WO2019099342A1 - Methods and systems for consistent thrombus formation and measurement thereof - Google Patents

Methods and systems for consistent thrombus formation and measurement thereof Download PDF

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Publication number
WO2019099342A1
WO2019099342A1 PCT/US2018/060602 US2018060602W WO2019099342A1 WO 2019099342 A1 WO2019099342 A1 WO 2019099342A1 US 2018060602 W US2018060602 W US 2018060602W WO 2019099342 A1 WO2019099342 A1 WO 2019099342A1
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WIPO (PCT)
Prior art keywords
cartridge
microfluidic
thrombus
biological sample
channel
Prior art date
Application number
PCT/US2018/060602
Other languages
French (fr)
Inventor
Sumit KHETARPALL
David N. Ku
Mick S. BAKER
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Georgia Tech Research Corporation
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Publication of WO2019099342A1 publication Critical patent/WO2019099342A1/en

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    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/56—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving blood clotting factors, e.g. involving thrombin, thromboplastin, fibrinogen
    • 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
    • B01L2300/0816—Cards, e.g. flat sample carriers usually with flow in two horizontal directions
    • 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
    • B01L2300/0864—Configuration of multiple channels and/or chambers in a single devices comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
    • 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
    • B01L2400/0457—Moving fluids with specific forces or mechanical means specific forces passive flow or gravitation

Definitions

  • the present invention relates generally to diagnostic systems and methods, and more specifically to the formation of thrombus in a biological sample and its characterization using an interrogation system.
  • Heart Attack is the leading cause of mortality all across the world. It results from the formation of blood clot inside a blood vessel, obstructing the flow of blood through the circulatory system. Plaque buildup and narrowing of arteries can take decades. However, once the constriction in the arteries becomes greater than fifty percent the risk of a major cardiac event becomes high. Arterial thrombosis - the process that involves going to full occlusion, only takes minutes. Therefore, the prevention of such an event with the optimal drug therapy becomes imperative.
  • Arterial thrombosis depends on two other key factors besides the greater than fifty percent stenosis in the artery. Firstly, presence of necessary bio-chemicals that is made available by a vascular injury in close proximity to the stenosis. The vascular injury can happen because of physical or emotional stress. Secondly, the blood of the patient plays a significant role. Some patient’s blood clots faster, while others with von Willebrand disease as an example, takes a long time.
  • Cardiovascular doctors use anti-platelet drugs like Aspirin, Plavix®, Brillinta®, or Effient® for the treatment of arterial thrombosis.
  • the challenge in creating an optimal therapy protocol is that not every patient responds to a given drug. Further, the drug dosage can be critical - too little drug may not make sufficient impact, while too much may lead to internal bleeding.
  • the present invention has identified several areas to improve repeatability and reproducibility associated with thrombus formation.
  • One of the target product quality requirements is to demonstrate low variability across multiple thrombi formed from a single patient blood draw.
  • An underlying principle of the present invention is to mimic arterial conditions necessary for thrombus formation, while avoiding or minimizing the noise sources.
  • One of the sources of variance in thrombus characterization comes from the pump that is used to generate flow of biological sample. Pump flows rates can vary both spatially and temporally. Further, there is pump-to-pump variation adding to the noise when comparing results of different patients using multiple devices.
  • the present invention uses gravity based pressure head to drive sample flow. Gravity provides the consistency that no pump can match.
  • a representative coronary artery is quarter of an inch in diameter. Reproducing a thrombus of that size under representative flow conditions would requires multiple liters of human blood. Realistic blood draw from a patient can only be in the order of milliliters. This requirement further dictates that the in-vitro arterial environment in the diagnostic device is scaled down via use of microfluidic elements. The challenge with using microfluidic elements is the associated part-to-part variation. The present invention accounts for this by measuring flow rates through each independent channel, and normalizing the final results appropriately.
  • An addition source of variance comes from the biological serums that are used to coat the microfluidic channels to simulate endothelial environment.
  • Some of the typical serums include collagen, fibrin, and vWF. These serums are coated onto the channels using a liquid deposition method, wherein the serum is inserted into the channels and left for a sufficient amount of time. This process leads to random and non-uniform coating.
  • the present invention uses an electrochemical process to deposit the serum inside the desired channel section.
  • the thrombus measurement approach also adds variance in final results.
  • the conventional approach is to use pressure sensors or direct microscopy. Pressure sensors only provide integrated point measurements, and exhibit sensor-to-sensor variation issues.
  • Direct microscopy is expensive, time consuming due to the associated post processing operation, and provides only surface measurements whereas thrombus formation is a volumetric phenomenon. Further, both pressure sensor and microscopy approaches don’t scale well for making measurements at multiple locations simultaneously.
  • the present invention uses an optical detection system involving laser light source and photodiodes.
  • the existing diagnostic devices focus on platelet activation.
  • the present invention tests whole blood in a simulated stenosed artery that leads to lumen occlusion.
  • the system quantifies the propensity of the blood of an individual to form an occlusive thrombus or self-lyse the clot, thus providing a prediction of future atherothrombotic occlusion in individual patients.
  • pathological high shear flow is required to flow over collagen surface.
  • low shear flow may be required to flow over alternate biochemical coating like fibrin.
  • the present invention covers both high and low shear flows with any biochemical coating.
  • the present invention includes systems and methods for testing a fluid sample in a simulated stenosed blood vessel that mimics conditions necessary for thrombus formation, while minimizing if not avoiding sources of error.
  • the present invention can quantify the propensity of the blood of an individual to form an occlusive thrombus or to self-lyse the clot, thus providing meaningful information on future atherothrombotic occlusion and enabling patient’s optimized drug therapy.
  • the present invention demonstrates low variability across multiple thrombi formed from a single patient blood draw.
  • the present invention provides improved repeatability and reproducibility associated with thrombus formation.
  • the present invention minimizes if not eliminates noise sources that if left unmitigated, would present unacceptable error levels in reliably characterizing a patient’s blood.
  • the present invention can use gravity-based pressure head for sample flow.
  • the present invention can measure flow rates through independent channels and normalize the final results using flow rate information.
  • the present invention can use an electrochemical process to deposit the serum inside a desired channel section.
  • the present invention can use an optical detection system involving laser light source and photodiodes.
  • the present invention can use a light blocking mask that allows light in selected regions of the channel where thrombus formation takes place.
  • a system useful in determining functional performance of a biological sample comprises a thrombosis unit in which thrombus formation takes place and an interrogation unit providing information related to the formation of the thrombus.
  • the thrombosis unit can comprise a disposable cartridge and the interrogation unit can comprise a cartridge reader that characterizes thrombus formation in the disposable cartridge.
  • the disposable cartridge can comprise three reservoirs, a fluid interfacing reservoir used to insert the biological sample into the disposable cartridge, an inlet reservoir, and an outlet reservoir, wherein the inlet and outlet reservoirs are used to provide identical gravity-based pressure head to multiple microfluidic channels of the disposable cartridge simultaneously.
  • the disposable cartridge can comprise a plurality of microfluidic channels, each inherently embodying a flow characteristic determined at least in part on the fabrication of each microfluidic channel, wherein the flow characteristic of each of the microfluidic channels is used in order to normalize final thrombus characterization results across each of the microfluidic channels upon operational use of the system.
  • the disposable cartridge can comprise a light blocking mask enabling precise characterization of thrombus formation where light passes through the thrombus and the transmission intensity is measured using a photo diode sensor.
  • the disposable cartridge can comprise a microfluidic cartridge and a cover slip for the microfluidic cartridge, wherein the cover slip comprises a conductive surface enabling the use of an electrochemical process for depositing a bio-chemical.
  • a system useful in determining functional performance of a biological sample comprises a thrombosis unit and an interrogation unit, wherein in the thrombosis unit, the propensity of the biological sample to form an occlusive thrombus is established, wherein the interrogation unit provides information related to the formation of the occlusive thrombus, wherein the information from the interrogation unit correlates to a prediction of future atherothrombotic occlusion, and wherein the thrombosis unit comprises one or more of a plurality of microfluidic channels, each inherently embodying a flow characteristic determined at least in part on the fabrication of each microfluidic channel, wherein the flow characteristic of each of the microfluidic channels is used in order to normalize final thrombus characterization results across each of the microfluidic channels upon operational use of the system, and/or three reservoirs, a fluid interfacing reservoir used to insert the biological sample into the thrombosis unit, an
  • a system useful in determining functional performance of a biological sample comprises a thrombosis unit comprising a plurality of microfluidic channels, each inherently embodying a flow characteristic determined at least in part on the fabrication of each microfluidic channel, wherein the flow characteristic of each of the microfluidic channels is used in order to normalize final thrombus characterization results across each of the microfluidic channels upon operational use of the system, three reservoirs, a fluid interfacing reservoir used to insert the biological sample into the thrombosis unit, an inlet reservoir, and an outlet reservoir, wherein the inlet and outlet reservoirs are used to provide identical gravity-based pressure head to the microfluidic channels simultaneously, a microfluidic cartridge and a cover slip for the microfluidic cartridge, wherein the cover slip comprises a conductive surface enabling the use of an electrochemical process for depositing a bio-chemical in a portion of the microfluidic channels, and a light blocking mask enabling precise characterization of
  • a method useful in determining functional performance of a biological sample comprises forming a thrombus in a thrombosis unit and interrogating the formation of the thrombus.
  • the thrombosis unit can comprise a disposable cartridge, and interrogating the formation of the thrombus can be via a cartridge reader that characterizes thrombus formation in the disposable cartridge.
  • an object of the present invention is to provide thrombosis characteristics for a given biological sample, in exemplary embodiments, a blood fluid sample, and in further exemplary embodiments, a whole blood sample. This is achieved by providing gravity-based pressure head to multiple independent calibrated channels, each of them incorporating a stenosis section. The thrombus formation is measured using a laser and photo-diode based optical system.
  • Another object of the present invention is to provide a method for characterizing thrombus formation using the system of the present invention.
  • FIG. 1 is an exploded view of the present invention according to an exemplary embodiment.
  • FIG. 2 illustrates the invention of FIG. 1 in another form, in an assembly of a cartridge reader containing a disposable cartridge, the cartridge reader designed to measure thrombus formation in the disposable cartridge.
  • FIG. 3 is an exploded view of the cartridge of the present invention according to an exemplary embodiment showing it formed of a pumping cartridge, a cover slip for the pumping cartridge, a microfluidic cartridge, a cover slip for the microfluidic cartridge, and a light blocking mask.
  • FIG. 4 is an isometric view of a pumping cartridge of FIG. 3.
  • FIG. 5 shows isometric views of the pumping cartridge and the cover slip for the pumping cartridge of FIG. 3.
  • FIG. 6 shows isometric views of the microfluidic cartridge of FIG. 3, and an enlarged view of a stenosis section of a channel of the microfluidic cartridge according to an exemplary embodiment of the present invention.
  • FIG. 7 shows isometric views of the microfluidic cartridge and the cover slip for the microfluidic cartridge of FIG. 3, the cover slip for the microfluidic cartridge having a bio- chemical coating according to an exemplary embodiment of the present invention.
  • FIG. 8 illustrates an exemplary way of depositing the bio-chemical coating on the cover slip of the microfluidic cartridge of FIG. 7.
  • FIG. 9 illustrates additional exemplary embodiments of the cartridge of the present invention.
  • Ranges may be expressed as from“about” or“approximately” or“substantially” one value and/or to“about” or“approximately” or“substantially” another value. When such a range is expressed, other exemplary embodiments include from the one value and/or to the other value. [0048] Similarly, as used herein,“substantially free” of something, or“substantially pure”, and like characterizations, can include both being“at least substantially free” of something, or“at least substantially pure”, and being“completely free” of something, or“completely pure”.
  • the term“identical” refers to values that are close to each other within the range of manufacturing tolerances.
  • “Comprising” or“containing” or“including” is meant that at least the named compound, element, particle, or method step is present in the composition or article or method, but does not exclude the presence of other compounds, materials, particles, method steps, even if the other such compounds, material, particles, method steps have the same function as what is named.
  • the characteristics described as defining the various elements of the invention are intended to be illustrative and not restrictive.
  • the material includes many suitable materials that would perform the same or a similar function as the material(s) described herein are intended to be embraced within the scope of the invention.
  • Such other materials not described herein can include, but are not limited to, for example, materials that are developed after the time of the development of the invention.
  • the system includes a thrombosis unit providing sample flow using identical gravity-based pressure head to multiple independent calibrated microfluidic channels simultaneously.
  • the thrombosis unit can comprise an inlet reservoir, an outlet reservoir, a fluid interfacing reservoir, and multiple unconnected channels that are in fluidic communication with either the top or the outlet reservoirs.
  • the thrombosis unit can further include a microfluidic cartridge comprising multiple flow channels allowing a flow loop to be completed between inlet and outlet reservoir.
  • One or more of the channels can feature a stenosis section that mimic narrowing of an artery due to plaque buildup.
  • a system for characterizing thrombus formation can include the thrombosis unit further comprising an integrated light blocking mask.
  • the microfluidic cartridge of the thrombosis unit features the multiple channels where the thrombus formation takes place.
  • the light blocking mask of the thrombosis unit enables precise characterization of thrombus formation in a dedicated setup where laser light passes through the thrombus and the transmission intensity is measured using a photo diode sensor.
  • a method for characterizing thrombus formation is disclosed. The method comprises delivering flow at identical gravity- based pressure head to multiple channels of the thrombosis unit simultaneously, and then using the light blocking mask that only allows light from the stenosis section present in the microfluidic channels.
  • thrombus formation of a fluid sample is investigated in a stenosis section of the invention.
  • a portion of the fluid sample is presented to the stenosis section via a stenosis section flow, and thrombus formation in the stenosis section over time is interrogated.
  • FIG. 1 A simplified schematic of the diagnostic system according to an exemplary embodiment of the present invention 100 is shown in FIG. 1, and comprises a thrombosis unit 200 and an interrogation unit 300.
  • the thrombosis unit 200 presents a system that mimics arterial conditions necessary for thrombus formation.
  • the interrogation system 300 provides precise characterization of thrombus formation in the thrombosis unit 200.
  • the thrombosis unit 200 can comprise a fluidic system capable of thrombosis formation of a patient’s fluid sample in the stenosis section.
  • the term“fluid sample” refers to a biological sample whose thrombosis characteristics are tested using the present invention.
  • the fluid sample can be a blood fluid sample, the blood fluid being whole blood, or platelets, or a biological sample that is an unprocessed fluid directly from a patient, or it might be processed and treated with other chemicals including, but not limited to, anti-coagulants and anti -platelet agents.
  • the thrombosis unit 200 can comprise a cartridge 1000 (preferably a disposable cartridge) having reservoirs and channels. Only a portion of the cartridge 1000 is shown in FIG. 1.
  • the cartridge 1000 comprises a pumping cartridge 1100, a cover slip 1200 for the pumping cartridge, a microfluidic cartridge 1300, a cover slip 1400 for the microfluidic cartridge, and a mask 1500. (See, FIG. 3).
  • the cartridge 1000 comprises three reservoirs in fluidic communication - a fluid interfacing reservoir 1110 where the fluid sample is initially contained, an inlet reservoir 1120 where the fluid sample is staged at the beginning of system operation, and an outlet reservoir 1130 where the fluid sample is collected at the end of system operation.
  • the cartridge 1000 comprises a plurality of inlet and outlet channels, for example, inlet channels 1121, 1122, 1123, and 1124 in fluidic communication with the inlet reservoir 1120 and outlet channels 1131, 1132, 1132, and 1134 in fluidic communication with the outlet reservoir 1130. (See, FIG. 4).
  • the interrogation system 300 can comprise an energy source and a detector.
  • the energy source can comprise a light source 2000 and the detector can comprise a light detection system 6000.
  • the present invention provides thrombus formation in the thrombosis unit 200 via fluid sample flow inside the cartridge 1000 from the inlet reservoir 1120 to the outlet reservoir 1130 through the independent and calibrated channels.
  • One or more of the independent channels has a stenosis section where thrombus formation takes place.
  • Thrombus formation is interrogated via interrogation system 300 preferably being measured using an optical detection system where light transmission intensity through the thrombus changes as thrombus formation evolves.
  • independent channel refers generally to a channel whose flow performance is independent of another channel. Specifically, in the context of the present invention it also implies that the all the independent channels have their own output and input ports.
  • the light source 2000 generates light 3000 that is passed through one or more optical elements 4000 to convert the light into a collimated sheet of light 5000.
  • Light transmission of the evolving thrombosis in the thrombosis unit 200 is measured using the light detection system that can be a photo sensing electronic circuit 6000.
  • the interrogation system 300 can be arranged within a housing 9000. As shown in FIG. 2, the light source 2000, optical element 4000, and light detection system 6000 can reside within cartridge reader 9000. Cartridge reader 9000 measures thrombus formation within the disposable cartridge 1000.
  • the cartridge 1000 can further comprise a reader alignment assembly to ensure proper alignment of the cartridge in the cartridge reader 9000.
  • the reader alignment assembly comprises notch 1090 and a cooperative alignment assembly of the cartridge reader 9000.
  • the notch 1090 for example, a triangular notch, allows the cartridge 1000 to be aligned within the cartridge reader device via the cooperative alignment assembly of the cartridge reader, for example, two rods. One of the rods fits the triangular notch, while the second rod balances the cartridge on the other side of the cartridge bottom plane.
  • Cartridge 1000 can further comprise a cartridge alignment system to align the various components of the cartridge 1000.
  • the pumping cartridge 1100, the cover slip 1200 for the pumping cartridge, the microfluidic cartridge 1300, the cover slip 1400 for the microfluidic cartridge, and the light blocking mask 1500 are aligned via the cartridge alignment system, for example, alignment holes 1271, 1371, 1471, 1571, and corresponding diametrically opposite alignment holes.
  • the cartridge alignment system can comprise alternative ways, and be achieved through other mechanical or optical means including computer vision and microscopy.
  • the thrombosis unit 200 enables thrombus formation in a fluid sample by providing at least one channel having a stenosis section with a gravity-based pressure head.
  • two or more channels having identical flow characteristics are presented with different portions of the fluid sample. Results of thrombus formation in more than one channel are then interrogated and results normalized.
  • the fluid interfacing reservoir 1110 where the fluid sample is presented
  • the inlet reservoir 1120 where the fluid sample is staged at the beginning of operation
  • the outlet reservoir 1130 where the fluid sample is collected at the end of operation.
  • the inlet reservoir is in fluidic communication with one or more inlet channels
  • the outlet reservoir is in fluidic communication with one or more outlet channels.
  • the pumping cartridge 1100 and the cover slip 1200 are separate components, at times the combination of 1100, 1200 may be considered the pumping cartridge.
  • the objective of the pumping cartridge is to provide identical gravity-based pressure head and identical flow resistance to multiple independent channels in the microfluidic cartridge 1300.
  • the element 1100 contains open channels, and the element 1200 acts as a cover slip to close the open channels. While as shown the pumping cartridge cannot function without its cover slip, for the sake of clarity and exposing details conveniently, some illustrations may refer to just the element 1100 as the pumping cartridge.
  • inlet reservoir 1120 is in fluidic communication with multiple inlet channels - 1121, 1122, 1123, and 1124.
  • the outlet reservoir 1130 is in fluidic communication with multiple outlet channels - 1131, 1132, 1132, and 1134.
  • the inlet and outlet channels are discontinuous in the pumping cartridge 1100. The fluidic continuity between inlet and outlet channels is established via the microfluidic cartridge 1300 as illustrated in FIG. 3.
  • the fluid sample is collected from the patient, for example, in a sample test tube with a stopper made of rubber or similar material.
  • the fluid sample can be loaded, for example, via pouring or injecting, into the fluid interfacing reservoir 1110.
  • the test tube filled with the fluid sample can be inserted into the fluid interfacing reservoir that can be cooperatively shaped as a cylindrical reservoir 1110 that houses at least one syringe needle which pierces through the rubber stopper into the fluid sample.
  • the base of the fluid interfacing reservoir 1110 is in communication with an aspiration channel 1141, such that the fluid sample can be routed from the reservoir 1110 to the inlet reservoir 1120 via the aspiration channel 1141.
  • the exemplary embodiment of the pumping cartridge as shown in FIG. 4 contains pressure channels to assist in routing of the fluid sample within the cartridge 1000.
  • a top pressure channel 1142 can be used to apply vacuum for routing fluid sample from the reservoir 1110 to the inlet reservoir 1120 via the aspiration channel 1141.
  • a bottom pressure channel 1145 can used to apply vacuum to initiate fluid sample flow from the inlet reservoir 1120 to the outlet reservoir 1130 via the assembly of the pumping cartridge 1100, the cover slip 1200 for the pumping cartridge, the microfluidic cartridge 1300, and the cover slip 1400 for the microfluidic cartridge.
  • Additional pressure channels, a second top channel 1143 and/or second bottom channel 1144 can be used to provide atmospheric pressure access to the top 1120 and outlet reservoir 1130, respectively.
  • FIG. 5 An exemplary embodiment of the pumping cartridge 1100 with its cover slip 1200 is shown in FIG. 5.
  • the cover slip 1200 encloses otherwise open channels and open reservoirs of the pumping cartridge 1200 while providing inlet ports - 1221, 1222, 1223, 1224 and outlet ports - 1231, 1232, 1233, and 1234.
  • the ports interface on the first hand, with the inlet and outlet ports of the pumping cartridge 1200, and on a second hand, with inlet and outlet of channels of the microfluidic cartridge 1300 as shown in FIG. 3.
  • the cover slip 1200 shown in FIG. 5 provides top pressure ports 1242 and 1243 for fluidic communication with the inlet reservoir 1120, and bottom pressure ports 1244 and 1245 for fluidic communication with the outlet reservoir 1130. The pressure applied on these ports helps route the fluid sample within the cartridge 1000.
  • the pumping cartridge 1100 and the cover slip 1200 for the pumping cartridge are separate components.
  • the split can be beneficial for fabricating closed channels.
  • Alignment of the two components 1100, 1200 to be bonded can be accomplished via alignment holes - 1112, 1113 on the pumping cartridge 1100 and their corresponding mating holes on the cover slip 1200.
  • Alternative alignment systems can be used, including another distinct feature on the pumping cartridge 1100 and a corresponding feature on the cover slip 1200 may be used for aligning.
  • a triangular notch 1115 and the bottom plane of the cartridge maybe used to align cartridge 1100 with cover slip 1200.
  • the alignment may also be achieved through other means including computer vision and microscopy.
  • microfluidic cartridge 1300 and the cover slip 1400 are separate components, at times the combination of 1300, 1400 may be considered the microfluidic cartridge.
  • the microfluidic cartridge contains multiple independent calibrated channels, each featuring their own stenosis section.
  • the element 1300 contains open channels, and the element 1400 acts as a cover slip to close the open channels. While as shown the microfluidic cartridge cannot function without its cover slip, for the sake of clarity and exposing details conveniently, some illustrations may refer to just the element 1300 as the microfluidic cartridge.
  • the microfluidic cartridge 1300 can comprise four independent calibrated channels 1310 that are flowed during device operation.
  • Each of the channels 1310 incorporates an input port 1311 that is used to receive fluid sample from the inlet channel of the pumping cartridge 1100, an output port 1317 that is used to send fluid sample back to the outlet channel of the pumping cartridge 1100, and a stenosis section of the channel comprising a raised stenosis portion 1314 that together with sloping sections 1313 and 1315 mimics the arterial constriction because of plaque buildup.
  • the stenosis section reduces the cross-sectional area of the channel 1310.
  • the channel can employ many shapes, including ovate and rectangular.
  • the depth of the channel at locations 1312, 1315 can be 180 microns, while the depth at the stenosis portion 1314 can be 70 microns.
  • the present invention can comprise a normalization process, where each channel 1310 is measured for flow rate, and the flow rates used to normalize the thrombus formation results so imperfections between each channel 1310 are not as large a factor, or no factor, in results expected in each channel 1310.
  • other methods like microscopy can be used to characterize microfluidic channel flow and used to normalize the thrombus formation results.
  • At least portions of the microfluidic channels 1310 as shown in FIG. 7 have bio- chemicals to simulate the arterial environment post vascular injury.
  • Bio-chemicals can include but are not limited to collagen and vWF.
  • the bio-chemicals can be provided via a coating along the length of entire channel 1310, or on only one or more portions of the channel.
  • a bio-chemical coating 1410 is provided in the stenosis sections including the stenosis portion 1314 together with sloping sections 1313 and 1315 of each channel 1310 via the alignment of the cover slip 1400 for the microfluidic cartridge.
  • the cover slip 1400 for the microfluidic cartridge comprises a conductive surface.
  • the conductive surface enables the use of an electrochemical process for depositing the bio-chemicals like collagen and vWF.
  • a setup for electrochemical deposition of the bio-chemicals is shown in FIG. 8.
  • the bio-chemicals to be coated are placed in a channel 8210 between two parallel conductive layers 1400 and 8100.
  • the cover slip 1400 to be coated with the bio-chemical forms the anode.
  • the opposing parallel surface 8100 forms the cathode.
  • the cathode has an inlet port 8110 and an outlet port 8120 to allow bio-chemical to flow through the channel 8210 where deposition takes place.
  • An insulating spacer 8200 separates the cathode from the anode.
  • the deposition process can be controlled using the magnitude of electrical voltage and time of voltage applied. In an exemplary embodiment 12V was used over a period of 60 minutes.
  • a laser light blocking mask 1500 is shown in FIG. 3.
  • the mask only allows the light from the light source 2000 to pass through the stenosis portion 1314, and the transition sections 1313 and 1315. Blocking the light everywhere except these regions increases the signal-to-noise ratio.
  • the light blocking mask 1500 is an integrated part of the cartridge 1000. In an alternate embodiment, the light blocking mask 1500 can be incorporated into the cartridge reader 9000.
  • the cartridge 1000 illustratively shows four vertical channels between the inlet reservoir 1120 and the outlet reservoir 1130.
  • the channels are designed to provide fluidic communication between the reservoirs 1120 and 1130, and paths to the stenosis sections. Thus, these channels need not be vertically parallel in layout.
  • FIG. 9 shows alternative routes for the channels, where for example, the stenosis section is in a horizontal configuration.
  • the cartridges in these embodiments provide identical gravity-based pressure head and identical flow resistance to the stenosis sections as the fully vertical channels. Further, the stenosis sections may exist as independent channels in a separate microfluidic cartridge similar to details provided in FIG. 3.

Abstract

Systems and methods to measure thrombus formation in a biological sample. The system can include at-least three reservoirs to enable gravity-based pressure head for sample flow, a microfluidic cartridge that contains at least one channel with a pre-measured flow rate, a conductive surface to enable deposition of biological coating via electrochemical process, and a light blocking mask that blocks light from selected regions in the microfluidic cartridge. Also provided are methods for using the system to measure thrombus formation.

Description

METHODS AND SYSTEMS FOR CONSISTENT THROMBUS FORMATION AND
MEASUREMENT THEREOF
BACKGROUND OF THE INVENTION
1. Field of the Invention
[0001] The present invention relates generally to diagnostic systems and methods, and more specifically to the formation of thrombus in a biological sample and its characterization using an interrogation system.
2. Description of Related Art
[0002] Heart Attack is the leading cause of mortality all across the world. It results from the formation of blood clot inside a blood vessel, obstructing the flow of blood through the circulatory system. Plaque buildup and narrowing of arteries can take decades. However, once the constriction in the arteries becomes greater than fifty percent the risk of a major cardiac event becomes high. Arterial thrombosis - the process that involves going to full occlusion, only takes minutes. Therefore, the prevention of such an event with the optimal drug therapy becomes imperative.
[0003] Arterial thrombosis depends on two other key factors besides the greater than fifty percent stenosis in the artery. Firstly, presence of necessary bio-chemicals that is made available by a vascular injury in close proximity to the stenosis. The vascular injury can happen because of physical or emotional stress. Secondly, the blood of the patient plays a significant role. Some patient’s blood clots faster, while others with von Willebrand disease as an example, takes a long time.
[0004] Cardiovascular doctors use anti-platelet drugs like Aspirin, Plavix®, Brillinta®, or Effient® for the treatment of arterial thrombosis. The challenge in creating an optimal therapy protocol is that not every patient responds to a given drug. Further, the drug dosage can be critical - too little drug may not make sufficient impact, while too much may lead to internal bleeding. [0005] There are existing diagnostic devices that characterize blood clot or related bio-markers in an in-vitro setting to further guide physicians in their decision-making process for optimal drug and dosage. However, randomized clinical trials utilizing such devices have not led to any improvements in patient mortality, bringing into question the repeatability and reproducibility of test results with such devices.
[0006] The present invention has identified several areas to improve repeatability and reproducibility associated with thrombus formation. One of the target product quality requirements is to demonstrate low variability across multiple thrombi formed from a single patient blood draw.
[0007] An underlying principle of the present invention is to mimic arterial conditions necessary for thrombus formation, while avoiding or minimizing the noise sources. One of the sources of variance in thrombus characterization comes from the pump that is used to generate flow of biological sample. Pump flows rates can vary both spatially and temporally. Further, there is pump-to-pump variation adding to the noise when comparing results of different patients using multiple devices. The present invention uses gravity based pressure head to drive sample flow. Gravity provides the consistency that no pump can match.
[0008] Another source of variances comes from inherent limitations of the fabrication processes. A representative coronary artery is quarter of an inch in diameter. Reproducing a thrombus of that size under representative flow conditions would requires multiple liters of human blood. Realistic blood draw from a patient can only be in the order of milliliters. This requirement further dictates that the in-vitro arterial environment in the diagnostic device is scaled down via use of microfluidic elements. The challenge with using microfluidic elements is the associated part-to-part variation. The present invention accounts for this by measuring flow rates through each independent channel, and normalizing the final results appropriately.
[0009] An addition source of variance comes from the biological serums that are used to coat the microfluidic channels to simulate endothelial environment. Some of the typical serums include collagen, fibrin, and vWF. These serums are coated onto the channels using a liquid deposition method, wherein the serum is inserted into the channels and left for a sufficient amount of time. This process leads to random and non-uniform coating. The present invention uses an electrochemical process to deposit the serum inside the desired channel section. [0010] The thrombus measurement approach also adds variance in final results. The conventional approach is to use pressure sensors or direct microscopy. Pressure sensors only provide integrated point measurements, and exhibit sensor-to-sensor variation issues. Direct microscopy is expensive, time consuming due to the associated post processing operation, and provides only surface measurements whereas thrombus formation is a volumetric phenomenon. Further, both pressure sensor and microscopy approaches don’t scale well for making measurements at multiple locations simultaneously. The present invention uses an optical detection system involving laser light source and photodiodes.
[0011] The existing diagnostic devices focus on platelet activation. The present invention tests whole blood in a simulated stenosed artery that leads to lumen occlusion. The system quantifies the propensity of the blood of an individual to form an occlusive thrombus or self-lyse the clot, thus providing a prediction of future atherothrombotic occlusion in individual patients. In some scenarios like arterial thrombosis, pathological high shear flow is required to flow over collagen surface. In other scenarios low shear flow may be required to flow over alternate biochemical coating like fibrin. The present invention covers both high and low shear flows with any biochemical coating.
[0012] Thus, it would be beneficial to develop a reliable diagnostic system that can characterize a patient’s blood for optimizing drug therapy, wherein such a system and methods can generate results with high repeatability and reproducibility. It is to such a diagnostic system and method that the present invention is directed.
SUMMARY OF THE INVENTION
[0013] The present invention includes systems and methods for testing a fluid sample in a simulated stenosed blood vessel that mimics conditions necessary for thrombus formation, while minimizing if not avoiding sources of error.
[0014] The present invention can quantify the propensity of the blood of an individual to form an occlusive thrombus or to self-lyse the clot, thus providing meaningful information on future atherothrombotic occlusion and enabling patient’s optimized drug therapy.
[0015] In some scenarios like arterial thrombosis, pathological high shear flow is necessary to flow over collagen surface. In other scenarios low shear flow may be necessary to flow over alternate bio-chemical coating like fibrin. The present invention can consider both high and low shear flows with a bio-chemical coating.
[0016] The present invention demonstrates low variability across multiple thrombi formed from a single patient blood draw. The present invention provides improved repeatability and reproducibility associated with thrombus formation.
[0017] The present invention minimizes if not eliminates noise sources that if left unmitigated, would present unacceptable error levels in reliably characterizing a patient’s blood.
[0018] To overcome conventional pump issues, the present invention can use gravity-based pressure head for sample flow.
[0019] To overcome issues related to conventional fabrication tolerances, the present invention can measure flow rates through independent channels and normalize the final results using flow rate information.
[0020] To overcome issues related to conventional liquid deposition of biological serums, the present invention can use an electrochemical process to deposit the serum inside a desired channel section.
[0021] To overcome issues related to use of pressure sensors or direct microscopy, the present invention can use an optical detection system involving laser light source and photodiodes.
[0022] To overcome issues related to measuring a light transmission signal, the present invention can use a light blocking mask that allows light in selected regions of the channel where thrombus formation takes place.
[0023] In an exemplary embodiment of the present invention, a system useful in determining functional performance of a biological sample comprises a thrombosis unit in which thrombus formation takes place and an interrogation unit providing information related to the formation of the thrombus. The thrombosis unit can comprise a disposable cartridge and the interrogation unit can comprise a cartridge reader that characterizes thrombus formation in the disposable cartridge.
[0024] The disposable cartridge can comprise three reservoirs, a fluid interfacing reservoir used to insert the biological sample into the disposable cartridge, an inlet reservoir, and an outlet reservoir, wherein the inlet and outlet reservoirs are used to provide identical gravity-based pressure head to multiple microfluidic channels of the disposable cartridge simultaneously.
[0025] The disposable cartridge can comprise a plurality of microfluidic channels, each inherently embodying a flow characteristic determined at least in part on the fabrication of each microfluidic channel, wherein the flow characteristic of each of the microfluidic channels is used in order to normalize final thrombus characterization results across each of the microfluidic channels upon operational use of the system.
[0026] The disposable cartridge can comprise a light blocking mask enabling precise characterization of thrombus formation where light passes through the thrombus and the transmission intensity is measured using a photo diode sensor.
[0027] The disposable cartridge can comprise a microfluidic cartridge and a cover slip for the microfluidic cartridge, wherein the cover slip comprises a conductive surface enabling the use of an electrochemical process for depositing a bio-chemical.
[0028] In another exemplary embodiment of the present invention, a system useful in determining functional performance of a biological sample comprises a thrombosis unit and an interrogation unit, wherein in the thrombosis unit, the propensity of the biological sample to form an occlusive thrombus is established, wherein the interrogation unit provides information related to the formation of the occlusive thrombus, wherein the information from the interrogation unit correlates to a prediction of future atherothrombotic occlusion, and wherein the thrombosis unit comprises one or more of a plurality of microfluidic channels, each inherently embodying a flow characteristic determined at least in part on the fabrication of each microfluidic channel, wherein the flow characteristic of each of the microfluidic channels is used in order to normalize final thrombus characterization results across each of the microfluidic channels upon operational use of the system, and/or three reservoirs, a fluid interfacing reservoir used to insert the biological sample into the thrombosis unit, an inlet reservoir, and an outlet reservoir, wherein the inlet and outlet reservoirs are used to provide identical gravity-based pressure head to multiple microfluidic channels of the thrombosis unit simultaneously, and/or a microfluidic cartridge and a cover slip for the microfluidic cartridge, wherein the cover slip comprises a conductive surface enabling the use of an electrochemical process for depositing a bio-chemical, and/or a light blocking mask enabling precise characterization of thrombus formation where light passes through the thrombus and the transmission intensity is measured using a photo diode sensor.
[0029] In another exemplary embodiment of the present invention, a system useful in determining functional performance of a biological sample comprises a thrombosis unit comprising a plurality of microfluidic channels, each inherently embodying a flow characteristic determined at least in part on the fabrication of each microfluidic channel, wherein the flow characteristic of each of the microfluidic channels is used in order to normalize final thrombus characterization results across each of the microfluidic channels upon operational use of the system, three reservoirs, a fluid interfacing reservoir used to insert the biological sample into the thrombosis unit, an inlet reservoir, and an outlet reservoir, wherein the inlet and outlet reservoirs are used to provide identical gravity-based pressure head to the microfluidic channels simultaneously, a microfluidic cartridge and a cover slip for the microfluidic cartridge, wherein the cover slip comprises a conductive surface enabling the use of an electrochemical process for depositing a bio-chemical in a portion of the microfluidic channels, and a light blocking mask enabling precise characterization of thrombus formation where light passes through the thrombus and the transmission intensity is measured using a photo diode sensor, and an interrogation unit, wherein in the thrombosis unit, the propensity of the biological sample to form an occlusive thrombus is established, wherein the interrogation unit provides information related to the formation of the occlusive thrombus, and wherein the information from the interrogation unit correlates to a prediction of future atherothrombotic occlusion.
[0030] In another exemplary embodiment of the present invention, a method useful in determining functional performance of a biological sample comprises forming a thrombus in a thrombosis unit and interrogating the formation of the thrombus. The thrombosis unit can comprise a disposable cartridge, and interrogating the formation of the thrombus can be via a cartridge reader that characterizes thrombus formation in the disposable cartridge.
[0031] Thus, an object of the present invention is to provide thrombosis characteristics for a given biological sample, in exemplary embodiments, a blood fluid sample, and in further exemplary embodiments, a whole blood sample. This is achieved by providing gravity-based pressure head to multiple independent calibrated channels, each of them incorporating a stenosis section. The thrombus formation is measured using a laser and photo-diode based optical system.
[0032] Another object of the present invention is to provide a method for characterizing thrombus formation using the system of the present invention.
[0033] These and other objects, features and advantages of the present invention will become more apparent upon reading the following specification in conjunction with the accompanying drawing figures.
BRIEF DESCRIPTION OF THE DRAWINGS
[0034] The accompanying figures, where like reference numerals refer to identical or functionally similar elements throughout the separate views, and which, together with the detailed description below, are incorporated in and form part of the specification, serve to further illustrate various embodiments and explain various principles and advantages, all in accordance with the present invention;
[0035] FIG. 1 is an exploded view of the present invention according to an exemplary embodiment.
[0036] FIG. 2 illustrates the invention of FIG. 1 in another form, in an assembly of a cartridge reader containing a disposable cartridge, the cartridge reader designed to measure thrombus formation in the disposable cartridge.
[0037] FIG. 3 is an exploded view of the cartridge of the present invention according to an exemplary embodiment showing it formed of a pumping cartridge, a cover slip for the pumping cartridge, a microfluidic cartridge, a cover slip for the microfluidic cartridge, and a light blocking mask.
[0038] FIG. 4 is an isometric view of a pumping cartridge of FIG. 3.
[0039] FIG. 5 shows isometric views of the pumping cartridge and the cover slip for the pumping cartridge of FIG. 3.
[0040] FIG. 6 shows isometric views of the microfluidic cartridge of FIG. 3, and an enlarged view of a stenosis section of a channel of the microfluidic cartridge according to an exemplary embodiment of the present invention. [0041] FIG. 7 shows isometric views of the microfluidic cartridge and the cover slip for the microfluidic cartridge of FIG. 3, the cover slip for the microfluidic cartridge having a bio- chemical coating according to an exemplary embodiment of the present invention.
[0042] FIG. 8 illustrates an exemplary way of depositing the bio-chemical coating on the cover slip of the microfluidic cartridge of FIG. 7.
[0043] FIG. 9 illustrates additional exemplary embodiments of the cartridge of the present invention.
DETAIL DESCRIPTION OF THE INVENTION
[0044] To facilitate an understanding of the principles and features of the various embodiments of the invention, various illustrative embodiments are explained below. Although exemplary embodiments of the invention are explained in detail, it is to be understood that other embodiments are contemplated. Accordingly, it is not intended that the invention is limited in its scope to the details of construction and arrangement of components set forth in the following description or illustrated in the drawings. The invention is capable of other embodiments and of being practiced or carried out in various ways.
[0045] As used in the specification and the appended Claims, the singular forms“a,”“an” and “the” include plural references unless the context clearly dictates otherwise. For example, reference to a component is intended also to include a composition of a plurality of components. References to a composition containing“a” constituent is intended to include other constituents in addition to the one named.
[0046] In describing exemplary embodiments, terminology will be resorted to for the sake of clarity. It is intended that each term contemplates its broadest meaning as understood by those skilled in the art and includes all technical equivalents that operate in a similar manner to accomplish a similar purpose.
[0047] Ranges may be expressed as from“about” or“approximately” or“substantially” one value and/or to“about” or“approximately” or“substantially” another value. When such a range is expressed, other exemplary embodiments include from the one value and/or to the other value. [0048] Similarly, as used herein,“substantially free” of something, or“substantially pure”, and like characterizations, can include both being“at least substantially free” of something, or“at least substantially pure”, and being“completely free” of something, or“completely pure”.
[0049] As used herein, the term“identical” refers to values that are close to each other within the range of manufacturing tolerances.
[0050]“Comprising” or“containing” or“including” is meant that at least the named compound, element, particle, or method step is present in the composition or article or method, but does not exclude the presence of other compounds, materials, particles, method steps, even if the other such compounds, material, particles, method steps have the same function as what is named.
[0051] The characteristics described as defining the various elements of the invention are intended to be illustrative and not restrictive. For example, if the characteristic is a material, the material includes many suitable materials that would perform the same or a similar function as the material(s) described herein are intended to be embraced within the scope of the invention. Such other materials not described herein can include, but are not limited to, for example, materials that are developed after the time of the development of the invention.
[0052] In an exemplary embodiment of the present invention, the system includes a thrombosis unit providing sample flow using identical gravity-based pressure head to multiple independent calibrated microfluidic channels simultaneously. The thrombosis unit can comprise an inlet reservoir, an outlet reservoir, a fluid interfacing reservoir, and multiple unconnected channels that are in fluidic communication with either the top or the outlet reservoirs. The thrombosis unit can further include a microfluidic cartridge comprising multiple flow channels allowing a flow loop to be completed between inlet and outlet reservoir. One or more of the channels can feature a stenosis section that mimic narrowing of an artery due to plaque buildup.
[0053] In another exemplary embodiment of the present invention, a system for characterizing thrombus formation is disclosed. The system can include the thrombosis unit further comprising an integrated light blocking mask. The microfluidic cartridge of the thrombosis unit features the multiple channels where the thrombus formation takes place. The light blocking mask of the thrombosis unit enables precise characterization of thrombus formation in a dedicated setup where laser light passes through the thrombus and the transmission intensity is measured using a photo diode sensor. [0054] In another exemplary embodiment of the present invention, a method for characterizing thrombus formation is disclosed. The method comprises delivering flow at identical gravity- based pressure head to multiple channels of the thrombosis unit simultaneously, and then using the light blocking mask that only allows light from the stenosis section present in the microfluidic channels.
[0055] In an exemplary embodiment of the present invention, thrombus formation of a fluid sample is investigated in a stenosis section of the invention. A portion of the fluid sample is presented to the stenosis section via a stenosis section flow, and thrombus formation in the stenosis section over time is interrogated.
[0056] A simplified schematic of the diagnostic system according to an exemplary embodiment of the present invention 100 is shown in FIG. 1, and comprises a thrombosis unit 200 and an interrogation unit 300. The thrombosis unit 200 presents a system that mimics arterial conditions necessary for thrombus formation. The interrogation system 300 provides precise characterization of thrombus formation in the thrombosis unit 200.
[0057] The thrombosis unit 200 can comprise a fluidic system capable of thrombosis formation of a patient’s fluid sample in the stenosis section. As used herein, the term“fluid sample” refers to a biological sample whose thrombosis characteristics are tested using the present invention. For example, the fluid sample can be a blood fluid sample, the blood fluid being whole blood, or platelets, or a biological sample that is an unprocessed fluid directly from a patient, or it might be processed and treated with other chemicals including, but not limited to, anti-coagulants and anti -platelet agents.
[0058] The thrombosis unit 200 can comprise a cartridge 1000 (preferably a disposable cartridge) having reservoirs and channels. Only a portion of the cartridge 1000 is shown in FIG. 1. In an exemplary embodiment, the cartridge 1000 comprises a pumping cartridge 1100, a cover slip 1200 for the pumping cartridge, a microfluidic cartridge 1300, a cover slip 1400 for the microfluidic cartridge, and a mask 1500. (See, FIG. 3).
[0059] In an exemplary embodiment, the cartridge 1000 comprises three reservoirs in fluidic communication - a fluid interfacing reservoir 1110 where the fluid sample is initially contained, an inlet reservoir 1120 where the fluid sample is staged at the beginning of system operation, and an outlet reservoir 1130 where the fluid sample is collected at the end of system operation. In an exemplary embodiment, the cartridge 1000 comprises a plurality of inlet and outlet channels, for example, inlet channels 1121, 1122, 1123, and 1124 in fluidic communication with the inlet reservoir 1120 and outlet channels 1131, 1132, 1132, and 1134 in fluidic communication with the outlet reservoir 1130. (See, FIG. 4).
[0060] The interrogation system 300 can comprise an energy source and a detector. In an exemplary embodiment, the energy source can comprise a light source 2000 and the detector can comprise a light detection system 6000.
[0061] In operation, the present invention provides thrombus formation in the thrombosis unit 200 via fluid sample flow inside the cartridge 1000 from the inlet reservoir 1120 to the outlet reservoir 1130 through the independent and calibrated channels. One or more of the independent channels has a stenosis section where thrombus formation takes place. Thrombus formation is interrogated via interrogation system 300 preferably being measured using an optical detection system where light transmission intensity through the thrombus changes as thrombus formation evolves.
[0062] As used herein, the term independent channel refers generally to a channel whose flow performance is independent of another channel. Specifically, in the context of the present invention it also implies that the all the independent channels have their own output and input ports.
[0063] In an exemplary embodiment, the light source 2000 generates light 3000 that is passed through one or more optical elements 4000 to convert the light into a collimated sheet of light 5000. Light transmission of the evolving thrombosis in the thrombosis unit 200 is measured using the light detection system that can be a photo sensing electronic circuit 6000.
[0064] The interrogation system 300 can be arranged within a housing 9000. As shown in FIG. 2, the light source 2000, optical element 4000, and light detection system 6000 can reside within cartridge reader 9000. Cartridge reader 9000 measures thrombus formation within the disposable cartridge 1000.
[0065] The cartridge 1000 can further comprise a reader alignment assembly to ensure proper alignment of the cartridge in the cartridge reader 9000. In an exemplary embodiment, the reader alignment assembly comprises notch 1090 and a cooperative alignment assembly of the cartridge reader 9000. The notch 1090, for example, a triangular notch, allows the cartridge 1000 to be aligned within the cartridge reader device via the cooperative alignment assembly of the cartridge reader, for example, two rods. One of the rods fits the triangular notch, while the second rod balances the cartridge on the other side of the cartridge bottom plane.
[0066] An exploded view of an exemplary embodiment of cartridge 1000 is shown in FIG. 3 by way of illustration and not limitation. Cartridge 1000 can further comprise a cartridge alignment system to align the various components of the cartridge 1000. The pumping cartridge 1100, the cover slip 1200 for the pumping cartridge, the microfluidic cartridge 1300, the cover slip 1400 for the microfluidic cartridge, and the light blocking mask 1500 are aligned via the cartridge alignment system, for example, alignment holes 1271, 1371, 1471, 1571, and corresponding diametrically opposite alignment holes. The cartridge alignment system can comprise alternative ways, and be achieved through other mechanical or optical means including computer vision and microscopy.
[0067] The thrombosis unit 200 enables thrombus formation in a fluid sample by providing at least one channel having a stenosis section with a gravity-based pressure head. In an exemplary embodiment, two or more channels having identical flow characteristics are presented with different portions of the fluid sample. Results of thrombus formation in more than one channel are then interrogated and results normalized.
[0068] In an exemplary embodiment of the pumping cartridge 1100 shown in FIG. 4, three reservoirs are incorporated, the fluid interfacing reservoir 1110 where the fluid sample is presented, the inlet reservoir 1120 where the fluid sample is staged at the beginning of operation, and the outlet reservoir 1130 where the fluid sample is collected at the end of operation. The inlet reservoir is in fluidic communication with one or more inlet channels, and the outlet reservoir is in fluidic communication with one or more outlet channels.
[0069] As used herein, while the pumping cartridge 1100 and the cover slip 1200 are separate components, at times the combination of 1100, 1200 may be considered the pumping cartridge. The objective of the pumping cartridge is to provide identical gravity-based pressure head and identical flow resistance to multiple independent channels in the microfluidic cartridge 1300. As shown, the element 1100 contains open channels, and the element 1200 acts as a cover slip to close the open channels. While as shown the pumping cartridge cannot function without its cover slip, for the sake of clarity and exposing details conveniently, some illustrations may refer to just the element 1100 as the pumping cartridge.
[0070] In an exemplary embodiment shown in FIG. 3, inlet reservoir 1120 is in fluidic communication with multiple inlet channels - 1121, 1122, 1123, and 1124. The outlet reservoir 1130 is in fluidic communication with multiple outlet channels - 1131, 1132, 1132, and 1134. The inlet and outlet channels are discontinuous in the pumping cartridge 1100. The fluidic continuity between inlet and outlet channels is established via the microfluidic cartridge 1300 as illustrated in FIG. 3.
[0071] The fluid sample is collected from the patient, for example, in a sample test tube with a stopper made of rubber or similar material. The fluid sample can be loaded, for example, via pouring or injecting, into the fluid interfacing reservoir 1110. In another embodiment, the test tube filled with the fluid sample can be inserted into the fluid interfacing reservoir that can be cooperatively shaped as a cylindrical reservoir 1110 that houses at least one syringe needle which pierces through the rubber stopper into the fluid sample. The base of the fluid interfacing reservoir 1110 is in communication with an aspiration channel 1141, such that the fluid sample can be routed from the reservoir 1110 to the inlet reservoir 1120 via the aspiration channel 1141.
[0072] The exemplary embodiment of the pumping cartridge as shown in FIG. 4 contains pressure channels to assist in routing of the fluid sample within the cartridge 1000. A top pressure channel 1142 can be used to apply vacuum for routing fluid sample from the reservoir 1110 to the inlet reservoir 1120 via the aspiration channel 1141. A bottom pressure channel 1145 can used to apply vacuum to initiate fluid sample flow from the inlet reservoir 1120 to the outlet reservoir 1130 via the assembly of the pumping cartridge 1100, the cover slip 1200 for the pumping cartridge, the microfluidic cartridge 1300, and the cover slip 1400 for the microfluidic cartridge. Additional pressure channels, a second top channel 1143 and/or second bottom channel 1144 can be used to provide atmospheric pressure access to the top 1120 and outlet reservoir 1130, respectively.
[0073] An exemplary embodiment of the pumping cartridge 1100 with its cover slip 1200 is shown in FIG. 5. The cover slip 1200 encloses otherwise open channels and open reservoirs of the pumping cartridge 1200 while providing inlet ports - 1221, 1222, 1223, 1224 and outlet ports - 1231, 1232, 1233, and 1234. The ports interface on the first hand, with the inlet and outlet ports of the pumping cartridge 1200, and on a second hand, with inlet and outlet of channels of the microfluidic cartridge 1300 as shown in FIG. 3. The cover slip 1200 shown in FIG. 5 provides top pressure ports 1242 and 1243 for fluidic communication with the inlet reservoir 1120, and bottom pressure ports 1244 and 1245 for fluidic communication with the outlet reservoir 1130. The pressure applied on these ports helps route the fluid sample within the cartridge 1000.
[0074] As shown in FIG. 5, the pumping cartridge 1100 and the cover slip 1200 for the pumping cartridge are separate components. The split can be beneficial for fabricating closed channels. Alternatively, it is possible to form cartridge 1000 in a single fabrication process and from a unitary component.
[0075] If in two components, they can be bonded together via thermal bonding process. Alternative techniques like laser assisted bonding or solvent assisted bonding may also be utilized. Essential to proper operation, pre- and post-bonding treatment processes cannot leave appreciable particulate matter or debris that may interfere with the thrombus formation process.
[0076] Alignment of the two components 1100, 1200 to be bonded can be accomplished via alignment holes - 1112, 1113 on the pumping cartridge 1100 and their corresponding mating holes on the cover slip 1200. Alternative alignment systems can be used, including another distinct feature on the pumping cartridge 1100 and a corresponding feature on the cover slip 1200 may be used for aligning. In an exemplary embodiment a triangular notch 1115 and the bottom plane of the cartridge maybe used to align cartridge 1100 with cover slip 1200. The alignment may also be achieved through other means including computer vision and microscopy.
[0077] As used herein, while the microfluidic cartridge 1300 and the cover slip 1400 are separate components, at times the combination of 1300, 1400 may be considered the microfluidic cartridge. The microfluidic cartridge contains multiple independent calibrated channels, each featuring their own stenosis section. The element 1300 contains open channels, and the element 1400 acts as a cover slip to close the open channels. While as shown the microfluidic cartridge cannot function without its cover slip, for the sake of clarity and exposing details conveniently, some illustrations may refer to just the element 1300 as the microfluidic cartridge.
[0078] Two views of an exemplary embodiment of the microfluidic cartridge 1300 are shown in FIG. 6. The microfluidic cartridge 1300 can comprise four independent calibrated channels 1310 that are flowed during device operation. Each of the channels 1310 incorporates an input port 1311 that is used to receive fluid sample from the inlet channel of the pumping cartridge 1100, an output port 1317 that is used to send fluid sample back to the outlet channel of the pumping cartridge 1100, and a stenosis section of the channel comprising a raised stenosis portion 1314 that together with sloping sections 1313 and 1315 mimics the arterial constriction because of plaque buildup. The stenosis section reduces the cross-sectional area of the channel 1310. The channel can employ many shapes, including ovate and rectangular. In an exemplary embodiment, the depth of the channel at locations 1312, 1315 can be 180 microns, while the depth at the stenosis portion 1314 can be 70 microns.
[0079] Due to the inherent limitations of the fabrication processes, there are variations in the depth of all channels, and importantly the stenosis section of the channel 1310 when compared from one channel 1310 to another. These variations can impact the final thrombus formation results. Thus, the present invention can comprise a normalization process, where each channel 1310 is measured for flow rate, and the flow rates used to normalize the thrombus formation results so imperfections between each channel 1310 are not as large a factor, or no factor, in results expected in each channel 1310. In alternate embodiments, other methods like microscopy can be used to characterize microfluidic channel flow and used to normalize the thrombus formation results.
[0080] At least portions of the microfluidic channels 1310 as shown in FIG. 7 have bio- chemicals to simulate the arterial environment post vascular injury. Bio-chemicals can include but are not limited to collagen and vWF. The bio-chemicals can be provided via a coating along the length of entire channel 1310, or on only one or more portions of the channel. In one embodiment shown in FIG. 7, a bio-chemical coating 1410 is provided in the stenosis sections including the stenosis portion 1314 together with sloping sections 1313 and 1315 of each channel 1310 via the alignment of the cover slip 1400 for the microfluidic cartridge.
[0081] In another exemplary embodiment of the present invention, the cover slip 1400 for the microfluidic cartridge comprises a conductive surface. The conductive surface enables the use of an electrochemical process for depositing the bio-chemicals like collagen and vWF. A setup for electrochemical deposition of the bio-chemicals is shown in FIG. 8. The bio-chemicals to be coated are placed in a channel 8210 between two parallel conductive layers 1400 and 8100. The cover slip 1400 to be coated with the bio-chemical forms the anode. The opposing parallel surface 8100 forms the cathode. The cathode has an inlet port 8110 and an outlet port 8120 to allow bio-chemical to flow through the channel 8210 where deposition takes place. An insulating spacer 8200 separates the cathode from the anode. The deposition process can be controlled using the magnitude of electrical voltage and time of voltage applied. In an exemplary embodiment 12V was used over a period of 60 minutes.
[0082] Completing the assembly of cartridge 1000, a laser light blocking mask 1500 is shown in FIG. 3. The mask only allows the light from the light source 2000 to pass through the stenosis portion 1314, and the transition sections 1313 and 1315. Blocking the light everywhere except these regions increases the signal-to-noise ratio. In an exemplary embodiment, the light blocking mask 1500 is an integrated part of the cartridge 1000. In an alternate embodiment, the light blocking mask 1500 can be incorporated into the cartridge reader 9000.
[0083] Returning to FIG. 1, the cartridge 1000 illustratively shows four vertical channels between the inlet reservoir 1120 and the outlet reservoir 1130. The channels are designed to provide fluidic communication between the reservoirs 1120 and 1130, and paths to the stenosis sections. Thus, these channels need not be vertically parallel in layout. FIG. 9 shows alternative routes for the channels, where for example, the stenosis section is in a horizontal configuration. The cartridges in these embodiments provide identical gravity-based pressure head and identical flow resistance to the stenosis sections as the fully vertical channels. Further, the stenosis sections may exist as independent channels in a separate microfluidic cartridge similar to details provided in FIG. 3.
[0084] The design and functionality described in this application is intended to be exemplary in nature and is not intended to limit the instant disclosure in any way. Those having ordinary skill in the art will appreciate that the teachings of the disclosure may be implemented in a variety of suitable forms, including those forms disclosed herein and additional forms known to those having ordinary skill in the art.
[0085] While certain embodiments of this disclosure have been described in connection with what is presently considered to be the most practical and various embodiments, it is to be understood that this disclosure is not to be limited to the disclosed embodiments, but on the contrary, is intended to cover various modifications and equivalent arrangements included within the scope of the appended claims. Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.
[0086] This written description uses examples to disclose certain embodiments of the technology and to enable any person skilled in the art to practice certain embodiments of this technology, including making and using any apparatuses or systems and performing any incorporated methods. The patentable scope of certain embodiments of the technology is defined in the claims, and may include other examples that occur to those skilled in the art. Such other examples are intended to be within the scope of the claims if they have structural elements that do not differ from the literal language of the claims, or if they include equivalent structural elements with insubstantial differences from the literal language of the claims.

Claims

What is claimed is:
1. A system useful in determining functional performance of a biological sample comprising: a thrombosis unit in which thrombus formation takes place; and
an interrogation unit providing information related to the formation of the thrombus.
2. The system of Claim 1, wherein the thrombosis unit comprises a disposable cartridge; and
wherein the interrogation unit comprises a cartridge reader that characterizes thrombus formation in the disposable cartridge.
3. The system of Claim 2, wherein the disposable cartridge comprises three reservoirs: a fluid interfacing reservoir used to insert the biological sample into the disposable cartridge;
an inlet reservoir; and
an outlet reservoir;
wherein the inlet and outlet reservoirs are used to provide identical gravity-based pressure head to multiple microfluidic channels of the disposable cartridge simultaneously.
4. The system of Claim 2, wherein the disposable cartridge comprises a plurality of microfluidic channels, each inherently embodying a flow characteristic determined at least in part on the fabrication of each microfluidic channel;
wherein the flow characteristic of each of the microfluidic channels is used in order to normalize final thrombus characterization results across each of the microfluidic channels upon operational use of the system.
5. The system of Claim 2, wherein the disposable cartridge comprises a light blocking mask enabling precise characterization of thrombus formation where light passes through the thrombus and the transmission intensity is measured using a photo diode sensor.
6. The system of Claim 2, wherein the disposable cartridge comprises:
a microfluidic cartridge; and
a cover slip for the microfluidic cartridge;
wherein the cover slip comprises a conductive surface enabling the use of an electrochemical process for depositing a bio-chemical.
7. A system useful in determining functional performance of a biological sample comprising:
a disposable cartridge in which thrombus formation takes place comprising:
a pumping cartridge; and
a microfluidic cartridge; and
a cartridge reader that characterizes thrombus formation in the disposable cartridge.
8. The system of Claim 7, wherein the pumping cartridge comprises three reservoirs:
a fluid interfacing reservoir used to insert the biological sample into the disposable cartridge;
an inlet reservoir; and
an outlet reservoir; wherein the inlet and outlet reservoirs are used to provide identical gravity-based pressure head to multiple calibrated microfluidic channels of the pumping cartridge simultaneously.
9. The system of Claim 7, wherein the pumping cartridge comprises a plurality of calibrated microfluidic channels each having a pre-use flow characteristic determined prior to operational use of the system via provision of identical initial flow conditions presented to each of the calibrated microfluidic channels;
wherein the pre-use flow characteristic of each of the microfluidic channels are used in order to normalize final thrombus characterization results across each of the calibrated microfluidic channels upon operational use of the system.
10. The system of Claim 7, wherein the disposable cartridge further comprises a light blocking mask enabling precise characterization of thrombus formation where light passes through the thrombus and the transmission intensity is measured using a photo diode sensor.
11. A system useful in determining functional performance of a biological sample comprising:
a thrombosis unit; and
an interrogation unit;
wherein in the thrombosis unit, the propensity of the biological sample to form an occlusive thrombus is established;
wherein the interrogation unit provides information related to the formation of the occlusive thrombus;
wherein the information from the interrogation unit correlates to a prediction of future atherothrombotic occlusion; and
wherein the thrombosis unit comprises one or more of:
(i) a plurality of microfluidic channels, each inherently embodying a flow characteristic determined at least in part on the fabrication of each microfluidic channel, wherein the flow characteristic of each of the microfluidic channels is used in order to normalize final thrombus characterization results across each of the microfluidic channels upon operational use of the system; and/or
(ii) three reservoirs, a fluid interfacing reservoir used to insert the biological sample into the thrombosis unit, an inlet reservoir, and an outlet reservoir, wherein the inlet and outlet reservoirs are used to provide identical gravity-based pressure head to multiple microfluidic channels of the thrombosis unit simultaneously; and/or
(iii) a microfluidic cartridge and a cover slip for the microfluidic cartridge, wherein the cover slip comprises a conductive surface enabling the use of an electrochemical process for depositing a bio-chemical; and/or (iv) a light blocking mask enabling precise characterization of thrombus formation where light passes through the thrombus and the transmission intensity is measured using a photo diode sensor.
12. The system of Claim 11, wherein determining functional performance of the biological sample comprises determining a patient’s drug therapy from the biological sample of the patient.
13. The system of Claim 11, wherein determining functional performance of the biological sample comprises determining a patient’s optimized drug therapy for the treatment of arterial thrombosis.
14. The system of Claim 11, wherein the thrombosis unit comprises at least element (i) - a plurality of microfluidic channels, each inherently embodying a flow characteristic determined at least in part on the fabrication of each microfluidic channel, wherein the flow characteristic of each of the microfluidic channels is used in order to normalize final thrombus characterization results across each of the microfluidic channels upon operational use of the system;
wherein a portion of the biological sample passes through each microfluidic channel, each microfluidic channel comprising a stenosis section that mimics narrowing of an artery due to plaque buildup by presenting a smaller cross-sectional area of the microfluidic channel than upstream of the stenosis section.
15. The system of Claim 14, wherein the stenosis section presents a smaller cross-sectional area of the microfluidic channel than downstream of the stenosis section.
16. The system of Claim 15, wherein each microfluidic channel has an inlet at a first pressure and an outlet at a second pressure;
wherein the stenosis section is located between the inlet and the outlet; and
wherein the pressure difference between the inlet and the outlet of each microfluidic channel at least partially imparts biological sample flow through the stenosis section prior to any occlusion at the stenosis section.
17. The system of Claim 16, wherein the thrombosis unit further comprises element (ii) - three reservoirs, a fluid interfacing reservoir used to insert the biological sample into the thrombosis unit, an inlet reservoir, and an outlet reservoir, wherein the inlet and outlet reservoirs are used to provide identical gravity-based pressure head to each microfluidic channel of the thrombosis unit simultaneously.
18. The system of Claim 14, wherein the thrombosis unit further comprises element (iii) - a microfluidic cartridge and a cover slip for the microfluidic cartridge, wherein the cover slip comprises a conductive surface enabling the use of an electrochemical process for depositing a bio-chemical;
wherein the bio-chemical is located at least in a portion of a stenosis section of the microfluidic channels.
19. A system useful in determining functional performance of a biological sample comprising:
a thrombosis unit comprising:
a plurality of microfluidic channels, each inherently embodying a flow characteristic determined at least in part on the fabrication of each microfluidic channel, wherein the flow characteristic of each of the microfluidic channels is used in order to normalize final thrombus characterization results across each of the microfluidic channels upon operational use of the system;
three reservoirs, a fluid interfacing reservoir used to insert the biological sample into the thrombosis unit, an inlet reservoir, and an outlet reservoir, wherein the inlet and outlet reservoirs are used to provide identical gravity-based pressure head to the microfluidic channels simultaneously;
a microfluidic cartridge and a cover slip for the microfluidic cartridge, wherein the cover slip comprises a conductive surface enabling the use of an electrochemical process for depositing a bio-chemical in a portion of the microfluidic channels; and
a light blocking mask enabling precise characterization of thrombus formation where light passes through the thrombus and the transmission intensity is measured using a photo diode sensor; and
an interrogation unit;
wherein in the thrombosis unit, the propensity of the biological sample to form an occlusive thrombus is established;
wherein the interrogation unit provides information related to the formation of the occlusive thrombus; and
wherein the information from the interrogation unit correlates to a prediction of future atherothrombotic occlusion.
20. A system useful in determining functional performance of a biological sample comprising:
a disposable cartridge comprising:
a pumping cartridge;
a cover slip for the pumping cartridge;
a microfluidic cartridge;
a cover slip for the microfluidic cartridge; and
a mask; and a cartridge reader;
wherein the pumping cartridge comprises one or more of:
a plurality of open microfluidic channels, each inherently embodying a flow characteristic determined at least in part on the fabrication of each microfluidic channel, wherein the flow characteristic of each of the microfluidic channels is used in order to normalize final thrombus characterization results across each of the microfluidic channels upon operational use of the system; and/or
three open reservoirs, a fluid interfacing reservoir used to insert the biological sample into the disposable cartridge, an inlet reservoir, and an outlet reservoir, wherein the inlet and outlet reservoirs are used to provide identical gravity-based pressure head to multiple microfluidic channels of the disposable cartridge simultaneously.
21. The system of Claim 20, wherein the cover slip for the microfluidic cartridge comprises a conductive surface enabling the use of an electrochemical process for depositing a bio-chemical.
22. The system of Claim 20, wherein the mask comprises a light blocking mask enabling precise characterization of thrombus formation in the disposable cartridge where light passes through the thrombus and the transmission intensity is measured using a photo diode sensor.
23. The system of Claim 20, wherein the biological sample is staged at the beginning of system operation in the inlet reservoir;
wherein the biological sample is collected at the end of system operation in the outlet reservoir; wherein each of the plurality of microfluidic channels is formed from a discontinuous inlet channel and outlet channel;
wherein each inlet channel is in fluidic communication with the inlet reservoir; and wherein each outlet channel is in fluidic communication with the outlet reservoir.
24. The system of Claim 23, wherein the cover slip for the pumping cartridge comprises: a plurality of inlet ports; and
a plurality of outlet ports;
wherein the cover slip for the pumping cartridge encloses otherwise open microfluidic channels and open reservoirs of the pumping cartridge while providing the inlet and outlet ports; and
wherein the inlet and outlet ports of the cover slip for the pumping cartridge are paired to an individual microfluidic channel of the pumping cartridge, such that an inlet port of the cover slip for the pumping cartridge interfaces with an attendant inlet channel of a microfluidic channel of the pumping cartridge, and an inlet port of the cover slip for the pumping cartridge interfaces with an attendant outlet channel of the same microfluidic channel of the pumping cartridge.
25. The system of Claim 24, wherein the microfluidic cartridge comprises a plurality of independent calibrated channels, each calibrated channel having an inlet and an outlet; and
wherein upon alignment and adjacent assembly of the pumping cartridge, the cover slip for the pumping cartridge, and the microfluidic cartridge, the inlet reservoir is in fluidic communication with the outlet reservoir.
26. The system of Claim 25, wherein upon alignment and adjacent assembly of the pumping cartridge, the cover slip for the pumping cartridge, and the microfluidic cartridge, the microfluidic channels of the pumping cartridge formed of a discontinuous inlet channel and outlet channel embody a continuous flow path from inlet reservoir to outlet reservoir via fluidic communication of the inlet reservoir to the inlet channel of the pumping cartridge, to the inlet port of the cover slip for the pumping cartridge, to the inlet of the independent calibrated channel of the microfluidic cartridge, to the outlet of the independent calibrated channel of the microfluidic cartridge, to the outlet port of the cover slip for the pumping cartridge, to the outlet channel of the pumping cartridge, to the outlet reservoir.
27. The system of Claim 25, wherein each calibrated channel of the microfluidic cartridge further comprises a stenosis section between the inlet and outlet, the stenosis section presenting a smaller cross-sectional area of the calibrated channel than upstream and downstream of the stenosis section.
28. The system of Claim 27, wherein each calibrated channel of the microfluidic cartridge further comprises an upstream sloping section and a downstream sloping section, the stenosis section located between the sloping sections, the sloping sections providing a gradual channel narrowing profile toward and away from the stenosis section.
29. The system of Claim 20, wherein the cartridge reader comprises:
a light source; and
a photo sensing electronic circuit.
30. The system of Claim 29, wherein the cartridge reader further comprises one or more optical elements.
31. The system of Claim 30, wherein the light source is capable of generating light that is passed through the one or more optical elements to convert the light into a collimated sheet of light that upon passing through one or more evolving thrombosis in the disposable cartridge is measured using the photo sensing electronic circuit.
33. A method useful in determining functional performance of a biological sample comprising:
forming a thrombus in a thrombosis unit; and
interrogating the formation of the thrombus.
34. The method of Claim 33, wherein the thrombosis unit comprises a disposable cartridge; and
wherein interrogating the formation of the thrombus is via a cartridge reader that characterizes thrombus formation in the disposable cartridge.
35. The method of Claim 34, wherein the disposable cartridge comprises three reservoirs: a fluid interfacing reservoir used to insert the biological sample into the disposable cartridge;
an inlet reservoir; and
an outlet reservoir;
wherein the method further comprises using the inlet and outlet reservoirs to provide identical gravity-based pressure head to multiple microfluidic channels of the disposable cartridge simultaneously.
36. The method of Claim 34, wherein the disposable cartridge comprises a plurality of microfluidic channels, each inherently embodying a flow characteristic determined at least in part on the fabrication of each microfluidic channel;
wherein the method further comprises using the flow characteristic of each of the microfluidic channels in order to normalize final thrombus characterization results across each of the microfluidic channels upon operational use of the system.
37. The method of Claim 34, wherein interrogating the formation of the thrombus comprises: using a light blocking mask of the disposable cartridge enabling precise characterization of thrombus formation;
passing light through the thrombus; and
measuring the transmission intensity using a photo diode sensor.
38. The method of Claim 34 further comprising using of an electrochemical process for depositing a bio-chemical;
wherein the disposable cartridge comprises:
a microfluidic cartridge; and
a cover slip for the microfluidic cartridge;
wherein the cover slip comprises a conductive surface enabling the use of the electrochemical process for depositing the bio-chemical.
39. A method useful in determining functional performance of a biological sample comprising:
examining the propensity of the biological sample to form an occlusive thrombus; and determining information related to the formation of the occlusive thrombus.
40. A method of determining a patient’s drug therapy comprising:
examining the propensity of a biological sample of the patient to form an occlusive thrombus;
determining information related to the formation of the occlusive thrombus;
correlating the information related to the formation of the occlusive thrombus to a prediction of future atherothrombotic occlusion; and
determining a patient’s drug therapy based upon the correlation.
41. The method of Claim 40, wherein examining the propensity of a biological sample of the patient to form an occlusive thrombus comprises:
passing different portions of the biological sample through a plurality of microfluidic channels; and
mimicking narrowing of an artery due to plaque buildup by passing the portions of the biological sample through a stenosis section of each microfluidic channel presenting a smaller cross-sectional area of the microfluidic channel than upstream of the stenosis section.
42. A method of determining a patient’s drug therapy comprising:
examining the propensity of a biological sample of the patient to form an occlusive thrombus;
determining information related to the formation of the occlusive thrombus;
correlating the information related to the formation of the occlusive thrombus to a prediction of future atherothrombotic occlusion;
minimizing error in the steps of determining and correlating; and
determining a patient’s drug therapy based upon the correlation.
43. The method of Claim 42, wherein minimizing error in the steps of determining and correlating comprises providing gravity-based pressure head for biological sample flow.
44. The method of Claim 42, wherein examining the propensity of a biological sample of the patient to form an occlusive thrombus comprises:
passing different portions of the biological sample through a plurality of microfluidic channels; and
mimicking narrowing of an artery due to plaque buildup by passing the portions of the biological sample through a stenosis section of each microfluidic channel presenting a smaller cross-sectional area of the testing channel than upstream of the stenosis section;
wherein minimizing error in the steps of determining and correlating comprises:
assigning a flow characteristic to each microfluidic channel determined at least in part on the fabrication of each microfluidic channel; and
normalizing the results from the use of the microfluidic channels.
45. The method of Claim 42, wherein examining the propensity of a biological sample of the patient to form an occlusive thrombus comprises:
passing different portions of the biological sample through a plurality of microfluidic channels; and
mimicking narrowing of an artery due to plaque buildup by passing the portions of the biological sample through a stenosis section of each microfluidic channel presenting a smaller cross-sectional area of the testing channel than upstream of the stenosis section;
wherein minimizing error in the steps of determining and correlating comprises using an electrochemical process to deposit a bio-chemical inside a desired location of the microfluidic channels.
46. The method of Claim 42, wherein minimizing error in the steps of determining and correlating comprises:
using a light blocking mask enabling precise characterization of thrombus formation; passing light through the thrombus; and
measuring the transmission intensity using a photo diode sensor.
47. A system as described herein.
48. The system of Claim 47, including each novel feature or combination of features disclosed herein.
49. A method as described herein.
50. The method of Claim 49, including each novel feature or combination of features disclosed herein.
51. A device as described herein.
52. The device of Claim 51, including each novel feature or combination of features disclosed herein.
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