EP3621733A1 - Devices and methods for multiplexing liquid in biosensor micro-chambers - Google Patents

Devices and methods for multiplexing liquid in biosensor micro-chambers

Info

Publication number
EP3621733A1
EP3621733A1 EP18724839.8A EP18724839A EP3621733A1 EP 3621733 A1 EP3621733 A1 EP 3621733A1 EP 18724839 A EP18724839 A EP 18724839A EP 3621733 A1 EP3621733 A1 EP 3621733A1
Authority
EP
European Patent Office
Prior art keywords
micro
biosensor
functional liquid
chambers
chamber
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP18724839.8A
Other languages
German (de)
French (fr)
Inventor
Giacomo SAVIOZZI
Juan Pablo SALVADOR VICO
Manuel LÓPEZ DE MIGUEL
Francisco PALACIO BONET
Raquel PRUNA MORALES
M. Pilar MARCO COLÁS
Cecilia Laschi
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Consejo Superior de Investigaciones Cientificas CSIC
Universitat de Barcelona UB
Centro de Investigacion Biomedica en Red CIBER
Scuola Superiore di Studi Universitari e di Perfezionamento SantAnna
Original Assignee
Consejo Superior de Investigaciones Cientificas CSIC
Universitat de Barcelona UB
Centro de Investigacion Biomedica en Red CIBER
Scuola Superiore di Studi Universitari e di Perfezionamento SantAnna
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Consejo Superior de Investigaciones Cientificas CSIC, Universitat de Barcelona UB, Centro de Investigacion Biomedica en Red CIBER, Scuola Superiore di Studi Universitari e di Perfezionamento SantAnna filed Critical Consejo Superior de Investigaciones Cientificas CSIC
Publication of EP3621733A1 publication Critical patent/EP3621733A1/en
Pending legal-status Critical Current

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502738—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 integrated valves
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0636—Integrated biosensor, microarrays
    • 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
    • B01L2300/0848—Specific forms of parts of containers
    • B01L2300/0851—Bottom walls
    • 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
    • B01L2400/06—Valves, specific forms thereof
    • B01L2400/0622—Valves, specific forms thereof distribution valves, valves having multiple inlets and/or outlets, e.g. metering valves, multi-way valves
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/06—Valves, specific forms thereof
    • B01L2400/0633—Valves, specific forms thereof with moving parts
    • B01L2400/0644—Valves, specific forms thereof with moving parts rotary valves
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N35/1095—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices for supplying the samples to flow-through analysers
    • G01N35/1097—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices for supplying the samples to flow-through analysers characterised by the valves

Definitions

  • the present invention relates to the field of continous flow biosensor devices, and to methods for supplying a functional liquid to biosensor micro-chambers in biosensor devices for multiple-analyte analysis.
  • Continuous flow biosensors devices are known in which micro-fluidic systems are present through different liquids are automatically inserted in the biosensor micro-chamber at costant velocity liquid flow.
  • Flooding a biosensor micro-chamber with different liquids is a natural and necessary operation within the biosensors devices both to perform the biorecognition event and to wash/regenerate, i.e. to restore, the biosensor for subsequent analysis.
  • Document CN 204631053 discloses a micro-fluidic box for automatic biosensor analysis, wherein a body is provided with a plurality of sets of thermal measuring needles and a plurality of reagent micro-chambers.
  • Document KR 20140045852 relates to a device for injecting liquid selectively into a reaction chamber, composed of a microfluidic system comprising a plurality of injection paths through which liquid is injected and a plurality of flow paths connected with the injection paths correspondingly, and which is bonded to a reaction chamber; a connection path connected with the flow paths; a connection groove able to rotate, and which is selectively connected with one of the flow paths along the rotation based on the central axis; and a flow path selecting means equipped with an outlet which is connected with the connection groove and the reaction container so as to discharge liquids, which flows in through the selected flow path and the connection groove, into the reaction chamber.
  • the object of the aforementioned solutions is the automatic insertion, at a later stage, in the biosensor chamber of different liquids to carry out the required measurements or restoring operations.
  • a microfluidic system comprises a supply circuit and a plurality of flow paths connecting the supply circuit with the biosensor micro-chamber.
  • the supply circuit is provided with a micro-pump.
  • the microfluidic circuit must be adapted to flood the micro- chambers of parallel biosensors with a few hundred predefined micro-liters of the same compound/liquid in a predetermined time (low flow rate) and in identical amounts for each micro-chamber.
  • the distribution of the liquid to the various micro-chambers is governed only by the geometry of the fluidic paths which are made symmetric with the highest degree of accuracy in order to assure that a same amount of liquid reaches each micro- chamber. Nevertheless, in such multi-analyte biosensor applications there is a problem that different amounts of the sample liquid enters the biosensor micro-chambers. Another problem of the above conventional continuous flow biosensor applications is that it is very difficult to supply the biosensor micro-chambers with a sufficiently precise low flow rate of liquid.
  • a pulsating liquid flow In the presence of a pulsating liquid flow, turbulence and rest periods alternate in the micro-chambers allowing a much better effectiveness of the functional liquid during typical biosensors operations such as antigens accumulation, electrodes washing and
  • Turbulence supports a true contact of the functional liquid with biosensor electrodes counteracting typical surface electrodes effects arising with a continuous, constant liquid flow.
  • predetermined functional time period being the amount of functional liquid supplied to each biosensor micro-chamber independently controlled.
  • a biosensors device for simultaneous multiple-analyte analysis as above outlined the flow of a same pressured functional liquid is selectively inlaid in the biosensor micro- chambers by one or more micro-valves controlled by an injection control unit so that the flow in each biosensor micro-chamber can be independently controlled with great accuracy.
  • the injection control unit operates the at least one micro-valve so that each biosensor micro chamber is flooded with the functional liquid according to a pulsating flow mode.
  • the above outlined device makes it possible to multiplex a functional liquid to a plurality of biosensor micro-chambers accordingly the pulsating flow mode.
  • the biosensor micro-chambers are supplied one by one in a rapid succession of flow pulsations with the same pulsing functional liquid so that it is possible to supply within a functional time period each biosensor micro-chamber with an identical amount of functional liquid and the biosensor behaves as in the continuous flow mode with very low flow rate even though a pulsating flow is effectively entering the biosensor micro-chamber.
  • Another aspect of the present invention relates to a method for supplying a functional liquid to biosensor micro-chambers in a device for analytes analysis by biosensors in which a biosensor is dipped in at least one biosensor micro-chamber flooded with a functional liquid supplied by a fluidic circuit and the functional liquid floods the biosensor micro-chamber for a functional time period, the functional liquid is pressured in the fluidic circuit and it is supplied to the biosensor micro-chamber according to a pulsating flow mode wherein the functional time period contains a plurality of flooding pulsations of the pulsing functional liquid.
  • the method of the invention allows to greatly reduce the total flow rate in a functional time period for a biosensor micro-chamber.
  • FIG. 1 is a schematic view of a micro-fluidic system of a device according to the present invention.
  • FIG. 2 is a time chart of the operation of the device of FIG. 1 according to the method of the present invention.
  • FTP functional time period.
  • FPCP flooding pulsation cycle period.
  • FPDT flooding pulsation duration time.
  • MC 1 to 6 biosensor micro-chambers.
  • FIG. 3 is a schematic view of an alternative layout of the micro-fluidic system of FIG.1.
  • FIG. 4 is a schematic view of a different embodiment of a micro-fluidic system of a device according to the present invention.
  • a device for simultaneous multiple-analyte analysis by biosensors dipped in biosensor micro-chambers comprises a fluidic system, 10, providing:
  • the device also comprises an injection control unit, 16, operating the micro-pump 12, the relief micro-valve 17 and the micro-valves 15 so that during a functional time period each biosensor micro chamber 13 is flooded with the functional liquid according to a pulsating flow mode, that is according to a sequence of pulses.
  • the micro-valves 13 are active isolation micro-valves and the injection control unit 16 is adapted to control the isolation micro-valves switch on/off independently from one another.
  • the layout of the fluidic circuit 1 1 is such that a linear path is designed between the micro pump 12 and the relief valve 17 and the injection paths 14 are spaced along the linear path.
  • FIG. 2 an example of a pulsating flow mode is shown in a time chart in which the flow of the functional liquid into each of the six biosensor micro-chambers 13 of the device of FIG. 2 is shown as a function of time.
  • Each biosensor micro-chamber is flooded with the functional liquid for a functional time period which can be an incubation, a detection period, a washing period, a regeneration period or any other specific functional period of the biosensors device.
  • the micro-valve 15 operably associated to a specific biosensor micro chamber 13 is controlled by the injection control unit 16 to open a predefined amount of times, each time for a flooding pulsation time.
  • the injection control unit is programmed to schedule opening times of each micro-valve 15 according to regular time intervals so that for each biosensor micro-chamber 13 a flooding pulsation duration time and a flooding pulsation cycle period are defined. Due to the fact that the fluidic circuit 1 1 , that is the portion of the micro-fluidic circuit comprised among the micro-pump 12, relief-valve 17 and micro-valves 15, is pressured, each time a micro-valve opens, the functional liquid flows through the injection path 14 into the associated biosensor micro-chamber 13 according to a steady flow rate so that the exact amount of functional liquid flowing in the biosensor micro- chamber 13 can be predetermined.
  • the fluidic circuit 1 1 that is the portion of the micro-fluidic circuit comprised among the micro-pump 12, relief-valve 17 and micro-valves 15 is pressured
  • a same flooding pulsation duration time and a same flooding pulsation cycle period are defined but the flooding pulsation times are subsequently offset of a flooding pulsation duration time and the flooding pulsation cycle period is defined as the sum of the flooding pulsation time periods of all the biosensor micro-chambers 13.
  • FIG. 3 a different layout of a fluidic circuit 1 1 ' is shown where injection paths 14' depart radially from a circular path of the fluidic circuit 1 1 ' symmetrically with respect to an inlet path, 1 10, and an outlet path, 1 1 1 , of the fluidic circuit 1 1 '.
  • a circular discharge fluidic circuit 18' is arranged for receiving the functional liquid by the biosensor micro-chambers 13.
  • the symmetric position of the injection paths 14' is not a prerequisite for the device functioning. Thanks to the presence of the micro-valves 15, the geometry of the fluidic circuit 1 1 and 1 1 ' do not affect the amount of functional liquid that flows in the biosensor micro- chambers 13.
  • a different embodiment of a micro-fluidic system, 10" in a device according to the present invention provides a single rotary distribution micro-valve, 15", controlled by the injection control unit 16 for distributing the functional liquid to the injection paths 14 and then to the biosensor micro-chambers 13.
  • the rotary distribution micro-valve 15" is adapted to allow the crossing of the functional liquid to the biosensor micro-chambers 13 one-by-one.
  • the injection control unit 16 is programmable to define the interval rotary times of the rotary distribution micro-valve 15" so that a pulsating flow mode according to FIG. 2 is achieved, in a way that in a functional time period each micro-chamber is flooded with the functional liquid a predefined amount of times, each time for a flooding pulsation time.
  • a discharge micro-valve (not shown in figures) is interposed between each biosensor micro-chamber 13 and the discharge un-pressured fluidic circuit 18.
  • the discharge micro-valve prevents the functional liquid from naturally flowing from biosensor micro-chambers 13 into the discharge fluidic circuit 18 so that the flow of the functional liquid in each biosensor micro- chamber 13 can be independently controlled by properly operating the respective biosensor micro-valve.
  • the same discharge micro-valve can be operated to control the micro-fluidic circuit pressure and, consequently, the flow and its peak in the micro- chambers.
  • a method for supplying a functional liquid to biosensor micro-chambers in a device for analytes analysis by biosensors in which a biosensor is dipped in at least one biosensor micro-chamber flooded with a functional liquid supplied by a pressured fluidic circuit and the functional liquid floods the biosensor micro-chamber for a functional time period provides that the functional liquid is supplied to the biosensor micro-chamber according to a pulsating flow mode wherein the functional time period contains a plurality of flooding pulsations of the pulsing functional liquid.
  • a method for supplying a functional liquid as above defined allow to supply the biosensor micro-chamber with a very low total amount of functional liquid in a functional time period.
  • a method according to the invention is applied to a biosensors device for simultaneous multiple-analyte analysis according to the present invention with a micro-fluidic system comprising:
  • biosensor micro-chambers 13, each equipped with a biosensor for revealing analytes present in a sample liquid which is one of a plurality of functional liquids;
  • the method of the invention provides that a unique functional liquid is kept at a substantially steady pressure in the fluidic circuit 1 1 and the functional liquid is multiplexed to the plurality of biosensor micro-chambers 13 by allowing it to flow selectively from the pressured fluidic circuit 1 1 into the plurality of biosensor micro- chambers 13, a biosensor micro-chamber at a time.
  • the functional liquid is multiplexed in a rapidly timed sequence of flooding pulsations in which, for each biosensor micro-chamber 13 are defined a same flooding pulsation duration time and a same flooding pulsation cycle period so that each biosensor micro-chamber 13 receives the same volume of the functional liquid in a functional time period.
  • a flooding pulsation duration time and a flooding pulsation cycle period are independently defined for each biosensor micro-chamber 13 so that each biosensor micro-chamber 13 receives its own volume of the functional liquid in a biosensor functional time period.
  • biosensors of multiple biosensor micro- chambers 13 work at their best when they receive the functional liquid with a different flow rate.
  • At least one biosensor micro-chamber 13 of the plurality of biosensor micro-chambers receives a single flooding pulsation of the functional liquid during the functional time period, for instance in case a biosensor works at his best when the biosensor micro-chamber is only initially flooded with functional liquid which is then kept motionless in the micro-chamber.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Dispersion Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Automatic Analysis And Handling Materials Therefor (AREA)

Abstract

A new method, the pulsating liquid flow mode, is proposed in biosensor devices for simultaneous multiple-analyte analysis. The biosensor micro-chambers are flooded at a time with a functional liquid flowing from a fluidic circuit with no symmetry prerequisite comprising micro-valves operably associated to micro-chambers for selectively allowing the liquid crossing. With the pulsating flow mode, turbulence and rest alternate in each micro-chamber so that the liquid function, such as accumulation, washing, regeneration, is greatly improved. The needed total liquid amounts in a micro-chamber can be greatly reduced and the unique entering liquid can be equally distributed without difficulties as the pressured functional liquid is easily multiplexed to biosensor micro-chambers. Related simple biosensor devices are proposed for such a pulsating liquid flow method.

Description

Devices and methods for multiplexing liquid in biosensor micro-chambers TECHNICAL FIELD The present invention relates to the field of continous flow biosensor devices, and to methods for supplying a functional liquid to biosensor micro-chambers in biosensor devices for multiple-analyte analysis.
BACKGROUND ART
Continuous flow biosensors devices are known in which micro-fluidic systems are present through different liquids are automatically inserted in the biosensor micro-chamber at costant velocity liquid flow. Flooding a biosensor micro-chamber with different liquids is a natural and necessary operation within the biosensors devices both to perform the biorecognition event and to wash/regenerate, i.e. to restore, the biosensor for subsequent analysis.
Document CN 204631053 discloses a micro-fluidic box for automatic biosensor analysis, wherein a body is provided with a plurality of sets of thermal measuring needles and a plurality of reagent micro-chambers.
Document KR 20140045852 relates to a device for injecting liquid selectively into a reaction chamber, composed of a microfluidic system comprising a plurality of injection paths through which liquid is injected and a plurality of flow paths connected with the injection paths correspondingly, and which is bonded to a reaction chamber; a connection path connected with the flow paths; a connection groove able to rotate, and which is selectively connected with one of the flow paths along the rotation based on the central axis; and a flow path selecting means equipped with an outlet which is connected with the connection groove and the reaction container so as to discharge liquids, which flows in through the selected flow path and the connection groove, into the reaction chamber.
The object of the aforementioned solutions is the automatic insertion, at a later stage, in the biosensor chamber of different liquids to carry out the required measurements or restoring operations.
Continuous flow multi-analyte biosensor cartridges are also known for the simultaneous measurement of biologically relevant analytes in a sample solution. In such biosensor applications a microfluidic system comprises a supply circuit and a plurality of flow paths connecting the supply circuit with the biosensor micro-chamber. The supply circuit is provided with a micro-pump. The microfluidic circuit must be adapted to flood the micro- chambers of parallel biosensors with a few hundred predefined micro-liters of the same compound/liquid in a predetermined time (low flow rate) and in identical amounts for each micro-chamber. The distribution of the liquid to the various micro-chambers is governed only by the geometry of the fluidic paths which are made symmetric with the highest degree of accuracy in order to assure that a same amount of liquid reaches each micro- chamber. Nevertheless, in such multi-analyte biosensor applications there is a problem that different amounts of the sample liquid enters the biosensor micro-chambers. Another problem of the above conventional continuous flow biosensor applications is that it is very difficult to supply the biosensor micro-chambers with a sufficiently precise low flow rate of liquid.
SUMMARY OF INVENTION It is an object of the present invention to propose a device and a method for simultaneous multiple-analyte analysis by biosensors dipped in biosensor micro-chambers which allow an identical amount of a functional liquid to flood a plurality of biosensor micro-chambers in a predetermined functional time period with a pulsating liquid flow. In the presence of a pulsating liquid flow, turbulence and rest periods alternate in the micro-chambers allowing a much better effectiveness of the functional liquid during typical biosensors operations such as antigens accumulation, electrodes washing and
regeneration. Turbulence supports a true contact of the functional liquid with biosensor electrodes counteracting typical surface electrodes effects arising with a continuous, constant liquid flow.
It is another object of the present invention to propose a method and a device for simultaneous multiple-analyte analysis by biosensors dipped in biosensor micro-chambers which supply a functional liquid to a plurality of biosensor micro-chambers in a
predetermined functional time period being the amount of functional liquid supplied to each biosensor micro-chamber independently controlled.
It is another object of the present invention to propose a method and a device for supplying a functional liquid to biosensor micro-chambers in a biosensors device which flood at least one biosensor micro-chamber in a functional time period at an accurate very low flow rate by using the fluidic circuit.
An aspect of the present invention relates to the provision of a device for simultaneous multiple-analyte analysis by biosensors dipped in biosensor micro-chambers comprises: a feeding fluidic circuit with a pressured functional liquid supplied by a micro-pump; a plurality of biosensor micro-chambers each equipped with a biosensor; a plurality of injection paths, each of them connecting the feeding fluidic circuit to a respective single biosensor micro-chamber of the plurality of biosensor micro-chambers; at least one micro- valve associated to the injection paths for selectively allowing the crossing of the functional liquid to a single biosensor micro-chamber at a time; and an injection control unit operating the at least one micro-valve.
In a biosensors device for simultaneous multiple-analyte analysis as above outlined the flow of a same pressured functional liquid is selectively inlaid in the biosensor micro- chambers by one or more micro-valves controlled by an injection control unit so that the flow in each biosensor micro-chamber can be independently controlled with great accuracy. Advantageously, the injection control unit operates the at least one micro-valve so that each biosensor micro chamber is flooded with the functional liquid according to a pulsating flow mode.
The above outlined device makes it possible to multiplex a functional liquid to a plurality of biosensor micro-chambers accordingly the pulsating flow mode. In fact, the biosensor micro-chambers are supplied one by one in a rapid succession of flow pulsations with the same pulsing functional liquid so that it is possible to supply within a functional time period each biosensor micro-chamber with an identical amount of functional liquid and the biosensor behaves as in the continuous flow mode with very low flow rate even though a pulsating flow is effectively entering the biosensor micro-chamber.
The fluidic circuit with a pressured functional liquid supplied by a micro-pump, and all its paths needed to feed one-by-one the biosensor micro-chambers, do not ask for any special geometrical symmetry overcoming the difficulties of current implementations to equally flood each biosensor micro-chambers.
Particular embodiments of the device of the invention are disclosed by the apparatus dependent claims. Another aspect of the present invention relates to a method for supplying a functional liquid to biosensor micro-chambers in a device for analytes analysis by biosensors in which a biosensor is dipped in at least one biosensor micro-chamber flooded with a functional liquid supplied by a fluidic circuit and the functional liquid floods the biosensor micro-chamber for a functional time period, the functional liquid is pressured in the fluidic circuit and it is supplied to the biosensor micro-chamber according to a pulsating flow mode wherein the functional time period contains a plurality of flooding pulsations of the pulsing functional liquid. The method of the invention allows to greatly reduce the total flow rate in a functional time period for a biosensor micro-chamber.
Particular embodiments of the method of the invention are disclosed by the method dependent claims.
Throughout the description and claims the word "comprise" and variations of the word, are not intended to exclude other technical features, additives, components, or steps.
Furthermore, the word "comprise" encompasses the case of "consisting of". Additional objects, advantages and features of the invention will become apparent to those skilled in the art upon examination of the description or may be learned by practice of the invention. The following examples and drawings are provided by way of illustration, and they are not intended to be limiting of the present invention.
BRIEF DESCRIPTION OF DRAWINGS
The constructional, functional features and the advantages of the device and method of the invention will be made clearer by the following detailed description, in which reference is made to the accompanying drawings which represent preferred and non-limiting embodiments thereof and in which:
FIG. 1 is a schematic view of a micro-fluidic system of a device according to the present invention.
FIG. 2 is a time chart of the operation of the device of FIG. 1 according to the method of the present invention. FTP = functional time period. FPCP = flooding pulsation cycle period. FPDT = flooding pulsation duration time. MC 1 to 6 = biosensor micro-chambers.
FIG. 3 is a schematic view of an alternative layout of the micro-fluidic system of FIG.1. FIG. 4 is a schematic view of a different embodiment of a micro-fluidic system of a device according to the present invention.
DESCRIPTION OF EMBODIMENTS With reference first to FIG. 1 , a device for simultaneous multiple-analyte analysis by biosensors dipped in biosensor micro-chambers comprises a fluidic system, 10, providing:
- a fluidic circuit, 1 1 , with a pressured functional liquid, eventually carrying analytes, supplied by a micro-pump, 12;
- a relief valve, 17, operably associated to the fluidic circuit 1 1 to control the pressure of the functional liquid in the fluidic circuit and/or to discharge the functional liquid;
- a plurality of biosensor micro-chambers, 13, each equipped with a biosensor for revealing analytes present in a liquid compound which is one of a plurality of functional liquids;
- a plurality of injection paths, 14, each of them connecting the fluidic circuit 1 1 to a respective single biosensor micro-chamber 13;
- a plurality of micro-valves, 15, associated to the injection paths 14 for selectively allowing the crossing of the functional liquid to a single biosensor micro-chamber 13 at a time;
- a discharge fluidic circuit, 18, for receiving the functional liquid from the biosensor micro- chambers 13.
The device also comprises an injection control unit, 16, operating the micro-pump 12, the relief micro-valve 17 and the micro-valves 15 so that during a functional time period each biosensor micro chamber 13 is flooded with the functional liquid according to a pulsating flow mode, that is according to a sequence of pulses.
The micro-valves 13 are active isolation micro-valves and the injection control unit 16 is adapted to control the isolation micro-valves switch on/off independently from one another.
The layout of the fluidic circuit 1 1 is such that a linear path is designed between the micro pump 12 and the relief valve 17 and the injection paths 14 are spaced along the linear path.
With reference to FIG. 2, an example of a pulsating flow mode is shown in a time chart in which the flow of the functional liquid into each of the six biosensor micro-chambers 13 of the device of FIG. 2 is shown as a function of time. Each biosensor micro-chamber is flooded with the functional liquid for a functional time period which can be an incubation, a detection period, a washing period, a regeneration period or any other specific functional period of the biosensors device. During a functional time period the micro-valve 15 operably associated to a specific biosensor micro chamber 13 is controlled by the injection control unit 16 to open a predefined amount of times, each time for a flooding pulsation time. In the embodiment of FIG. 2 the injection control unit is programmed to schedule opening times of each micro-valve 15 according to regular time intervals so that for each biosensor micro-chamber 13 a flooding pulsation duration time and a flooding pulsation cycle period are defined. Due to the fact that the fluidic circuit 1 1 , that is the portion of the micro-fluidic circuit comprised among the micro-pump 12, relief-valve 17 and micro-valves 15, is pressured, each time a micro-valve opens, the functional liquid flows through the injection path 14 into the associated biosensor micro-chamber 13 according to a steady flow rate so that the exact amount of functional liquid flowing in the biosensor micro- chamber 13 can be predetermined. In the embodiment of FIG. 2, for each biosensor micro-chamber 13 a same flooding pulsation duration time and a same flooding pulsation cycle period are defined but the flooding pulsation times are subsequently offset of a flooding pulsation duration time and the flooding pulsation cycle period is defined as the sum of the flooding pulsation time periods of all the biosensor micro-chambers 13. With the above pulsating flow mode the crossing of the functional liquid is selectively allowed to a single biosensor micro-chamber 13 one-by-one and the micro-valves 15 are never all simultaneously closed, so that in a functional time period a same pulsing functional liquid is multiplexed to the plurality of biosensor micro-chambers 13. In the functional time period each biosensor micro- chamber 13 receives an identical amount of functional liquid. In fact, the liquid
fragmentation obtained by switching on/off each micro-valve 15 (which causes the flooding pulsation of functional liquid in the micro-chambers from a pressured circuit) one at a time will certainly assure the same quantity of flux per channel / micro-chamber (almost impossible with a simple geometrical subdivision of a flow of a single functional liquid from an upstream single path to the multiple paths of the plurality of micro- chambers).
In addition, the adoption in a functional time period of a pulsating flow mode by switching on/off each micro-valve 15 a plurality of times creates turbulence in the micro-chambers as opposed to a single pulse or a continuous flow mode. The alternation of turbulence and rest interval times assured by the pulsating flow mode allows a much better accumulation of antigens onto biosensor electrodes and therefore more consistent and easy
measurements by an electronic board. Similar improvements are obtained for other typical biosensor operations such as electrodes washing and regeneration.
With reference to FIG. 3 a different layout of a fluidic circuit 1 1 ' is shown where injection paths 14' depart radially from a circular path of the fluidic circuit 1 1 ' symmetrically with respect to an inlet path, 1 10, and an outlet path, 1 1 1 , of the fluidic circuit 1 1 '. A circular discharge fluidic circuit 18' is arranged for receiving the functional liquid by the biosensor micro-chambers 13. Obviously, in the present invention, the symmetric position of the injection paths 14' is not a prerequisite for the device functioning. Thanks to the presence of the micro-valves 15, the geometry of the fluidic circuit 1 1 and 1 1 ' do not affect the amount of functional liquid that flows in the biosensor micro- chambers 13.
With reference to FIG. 4, a different embodiment of a micro-fluidic system, 10", in a device according to the present invention provides a single rotary distribution micro-valve, 15", controlled by the injection control unit 16 for distributing the functional liquid to the injection paths 14 and then to the biosensor micro-chambers 13. The rotary distribution micro-valve 15" is adapted to allow the crossing of the functional liquid to the biosensor micro-chambers 13 one-by-one. Advantageously, the injection control unit 16 is programmable to define the interval rotary times of the rotary distribution micro-valve 15" so that a pulsating flow mode according to FIG. 2 is achieved, in a way that in a functional time period each micro-chamber is flooded with the functional liquid a predefined amount of times, each time for a flooding pulsation time.
In a specific embodiment of the device of the invention, a discharge micro-valve (not shown in figures) is interposed between each biosensor micro-chamber 13 and the discharge un-pressured fluidic circuit 18. The discharge micro-valve prevents the functional liquid from naturally flowing from biosensor micro-chambers 13 into the discharge fluidic circuit 18 so that the flow of the functional liquid in each biosensor micro- chamber 13 can be independently controlled by properly operating the respective biosensor micro-valve. The same discharge micro-valve can be operated to control the micro-fluidic circuit pressure and, consequently, the flow and its peak in the micro- chambers.
A method for supplying a functional liquid to biosensor micro-chambers in a device for analytes analysis by biosensors in which a biosensor is dipped in at least one biosensor micro-chamber flooded with a functional liquid supplied by a pressured fluidic circuit and the functional liquid floods the biosensor micro-chamber for a functional time period, provides that the functional liquid is supplied to the biosensor micro-chamber according to a pulsating flow mode wherein the functional time period contains a plurality of flooding pulsations of the pulsing functional liquid. In a device for analytes analysis by biosensors with a single biosensor micro-chamber a method for supplying a functional liquid as above defined allow to supply the biosensor micro-chamber with a very low total amount of functional liquid in a functional time period. By properly setting a flooding pulsation duration time and a flooding pulsation cycle period the biosensor behaves as if it receives a continuous flow of functional liquid during the functional time period but the functional liquid is effectively supplied in a pulsating flow mode so that the total flow rate can be greatly reduced with respect to conventional continuous flow micro-fluidic systems of biosensors devices.
In addition, the adoption in a functional time period of a pulsating flow mode creates turbulence in the micro-chambers as opposed to a single pulse or a continuous liquid flow mode. The alternation of turbulence and rest interval times assured by the pulsating flow mode allows a much better accumulation of antigens onto biosensor electrodes and therefore more consistent and easy measurements by an electronic board. A method according to the invention is applied to a biosensors device for simultaneous multiple-analyte analysis according to the present invention with a micro-fluidic system comprising:
- a fluidic circuit, 1 1 , pressured with a functional liquid supplied by a micro-pump, 12;
- a relief (or discharge) micro-valve, 17, operably associated to the fluidic circuit 1 1 ;
- a plurality of biosensor micro-chambers, 13, each equipped with a biosensor for revealing analytes present in a sample liquid which is one of a plurality of functional liquids;
- a plurality of injection paths, 14, each of them connecting the fluidic circuit 1 1 to a respective single biosensor micro-chamber 13;
- a plurality of micro-valves, 15, associated to the injection paths 14 for selectively allowing the crossing of the functional liquid to a single biosensor micro-chamber 13 at a time;
- a discharge fluidic circuit, 18, for receiving the functional liquid from the biosensor micro- chambers 13.
In such a device, the method of the invention provides that a unique functional liquid is kept at a substantially steady pressure in the fluidic circuit 1 1 and the functional liquid is multiplexed to the plurality of biosensor micro-chambers 13 by allowing it to flow selectively from the pressured fluidic circuit 1 1 into the plurality of biosensor micro- chambers 13, a biosensor micro-chamber at a time.
In a preferred embodiment of the method of the present invention the functional liquid is multiplexed in a rapidly timed sequence of flooding pulsations in which, for each biosensor micro-chamber 13 are defined a same flooding pulsation duration time and a same flooding pulsation cycle period so that each biosensor micro-chamber 13 receives the same volume of the functional liquid in a functional time period.
In an alternative embodiment of the method of the present invention a flooding pulsation duration time and a flooding pulsation cycle period are independently defined for each biosensor micro-chamber 13 so that each biosensor micro-chamber 13 receives its own volume of the functional liquid in a biosensor functional time period.
The above embodiment is preferable where the biosensors of multiple biosensor micro- chambers 13 work at their best when they receive the functional liquid with a different flow rate.
In a further alternative embodiment of the method of the invention at least one biosensor micro-chamber 13 of the plurality of biosensor micro-chambers receives a single flooding pulsation of the functional liquid during the functional time period, for instance in case a biosensor works at his best when the biosensor micro-chamber is only initially flooded with functional liquid which is then kept motionless in the micro-chamber.
Although the invention has been described above with particular reference to some specific embodiments, numerous modifications and variations will be apparent to a person skilled in the art in view of the above description. The present invention intends, therefore, to embrace all the embodiments that fall within the ambit of protection defined by the following claims.

Claims

1 . Method for supplying a functional liquid to biosensor micro-chambers in a device for simultaneous multiple analytes analysis by biosensors dipped in respective biosensor micro-chambers (13) flooded with a unique functional liquid supplied by a fluidic circuit (1 1 ) and said functional liquid floods said biosensor micro-chambers (13) for a functional time period, characterized in that said functional liquid is multiplexed by allowing it to flow selectively from said fluidic circuit (1 1 ) into said plurality of biosensor micro-chambers (13), a biosensor micro-chamber at a time and in that said functional liquid is supplied to said biosensor micro-chambers (13) according to a pulsating flow mode wherein said functional time period contains a plurality of flooding pulsations of the pulsing functional liquid.
2. Method according to the previous claim characterized in that said unique functional liquid is kept at a substantially steady pressure in said fluidic circuit (1 1 ).
3. Method according to claim 1 or 2 characterized in that for each biosensor micro- chamber (13) are defined a same flooding pulsation duration time and a same flooding pulsation cycle period so that each biosensor micro-chamber (13) receives a same volume of said functional liquid in said functional time period.
4. Method according to claim 1 or 2 characterized in that a flooding pulsation duration time and a flooding pulsation cycle period are independently defined for each biosensor micro- chamber (13) so that each biosensor micro-chamber (13) receives its own volume of said functional liquid in said functional time period.
5. Device for simultaneous multiple analytes analysis by biosensors dipped in biosensor micro-chambers comprising:
- a fluidic circuit (1 1 ) with a pressured functional liquid supplied by a micro-pump (12); - a plurality of biosensor micro-chambers (13) each equipped with a biosensor;
- a plurality of injection paths (14), each of them connecting said fluidic circuit (1 1 ) to a respective single biosensor micro-chamber (13) of said plurality of biosensor micro- chambers;
- at least a micro-valve (15) associated to said injection paths (14);
- an injection control unit (16) operating said at least a micro-valve (15, 15"),
characterized in that said injection control unit (16) is configured to operate said at least a micro-valve (15, 15") so that each biosensor micro-chamber (13) is flooded with said functional liquid according to a pulsating flow mode and for selectively allowing the crossing of said functional liquid to a single biosensor micro-chamber (13) at a time.
6. Device according to claim 5 characterized in that said at least a micro-valve (15) is a plurality of isolation micro-valves, an isolation micro-valve for each injection path (14), said injection control unit (16) being adapted to control the isolation micro-valves switch on/off.
7. Device for simultaneous multiple analytes analysis by biosensors according to the previous claim characterized in that said injection control unit (16) is programmable to schedule opening times of each micro-valve (15) so that for each biosensor micro- chamber (13) are defined a flooding pulsation duration time and a flooding pulsation cycle period.
8. Device for simultaneous multiple analytes analysis by biosensors according to claim 5 characterized in that said at least a micro-valve (15") is a rotary distribution micro-valve controlled by said injection control unit (16).
9. Device for simultaneous multiple analytes analysis by biosensors according to the previous claim characterized in that said injection control unit (16) is programmable to define the interval rotary times of said rotary distribution micro-valve (15").
10. Device according to any claim 5 to 9 characterized in that said fluidic circuit (1 1 ) is provided with a relief valve (17) to control the pressure of said functional liquid in the fluidic circuit (1 1 ) and/or to discharge the functional liquid.
1 1 . Device according to any claim 5 to 10 characterized in that it comprises a discharge fluidic circuit (18) for receiving said functional liquid from said biosensor micro-chambers (13), a discharge micro-valve being interposed between each biosensor micro-chamber (13) and said discharge fluidic circuit (18).
EP18724839.8A 2017-05-12 2018-05-11 Devices and methods for multiplexing liquid in biosensor micro-chambers Pending EP3621733A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP17382269.3A EP3401013A1 (en) 2017-05-12 2017-05-12 Devices and methods for multiplexing liquid in biosensor micro-chambers
PCT/EP2018/062230 WO2018206775A1 (en) 2017-05-12 2018-05-11 Devices and methods for multiplexing liquid in biosensor micro-chambers

Publications (1)

Publication Number Publication Date
EP3621733A1 true EP3621733A1 (en) 2020-03-18

Family

ID=59009637

Family Applications (2)

Application Number Title Priority Date Filing Date
EP17382269.3A Withdrawn EP3401013A1 (en) 2017-05-12 2017-05-12 Devices and methods for multiplexing liquid in biosensor micro-chambers
EP18724839.8A Pending EP3621733A1 (en) 2017-05-12 2018-05-11 Devices and methods for multiplexing liquid in biosensor micro-chambers

Family Applications Before (1)

Application Number Title Priority Date Filing Date
EP17382269.3A Withdrawn EP3401013A1 (en) 2017-05-12 2017-05-12 Devices and methods for multiplexing liquid in biosensor micro-chambers

Country Status (2)

Country Link
EP (2) EP3401013A1 (en)
WO (1) WO2018206775A1 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022242549A1 (en) * 2021-05-16 2022-11-24 深圳市品学优技术有限公司 Reaction device and reaction driving device

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050176135A1 (en) * 2004-02-06 2005-08-11 Brian Jones Cassette for isolation, amplification and identification of DNA or protein and method of use
EP1979079A4 (en) * 2006-02-03 2012-11-28 Integenx Inc Microfluidic devices
JP2009533656A (en) * 2006-04-07 2009-09-17 コーニング インコーポレイテッド Closed flow-through microplate and methods of use and manufacturing thereof
US20130196343A1 (en) * 2010-03-31 2013-08-01 Carlos H. Mastrangelo Signal locking label free biosensing
US20110301049A1 (en) * 2010-06-04 2011-12-08 The Government Of The United States Of America, As Represented By The Secretary Of The Navy Fluid Flow Contour Control Using Flow Resistance
KR101414048B1 (en) 2012-10-09 2014-07-02 디지탈 지노믹스(주) Device for injecting liquid selectively into reaction chamber and biosensor comprising the same
CN105026932B (en) * 2013-03-15 2017-06-13 西门子医疗保健诊断公司 Micro-fluidic distributing equipment
DE102014206140A1 (en) * 2014-04-01 2015-10-01 Robert Bosch Gmbh A microfluidic device and method for analyzing a sample of biological material
CN204631053U (en) 2015-05-07 2015-09-09 江西恒盛晶微技术有限公司 A kind of micro-fluidic box automatic analysis biochip

Also Published As

Publication number Publication date
WO2018206775A1 (en) 2018-11-15
EP3401013A1 (en) 2018-11-14

Similar Documents

Publication Publication Date Title
US7431884B2 (en) Chemical assays
CN101384354B (en) Method for controlling the progress of a liquid in a microfluid component
US20130296174A1 (en) Microfluidics System for Sequencing
US9592483B2 (en) Fluid mixing and rinsing system for a flow cytometer
WO2010005467A3 (en) Analytical cartridge with fluid flow control
US10107403B2 (en) Pinch flow regulator
US3195558A (en) Proportioning apparatus
JP2012528328A5 (en)
RU99100385A (en) PUMP FOR DELIVERY OF LIQUID FROM THE CONTAINER TO THE PATIENT, SYSTEM OF THE PUMP FOR DELIVERY OF LIQUID, METHOD OF DELIVERY OF LIQUID
JP2011524008A5 (en)
US10926258B2 (en) Microfluidic probe head for processing a sequence of liquid volumes separated by spacers
US4162689A (en) Time division flow control
EP3621733A1 (en) Devices and methods for multiplexing liquid in biosensor micro-chambers
EP1359998A1 (en) Microchannel device
US7379832B2 (en) Multi-channel metering apparatus with automatic calibration
US20190083982A1 (en) Method and Dispensing Device for Dispensed Fluid Output
KR101897584B1 (en) Apparatus and method for mixing
US9492821B2 (en) Liquid feeder and chemical analyzer including same
US9339814B2 (en) Liquid supply drive mechanism using osmotic pump and microchip having the liquid supply drive mechanism
Saviozzi et al. Devices and methods for multiplexing liquid in biosensor micro-chambers
EP3285936B1 (en) Method, device and apparatus for coating an inner surface of a substrate
CN219911849U (en) Rotary valve and fluid path system
US20070274840A1 (en) Magnetohydrodynamic pump
US6350110B1 (en) Multiport metering pump
US20210138458A1 (en) Sequenced droplet ejection to deliver fluids

Legal Events

Date Code Title Description
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: UNKNOWN

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE

PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE

17P Request for examination filed

Effective date: 20191212

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

AX Request for extension of the european patent

Extension state: BA ME

DAV Request for validation of the european patent (deleted)
DAX Request for extension of the european patent (deleted)