WO2010077784A1 - Cartouches d'essai pour des essais en flux et procédés pour leur utilisation - Google Patents

Cartouches d'essai pour des essais en flux et procédés pour leur utilisation Download PDF

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Publication number
WO2010077784A1
WO2010077784A1 PCT/US2009/067742 US2009067742W WO2010077784A1 WO 2010077784 A1 WO2010077784 A1 WO 2010077784A1 US 2009067742 W US2009067742 W US 2009067742W WO 2010077784 A1 WO2010077784 A1 WO 2010077784A1
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WO
WIPO (PCT)
Prior art keywords
test
well
cartridge
inlet
channel
Prior art date
Application number
PCT/US2009/067742
Other languages
English (en)
Inventor
Gillian Stephens
Patrick Andre
Richard Lumpkin
Jeffery J. Christian
Original Assignee
Portola Pharmaceuticals, Inc.
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 Portola Pharmaceuticals, Inc. filed Critical Portola Pharmaceuticals, Inc.
Publication of WO2010077784A1 publication Critical patent/WO2010077784A1/fr

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502715—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
    • 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
    • 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
    • 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
    • B01L2300/0877—Flow chambers
    • 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
    • B01L2300/0887—Laminated structure
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
    • B01L2400/049—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics vacuum
    • 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
    • B01L3/502723—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 venting arrangements

Definitions

  • the present invention relates generally to devices and methods for performing flow- based biological assays.
  • the present invention relates to a device and method for performing assays on blood or other biological materials flowing through a test cartridge from a source well, through a test channel, and into a collection well.
  • Published PCT Patent Application WO 2006/065739 describes devices and methods for imaging and analyzing thrombosis formation in a test cartridge over time.
  • Figs. 31-3 M describe the use of test cartridges having multiple cylindrical test channels in an optically clear test block or body. In use, the test cartridge is connected to a source of blood, and the blood is drawn through the test cartridge into individual syringes temporarily coupled to each channel.
  • test cartridges described in this commonly owned PCT application are not adapted to safely contain a blood or other biological sample for disposal.
  • the coupling of the test cartridge to the source of test sample and the syringes can in rare instances increase the risk of introducing bubbles or other contaminants into the sample as it passes through the test channels.
  • Another test system for monitoring blood clotting and other biological processes under flow conditions is the BioFluxTM 200 microfluidic flow system for live cell assays, commercially available from Fluxion, South San Francisco, California 94080 (www.fluxionbio.com).
  • the BioFluxTM 200 system uses a microwell plate of generally conventional construction, where a serpentine flow path is micromolded in polydimethylsiloxane (PDMS) onto the plate bottom, connecting pairs of adjacent wells and forming a test channel.
  • PDMS polydimethylsiloxane
  • a microscope coverslip is then secured over the molded flowpath, and blood or other fluids to be tested can be introduced into one well using a pipette or liquid handling workstation and drawn into the other connecting well using an electropneumatic pump.
  • a light source is placed above the plate and a microscope objective is positioned below the plate to view flow through the flowpath. While functional, the design of the micro well test plate has certain disadvantages.
  • test plate presents different thicknesses and shapes for the illumination path necessary to illuminate the sample for quantitative microscopy.
  • product descriptions illustrate open sample and waste wells which make sample containment and disposal more difficult.
  • test cartridges for these reasons, it would be desirable to provide improved test cartridges, test systems, and methods for testing biological fluids by flowing said fluids through test channels.
  • test cartridges which allow for convenient initial introduction of blood or other test fluid into the cartridge with minimum risk of initiating flow until desired.
  • test cartridges which permit the convenient and secure containment of the test sample within the cartridge at all times after initial introduction of the test fluid, including both during the test and after the test is complete.
  • test cartridges which are designed to minimize the risk of bubble and artifact creation within the test fluid as the test is performed. At least some of these objectives will be met by the inventions described hereinbelow.
  • WO 2006/065739 has been described above.
  • the BioFluxTM 200 and its use are described in a Product Data Sheet ⁇ 2008 Fluxion Biosciences, Inc.; a white paper entitled "BioFlux System for Cellular Interactions: Microfluidic Flow System for Live Cell Assays” ⁇ 2008 Fluxion Biosciences, Inc.; and an Application Note entitled Platelet Adhesion: Platelet Aggregation Assays Under Controlled Shear Flow, ⁇ 2008 Fluxion Biosciences, Inc.
  • a platelet thrombosis adhesion assay is described in an Application Note ClOO published by Cellix Ltd., Dublin, Ireland ⁇ 2007 Cellix Ltd., which cites Williams et al. (2002), JAssoc.
  • the Cellix VenaFluxTM system (commercially available from Cellix Ltd.) measures cell adhesion to antibody-coated or endothelial-cell cultured microcapillaries under shear stress conditions mimicking physiological flow.
  • the system requires manual assembly of the plastic fluidic device, and blood and buffer samples are delivered to the device from external reservoirs via connecting tubing. Kantak, et al. (2002, 2003); Martin, et al. (2007); Kastrup, et al. (2007); Shen, et al. (2008a); Shen, et al. (2008b); Runyon, et al. (2008); Gutierrez, et al.
  • Ismagilov R. (2008a), Using microfluidics to understand the effect of spatial distribution of tissue factor on blood coagulation. Thromb Res. 122 Suppl 1 :S27-30; Shen F., Kastrup C, Liu Y., and Ismagilov R. (2008b), Threshold Response of Initiation of Blood Coagulation by Tissue Factor in Patterned Microfluidic Capillaries Is Controlled by Shear Rate. Arterioscler Thromb Vase Biol. 28:000-000 (November 2008 issue).
  • the present invention provides improved test cartridges, systems, methods, and protocols for performing flow-based assays on blood and other biological fluids. While the cartridges and methods are particularly suitable for performing analysis of the kinetics of thrombosis and coagulation, including platelet adhesion, thrombus growth, stability, reversal, and the like, they are also suitable for testing the behavior of constituents of other biological samples, including blood constituents, such as leukocytes, fibrin, and the like, as well as circulating tumor cells. The test cartridges and methods will also be useful for obtaining clinically relevant information for characterizing, analyzing, and predicting the utility of response modifiers as affected by genetic, experimental, and/or pharmacological modulation and/or va ⁇ ation
  • a test cartridge comp ⁇ ses a body having at least one mlet well and at least one collection well
  • a test channel is present in the body and extends from the inlet well to the collection well, thus providing a flow path or a channel suitable for testing and observing the flow of fluids from the mlet well to the collection well, where the flow is typically initiated by a pressure difference initiated between said wells
  • at least one of the wells will be adapted to couple to a pressure source to induce transfer or flow of the test fluid from the mlet well, through the test channel, to the collection well
  • the pressure source may be negative, i e a vacuum or partial vacuum, where it is applied to the collection well to draw the test fluid through the test channel
  • the pressure source may be a positive pressure source where it is applied to the inlet well to cause the test fluid to flow from the inlet well toward the collection well
  • use of the negative pressure will be preferred since it reduces the risk of inadvertent loss of test fluid from the
  • the test cartridge is provided with sealable mlet and collection wells That is, a cover, cap, seal, valve, or other structure is provided which can cover the well to prevent loss of fluid
  • the cover or other structure will be removable du ⁇ ng a portion of the testing procedure
  • the cover over the mlet wells may be removed while the test fluid(s) are transferred into the mlet wells, typically using pipettes, syringes, tubes, pumps, liquid-handling robotics, or other conventional laboratory fluid transfer devices and techniques
  • the mlet wells may also be open or opened du ⁇ ng the fluid transfer phase of the test or protocol to allow connection to a positive pressure source, as discussed above
  • the fluid collection wells may be open du ⁇ ng the fluid transfer portion of a test protocol, typically to allow connection to a negative pressure source, such as a pump, sy ⁇ nge, or the like In most or all cases, however, it will be desirable that after the procedure is complete, both the test in
  • the test cartridges of the present invention may include only a single sealable inlet well, a single sealable collection well, and a single test channel there between. In other embodiments, however, two, three, four, five, or more test channels may be provided in the test cartridge body. Such multiple test channels may be connected to a single, common inlet well and/or common outlet well, but will more usually be connected to individual inlet wells and/or individual outlet wells, where each test channel is connected to one and only one inlet well and/or to one and only one collection well.
  • the cartridge body will be composed at least partially of an optically transparent polymer or other material so that fluid flow through the test channels may be observed using a microscope or other optical equipment.
  • Suitable polymeric materials include polycarbonates, polystyrenes, polyacrylates, and the like.
  • Suitable non-polymeric materials include siliconized glass, and the like.
  • the bodies may be formed from a base and a cover, each having surfaces where the test channels are formed in either or both of the surfaces by machining, etching, laser etching, embossing, molding, or the like.
  • the cover seals against the base thus forming the test channels between the opposed surfaces.
  • seals may be provided around the test channels in order to further prevent leakage.
  • the test channels have surfaces which may be coated, treated, or otherwise formed to interact with the test fluid in some desired manner. For example, for thrombosis testing, the surfaces of the test channels may have an immobilized thrombotic or other substance thereon to initiate thrombosis as blood flows there through.
  • the test channels may have any cross-sectional geometry or size, but will typically have a rectangular cross-section with a width in the range from 0.25 mm to 1.5 mm and a depth in the range from 25 ⁇ m to 500 ⁇ m.
  • the cross-sectional geometry of the test channels will depend on a number of factors including the materials and fabrication techniques. For example, the isotropic etching of glass will typically produce a U-shaped channel, while the anisotropic etching of a negative mold, such as a silicon mold, can provide rectangular channels. Wide, rectangular channels are generally desirable to improve optics as curved surfaces can introduce optical distortion.
  • the wide, shallow channel dimensions above are desirable as they provide a relatively flat velocity profile across the width of the channel when the test fluid flows as a result of a pressure differential.
  • the test channels are preferably linear between their inlet end and their outlet end. Further, the width of the test channel is typically much greater than the depth. Linear channels that are aligned in parallel are preferable since they facilitate optical imaging, and linear, rectangular channels which are wide and shallow in the region of the viewing window are particularly desirable as the resulting fluidics help assure that the volumetric flow rates through the channels are the same and that the flow velocity profile is flat across the width of each channel.
  • the test channels will be formed in an upper surface of the body of the test cartridge where the inlet and outlet wells extend downwardly from the upper surface.
  • the wells are connected to the inlet and outlet ends of the test channels by vertical “chimneys" which connect to the lower end of the inlet well and allow fluid to flow upward to the inlet end of the test channel under a pressure gradient.
  • the outlet end of the test well is connected to the lower end of the collection well by another vertical chimney, allowing the test fluid to flow downwardly through the chimney into the bottom of the collection well.
  • the flow and testing can begin when the differential pressure is applied to the test cartridge above some threshold level as described in more detail below.
  • Test systems comprise test cartridges, generally as described above, in combination with a pressure source having connector(s) capable of being coupled to each of the collection wells and/or inlet wells, preferably where the pressure sources can independently apply a pressure differential across each of the test channels.
  • the pressure source will comprise a plurality of syringes, where the syringes may be connectable to the collection wells in order to draw fluid through the test channels and/or to the inlet wells in order to push fluid through the test channels.
  • the detection system will further include optical or other systems for observing and monitoring the blood or other biological fluids which flow through the test channels. In an exemplary embodiment, the detection systems will include one or more cameras for viewing each test channel.
  • each test channel will have an individual or dedicated camera arranged to observe flow in that channel.
  • the detection system will employ a single camera with optics which allow for observing each of the test channels, usually by sequentially viewing each test channel.
  • Exemplary embodiments usually further include fluorescent emission detection means for observing the accumulation of fluorescent markers within the assay system.
  • methods for analyzing a biological fluid sample comprise introducing at least one sample into an inlet well in a test cartridge.
  • a pressure differential is applied to a first test channel in the test cartridge, where the pressure differential induces flow of the biological fluid sample from the inlet well through the test channel into a collection well in the test channel.
  • the biological fluid sample is analyzed as it flows through and is contained within the test channel, and the test cartridge is disposed of while the biological fluid remains contained and sealed within the test cartridge.
  • the differential pressure may be applied to draw a plurality of fluid samples through a plurality of test channels and into a plurality of collection wells.
  • the differential pressure may be applied at different levels to at least some of said plurality of test channels in order to draw the fluid biological sample at different rates and/or in different amounts through at least some of the test channels.
  • the sample may be initially contained in a single inlet well, but will more usually be provided in different inlet wells, more usually in inlet wells dedicated to particular test channels. In that way, different samples and/or different sample preparations may be provided for each of the test channels.
  • the biological fluid sample may comprise any biological specimen which is a fluid or which may be turned into a fluid or a solution, but will most usually comprise blood or a blood product. The most common analytical test performed will be for thrombus formation, blood coagulation, inflammatory response detection, circulating tumor cell recruitment, and the like.
  • Thrombus formation and blood coagulation may be detected by monitoring platelet accumulation on the thrombogenic surface of the test channel.
  • the analysis will be performed before, during, or after the biological material is caused to flow through the test channel, but will usually be analyzed at least partially while the sample is flowing through the test channel.
  • FIG. 1 is a perspective view of a cartridge for testing biological fluid samples constructed in accordance with the principles of the present invention.
  • Fig 2 is a cross-sectional view of the test cartridge of Fig 1 , taken along line 2-2 of Fig 1, shown with the mlet and collection well seals removed
  • Fig 3 is a top view of the test cartridge of Fig 2, shown with the mlet and collection well seals removed
  • Fig 4 is a cross-sectional view taken along line 4A-4A of Figs 2 and 3, illustrating the cross-sections of the test channels
  • Fig 4A is a detailed view of a single test channel taken along line 4A-4A of Fig 4
  • Fig 5 is a schematic view illustrating the optics of a test system of the present invention DETAILED DESCRIPTION OF THE INVENTION
  • a test cartridge 10 constructed in accordance with the principles of the present invention comp ⁇ ses a body 12 having a central bridge section 14, an mlet well section 16, and a collection well section 18
  • the test cartridge 10 includes three generally axial test channels 20a, 20b, and 20c, respectively
  • Each of the test channels 20 is connected to a single mlet well 22 formed in the mlet well section 16, with a cross-section of mlet well 22b illustrated in Fig 2
  • each of the test channels 20 will be connected to a single collection well 24, with collection well 24b connected to test channel 20b illustrated in the cross-sectional view of Fig 2
  • the test cartridge body 12 may have an lnverted-U profile with the central b ⁇ dge section 14 defining an upper surface 30 with the test channels 20 being exposed through an optically transparent portion thereof
  • the mlet wells 22 depend from one side of the central b ⁇ dge section 14 in the vertical mlet well section 16
  • the collection wells 24 depend from the central b ⁇ dge section 14 m the vertical collection well section 18
  • the top surface 30 of the central b ⁇ dge section 14 is best illustrated in Fig 3 where the test channels 20 are observable through the optically transparent window
  • the mlet openings 32 provide an open passage, as best seen in Fig 2, for allowing the blood or other biological test fluid to be introduced into the well, typically using a sy
  • sealable tops allow the openings 32 to be sealed to prevent loss to the biological fluid after the fluid has been introduced.
  • the tops will be vented to allow air to enter the inlet wells 22 as the fluid is being drawn through the test channels using a vacuum applied to the collection wells, as described below.
  • the sealable cap will be open to allow a syringe or other positive pressure source to be connected to the inlet openings 32 in order to drive or push the test fluid through the system.
  • the outlet openings 34 will typically be configured to facilitate coupling to a bank of three syringes or other vacuum source. As best illustrated in Fig. 2, the openings 34 may comprise a conical passage through the top surface 30 as well as a raised flange portion 38 surrounding the conical opening.
  • Fluid from the inlet wells 22 passes through the test channels 20 through a vertical "chimney" 40, as best seen in Fig. 2 where chimney 40b is connected to inlet well 22b by a short horizontal passage 42b.
  • chimney 40b is connected to inlet well 22b by a short horizontal passage 42b.
  • Similar chimneys 40 will be provided for each of the inlet wells (although for ease of illustration, they are not shown in the drawings).
  • the collection wells 24 are similarly connected to the test channels 20 by vertical chimneys 50, as best shown in Fig. 2.
  • vertical chimney 50b is connected to the bottom of collection well 24b by the short horizontal connector passage 52b and into the bottom of the collection well 24b.
  • the fluids will be drawn through the system by applying a vacuum to the openings 34 of the test cartridge body 12.
  • the use of the vertical chimneys allows the fluid initially held in the inlet wells 32 to be transferred from the bottom of the wells to the test channels which are generally aligned with the tops of the wells. Taking the fluids from the bottoms of the wells reduced the risk of bubbles entering the test channel and maximizes the amount of the fluid which may actually be accessed and drawn into the test wells.
  • Coupling elements 60a, 60b, and 60c may be provided on the outlet openings 34 of the collection wells 24.
  • the couplings may take a variety of configurations, and in some instances may be included in a single pad or foot structure, in order to facilitate interface of the collection wells with a syringe or other vacuum structure for drawing blood or other test fluids into the test channels.
  • the syringes may be independently controlled, in which case different volumes and/or flow rates may be drawn through the test channels.
  • the syringes may be ganged together so that the flow volumes and flow rates are identical in each test channel.
  • the test channels 20 may be formed by etching, machining, molding or otherwise shaping into a surface of the test body cartridge 12.
  • the central bridge section 14 may comprise a lower component 66 and an upper plate 68.
  • Test channels may be formed in an upper surface of the lower portion 66, where the test channels are then closed by placement of the upper plate 68.
  • the test channels are preferably shallow with a width w in the range from 0.25 mm to 1.5 mm and a depth d in the range from 25 ⁇ m to 500 ⁇ m. The use of wide, shallow test channels is advantageous in a number of respects, as discussed above.
  • the inlet wells will typically have a maximum volume in the range from 2 ml to 5 ml, and the collection wells will typically have a volume in the range from 2 ml to 5 ml.
  • the actual volumes of the inlet wells and outlet wells will be determined by the particular assay protocol. For example, a specific volumetric flow rate may be needed to achieve a desired shear rate, so that the volume requirements are dependent on the duration of the assay and the dimensions of the test channels as well as the flow rates, which in turn are dependent on the magnitude of the applied pressure differential.
  • test channels are disposed in an axial direction and are visible from the upper surface of the central bridge section 14. Often, reinforcing structure will be provided on the lower surface of the central bridge section 14, although no reinforcement is illustrated for the sake of simplicity in the drawings.
  • FIG. 5 an optical system 70 for observing a fluorescent tag being accumulated within a test channel 20 in the test cartridge 10 of the present invention will be described.
  • the test cartridge 10, a portion of which is shown in Fig. 5, is held in a movable relationship with the remainder of the optical test system 70.
  • the test cartridge 10 will be held on a movable X-Y platform so that it can be positioned relative to an objective lens 72 of the system 70.
  • a light source such as a light-emitting diode (LED) 74 (or laser source) is arranged to deliver light through the objective lens 72, for example by focusing the light through a plano-convex lens 76, then through an excitation filter 78 and then onto a dichroic mirror, which reflects the light downward through the objective lens 72 and illuminates the fluorescent label which has accumulated in the test channel 20, typically as a result of a biological action such as incorporation of fluorescently labeled materials flowing through the test channel into a thrombus as it forms on the surface of the test channel.
  • the light excites the fluorescent label, causing the fluorescent label to fluoresce and emit longer wavelength photons which are collected by the objective lens 72 and passed through the dichroic mirror 80.
  • the fluorescent light may be detected by a camera 82 after it passes through an emission filter 84. Suitable systems and chemistries employing these general principles are described in commonly-owned PCT application WO 2006/065739, which has been previously incorporated herein by reference.

Abstract

La présente invention concerne une cartouche d'essai pour effectuer des essais en flux contrôlé ou autres sur du sang ou d'autres échantillons biologiques qui comprend un corps ayant au moins un puits d'entrée scellable et au moins un puits de collecte scellable. Un canal d'essai s'étend entre le puits d'entrée scellable et le puits de collecte scellable, et l'un ou les deux puits est/sont adapté(s) pour couplage à une source de pression pour transférer un fluide d'essai du puits d'entrée vers le puits de collecte. La cartouche d'essai peut être utilisée dans des systèmes pour surveiller des réactions biologiques telles que la coagulation sanguine ou la formation de thrombose qui se produisent dans le canal d'essai lorsque le fluide biologique s'écoule dans celui-ci.
PCT/US2009/067742 2008-12-15 2009-12-11 Cartouches d'essai pour des essais en flux et procédés pour leur utilisation WO2010077784A1 (fr)

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US12263408P 2008-12-15 2008-12-15
US61/122,634 2008-12-15

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WO2015102726A3 (fr) * 2013-10-16 2015-10-29 President And Fellows Of Harvard College Dispositif microfluidique pour la surveillance clinique en temps réel et l'évaluation quantitative de la coagulation du sang total
US11042018B2 (en) 2017-07-20 2021-06-22 The University Of Bristol Microfluidics analysis system
AT525192A1 (de) * 2021-06-15 2023-01-15 Genspeed Biotech Gmbh Mikrofluidischer chip

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