on it.
  • the membrane can be bonded to the structure using a pressure sensitive adhesive or a heat seal. As current is passed through a track a selected high resistance area heats up and melts the adhesive and laminate to make a hole. Down the length of the chip there are a number of vents 560.
  • the device can be used in combination with a reader, the reader being configured to supply the necessary current for melting the membrane at predetermined times once the sample has been applied to the assay device.
  • Each channel of the chip includes three discrete zones, a deposition and resuspension zone 570, a detection zone 580, and a sink 590.
  • the channel can include a control zone.
  • a control zone generally operates like a detection zone, but is configured to produce a detectable change regardless of the presence of analyte in the sample. In this way, a user can be assured that the assay operated correctly, even when no detectable change is observed in the detection zone.
  • Windows 520 are located above detection zone 580.
  • An affinity reagent is deposited in deposition and resuspension zone 570. The pillars help to distribute the reagent homogeneously through the zone when it is deposited. Once the fluid moves into the zone on opening of the vent it moves up to the next vent position. The fluid then resides in the zone until the reagent is resuspended.
  • the subsequent vent is opened and the fluid fills the detection zone 580, taking with it the resuspended reagent.
  • the subsequent vent is opened and the fluid fills the detection zone 580, taking with it the resuspended reagent.
  • portions of the walls at either side of the chamber are removed.
  • the reader takes a measurement. Once the measurement is completed the final vent is opened and the fluid moves to fill the sink 590, washing all the unbound reagent into the well.
  • the analysis region can include a reagent for detecting an analyte in the liquid sample.
  • the reagent can bind to or react with the analyte to produce a detectable change.
  • the detectable change can be, for example, a change in optical properties such as an absorption or emission of light.
  • the detectable change can be a change in color.
  • the reagent can include an affinity molecule that binds to the analyte.
  • the affinity molecule preferably binds tightly and specifically to the analyte. In other words, the affinity molecule has a large association constant for the analyte, while having a much lower (for example, by one or more orders of magnitude) association constants for other components present in the liquid sample.
  • the affinity molecule can be, for example, a protein, a peptide, an antibody, a nucleic acid, or a small molecule.
  • the analyte can be, for example, a protein, a peptide, an antibody, a nucleic acid, or a small molecule.
  • the affinity molecule can be selected for its affinity and selectivity towards its ligand.
  • whole blood e.g., 1-5 microliters of whole blood
  • Liquid travels unidirectionally through the various sections of the device, being passively driven by capillary forces.
  • the whole blood is transported to the blood separation zone and the red blood cells are sedimented from plasma using changes in capillary force and its rouleaux-forming properties. Any remaining cells are captured and removed through microstructures of grooved surfaces and pillars.
  • the gaps between the grooved surfaces or pillars can have a width of, for example, 2 microns.
  • the affinity molecule can be linked to a support, such as a bead or a surface of the device.
  • the link can include a spacer and an anchor.
  • the anchor can chemically or physically attach the affinity molecule to the bead or surface.
  • the support can be linked to multiple affinity molecules.
  • the link can be formed by combining the affinity molecule, the anchor, and a spacer precursor.
  • the spacer precursor can include one or more reactive functional groups that associate with the affinity molecule or the anchor.
  • the link can be formed by modifying the affinity molecule, the anchor, or both, to introduce a reactive functional group. A reaction between the introduced reactive functional group or groups can form the link.
  • the spacer precursor can be, for example, a bifunctional compound, i.e., a compound having two reactive functional groups.
  • the reactive functional groups can be the same or different.
  • a reactive functional group can react with, for example, an amino group or a sulfhydryl group.
  • Examples of spacer precursors include, for example, disuccinimidyl suberate (DSS), or a di-(N-hydroxysuccinimidyl)polyethylene glycol.
  • the length of a di-(N-hydroxysuccinimidyl)polyethylene glycol can vary, depending on the number of ethylene glycol units in the polyethylene glycol.
  • the di-(N- hydroxysuccinimidyl)polyethylene glycol can have a molecular weight of less than 3 kDa, approximately 3 kDa, approximately 6 kDa, or higher.
  • the anchor, affinity molecule, or both can be modified to introduce a reactive functional group.
  • the reactive functional groups can be selected to be complementary; in other words, the reactive functional groups are selected to react with one another to form the link.
  • the anchor or affinity molecule can be modified with a bifunctional reagent, such as, for example, NHS-PEG-maleimide, SMCC, NHS-PEG- vinylsulfone to introduce a group that reacts with a sulfhydryl group.
  • a sulfhydryl group can be introduced by a reagent such as, for example, SATP, SATA 5 SDPP, or SAMSA.
  • the anchor facilitates the attachment of the affinity molecule to the support (e.g., a surface of the device).
  • the anchor can include, for example, a polysaccharide (e.g., a dextran), or a protein, for example an albumin such as bovine serum albumin (BSA).
  • BSA bovine serum albumin
  • the anchor can include a surfactant or detergent such as, for example, a TWEEN, a PLURAFAC, or a PLURONIC.
  • the anchor can be associated with a surface of the device by physical or chemical interactions. A surface of the device can be modified to bind the anchor to hold the anchor to the surface.
  • a plastic surface such as, for example, polystyrene or polycarbonate
  • a plasma i.e., an ionized gas
  • the anchor can bind directly to an unmodified surface, such as plastic or glass.
  • the assay can produce a detectable change.
  • the detectable change can be an optical change, for example, a change in chemiluminescence, a fluorescence (including fluorescent energy transfer and time resolved fluorescence), a change of resonant light scattering, or a change in absorption of light.
  • the extent of detectable change is proportional to amount or concentration of analyte in the sample, such that a quantitative measurement of the amount or concentration of analyte can be made.
  • a detectable change in chemiluminescent signal can be produced when an analyte molecule in a sample brings two particles (or beads) together in close proximity.
  • a cascade of chemical reactions that depends on the proximity of the beads (and therefore on the presence of the analyte) can produce greatly amplified signal. Detection of an analyate at attomolar (i.e., on the order of 10 " molar) concentrations is possible.
  • Photosensitizer particles (donor particles) including phthalocyanine can generate singlet oxygen when irradiated with light having a wavelength of 680 nm.
  • the singlet oxygen produced has a very short half-life - about 4 microseconds - and hence it decays rapidly to a ground state. Therefore it can only diffuse to a distance of a few hundred microns from the surface of the particles before it decays to ground state. However, the singlet state can survive long enough to enter any paired adjacent particle.
  • the paired adjacent particles (acceptor particles) include a dye that is activated by singlet oxygen to produce chemiluminescent emission.
  • This chemiluminescent emission can activate further fluorophores contained in the same bead, subsequently causing emission of light at 520-620 nm. See, for example, Proc. Natl. Acad. ScL 91 :5426-5430 1994; and U.S. Patent No. 6,143,514, each of which is incorporated by reference in its entirety.
  • the reagents will be lyophilized in a well with an optimized geometry and reflection design to insure maximum excitation and light capturing efficiency.
  • the reagent can include a bead linked to an antibody.
  • the bead can include a polymeric material, for example, latex or polystyrene.
  • the bead can include a light- absorbing or light-emitting compound.
  • a latex bead can include a dye or a fluorescent compound.
  • the reagent can include a plurality of beads. The beads in the plurality can be linked to one or more distinct antibodies. A single bead can be linked to two or more distinct antibodies, or each bead can have only one distinct antibody linked to it.
  • the reagent can have more than one distinct antibody each capable of binding to the same ligand, or antibodies that recognizes different ligands. The intensity of the emitted light can be measured and correlated to the concentration of ligand in the sample.
  • a detectable change can be produced by the enzyme multiplied immunoassay technique (EMIT).
  • EMIT enzyme multiplied immunoassay technique
  • the label used is an enzyme-analyte conjugate.
  • One reagent can include an antibody specific for the analyte, an enzyme substrate, and (optionally) a coenzyme.
  • a second reagent can include an analyte linked to an enzyme (the labeled analyte).
  • the enzyme can be a glucose-6-phosphate dehydrogenase (G-6-PDH).
  • G-6-PDH can catalyze the reaction of glucose-6-phosphate with NAD(P) to yield 6-phosphoglucono- d-lactone and NAD(P)H.
  • NAD(P)H absorbs light with a wavelength of 340 nm, whereas NAD(P) does not.
  • a change in absorption of 340 nm light as a result of the G-6- PDH catalyzed reaction can be a detectable change.
  • CEDIA cloned enzyme donor immunoassay
  • CEDIA is a homogeneous immunoassay based on the bacterial enzyme ⁇ -galactosidase of E. coli which has been genetically engineered into two inactive fragments. These two inactive fragments can recombine to form an active enzyme, one fragment consists of an analyte-fragment conjugate, and the other consists of an antibody-fragment conjugate.
  • the amount of active enzyme that generates the signal is proportional to the analyte concentrations. See, for example, Khanna, PX. and Coty, W.A. (1993) In: Methods of Immunological Analysis, volume 1 (Masseyeff, R.F., Albert, W.H., and Staines, N.A., eds.) Weinheim, FRG: VCH Verlagsgesellschaft MbH, 1993: 416-426; Coty, W. A., Loor, R., Powell, M., and Khanna, P.L. (1994) J. CHn. Immunoassay 17(3): 144-150; and Coty, W.A., Shindelman, J., Rouhani, R. and Powell, MJ. (1999) Genetic Engineering News 19(7), each of which is incorporated by reference in its entirety.
  • a reagent for producing a detectable change can include a metal nanoparticle, for example, a gold or silver nanoparticle. Such particles can take advantage of resonance light scattering (RLS) to produce a detectable change.
  • RLS resonance light scattering
  • a metal nanoparticle When a metal nanoparticle is excited by light (for example, polychromatic light), it electrons can resonate in phase with the incident light forming an electromagnetic dipole that emits energy. This energy produces collective conduction resonance of the electrons and therefore intense monochromatic light is scattered with remarkable efficiency.
  • the signal intensity can achieve a sensitivity greater than any fluorophore, by a factor of 10,000.
  • the wavelength of the scattered light is a function of the particle size and the particle composition.
  • the gold or silver nanoparticles generate signals of predetermined color and intensity, typically 20-160 nm in diameter and the color changes from green to red as the size changes from 40 to 160 nm, more specifically the different particle diameters are: 40 nm green, 80 nm greenish yellow, 120 nm orange and 160 nm red.
  • the RLS signal does not photo-bleach, quench or decay. The lack of signal degradation allows multiple measurements of RLS-based assays, enabling the use of different exposure times without risk of pignal degradation
  • An RLS homogenous assay can take advantage of changes in the scattering spectrum when two are more particles are brought together at distances less than 2 particle diameters. These spectral changes are due to particle-particle perturbations and depend on the exact distance between the particles.
  • each analyte molecule has several antibody sites such that it can bind two or more antibody-linked metal nanoparticles.
  • This change can be used to detect and quantify the analyte concentration.
  • the detectable change can be a change in fluorescence or absorbance (e.g., color).
  • a fluorescent signal or fluorescence can be measured in a time-resolved fashion.
  • Homogeneous assays can be noncompetitive or competitive, hi the noncompetitive, or direct, homogeneous assay, the analyte has multiple antibody binding sites and one analyte molecule can directly crosslink at least two RLS particles as described above.
  • the competitive assay is used for analytes that have only one antibody binding site and therefore cannot crosslink antibody-linked metal nanoparticles.
  • These types of analytes are usually substances of low molecular weight, such as, for example, drugs of abuse.
  • an antigen analog in which two antigen molecules are linked covalently is synthesized.
  • This analog is added to the suspension of gold-antibody particles at a concentration where it cross links some of the metal-antibody particles resulting in a significant change in the light scattering spectrum of the suspension.
  • the single site (i.e., single epitope) analyte is added to the latter mixture, it competes with the two site analyte analog for binding to the metal particles and prevents particle-particle aggregation. This decreases particle aggregation, changing the light scattering spectrum of the mixture. The single site analyte can thus be detected and quantified from these spectral changes.
  • the analysis region can be washed after the sample has been allowed to interact with a reagent in the analysis region.
  • the washing can remove unbound analyte and other components that can interfere with measurement of the detectable change.
  • the wash reagent can be mixed with the liquid sample after the sample has been applied to the device.
  • the removal reagent can be stored in a well 140 in fluid communication with the analysis region 120 of the device. See FIG. 16, which depicts device 10, including base 20, cover 30, inlet 40, analysis region 120 and well 140.
  • the well includes a port at a lower end, the port being fluidly connected to the analysis region.
  • the wash reagent may be stored in a pouch or added separately.
  • the wash reagent can be a liquid that remains in the well by virtue of its surface tension at the port. When a sample is applied to the sample inlet, the liquid flows through the device to the analysis region as described above. Upon reaching the port, the flowing liquid meets the liquid in the well, breaking the surface tension.
  • Hydrostatic pressure e.g., from a pouch of liquid that is pushed or squeezed
  • the liquid in the well including the wash reagent
  • Hydrostatic pressure can then cause the liquid in the well, including the wash reagent, to flow into the analysis region of the device.
  • the wash reagent Once the wash reagent reaches the analysis region, it washes plasma, including unbound analyte and other plasma components, out of the analysis region.
  • Reagents can be deposited on the base by lyopbilization, glazing, or spray-drying. Lyophilization (or freeze drying) can stably preserve the function of biological reagents while allowing easy reconstitution with aqueous buffers and plasma. Direct deposition of reagents onto the chip with positive displacement micro-pumps and lyophilization of the entire chip can also be used. Lyophilized pellets and pucks can be prepared separately from the base of the device and physically deposited on the device. Pellets are produced by dispensing a solution of reagent into a pool of liquid nitrogen, freezing the droplet into a sphere. The frozen sphere then drops to the bottom of the pool and can later be removed and freeze dried. A puck is produced by dispensing the frozen droplet onto a frozen tray which is later loaded into the freeze dryer.
  • a larger amount of reagent can be placed in the chip using a puck due to the puck's reduced height compared to the spherical pellet.
  • Using a puck or pellet instead of directly dispensing onto the chip requires less freeze dryer space to be used for a given number of devices, since the actual device is not loaded into the freeze dryer. Pellets and pucks can be placed into the chip using an automated system after undergoing the freeze drying process.
  • Spray-drying and glazing can be less time consuming than a relatively lengthy lyophilization cycle and this could outweigh the potential reduction in stability or ease of rehydration.
  • the reagent can be arranged on a porous or nonporous capture material in the device such that the binding or detection zone develops uniformly.
  • the reagent is arranged to provide concentrations of particles and required particle distribution into the analysis to give precise of measurements.
  • the first reaction zone The reagent and the reaction zone are optimized such that when the reagent is reconstituted promptly upon contact with the sample or the fluid entering the device.
  • the resultant reaction mixture delivers a uniform reagent suspension.
  • the device can be used in combination with a reader configured to measure the detectable change.
  • the reader can include an optical system to detect light from the analysis region.
  • the light to be detected can be, for example, emitted, transmitted, reflected, or scattered from the analysis region. Emitted light can result from, for example, chemiluminescent or fluorescent emission.
  • the optical system can include an illumination source, for example, to be used in the detection of a change in fluorescence, absorbance, or reflection of light.
  • the system can include a cartridge and cartridge reader.
  • the system can be used to assay a variety of analytes or markers.
  • the cartridge can be prepared to detect one or more of a variety of analytes or markers.
  • the user can select a cartridge that is sensitive to the desired analyte or marker, and obtain a measurement of that analyte with the system.
  • the reader can also include an output display configured to display the results of the measurement to a user.
  • system 1000 includes cartridge reader 1050 and cartridge 1100.
  • Cartridge 1100 is held in reader 1050 by tray 1150.
  • Reader 1050 includes display 1200.
  • Tray 1150 can be positioned in an open or closed position. The open position allows a new cartridge to be inserted or a used cartridge to be removed. The tray is held in the closed position when an assay is performed. When held in the closed position, the cartridge is aligned with instrumentation internal to reader 1050. Proper alignment can be important for obtaining accurate results.
  • Display 1200 can also be used to display text messages, help messages, instructions, queries, test results, and various information to users Additionally, display 1200 may be used to display images in various formats, for example, joint photographic experts group (JPEG) format, tagged image file format (TIFF), graphics interchange format (GIF), or bitmap.
  • Display 1200 can optionally provide input region 1400 including keys 1600.
  • input region 1400 can be implemented as symbols displayed on the display 1200, for example when display 1200 is a touch- sensitive screen.
  • User instructions and queries are presented to the user on display 1200. The user can respond to the queries via the input region.
  • Display 1200 can provide a user with an input region 1400.
  • Input region 1400 can include keys 1600.
  • input region 1400 can be implemented as symbols displayed on the display 1200, for example when display 1200 is a touch-sensitive screen.
  • User instructions and queries are presented to the user on display 1200. The user can respond to the queries via the input region.
  • Reader 1000 also includes a cartridge reader, 1050, which accepts diagnostic test cartridges 1100 for reading.
  • the cartridge reader 1050 can measure the level of a biomarker based on, for example, the magnitude of a color change or other detectable change that occurs on a test cartridge 4000.
  • Cartridge reader 1800 includes optical systems for measuring the detectable change, for example, a light source, filter, and photon detector, e.g., a photodiode, photomultiplier, or Avalance photo diode.
  • Device 1000 can further include a communication port 2200.
  • Communication port 2200 can be, for example, a connection to a telephone line or computer network.
  • Device 1000 can communicate the results of a measurement to an output device, remote computer, or to a health care provider from a remote location.
  • the cartridge can include two testing zones.
  • a cartridge can include 1 , 2, 3, 4, or 5 or more testing zones.
  • Each testing zone can test the level of a biomarker.
  • Each testing zone includes a sample input, and a result window.
  • the cartridge can include a microfluidic assay.
  • a health care provider or other user can use device 1000 for testing and recording the levels of various analytes, such as, for example, a biomarker, a metabolite, or a drug of abuse.
  • diagnostic device 1000 may access programs and/or data stored on a storage medium (e.g., video cassette recorder (VCR) tape or digital video disc (DVD); compact disc (CD); or floppy disk). Additionally, various implementations may access programs and/or data stored on another computer system through a communication medium including a direct cable connection, a computer network, a wireless network, a satellite network, or the like.
  • the software controlling the reader can be in the form of a software application running on any processing device, such as, a general-purpose computing device, a personal digital assistant (PDA), a special-purpose computing device, a laptop computer, a handheld computer, or a network appliance.
  • the reader may be implemented using a hardware configuration including a processor, one or more input devices, one or more output devices, a computer-readable medium, and a computer memory device.
  • the processor may be implemented using any computer processing device, such as, a general-purpose microprocessor or an application- specific integrated circuit (ASIC).
  • the processor can be integrated with input/output (I/O) devices to provide a mechanism to receive sensor data and/or input data and to provide a mechanism to display or otherwise output queries and results to a service technician.
  • I/O input/output
  • Input device may include, for example, one or more of the following: a mouse, a keyboard, a touch-screen display, a button, a sensor, and a counter.
  • the display 1200 may be implemented using any output technology, including a liquid crystal display (LCD), a television, a printer, and a light emitting diode (LED).
  • the computer-readable medium provides a mechanism for storing programs and data either on a fixed or removable medium.
  • the computer-readable medium may be implemented using a conventional computer hard drive, or other removable medium.
  • the system uses a computer memory device, such as a random access memory (RAM), to assist in operating the reader.
  • RAM random access memory
  • Implementations of the reader can include software that directs the user in using the device, stores the results of measurements.
  • the reader 1000 can provide access to applications such as a medical records database or other systems used in the care of patients, hi one example, the device connects to a medical records database via communication port. 2200.
  • Device 1000 may also have the ability to go online, integrating existing databases and linking other websites.
  • the cartridge reader 1050 can optionally include a communication port.
  • the communication port can be, for example, a connection to a telephone line, computer, or computer network.
  • System 1000 can communicate the results of a measurement to an output device (e.g., a printer), a remote computer, or to a health care provider from a remote location via the communication port.
  • FIG. 18 shows a partial cutaway view of system 1000.
  • the cutaway view reveals motor assembly 1600 for driving tray 1150, and system electronics 1700.
  • System electronics 1700 includes the necessary circuitry for powering reader 1100, operating display 1200, performing the measurement of the analyte and communicating the result to the user.
  • the reader instrument can operate with a high degree of automation in order to reduce the possibility of user error.
  • Partially or fully automated features of the system include control of sample volume; detection of a cartridge in the tray; and location detection of the tray (i.e., whether the tray is in the open or closed position). Detection of a successful assay run is made by detection of fluid at the end of the flow path. Timing of fluid flow through different regions of the flow path can be controlled.
  • the instrument can use a fully automated tray. Once the cartridge is inserted into the tray, a strip-in switch detects the presence of the cartridge. The tray is then moved to the closed position, pulling the cartridge into the reader so that the batch code on the cartridge can be checked against the instrument ROM key.
  • a ROM key is used to store cartridge batch calibration information, avoiding the requirement for high and low controls on chip.
  • the batch code can be stored as a bar code.
  • FIG. 19 is a schematic diagram illustrating the optical system for measuring the assay result.
  • a sample When a sample is applied to the cartridge, fluid flows from the sample inlet along the flow path.
  • a detectable change occurs in the detection window.
  • the optical system measures the extent of the detectable change.
  • the detectable change can be a change in absorption or transmittance of light, or of emission of light (e.g., a fluorescent or chemiluminescent emission).
  • the detection window 2600 is located between light source 1800 and detector 1900.
  • Light source 1800 can be, for example, a lamp or LED.
  • the light source can be a 375 nm LED collimated through a 5 mm tube.
  • Detector 1900 can be, for example, a photodiode.
  • Optional illumination filter 2000 is located between light source 1800 and cartridge 1100 and can select a desired wavelength or range of wavelengths to pass to the cartridge.
  • Filter 2000 can include, for example a bandpass filter.
  • Optional detection filter 2100 is located between cartridge 1100 and detector 1900.
  • Detection filter 210 can include a thin film filter.
  • the filters can be configured with a terry red filter before two UV filters.
  • FIGS. 20-26 schematically illustrate cartridge 1100.
  • Cartridge 1100 features sample inlet 2500, detection windows 2600, and orientation features 2700, 2800, and 2900.
  • Tray 1150 can include features complementary to orientation features 2700, 2800, and 2900, to ensure that the cartridge can be placed in the tray only in the correct orientation.
  • Sample inlet 2500 is designed to take 20 ⁇ L of whole blood from any finger. Blood can be applied directly to the inlet in any volume. Too much sample cannot be applied as the device has a fixed volume. If too little sample is applied, the device will not run as the instrument will detect (e.g., optically) the volume of blood in the inlet channel. Once sufficient volume has been applied, the instrument informs the user to stop applying sample and the cartridge will be drawn into the instrument (i.e., the tray will close) and the measurement will start. Sample inlet 2500 optionally includes EDTA, an anti-coagulant, or both. For embodiments where sample application and blood separation take more than one minute, it can be preferable to prepare the cartridge with EDTA and an anti-coagulant in the sample inlet.
  • cartridge 1100 can include a sample inlet fluidly connected to a microstructured flow path.
  • the microstructured flow path can provide capillary forces to the sample. The capillary forces move the fluid from the sample inlet and along the flow path.
  • Cartridge 1100 can be manufactured by forming a base 3000 and lid 3500 which are attached or sealed together.
  • FIGS. 21-25 illustrate elements of cartridge 1100.
  • the base shown in FIG. 21 includes sample inlet 2500 connected to intake region 3050. When the sample is applied to the sample inlet 2500, fluid flows from inlet along intake region 3050, bifurcating into dual flow paths.
  • the flow paths pass into the lid 3500 to carry the fluid to a filter 3600 configured to remove cells from the sample.
  • the filter is located above filter support regions 3100 and 3120.
  • FIG. 22 illustrates schematically how filter 3600 is positioned relative to base 3000, lid 3500 and filter support regions 3100 and 3120 to filter cells from the sample.
  • the curved arrow in FIG. 22 illustrates the direction of fluid flow.
  • Filter support regions 3100 and 3120 include microstructure in direct physical contact with the membrane, which helps to improve the efficiency of separation.
  • FIG. 23 shows a close up view of filter support region 3100.
  • FIG. 23 illustrates that the microstructure of filter support region 3100 includes pillars 3700 having V-shaped grooves which run from their base to their top. At the base of these pillars are very small scale grooves, having a dimension on the order of 10 ⁇ m.
  • Plasma is initially drawn through the membrane, down the V-shaped grooves in the pillars and immediately starts to wet the base of the chamber through the small grooves in the base of the chamber. A connection is thus made between the base and the membrane which yields a fluid meniscus which has a high surface energy. As the fluid tries to reduce the surface energy in the meniscus, it draws more fluid through the membrane. A plasma pool is • created with sufficient volume to start to run the assay. Eventually the blood separation zone is full and ready to be run through the assay.
  • Base 3000 is shown in FIG. 21 with two assay channels, 3200 and 3220.
  • Assay channels 3200 and 3220 are flanked by energy directors 3300 which can form an airtight seal with the Hd, when the lid and base are sealed together, for example by an ultrasonic weld.
  • Base 3000 also includes divots 3400. Divots 3400 can align with matching pins on the lid to assure correct orientation and alignment of base and lid when the cartridge is assembled. Alternatively, pins can be located on the base with matching divots on the lid.
  • Each assay channel includes a deposition and resuspension zone 3800, detection zone 3900 and sink 4000. Timing of fluid flow in the device is controlled, for example, by selective venting.
  • the channel after the blood separator is a sealed volume once the plasma fills the blood separator. Fluid cannot enter the channel as the capillary force cannot overcome the pressure which builds up as the fluid tries to fill the channel, unless a vent is opened.
  • a membrane having fusible tracks printed on it can be applied to the cartridge. The membrane can be bonded to the structure using a pressure sensitive adhesive or a heat seal. As current is passed through a track a selected high resistance area heats us and melts the adhesive and laminate to make a hole. The cartridge reader 1050 can deliver the necessary current. Down the length of the chip there are a number of vents 4100.
  • the reader can be configured to supply the necessary current for melting the membrane at predetermined times once the sample has been applied to the assay device.
  • Each channel of the cartridge includes three discrete zones, a deposition and resuspension zone 3800, a detection zone 3900, and a sink 4000.
  • FIG. 24 shows one channel of the cartridge in greater detail.
  • Sink 4000 can have a volume approximately 5 times the volume of the detection zone.
  • the channel can include a control zone.
  • Detection windows 2600 are located above detection zones 3900.
  • An affinity reagent is deposited in deposition and resuspension zone 3800. Microstructured pillars in deposition and resuspension zone 3800 help to distribute the reagent homogeneously through the zone when it is deposited.
  • the fluid moves into the zone on opening of the vent it moves up to the next vent position.
  • the fluid then resides in the zone until the reagent is resuspended.
  • the walls of the deposition and resuspension zone 3800 and detection zone 3900 have been removed (wall-removed regions 3850, 3950) to prevent label from sticking to the walls. Removing the walls at these locations can reduce the background in the detection zone and hence improve the signal-to-noise ratio in the measurement of the detectable change.
  • the cartridge can be configured to perform a sandwich assay.
  • the deposition and resuspension zone 3800 can include a first antibody.
  • the first antibody can bind specifically to the analyte and is labeled, for example with a color.
  • the color can be, for example, a colored latex particle or metal nanoparticle.
  • the detection zone 3900 can include a second antibody immobilized on the base.
  • the second antibody can also bind specifically to the analyte. Analyte present in the sample is bound by both the immobilized second antibody and the first labeled antibody.
  • the labeled antibody becomes immobilized in the detection zone in amount related to the amount of analyte in the sample, while unbound labeled antibody flows past the detection zone.
  • the degree of color change observable in detection zone 3900 is thus related to the amount of analyte in the sample.
  • the subsequent vent is opened and the fluids fills the detection zone 3900, taking with it the resuspended reagent.
  • the subsequent vent is opened and the fluids fills the detection zone 3900, taking with it the resuspended reagent.
  • portions of the walls at either side of the chamber are removed.
  • the reader takes a measurement. Once the measurement is completed the final vent is opened and the fluid fills the sink 4000, washing all the unbound reagent into the well.
  • the assay can be run in a discrete-volume format or in a constant-flow format.
  • discrete amount of fluid would incubate in the detection zone.
  • complexes of the first antibody bound to the analyte would, through diffusion, bind to the functional surface.
  • a downstream vent would be opened and the detection zone would be washed clear of unbound material.
  • constant-flow format fluid would not be incubated in the detection zone, but instead would be allowed to wash past the detection zone slowly.
  • the constant-flow format is similar to traditional lateral flow assays, for example, those using nitrocellulose strips.
  • a functional surface is created on the base 3000, lid 3500, or both.
  • a quasi three-dimensional structure increases the binding capacity of the surface, to allow a lower sensitivity detection scheme to be used.
  • a pillared array is included which increases the surface area in the detection zone, beneficial for the functional surface as well as increasing mixing through lamination.
  • the instrument can determine the quantity of analyte present in the sample using an optical detection of a label.
  • the label is part of the detection reagent.
  • the cartridge can be interrogated in a dark environment to avoid background interference.
  • high gain operational amplifiers can amplify the signal from the optical detector. Any external EMC or RF sources can lead to an increase in noise and a reduction in sensitivity of the signal from the amplification circuit. Therefore, the cartridge can be read in an environment screened from such interference.
  • FIG. 25 shows a bottom view of cartridge 1100, illustrating vent holes 4100 and orientation groove 4200.
  • Orientation groove 4200 can be used in aligning the cartridge
  • FIG. 26 is a photomicrograph showing a portion of assay channel 3200. In particular, portions of deposition and resuspension zone 3800 and detection zone 3900 can be seen, along with wall-removed regions 3850 and 3900.
  • the detection limits of the system can be affected by amplifier gain, noise, and choice of filters; detector area; index matching the lid and detector; use of off-line detection; digital signal processing; the sampling rate during measurement; and the optical design and surface are of the detection zone.
  • FIG. 27 shows time resolved fluorescence results obtained by pipetting 1 ⁇ L of reagent including various amounts of fluorescent beads directly onto a cartridge and closing with a 0.7 mm polymer lid.
  • the detection limit achieved was 10 4 beads above background.

Abstract

La présente invention a trait à un système pour la détection d'un analyte comportant une cartouche de système et une cartouche de lecteur. Le système est automatisé pour une utilisation simple par un patient à domicile, ou par un professionnel de soins de santé. Le système assure la mesure rapide et précise d'un analyte. La cartouche peut comporter un dispositif de dosage microfluidique et peut mesurer un analyte dans un faible volume d'un échantillon liquide.
PCT/US2006/011322 2005-03-29 2006-03-28 Dispositifs de dosage et procedes WO2006105110A2 (fr)

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US66586705P 2005-03-29 2005-03-29
US60/665,867 2005-03-29
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FR2931244A1 (fr) * 2008-05-13 2009-11-20 Biosynex Sarl Dispositif de detection d'au moins un element contenu dans une solution sanguine
EP2304445B1 (fr) * 2008-07-09 2020-06-10 Micropoint Bioscience Inc Cartouche analytique avec réglage du débit de fluide
WO2010070521A1 (fr) 2008-12-18 2010-06-24 Koninklijke Philips Electronics N.V. Dispositif de mesure pour mesurer un fluide
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US9180449B2 (en) 2012-06-12 2015-11-10 Hach Company Mobile water analysis
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WO2014049371A3 (fr) * 2012-09-28 2014-05-22 Agplus Diagnostics Ltd Dispositif d'essai et porte-échantillons
USD768872S1 (en) 2012-12-12 2016-10-11 Hach Company Cuvette for a water analysis instrument
JP2014173934A (ja) * 2013-03-07 2014-09-22 Toshiba Corp 半導体マイクロ分析チップ及びその製造方法
US10073091B2 (en) 2014-08-08 2018-09-11 Ortho-Clinical Diagnostics, Inc. Lateral flow assay device
WO2019025610A1 (fr) * 2017-08-03 2019-02-07 Fibrotx Oü Dosage d'écoulement latéral et dispositif pour application de soins de la peau
JP2020529606A (ja) * 2017-08-03 2020-10-08 フィブロテックス エー スキンケアに適用するための側方流動分析及び装置
EP3714787A4 (fr) * 2017-11-21 2021-03-17 BBB Inc. Biocapteur
CN113167760A (zh) * 2018-12-02 2021-07-23 聚合物技术系统公司 用于电化学测试条带中的电子门特征的系统和方法
CN113167760B (zh) * 2018-12-02 2023-11-03 聚合物技术系统公司 用于电化学测试条带中的电子门特征的系统和方法
US20220155226A1 (en) * 2019-03-28 2022-05-19 National University Corporation Ehime University Optical analysis chip
WO2021146350A3 (fr) * 2020-01-13 2021-11-04 Lumiradx Uk Ltd. Régulation des fluides dans des dispositifs microfluidiques
GB2611504A (en) * 2020-01-13 2023-04-12 Lumiradx Uk Ltd Fluid control in microfluidic devices

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