WO2014049116A1 - Système microfluidique et procédé pour la distribution d'un échantillon d'un fluide corporel à un système d'analyse en utilisant le système microfluidique - Google Patents

Système microfluidique et procédé pour la distribution d'un échantillon d'un fluide corporel à un système d'analyse en utilisant le système microfluidique Download PDF

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Publication number
WO2014049116A1
WO2014049116A1 PCT/EP2013/070180 EP2013070180W WO2014049116A1 WO 2014049116 A1 WO2014049116 A1 WO 2014049116A1 EP 2013070180 W EP2013070180 W EP 2013070180W WO 2014049116 A1 WO2014049116 A1 WO 2014049116A1
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Prior art keywords
chamber
opening
microfluidic
channel
blood
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PCT/EP2013/070180
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English (en)
Inventor
Piotr Garstecki
Marcin Izydorzak
Kamil PRUSAK
Adam WARCHULSKI
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Pz Cormay S.A.
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Publication of WO2014049116A1 publication Critical patent/WO2014049116A1/fr

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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
    • B01L3/502753—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/483—Physical analysis of biological material
    • G01N33/487—Physical analysis of biological material of liquid biological material
    • G01N33/49—Blood
    • G01N33/491—Blood by separating the blood components
    • 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
    • B01L2400/0409—Moving fluids with specific forces or mechanical means specific forces centrifugal forces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics

Definitions

  • the invention relates to a microfluidic system and a method for delivery of a sample of body fluid to an analysing system with the use of the microfluidic system.
  • the invention relates especially to biochemical analyses of blood.
  • a blood sample is collected from a patient, then blood cells are separated from the blood plasma (or serum) and finally appropriate biochemical testing is performed on serum/plasma samples.
  • Typical ampoule lengths are in the order of a few centimetres with typical volumes ranging from a few hundred microliters to a few mililiters. It is also possible to collect a drop of capillary blood from a patient's finger to a capillary wetted by blood. In such a procedure, blood is collected to the capillary and fills appropriately prepared ampoule connected to the capillary. The ampoule is disconnected after the collection is completed.
  • Blood cells are usually separated from blood plasma/serum by centrifugation of blood contained in a vessel (e.g., in the ampoule mentioned in the paragraph above) in a centrifuge. As a result of centrifugation, blood cells are deposited at the bottom of the ampoule, with plasma (or serum) above.
  • the ampoules so prepared are placed in stands, essentially in vertical position, and analysed in biochemistry analysers.
  • Biochemistry analysers are advanced automated devices used to examine chemical composition of blood. For instance, contents of glucose, lipids (e.g. cholesterol, triglycerides), enzymes or ions in blood are assayed this way. Using a set of needles and a specially designed mechanism, an analyser collects from an ampoule a serum (or plasma) sample, mixes it with appropriately selected reagents in a small cuvette, as a result of which the concentration of components of interest in blood can be determined, for example by means of a photometrical analysis of the products of chemical reaction.
  • lipids e.g. cholesterol, triglycerides
  • enzymes or ions in blood are assayed this way.
  • an analyser collects from an ampoule a serum (or plasma) sample, mixes it with appropriately selected reagents in a small cuvette, as a result of which the concentration of components of interest in blood can be determined, for example by means of a photometrical analysis of the products of chemical reaction.
  • Biochemistry analysers so constructed and capable of carrying out the above- described test procedure are commercially available (e.g., Flexor XL analysers manufactured by ELITech), and disclosed in numerous patents and patent applications (e.g., publications JP 8211072 A, JP 10132735 A, JP 201033924 A, US 20040185549 Al, US20050014274 Al, US 4808380, US 6162399 or US20070065945 Al).
  • a separate group of analysers are instruments operated with disposable discs (e.g., commercially available Abaxis Piccolo ® Xpress and Samsung IVD-A10A models), containing reagents that are necessary to carry out diagnostic reactions (existing instruments use reagents in a freeze-dried form) and allowing for the use of the necessary biological material.
  • disposable discs e.g., commercially available Abaxis Piccolo ® Xpress and Samsung IVD-A10A models
  • reagents that are necessary to carry out diagnostic reactions existing instruments use reagents in a freeze-dried form
  • diagnostic reactions existing instruments use reagents in a freeze-dried form
  • Dispensing in these devices is performed with the use of centrifugal force which is supposed to assure uniform spreading out of biological material and solvent to places where the freeze-dried reagents are stored and reactions take place.
  • the fundamental limitations of the accuracy of measurements performed with the disc technology-based analysers are related to reproducibility of manufacture of small portions of reagents in the freeze-dried form and reproducibility of dispensing.
  • the process of obtaining blood serum (or blood plasma) needed for medical tests and subsequent delivery of the serum (or plasma) to an analyser is complex, quite invasive for a patient and requires many operations, including filling an ampoule, ampoule centrifugation, and finally multiple collections of serum (or plasma) from the ampoule for consecutive assays.
  • the presently existing procedures require performing several operations related to transporting of blood and serum (or plasma) samples between different vessels and/or hydraulic tubing, often involving manual or mechanical operations needed to make hydraulic connections between these vessels and/or tubing.
  • Reduction of the number of operations on blood and serum (or blood plasma) samples would be extremely advantageous in view of simplification of the procedure and minimisation of possibility of errors and contaminations.
  • Simplification of the procedure of transfer of blood plasma (or serum) to the analyser results also in shortening of time from blood collection from the patient to appropriate analysis.
  • a microfluidic system for delivery of a sample of body fluid, in particular blood, to an analysing system comprising the first chamber connected to the first opening and the second chamber, whereas the chambers are interconnected and connected to the third channel having the third opening, and in addition the first chamber is connected to the fourth channel and the fifth channel, having the fourth and the fifth openings, while the second chamber is connected to the second channel having the second opening, is characterized in that the connection between the first chamber and the second chamber is a constriction.
  • the constriction between the first chamber and the second chamber is a microfluidic channel.
  • the third channel is connected to the constriction, preferably to a microfluidic channel, between the first chamber and the second chamber.
  • the second, third, and fourth openings are arranged so that after placing correctly the microfluidic system in a centrifuge rotor, they are closer to the axis of rotation of the centrifuge rotor than the first opening.
  • connection between the fourth and the fifth channels is closer to the centrifugation axis than the first opening.
  • the third and the fourth channels connect to the first chamber or to the constriction at an acute angle with respect to the direction of the centrifugal force F cen tr, which centrifugal force appears when the microfluidic system is placed correctly in a centrifuge rotor and centrifuged.
  • the widths of the second, third, fourth, fifth, and microfluidic channels are from 0.4 mm to 1 mm, preferably 1 mm.
  • the cross-sections of the second, third, fourth, fifth, and microfluidic channels have the shape of a closed figure, preferably a trapezium, square, rectangle, circle or ellipse.
  • the first chamber and the second chamber are arranged so that after placing correctly the microfluidic system in the centrifuge rotor, the first chamber is closer to the rotation axis, and the second chamber is further away from the rotation axis.
  • the ratio of the volume of the first chamber to that of the second chamber is 2:1, 3:2, 1:1, 2:3, or 1:2.
  • the chamber volume ratio may be set arbitrarily, depending on the population's biological variability and the intended yield.
  • the second, third, fourth, fifth, and microfluidic channels comprise internal surfaces coated with substances which interact with a blood component, preferably selected from the group including: EDTA (salt of the ethylenediaminetetraacetic acid), trisodium citrate, heparinates (preferably in the form of sodium, lithium or ammonium salt), heparin, hirudin, potassium oxalate, sodium fluoride, iodoacetate, thrombin inhibitors (preferably ppack or agratroban), silica, kaolin, glass particles, diatomaceous earth, thrombin-based agents, and ellagic acid.
  • EDTA salt of the ethylenediaminetetraacetic acid
  • trisodium citrate heparinates (preferably in the form of sodium, lithium or ammonium salt)
  • heparinates preferably in the form of sodium, lithium or ammonium salt
  • heparin preferably in the form of sodium, lithium or ammonium salt
  • the present invention relates also to a method for delivery of a body fluid sample to an analysing system, with the use of the microfluidic system described above, the method characterised in that it comprises the following steps: a) body fluid is collected to the first chamber and the second chamber of the microfluidic system, b) optionally the body fluid is centrifuged in the microfluidic system, which results in its separation into constituents, and c) a constituent is pushed out through an opening in a centrifuge rotor to an analyser by pumping oil through the microfluidic system.
  • the above-described microfluidic system which has a volume from 50 ⁇ to 150 ⁇ , preferably 50 ⁇ , 100 ⁇ or 150 ⁇ .
  • the centrifugation speed ranges from 3000 to 6000 rpm, preferably 3000, 4000 or 6000 rpm.
  • the centrifugation lasts from 30 to 90 seconds, preferably 30, 60 or 90 seconds.
  • volume ratio of both chambers can be changed arbitrarily depending on patient's sex and age (hematocrit varies with age and sex).
  • Fig. 1 shows schematically a microfluidic system (chip) with the volume of 50 ⁇ , according to the invention in a preferred embodiment
  • Fig. 2 shows schematically the upper layer of the microfluidic system shown in Fig. 1;
  • Fig. 3 shows schematically the lower layer of the microfluidic system shown in Fig. 1;
  • Fig. 4 shows a scheme of the microfluidic system from Fig. 1 filled with a blood sample
  • Fig. 5 shows a scheme of the microfluidic system from Fig. 4 after completed centrifugation and separation of the blood cells from serum;
  • Fig. 6 shows schematically a microfluidic system with the volume of 100 ⁇ , according to the invention in a preferred embodiment
  • Fig. 7 shows a scheme of the microfluidic system from Fig. 6 filled with a blood sample
  • Fig. 8 shows a scheme of the microfluidic system from Fig. 7 after completed centrifugation and separation of the blood cells from serum;
  • Fig. 9 shows schematically a microfluidic system with the volume of 150 ⁇ , according to the invention in a preferred embodiment
  • Fig. 10 shows a scheme of the microfluidic system from Fig. 9 filled with a blood sample
  • Fig. 11 shows a scheme of the microfluidic system from Fig. 10 after completed centrifugation and separation of the blood cells from serum.
  • an arrow indicates the direction of action of the centrifugal force F centr , 01 - the first opening used for introducing blood into the system, 02, 03- the third and fourth (venting) openings used also for introducing the fluid which pushes out the serum (or plasma) from the first chamber, 05 - the fifth opening for delivering serum (or plasma) to an analyser, 04 - the second opening venting the chambers, Zl- the first chamber, Z2- the second chamber, Kl- the first channel, connecting the first opening 01 with the first chamber Zl, K3- the third channel, connecting the first and the second chamber with the third opening, K4 - the fourth channel, connecting the first chamber with the fifth channel, K5- the fifth channel, connecting the fourth channel with the fourth and fifth openings, K6 - the second channel, connecting the second opening with the second chamber, K2- the microfluidic channel connecting the first chamber with the second chamber.
  • Example 1 sample delivery using a microfluidic system (chip) with a volume of 50 ⁇
  • a two-layer microfluidic chip shown in Fig. 1 - 5 has been manufactured, with the lower layer comprising a channel draining the blood plasma (serum) from the system to an analyser through the fifth opening 05, other channels and chambers - the first chamber Zl and the second chamber Z2 - for blood sample.
  • the upper plate comprises four openings: the first 01, the third 02, the fourth 03, and the second 04, including the first opening 01 allowing for the supplying of blood sample directly from a patient's finger.
  • the first opening 01 can be adapted to install capillaries or a funnel to make blood collection easier.
  • the fifth opening 05 used to supply plasma (or serum) to an analyser, can optionally be adapted to install capillaries to facilitate the process of fluid transfer.
  • each of the openings can be used for blood sample collection, in addition each of the openings can be arranged both in the upper and the lower plate, as well as at the external edge formed by joined plates. It is important that the third opening 02, together with the third channel K3, are used after centrifugation is completed to push out the serum (or plasma) obtained in centrifugation, and that the fifth opening 05, together with the fifth channel K5, are used to supply the serum to an analyser - at that time the first opening 01 and the second opening 04 should remain closed.
  • the inlet of the microfluidic channel K2 to the second chamber Z2 should take into account biological variability of the material being collected so that after separation the blood cells are below that inlet. Both layers of the said system have been joined permanently so as to maintain the necessary tightness of the entire microfluidic system.
  • the volume of the microfluidic system in the discussed embodiment is about 50 ⁇ , and the third K3, fourth K4, fifth K5, second K6 and microfluidic K2 channels inside the chip are 1 mm wide. It is to be noted, however, that channels with the widths of 0.9 mm, as well as 0.8 mm, as well as 0.7 mm, as well as 0.6 mm, as well as 0.5 mm, and also 0.4 mm will fulfil their roles in the system.
  • the channels' cross sections can have the shape of any closed figure, and in particular of a trapezium, square, rectangle, circle, ellipse, etc.
  • Channels of the chip may comprise internal surfaces coated with substances which are desirable in view of the final outcome of the blood separation.
  • substances may be used to counteract blood coagulation and they include, by way of example and in a non- limiting manner: EDTA (salt of the ethylenediaminetetraacetic acid), trisodium citrate, heparinates (sodium, lithium or ammonium salt of heparin), hirudin, potassium oxalate, sodium fluoride, iodoacetate or thrombin inhibitors, (e.g., ppack or agratroban).
  • the substances may also be used to activate blood coagulation, for instance they may be: silica, kaolin, glass particles, diatomaceous earth, thrombin-based agents or ellagic acid.
  • microfluidic system - according to the invention - is used as follows:
  • the system discussed here can be filled by an adult person in about 30 seconds, whereas the amount of blood so collected and sufficient for performing analyses is as little as 50 ⁇ , i.e. about ten times less than for traditional syringes or blood collection ampoules mentioned in the introduction.
  • the centrifuge rotor comprises a well that is adapted to the size and shape of the chip.
  • the rotor is made of aluminium (or other suitable material - e.g., ABS, polyacetal (POM), polystyrene or polyamide 66 with glass fibre filler).
  • the blood separates into plasma or serum, and blood cells, whereas the plasma/serum is located closer to the axis of rotation in the first chamber Zl with the volume of 20 ⁇ , and the blood cells - further away from the axis of rotation in the second chamber Z2 with the volume of 30 ⁇ .
  • the direction of the centrifugal force during centrifugation is indicated with an arrow in Fig. 1 - 3.
  • the ratio of the chambers' volumes can be changed arbitrarily, depending on patient's sex and age (the hematocrit varies depending on age and sex).
  • the rotor and the chip comprise openings allowing for the supplying of oil through a hydraulic system that is used for dispensing plasma portions to an analysing system. Pumping oil into the microfluidic system (e.g., through the third opening 02 or the fourth opening 03) results in pushing out a portion of plasma therefrom. The plasma flows out through an opening in the rotor.
  • Centrifugation should be performed setting appropriate time and rotational speed values. Centrifugation parameters - time and rotational speed, e.g., 3 minutes at 8000 RPM (revolutions per minute) - are known to persons skilled in the art of diagnostic testing.
  • Example 2 sample delivery using a microfluidic system (chip) with the volume of 100 ⁇
  • a two-layer microfluidic chip shown in Fig. 6 - 8 has been manufactured, with the lower layer comprising a channel draining the blood plasma (serum) from the system to an analyser through the fifth opening 05, other channels and chambers - the first chamber Zl and the second chamber Z2 - for blood sample.
  • the upper plate comprises four openings (the first 01, the third 02, the fourth 03, and the second 04), including the first opening 01 allowing for the supplying of blood sample directly from a patient's finger.
  • the first opening 01 can be adapted to install capillaries or a funnel to make blood collection easier.
  • the fifth opening 05 used to supply plasma (or serum) to an analyser, can optionally be adapted to install capillaries to facilitate the process of fluid transfer.
  • each of the openings can be used for blood sample collection, in addition each of the openings can be arranged both in the upper and the lower plate, as well as at the external edge formed by joined plates. It is important that the third opening 02, together with the third channel K3, are used after centrifugation is completed to push out the serum (or plasma) obtained in centrifugation, and that the fifth opening 05, together with the fifth channel K5, are used to supply the serum to an analyser - at that time the first opening 01 and the second opening 04 should remain closed.
  • the inlet of the microfluidic channel K2 to the second chamber Z2 should take into account biological variability of the material being collected so that after separation the blood cells are below that inlet. Both layers of the said system have been joined permanently so as to maintain the necessary tightness of the entire microfluidic system.
  • the volume of the microfluidic system in the discussed embodiment is 100 ⁇ , and the third K3, fourth K4, fifth K5, second K6 and microfluidic K2 channels inside the chip are 1 mm wide. It is to be noted, however, that channels with the widths of 0.9 mm, as well as 0.8 mm, as well as 0.7 mm, as well as 0.6 mm, as well as 0.5 mm, and also 0.4 mm will fulfil their roles in the system.
  • the channels' cross sections can have the shape of any closed figure, and in particular of a trapezium, square, rectangle, circle, ellipse, etc.
  • Channels of the chip may comprise internal surfaces coated with substances which are desirable in view of the final outcome of the blood separation.
  • substances may be used to counteract blood coagulation and they include, by way of example: EDTA (salt of the ethylenediaminetetraacetic acid), trisodium citrate, heparinates (sodium, lithium or ammonium salt of heparin), hirudin, potassium oxalate, sodium fluoride, iodoacetate or thrombin inhibitors, (e.g., ppack or agratroban).
  • the substances may also be used to activate blood coagulation, for instance they may be: silica, kaolin, glass particles, diatomaceous earth, thrombin-based agents or ellagic acid.
  • microfluidic system - according to the invention - is used as follows:
  • Blood is aspirated into the system, and in particular to the first chamber Zl and the second chamber Z2, spontaneously, due to capillary forces.
  • the system discussed here can be filled by an adult person in about 30 seconds, whereas the amount of blood so collected and sufficient for performing analyses is as little as 100 ⁇ , i.e., several times less than for traditional syringes or blood collection ampoules mentioned in the introduction.
  • the centrifuge rotor comprises a well that is adapted to the size and shape of the chip.
  • the rotor is made of aluminium (or other suitable material - e.g., ABS, polyacetal (POM), polystyrene or polyamide 66 with glass fibre filler).
  • the blood separates into plasma or serum, and blood cells, whereas the plasma/serum is located closer to the axis of rotation in the first chamber Zl with the volume of 40 ⁇ , and the blood cells - further away from the axis of rotation in the second chamber Z2 with the volume of 60 ⁇ .
  • the direction of the centrifugal force during centrifugation is indicated with an arrow in Fig. 6.
  • the ratio of the chambers' volumes can be changed arbitrarily, depending on patient's sex and age (the hematocrit varies depending on age and sex).
  • the rotor and the chip have openings allowing for the supplying of oil through a hydraulic system that is used for dispensing plasma portions to an analysing system. Pumping oil into the microfluidic system (e.g., through the third opening 02 or the fourth opening 03) results in pushing out a portion of plasma therefrom. The plasma flows out through an opening in the rotor.
  • Centrifugation should be performed setting appropriate time and rotational speed values. Centrifugation parameters - time and rotational speed, e.g., 5 minutes at 6000 RPM - are known to persons skilled in the art of diagnostic testing.
  • Example 3 sample delivery using a microfluidic system (chip) with the volume of
  • a two-layer microfluidic chip shown in Fig. 9 - 11 has been manufactured, with the lower layer comprising a channel draining the blood plasma (serum) from the system to an analyser through the fifth opening 05, other channels and chambers - the first chamber Zl and the second chamber Z2 - for blood sample.
  • the upper plate comprises four openings (the first 01, the third 02, the fourth 03, and the second 04), including the first opening 01 allowing for the supplying of blood sample directly from a patient's finger.
  • the first opening 01 can be adapted to install capillaries or a funnel to make blood collection easier.
  • the fifth opening 05 used to supply plasma (or serum) to an analyser, can optionally be adapted to install capillaries to facilitate the process of fluid transfer.
  • each of the openings can be used for blood sample collection, in addition each of the openings can be arranged both in the upper and the lower plate, as well as at the external edge formed by joined plates. It is important that the third opening 02, together with the third channel K3, are used after centrifugation is completed to push out the serum (or plasma) obtained in centrifugation, and that the fifth opening 05, together with the fifth channel K5, are used to supply the serum to an analyser - at that time the first opening 01 and the second opening 04 should remain closed.
  • the inlet of the microfluidic channel K2 to the second chamber Z2 should take into account biological variability of the material being collected so that after separation the blood cells are below that inlet. Both layers of the said system have been joined permanently so as to maintain the necessary tightness of the entire microfluidic system.
  • the volume of the microfluidic system in the discussed embodiment is 150 ⁇ , and the third K3, fourth K4, fifth K5, second K6 and microfluidic K2 channels inside the chip are 1 mm wide. It is to be noted, however, that channels with the widths of 0.9 mm, as well as 0.8 mm, as well as 0.7 mm, as well as 0.6 mm, as well as 0.5 mm, and also 0.4 mm will fulfil their roles in the system.
  • the channels' cross sections can have a shape of any closed figure, and in particular of a trapezium, square, rectangle, circle, ellipse, etc.
  • Channels of the chip may comprise internal surfaces coated with substances which are desirable in view of the final outcome of the blood separation.
  • substances may be used to counteract blood coagulation and they include, by way of example: EDTA (salt of the ethylenediaminetetraacetic acid), trisodium citrate, heparinates (sodium, lithium or ammonium salt of heparin), hirudin, potassium oxalate, sodium fluoride, iodoacetate or thrombin inhibitors, (e.g., ppack or agratroban).
  • the substances may also be used to activate blood coagulation, for example and in a non-limiting manner they include: silica, kaolin, glass particles, diatomaceous earth, thrombin-based agents or ellagic acid.
  • microfluidic system - according to the invention - is used as follows:
  • Blood is aspirated into the system, and in particular to the first chamber Zl and the second chamber Z2, spontaneously, due of capillary forces.
  • the system discussed here can be filled by an adult person in about 30 seconds, whereas the amount of blood so collected and sufficient for performing analyses is as little as 150 ⁇ , i.e., several times less than for traditional syringes or blood collection ampoules mentioned in the introduction.
  • the centrifuge rotor comprises a well that is adapted to the size and shape of the chip.
  • the rotor is made of aluminium (or other suitable material - e.g., ABS, polyacetal (POM), polystyrene or polyamide 66 with glass fibre filler).
  • the blood separates into plasma or serum, and blood cells, whereas the plasma/serum is located closer to the axis of rotation in the first chamber Zl with the volume of 60 ⁇ , and the blood cells - further away from the axis of rotation in the second chamber Z2 with the volume of 90 ⁇ .
  • the direction of the centrifugal force during centrifugation is indicated with an arrow in Fig. 9.
  • the ratio of the chambers' volumes can be changed arbitrarily, depending on patient's sex and age (the hematocrit varies depending on age and sex).
  • the rotor and the chip have openings allowing for the supplying of oil through a hydraulic system that is used for dispensing plasma portions to an analysing system. Pumping oil into the microfluidic system (e.g., through the third opening 02 or the fourth opening 03) results in pushing out a portion of plasma therefrom. The plasma flows out through an opening in the rotor.
  • Centrifugation should be performed setting appropriate time and rotational speed values. Centrifugation parameters - time and rotational speed, e.g., 8 minutes at 2000 RPM - are known to persons skilled in the art of diagnostic testing.
  • microfluidic channel K2 interconnecting the first chamber Zl and the second chamber Z2, and at the same time representing a constriction between the first chamber Zl and the second chamber Z2, restricting free flow between these chambers Zl and Z2, blood separation is attained at lower rotational speeds and in shorter time compared with the situation where such constriction is absent.
  • the constriction (microfluidic channel K2) guarantees also that the separated blood cells do not spontaneously mix with the serum (plasma).
  • the third channel K3 allows for obtaining serum (plasma) without necessarily pushing blood cells, as it was the case in systems disclosed in the patent application publication no. WO 2013045695 A2. It allows for the maintaining of full purity of material needed for analyses. While analysing the architecture of the systems presented in examples 1-3 it should be noted that the components of the microfluidic system indicated below should be so arranged in the microfluidic system that after placing correctly the microfluidic system in the centrifuge rotor:
  • the third opening 02, the fourth opening 03, and the second opening 04 are closer to the rotation axis than the first opening (01); it aims at preventing possible material leak from the system;
  • connection between the fourth channel K4 and the fifth channel K5 is closer to the centrifugation axis than the first opening 01 - to prevent filling the above- mentioned channels with non-centrifuged material.
  • the microfluidic system according to the invention When the microfluidic system according to the invention is in use, it is placed in a centrifuge rotor and afterwards it is centrifuged, as the result of which a centrifugal force appears.
  • the centrifugal force When used properly according to the present invention, the centrifugal force will extend in the direction essentially along the first Zl and second Z2 chambers and - at the same time - essentially perpendicular to the plane in which the constriction between chambers Zl and Z2 is formed. , as illustrated by arrows F cen tr in the attached drawings.
  • the third channel K3 and the fourth channel K4 connect to the first chamber Zl or to the constriction at an acute angle with respect to the direction of the centrifugal force.
  • connection of the microfluidic channel K2 with the third channel K3 should always be arranged between the first chamber Zl and the second chamber Z2 - it allows for pushing serum (plasma) to an analyser without necessarily pushing blood cells through, as it was the case in systems disclosed in the patent application publication no. WO 2013045695 A2.

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Abstract

L'invention concerne un système microfluidique pour la distribution d'un échantillon de fluide corporel, en particulier du sang, à un analyseur, comprenant la première chambre (Z1) raccordée à la première ouverture (O1) et la deuxième chambre (Z2), ces chambres (Z1, Z2) étant interconnectées et raccordées au troisième canal (K3) comprenant la troisième ouverture (O2) et, en outre, la première chambre (Z1) étant raccordée aux quatrième (K4) et cinquième (K5) canaux, comprenant les quatrième (O3) et cinquième (O5) ouvertures, et la deuxième chambre (Z2) étant raccordée au deuxième canal (K6) comprenant la deuxième ouverture (O4), caractérisé en ce que le raccordement entre la première (Z1) et la deuxième chambre (Z2) est un étranglement. En outre, l'invention comprend un procédé de distribution d'un échantillon de fluide corporel à un analyseur en utilisant le système microfluidique décrit ci-dessus.
PCT/EP2013/070180 2012-09-27 2013-09-27 Système microfluidique et procédé pour la distribution d'un échantillon d'un fluide corporel à un système d'analyse en utilisant le système microfluidique WO2014049116A1 (fr)

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PL400953A PL400953A1 (pl) 2012-09-27 2012-09-27 Układ mikroprzepływowy i sposób dostarczania próbki płynu ustrojowego do układu analizującego z zastosowaniem układu mikroprzepływowego

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WO2017027643A1 (fr) * 2015-08-10 2017-02-16 Essenlix Corp. Dispositifs et procédés de dosages biochimiques pour des applications à étapes simplifiées, sur petits échantillons, à vitesse accélérée et faciles à utiliser
KR20180059828A (ko) * 2015-09-14 2018-06-05 에센릭스 코프. 샘플 특히 혈액샘플을 분석하기 위한 장치와 시스템 및 그 사용 방법
WO2018178895A1 (fr) * 2017-03-31 2018-10-04 Pz Cormay S.A. Système microfluidique
WO2021146350A3 (fr) * 2020-01-13 2021-11-04 Lumiradx Uk Ltd. Régulation des fluides dans des dispositifs microfluidiques
US12076720B2 (en) 2016-06-30 2024-09-03 Lumiradx Uk Ltd. Fluid control

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KR20180059828A (ko) * 2015-09-14 2018-06-05 에센릭스 코프. 샘플 특히 혈액샘플을 분석하기 위한 장치와 시스템 및 그 사용 방법
US12076720B2 (en) 2016-06-30 2024-09-03 Lumiradx Uk Ltd. Fluid control
WO2018178895A1 (fr) * 2017-03-31 2018-10-04 Pz Cormay S.A. Système microfluidique
WO2021146350A3 (fr) * 2020-01-13 2021-11-04 Lumiradx Uk Ltd. Régulation des fluides dans des dispositifs microfluidiques
GB2611504A (en) * 2020-01-13 2023-04-12 Lumiradx Uk Ltd Fluid control in microfluidic devices
GB2611504B (en) * 2020-01-13 2024-08-07 Lumiradx Uk Ltd Fluid control in microfluidic devices

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