EP2836302B1 - Procédé et dispositif de commande de processus ciblée dans un processeur microfluidique doté d'éléments actifs intégrés - Google Patents

Procédé et dispositif de commande de processus ciblée dans un processeur microfluidique doté d'éléments actifs intégrés Download PDF

Info

Publication number
EP2836302B1
EP2836302B1 EP13717014.8A EP13717014A EP2836302B1 EP 2836302 B1 EP2836302 B1 EP 2836302B1 EP 13717014 A EP13717014 A EP 13717014A EP 2836302 B1 EP2836302 B1 EP 2836302B1
Authority
EP
European Patent Office
Prior art keywords
liquid
microfluidic
active
reaction chamber
active elements
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.)
Active
Application number
EP13717014.8A
Other languages
German (de)
English (en)
Other versions
EP2836302A1 (fr
EP2836302B8 (fr
Inventor
Andreas Prof. Dr. RICHTER
Rinaldo Greiner
Merle ALLERDIßEN
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.)
Technische Universitaet Dresden
Original Assignee
Technische Universitaet Dresden
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 Technische Universitaet Dresden filed Critical Technische Universitaet Dresden
Publication of EP2836302A1 publication Critical patent/EP2836302A1/fr
Publication of EP2836302B1 publication Critical patent/EP2836302B1/fr
Application granted granted Critical
Publication of EP2836302B8 publication Critical patent/EP2836302B8/fr
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

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
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0621—Control of the sequence of chambers filled or emptied
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/04—Closures and closing means
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—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
    • 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
    • B01L2300/0867—Multiple inlets and one sample wells, e.g. mixing, dilution
    • 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
    • B01L2300/0874—Three dimensional network
    • 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
    • B01L2400/0672—Swellable plugs
    • 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
    • B01L2400/0677—Valves, specific forms thereof phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00—Chemistry: analytical and immunological testing
    • Y10T436/25—Chemistry: analytical and immunological testing including sample preparation
    • Y10T436/2575—Volumetric liquid transfer

Definitions

  • the invention relates to a microfluidic microchemomechanical system with integrated active elements and a method for microfluidic process control in a microfluidic microchemomechanical system.
  • Microfluid processors are mainly used today in biological, biochemical and chemical processes, with their use as “labs on chips” (LOC), “chip laboratories” or “micro-total analysis systems” ( ⁇ TAS) in focus scientific developments.
  • LOC las on chips
  • ⁇ TAS micro-total analysis systems
  • the LOC concept offers a variety of advantages.
  • the reduction in fluid volumes enables the analysis of the smallest amounts of samples and an economical use of reagents and samples, which are often valuable, rare, harmful or dangerous.
  • higher throughputs can also be achieved, since, due to the small quantities, shortened preparation, mixing and reaction times are required with minimized energy requirements.
  • Process control can also be easier due to lower system response times.
  • LOC structures enable significant process rationalizations by considerably shortening the process time and thus increasing the possible throughput as well as reducing the quantities of media required (test subjects, analytes, reagents, auxiliary media).
  • Microfluidic systems with active elements are known in the prior art.
  • Active fluidic elements based on solid-state actuators such as piezo actuators [ U.S. 5,224,843 , US 2003/0143122 ] and shape memory actuators [ U.S. 5,659,171 ] described. Although they can be easily miniaturized as individual elements, they have a complicated structure, are based on certain, mostly non-plastic-based, materials and must therefore be manufactured separately. A possible hybrid integration (e.g. sticking the elements on the LOC) is usually uneconomical.
  • Converter elements that are based on changes in the state of aggregation can be integrated into the layout of the channel structure carrier, sometimes with minor interventions, and are therefore mostly compatible with the manufacturing process of the plastic molded parts of the channel structure carrier.
  • fusible elements R. Pal et al. ', Anal. Chem. 16 (2004) 13, pp. 3740-3748
  • freezing elements U.S. 6,536,476
  • thermal bubble generators U.S. 6,283,718
  • the DE 101 57 317 A1 discloses a basic element of a microfluidic processor which is electronically compatible by controlling the degree of swelling of swellable polymer networks with volume phase transition behavior, in particular hydrogels, via an electrically or electronically controllable interface variable.
  • the controllable environmental variables or interface variables are preferably physical variables which can be easily generated by electronic or electrical means and which trigger volume phase transitions in swellable polymer networks.
  • a control variable that can be generated electrically in a very simple manner is temperature.
  • hydrogel-based active elements The main disadvantage of these hydrogel-based active elements is the need to use electrically generated control variables to generate volume phase transitions, so that the operation of such microfluidic systems is necessarily linked to electrical components. Autarkic use of microfluidic systems is therefore excluded.
  • the US 2011/0126913 A1 discloses a microfluidic system with at least two delimited chambers or the like, a pressure-sensitive valve being provided in the channel system between these chambers, which valve dissolves when the pressure increases. It can also be seen from the publication that an active element is provided in a channel system, which changes in volume when it comes into contact with the liquid and can thus completely block the channel. However, the described chambers do not have a common overlap area and are not limited by the active elements in such a way that a common reaction chamber with a defined volume is formed. The publication also does not reveal the timing of the targeted selection of materials and their dimensions.
  • the US 2008/0069729 A1 discloses a device for mixing defined volumes of liquid with defined amounts of reagents.
  • a liquid reservoir is filled with a liquid and this is introduced into corresponding reaction chambers by means of a suction ball-like object, the air present in the system being discharged. The pressure is maintained until the reaction chambers are completely filled.
  • the reaction chambers also have valves that prevent the liquid from flowing back into the ball, for example.
  • the valves can be designed in such a way that they swell or dissolve on contact with the liquid.
  • other active elements are described that act as barriers that can slow down or prevent the flow of liquid.
  • the WO 2008/049413 discloses a microfluidic system with active elements which can be controlled without auxiliary energy.
  • active elements which enable a volume phase transition depending on temperature or solvent.
  • the active elements bring about an active function by changing the degree of swelling or the mechanical properties.
  • swelling agent barriers are disclosed which swell when the solvent is absorbed and consequently limit the supply of swelling agent.
  • auxiliary energy-free active elements allows a largely self-sufficient use of microfluidic systems, especially in diagnostics, whereby the establishment of single-use analysis systems would be favored by dispensing with external electrical energy sources and using chemical energy sources.
  • the object of the present invention is therefore to specify a microfluidic microchemomechanical system which has active elements operated without auxiliary energy and is thereby capable of carrying out volumetrically defined mixing reactions in defined time sequences.
  • microfluidic microchemomechanical system according to claim 1.
  • Advantageous refinements are given in the dependent claims.
  • the microfluidic system comprises integrated active elements which can be activated without auxiliary energy by influencing environmental variables and which have active functions by changing their swelling state or their mechanical properties causing, are carried out.
  • the microfluidic microchemomechanical system comprises at least one structural support with at least one first channel, which as a rule belongs to a first channel system with a first process medium. It also includes at least one cover that at least partially covers the structural support and at least one second channel of a second channel system, which is either integrated on the structural carrier, which already carries the first channel of a first channel system, or is integrated into the cover.
  • the first and the second channel have reservoir spaces in a common overlap area. The reservoir spaces are limited by active elements and are able to form a common reaction chamber.
  • the active elements are designed as swelling agent barriers or liquid-soluble barriers.
  • an increase in the volume of the active element would occur due to the absorption of liquid, whereby the channel containing the active element is narrowed further and further until, as a result of the channel cross-section being completely filled, there is a flow separation in the channel and consequently the flow is blocked.
  • the active element designed as a swelling agent barrier is introduced into the channel of the microfluidic micromechanical system in a dried state. After the swelling agent barrier has increased in volume through absorption of liquid, the swelling agent barrier remains in the swollen state.
  • the swelling agent barrier is designed as a closing element, for example in order to seal off the reaction chamber from flowing liquids.
  • the active elements are designed as a liquid-soluble barrier
  • this barrier is dissolved by wetting the barrier with the liquid in the channel.
  • the progressive dissolution of the barrier leads to an increase in the flow through the channel cross-section and consequently to the formation of a flow of the liquid through the channel.
  • the basis that a dissolving element is perceived as an active element is based on its functional principle.
  • the load-bearing capacity or mechanical flexibility of a component can be changed by changing (a) the modulus of elasticity of the component material or (b) its cross-section.
  • (b) is used as the basis of the active function.
  • the dissolvable active element fulfills the function of an opening valve as soon as the control signal "liquid" is applied.
  • an active membrane is arranged between the first and second channel, whereby the common Reaction chamber is divided into a first reservoir space and a second reservoir space.
  • Due to a slowed flow, for example due to a blockage the second liquid could enter the first channel via the common reaction chamber, whereby an undefined mixing of the first and second liquid would not take place as desired in the common reaction chamber, but already in the first channel.
  • the volumetrically undefined mixtures thus produced would be insufficient for analysis purposes.
  • auxiliary energy is understood to mean dispensing with the supply of energy from an external electrical or thermal energy source to the active elements according to the invention.
  • microfluidic elements are known that can be activated by electrical and thermal energy; thermally or electrically switchable hydrogels may be mentioned here as examples.
  • an overlapping area is understood to mean the part between two connectable reservoir spaces that has a common wall.
  • the first and second liquids which flow into the reaction chamber are thoroughly mixed within this mixing zone.
  • an active element or an active function is understood to mean an active mechanical element or an active mechanical function.
  • the swelling agent barriers or liquid-soluble barriers are designed as valves.
  • the active elements can perform valve functions within the microfluidic microchemomechanical system.
  • the valves can perform both opening (liquid-soluble barriers) and closing functions (swelling agent barriers). Due to the time-definable and auxiliary energy-free function, such valves are particularly suitable for use in self-sufficient microfluidic systems.
  • All active components that fulfill the function of an opening valve are understood as opening elements. This can be done by (a) lowering the modulus of elasticity in crosslinked, swellable polymers and (b) dissolving in liquid-soluble materials.
  • the dissolving membranes are also seen as opening elements
  • the cover is designed as an upper structural carrier in an arrangement of at least two structural carriers.
  • the membrane between the first and second reaction spaces is made of a liquid-soluble material.
  • the membrane can be dissolved after the first and second reservoir chambers have been filled with the two liquids, whereby the reservoir chambers are connected to form the common reaction chamber and the liquids can be mixed in this as intended.
  • the further active elements which delimit the reaction chamber and are designed as swellable swelling agent barriers, prevent the liquids from flowing out of the channels into the reaction chamber.
  • the swelling of the swelling agent barriers creates a hermetically sealed reaction chamber, which is characterized by defined liquid volumes in the reservoir spaces, which are then connected to one another by the subsequent dissolution of the membrane so that their contents can mix with one another.
  • the membrane can be configured according to the needs of the application in such a way that the temporal course of the dissolution enables the liquids in the reaction chamber to be mixed at the desired point in time.
  • the temporal dissolution behavior of the membrane when it comes into contact with liquid can be structurally adjusted both through the selection of the material and through the thickness of the membrane. This is particularly advantageous because, in the event of flow slowdowns occurring in one of the two channels and an associated retarded flow into the reaction chamber, an undefined displacement of the liquids can be avoided.
  • more than two channel systems can also be connected to one another as described in order to carry out mixing processes with more than two liquids.
  • the active element in the bottom area of the second reservoir space of the reaction chamber is designed as a delivery system for active substances and other substances.
  • active substances and / or other substances can be embedded or fixed in the active element, these active substances and / or other substances being released by the activating environmental variable.
  • active substances and / or other substances such as enzymes, substrates, precursors, etc., can be immobilized in advance in the reaction chamber and mobilized when the liquid is present, with the time release of the active substances and / or other substances in turn being adapted to the needs of the user be able.
  • a release is possible after activation of the active elements delimiting the reaction chamber, so that the active substances and / or other substances are released into the volume defined by the reaction chamber. It is also conceivable that the release takes place before the membrane is dissolved.
  • the first and second liquids would mix in the reaction chamber, with the second liquid already containing the active substances and / or other substances.
  • the release into the reaction chamber would only take place after the first and second liquids have been mixed. This would be advantageous if the first and second liquids are to carry out a reaction first and the addition of a substrate, etc. is only possible after this reaction has ended.
  • the targeted immobilization of the active substances and / or other substances opens up a wide range of application of the microfluidic micromechanical system in analysis.
  • the delivery system for active substances and other substances is designed, for example, as a depot or storage facility, which is activated by the presence of liquid. That is why it can be called an active element.
  • Such a storage element could also be designed as a polymer network. During the deswelling process or dissolution process through the presence of liquid, it releases the swelling agent and the substances it contains.
  • the active elements are designed to be activated by the presence of liquid as an environmental variable. Both a change in the state of swelling due to the absorption of liquid and a dissolution of the active element as a result of the contact with the liquid are conceivable.
  • the active elements are designed to determine the time sequence and the time behavior of the mixing of the first and second liquids.
  • the time behavior of the active elements can be directly influenced.
  • the time behavior of the active elements can be controlled, for example, through a suitable selection of materials.
  • the timing can also be influenced by the dimensioning of the active elements. For example, larger-sized active elements, which experience an increase in volume due to the activating environmental size, can achieve a faster suppression of the liquid flow than comparably smaller-sized active elements.
  • a slowed-down dissolution due to the larger dimensioning of the active element can be set in a targeted manner.
  • the time sequence can be controlled both as a function of the material and as a function of the dimensions.
  • the active elements consist of hydrogels which are chemically crosslinked and / or physically crosslinkable.
  • hydrogels are a water-containing but water-insoluble polymer understood, the molecules of which chemically, e.g. B. by covalent bonds, or physically, e.g. B. by entangling the polymer chains, are linked to form a three-dimensional network. Due to built-in hydrophilic polymer components, they swell in liquids with a considerable increase in volume, but without losing their material cohesion. It is essential here that the hydrogels are designed in such a way that they remain in the swollen state after contact with liquids.
  • the active elements consist of hydrogels which are selected from a group which includes, for example, polyacrylamides, polyvinyl alcohols, polyacrylates, hydroxycellulose, polyvinyl pyridines or polyglycols (e.g. polyethylene glycol, polypropylene glycol) and their derivatives.
  • hydrogels which are selected from a group which includes, for example, polyacrylamides, polyvinyl alcohols, polyacrylates, hydroxycellulose, polyvinyl pyridines or polyglycols (e.g. polyethylene glycol, polypropylene glycol) and their derivatives.
  • the active elements are made from uncrosslinked polymers, salts or organic natural substances such as saccharides. This is the case when the active elements are designed as liquid-soluble barriers. All materials can be used which, when dried, form a solid, sol-gel or the like and dissolve on contact with a liquid.
  • the material basis of the uncrosslinked polymers can in principle be the same as for the crosslinked polymers. While the polymers crosslinked to form a three-dimensional network serve as swellable swelling agent barriers, the same polymers dissolve in the liquid if they are uncrosslinked, since the polymer chains that are not connected to one another can dissolve.
  • the present invention also relates to a method for microfluidic process control in a microfluidic micromechanical system, wherein a first liquid is introduced into a first channel, a second liquid is introduced into a second channel and the first and second liquid are mixed in a reaction chamber which is formed in the overlapping area of the first and second channels, the time sequence of the mixing of the first and second liquids in the reaction chamber being determined by active elements.
  • the method steps described above are particularly advantageous for controlling the timing of the mixing of two liquids in a microfluidic system.
  • the respectively desired temporal sequence of process steps such as mixing, dissolution of barriers, closure of desired channel sections by means of swelling agent barriers, release of active substances and / or other substances can be achieved in a user-specific manner.
  • the time sequence of the mixing the first and second liquid in the reaction chamber by the active elements which are liquid-soluble or designed as a swelling agent barrier. In this way, both a suppression of the flow and an opening of channel sections for the flow through with the first or second liquid can be realized.
  • the method further comprises the dissolution of a liquid-soluble membrane, which divides the reaction chamber into a first reservoir space and a second reservoir space, by the first and second liquids before the first and second liquids are mixed.
  • the dissolution of the membrane removes the division of the reaction chamber into a first and a second reaction chamber, so that the first and second liquids which are present in the first and second reservoir chambers are mixed.
  • the microfluidic microchemomechanical system is used to carry out processes based on antigen-antibody reactions, carry out processes based on the culture method, control and / or detect processes based on a polymerase chain reaction and detect the enzyme activity of a biochemical process. Further applications based on chemical or biochemical mixed reactions are conceivable.
  • the microfluidic microchemomechanical system according to the invention is characterized in that it enables a first and a second liquid to be mixed in a reaction chamber with a defined volume and in a time-controllable manner without auxiliary energy.
  • immobilized active substances and / or other substances can be released in a time-controlled manner and thus enable reactions in the reaction chamber.
  • Fig. 1 an inventive microfluidic microchemomechanical system is shown, which is designed as an autarkic and automatically working microfluidic processor for equidistant long-term investigations.
  • the microfluidic processor in Fig. 1 carries out long-term studies that consist of identical analytical or other mixture reactions and that are repeated according to a defined schedule. Equidistant examinations are among the most common methods in science and technology. Among other things, they are used to control critical parameters, e.g. B. the monitoring of bioreactors, for enzyme analysis, the analysis of growth factors or the quality control of chemical and biological products.
  • the microprocessor in Fig. 1 is divided into 192 serially connected, structurally identical levels 1 and comprises a total of 2096 active elements 7 and 384 reservoir spaces 9, 10.
  • a stage 1 ( Figures 2a, 2b ) of the microfluidic processor through all steps of sampling, sample preparation and the initiation of the mixing reaction completely independently and energy self-sufficient.
  • How level 1 works ( Fig. 2a ) is as follows.
  • the liquids 13 and 14 of the two channels 3 and 4 reach level 1, so that the binary concentration switches from 0 to 1.
  • This chemical signal activates the integrated active elements 7 and stimulates them to release their stored chemical energy in the form of a defined fluidic function in a by the fluidic interconnection predefined time sequence.
  • the closing elements 7a for example consisting of the hydrogel sodium acrylate, close the inlets and outlets of the reservoir spaces 9, 10 and thereby separate and dose them Liquids 13.14.
  • the closing time of the closing elements 7a is selected such that the reservoir spaces 9, 10 are most likely completely filled with the liquids 13, 14. It can be, for example, 45 s (ratio of volume V gel of the sodium acrylate actuator to the volume of the reaction chamber 6 VK 1: 5.6, see also Figure 6c ).
  • the desired reaction can now take place by mixing the liquids 13, 14.
  • the membrane 7e which is designed as an active membrane, for example, must be mechanically stable enough that it is not significantly deflected when the reservoir spaces 9, 10 are flooded. In addition, their opening time or opening time must not be too short in order to avoid unwanted, premature mixing.
  • an active membrane 7e made of uncrosslinked polyvinyl alcohol for example 70 ⁇ m thick, a corresponding dimensional stability can be achieved with an opening time of 7 minutes (see also FIG Figure 7a ).
  • the opening elements 7b which for example. made of polyethylene glycol (PEG) 6000, closed in the chamber bypasses.
  • PEG polyethylene glycol
  • the opening elements 7b are essential elements for sequential circuits with many stages or cascades. Without this, the fluidic resistances of the bypass channels would have to be chosen to be much higher than the fluidic resistances of the channels leading to the reservoir spaces. This would mean that the bypass resistors that add up due to the series connection would limit the number of stages that can be switched in series to 3 or 4.
  • the opening element 7d defines the time until the activation of the next stage. After the opening elements 7d have been triggered, the liquids 13, 14 flood the next stage. At this moment the closing elements 7c close the bypasses to the in Fig. 1 apparent circulation channels 12. Even with the circuit combination of the elements 7c and 7d, it is possible to use the pressure increase over the opening element 7d as a result of the closure of 7c to open 7d.
  • the in Figure 1 The microprocessor shown is capable of mixing reactions in time intervals of 2 minutes (opening elements 7d made of polyethylene glycol 6000 and an element length of 400 ⁇ m, see also Figure 7c ) to be carried out independently and automatically, but it can also work for up to 16 days with self-sufficient and automatic implementation of mixing reactions at two-hour intervals (opening elements 7d made of PEG 35000 and 1.2 mm in length).
  • the microfluidic microchemomechanical system in Fig. 1 has a two-level architecture (see Figure 2b ).
  • the upper structural carrier 2a which also functions as a cover, for example, contains the channel structure of the channel 3 for the liquid 13, while the lower channel structure carrier 2b carries the channel structure of the channel 4 for the liquid 14.
  • Both structural supports have a comparable design, for example, which can essentially be mirrored.
  • Channels 3 and 4 are for the in Fig. 1 Example shown 800 ⁇ m wide and 140 ⁇ m high.
  • the bypass channels 8 are 400 ⁇ m wide and 140 ⁇ m high.
  • the square diamonds for the closing elements have a volume of 1000 x 1000 x 140 ⁇ m 3 (7a) or 800 x 800 x 140 ⁇ m 3 (7c).
  • the configuration of the active elements for the arrangements in the Figures 1 and 2 is as follows: the thickness of the active membrane made of uncrosslinked polyvinyl alcohol is 70 ⁇ m.
  • the length of the opening elements 7b (PEG 6000) is 400 ⁇ m, the length of the opening elements 7d (PEG 6000) is 800 ⁇ m.
  • the in Fig.1 microfluidic microchemomechanical system shown in Fig. 1 realized with only one structural support 2 and an unstructured cover 2a. Both channel systems 3, 4 are located on the same structural support 2, that is, in one plane. In the overlapping area of the channels 3, 4 an opener element, which in principle is designed like the opener elements 7b, 7d, is arranged between the reservoir spaces 9,10, which connects the two reservoir spaces 9,10 to the reaction chamber 6 after it has been dissolved.
  • the monolithic microchips of the microfluidic microchemomechanical systems ( Fig. 1 ) made entirely of polymers.
  • the structural supports 2, which contain the channel networks, consist, for example, of polydimethylsiloxane (PDMS) and were made using multilayer soft lithography [ DC Duffy, JC McDonald, OJA Schueller, GM Whitesides, Anal. Chem. 70: 4974-4984 (1998) ] using a large-area replication technology with masters made from solid resists [ A., Richter, G. Paschew, Adv. Mater. 21 (2009), 979-983 ] manufactured.
  • the multilayer soft lithography using PDMS is primarily suitable for research and demonstrator construction.
  • thermoplastic polymers which may include, for example, polystyrene, polycarbonate, olefins such as cycloolefin, polyesters such as polyethylene terephthalate.
  • phase-changeable polymers are used, for example, which can be integrated into the microchip using simple microtechnical methods.
  • Polyethylene glycols are microstructured photolithographically using stencil printing and sodium acrylate actuators.
  • the active membranes made of polyvinyl alcohol can be integrated with pick-and-place technology, for example.
  • the microstructuring of the sodium acrylate actuators is carried out by photolithographic polymerization.
  • An exemplary production procedure is based on a mixture of 2 g of sodium acrylate, 0.04 g of the crosslinking agent N, N-methylenebisacrylamide (BIS), and 0.04 g of the photoinitiator 2-hydroxy-4 '- (2-hydroxyethoxy) -2-methylpropiophenone, everything dissolved in 14 ml of distilled water. This solution is stirred for 24 hours under an argon protective gas atmosphere.
  • this stock solution is referred to as c 0 .
  • the photopolymerization also takes place under an argon protective gas atmosphere either directly in the channel structures or in a photopolymerization chamber.
  • the quality and crosslinking properties of the sodium acrylate actuators depend on the polymerization time, the distance to the exposure source, the type of exposure source and the height of the polymerization chamber.
  • meltable polyethylene glycol which can be structured using a stencil printing technology, is used for the opener elements 7b and 7d.
  • a structured copper mask with a thickness of 20 ⁇ m was placed on the structure carriers 2a, b in such a way that their openings were at the desired positions of the opening elements 7b, d.
  • the melted PEG is placed on the copper mask and distributed with a metal squeegee so that the opening elements 7b, 7d are created in the structure carriers 2a, 2b in the mask openings.
  • the PEG touches the structural support it cools and hardens.
  • the generated opener elements already have their geometric dimensions, but do not yet seal the channels.
  • Hermetically sealed opening valves are achieved in a final microchip manufacturing step by briefly heating the already fully assembled microchip slightly above the melting temperature of the PEG. The PEG structures melt and seal the channels tightly.
  • a 5% strength polymer solution is poured into a mold and then dried.
  • the height of the membrane produced in this way can be determined by the filling quantity and thus the height of the solution in the casting mold.
  • a microfluidic microchemomechanical system is in the Figures 3a and 3b shown. These show the stage of a further microprocessor, which also consists of sequentially switched stages. Here the stages have the task of carrying out several mixing reactions with different ratios at the same time.
  • the simultaneous execution of investigations with different volume ratios of sample and analyte or simply two chemicals enables, among other things, the determination of reaction kinetics, for example the determination of an enzyme activity.
  • the mode of operation of the Figures 3a and 3b The stage shown is explained on the basis of the investigation of enzyme kinetics.
  • a liquid containing the enzyme of interest for example laccase, a polyphenol oxidase of the fungus Trametes versicolor, is fed through the feed channel 3, which is, for example, 800 ⁇ m wide and 140 ⁇ m high.
  • the initially closed opening element 7d forces the medium to flood the five parallel channel structures, which have a width of 400 mm, height 140 ⁇ m, for example, with reservoir spaces 9.
  • the process medium first flows via the bypass 8 in the direction of the circulation channel 12 functioning as a drain. This continues until the opening element 7d, which is made from PEG 6000 and has a length of, opens and the medium in channel 3 can flow to the next stage.
  • Each of the now hermetically closed reaction chambers now contains a volume of enzyme-containing process medium corresponding to the size of the reservoir space 9.
  • each reservoir space 9 has a depot 11 in the floor space, in which an analyte in the form of a dried, liquid-soluble active element 7f was introduced during the microchip production.
  • the analyte-containing active element 7f consists for example of dried, immobilized substrate 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) in a malonate buffer. The presence of the aqueous process medium causes the substrate to dissolve and the mixing reactions start.
  • the reservoir spaces 9 and the depots 11 located therein represent, for example, volume ratios of sample to analyte of 1: 3, 1: 2, 3: 1, 2: 1 and 1: 1.
  • a microfluidic microchemomechanical system which, as an autarkic and automatically operating, highly parallel microfluidic [NxM] matrix processor, realizes any possible combination of N chemicals organized in rows with M chemicals organized in columns.
  • Fig. 1 is a [48x48] matrix processor is shown.
  • An exemplary application scenario of such a [48x48] matrix processor is the parallel examination of 48 samples for 48 parameters, for example for screening purposes.
  • the advantage of such matrix processors is that all examinations are carried out at the same time under exactly the same conditions.
  • the massively parallel execution of the tests also brings the advantages of high integration to bear, so that test series that typically last days or weeks can be carried out in hours.
  • the [48x48] matrix processor carries out 2304 examinations simultaneously and fully automatically. It has a total of 2401 closing elements 7a and 2304 active membranes 7e. Its mode of operation is explained below using the Figures 5a and 5b for a [4 x 4] matrix section and an example configuration.
  • Liquids are introduced into the row channels 15 and 16 as well as the column channels 17 and 18 at the same time and at the same flow rate;
  • the liquids of the row channels 15, 16 flood the reservoir spaces 9, the liquids of the column channels 17, 18 simultaneously flood the reservoir spaces 10.
  • Figure 5c shows that by inserting further fluidic levels in each matrix point, more than two liquids can be mixed with one another.
  • a further central structural support 2c is integrated into the overall structure, which has a configuration of active elements 7a, 7e of the same type as the other two structural supports 2a, 2b.
  • This simple stacking of three structural supports makes it possible to combine three reservoir chambers 9, 10, 19, which are fed through different channels 16, 18, 21 to form a reaction chamber 6 and thus to mix three liquids with one another in one matrix point.
  • Figures 6a , 6b and 6c Opportunities to predefine the parameters of the closing elements 7a, 7c, in particular the closing time and the pressure resistance, through the choice of material and design parameters.
  • Figure 6a shows that the closing time is due to the hydrophilicity or the cooperative Diffusion coefficient of the selected material can be preset.
  • Two types of hydrogels can be distinguished, neutral hydrogels and polyelectrolytic hydrogels.
  • Neutral hydrogels such as cross-linked polyacrylamide, poly ( N -isopropyl acrylamide), polymethyl vinyl ether, polyvinyl alcohol or polyethylene glycol have cooperative diffusion coefficients D coop of the order of 10 -7 cm 2 s -1 .
  • hydrogels are predestined as a material basis for relatively slow closing elements with closing times in the range of minutes or hours.
  • Polyelectrolytic hydrogels which contain ionizable groups, for example acid or base groups, have cooperative diffusion coefficients of the order of 10 -7 to 10 -5 cm 2 s -1 due to additional inter- and intramolecular electrostatic interactions which have an expansive effect.
  • Polyelectrolytic hydrogels, which are used as superabsorbents have the largest D coop .
  • Sodium acrylate hydrogel is one of them. As Figure 6a shows, D coop of sodium acrylate depends on the crosslinking conditions.
  • the higher crosslinker density leads to mechanically more stable hydrogels, so that the compressive strength of the closing elements increases with increasing crosslinker content or increasing crosslinking density of the sodium acrylate actuators.
  • the closing time of the closing elements can also be set using a structural variable, namely the ratio of the dry volume of the sodium acrylate hydrogel actuator to the reaction chamber volume of the closing element seat ( Figure 6c ).
  • opening times of opening elements 7b, 7d and 7e can also be preset by choosing the material ( Figure 7a ). The following applies: the more hydrophilic the selected water-soluble polymer, the faster the active element dissolves.
  • One constructive parameter is of great importance for the opening time: the thickness of the active membrane ( Figure 7a ) or the length of the opening elements ( Figure 7b ).
  • opening elements 7b and 7d which are not subject to deflection
  • significantly softer materials can also be used, such as polyethylene glycol.
  • the flow rate of the liquid flowing past is the same significant influence on the opening time of opening elements.
  • Figure 7b clearly shows that opening elements open very slowly when the liquid is stagnant. In this case, saturation zones of dissolved polymer can form in front of the opening element, which impair the further dissolution process of the polymer. As the flow rate increases, these saturation zones are destroyed and the polymer dissolves more quickly.
  • Figure 6c demonstrates using the example of a PEG 6000 opening element that the standard deviation of active elements 7 can be kept very low even with simple microtechnical laboratory manufacturing methods.
  • uric acid content in serum or urine provides information about the breakdown of purine bases and is used, for example, in cases of suspected gout, monitoring of cell-destroying processes and stone diseases.
  • the recommended upper limit for men is 416 ⁇ mol / l.
  • the test is carried out as a coupled enzyme test in which uric acid is oxidized by the uricase. This creates hydrogen peroxide, which can be detected with a peroxidase (HRP).
  • HRP peroxidase
  • the substrate Amplex Red (5 mM in DMSO) is first mixed with 99 times the volume of an enzyme solution (0.1 M Tris / HCl, pH 7.4; 0.2 U / ml Uricase; 0.2 U / ml HRP) and introduced into a stage 1 of the microfluidic, microchemomechanical system.
  • the resulting reaction solution is then introduced into the second reservoir space via the second channel 4, while the first reservoir space 9 is filled with the same volume of the sample to be examined (contains 0-100 ⁇ M uric acid) via the first channel 3.
  • the soluble membrane 7e which separates the two reservoir spaces 9, 10 from one another, dissolves as a result of the contact with the liquid, as a result of which the reaction chamber 6 is formed and the reactants are mixed.
  • the enzymatic conversions now take place at a reaction temperature of 37 ° C. After a reaction time of 5 min, fluorescence at 590 nm can be detected after excitation with light (530 nm). The concentration can be calculated from the intensity of the fluorescence using a corresponding calibration.
  • a protein detection with ortho-phthalaldehyde is described.
  • the protein with the detection reagent OPA is involved a thiol-containing component, such as ⁇ - mercaptoethanol, implemented. This creates a fluorophore that can easily be detected.
  • 100 ⁇ l of the detection reagent (6 ⁇ g / ml OPA; 0.1 M phosphate buffer, pH 7.4; 0.05 vol% ⁇ -mercaptoethanol) are filled into the first reservoir space 9, while 100 ⁇ l of the sample to be examined, such as a 5-fold diluted serum, is passed into the other reservoir space 10.
  • Both reservoir spaces 9, 10 have the same volume.
  • a reaction occurs.
  • the resulting signal can be read from the reaction chamber 6 after 2-3 minutes.
  • the Figure 8a and 8b show the time-dependent course of the detected fluorescence intensity of four samples at a wavelength of 455 nm.
  • the corresponding protein concentration ( Figure 8a ) can be derived from a calibration line ( Figure 8b ) can be determined on the basis of BSA as reference protein.
  • Figure 9a and 9b describes the detection of myoglobin in blood.
  • Antibodies are immobilized in the reaction chamber 6.
  • the sample is then introduced into the reaction chamber 6 coated with an antibody (anti-myoglobin) and incubated for 1 hour at RT (room temperature). It is then washed with a washing solution (137 mM sodium chloride, 2.7 mM potassium chloride, 12 mM phosphate buffer, pH 7.4, 0.05% Tween 20).
  • a washing solution 137 mM sodium chloride, 2.7 mM potassium chloride, 12 mM phosphate buffer, pH 7.4, 0.05% Tween 20.
  • the antibody solution anti-myoglobin-HRP or anti-myoglobin-GFP
  • the signal can be read out directly (GFP 475/530 nm) or the reaction solution (0.1 M Tris / HCl, pH 7.5, 10 ⁇ M H 2 O 2 , 50 ⁇ M Amplex Red) must now be added to the read out the resulting signal (Amplex Red Ex: 530 nm Em: 590 nm).
  • the washing solution can be supplied via the first or second channel 3, 4.
  • HSA human serum albumin
  • the Figure 9a shows the time-dependent course of the detected fluorescence intensity from a triplicate determination of an HSA sample (0.3 mg / ml) at a wavelength of 423 nm can be determined from a calibration line based on BSA as a reference protein.
  • the Figure 9a shows the determined fluorescence intensity in the case of the detection of HSA as a triplicate determination at 423 nm after mixing with a detection reagent over time (gain: 178).
  • the corresponding protein concentration of the sample can be determined using a calibration line ( Figure 9b ) can be determined (gain: 100).
  • protein detection is described using the example of bovine serum albumin (BSA) with fluorescamine.
  • BSA bovine serum albumin
  • Fluorescamine reacts with amino acids to form pyrolinone derivatives, which can be excited at a wavelength of 395 nm, whereby a fluorescence maximum can be detected at 470 nm.
  • the Fig. 10 shows the concentration-dependent fluorescence intensity determined from a triplicate determination of a BSA sample at an excitation wavelength of 395 nm.
  • the fluorescence intensity increases as the concentration of BSA increases.
  • Bovine serum albumin (BSA) is detected as a triple determination at 470 nm after mixing with fluorescamine (gain: 80).

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)
  • Physical Or Chemical Processes And Apparatus (AREA)

Claims (14)

  1. Système microchimiomécanique microfluidique destiné au mélange volumétriquement défini d'un premier liquide avec au moins un second liquide dans des séquences temporelles définies comportant des éléments actifs (7) intégrés, lesquels sont conçus de manière à pouvoir être activés sans énergie auxiliaire par des grandeurs d'environnement influençables et à provoquer des fonctions actives par le changement de leur état de gonflement ou de leurs propriétés mécaniques, dans lequel les éléments actifs (7) sont conçus sous la forme de barrières d'agent gonflant ou barrières solubles dans un liquide, comprenant
    - au moins un support structural (2) comportant au moins un premier canal (3) pour le premier liquide,
    - un couvercle (2a), lequel recouvre au moins partiellement le support structural (2), et
    - au moins un second canal (4) pour le second liquide, dans lequel le second canal (4) est disposé sur le support structural (2) ou le couvercle (2a),
    dans lequel les canaux (3, 4) forment respectivement des espaces de réservoir (9, 10, 19) délimités par des éléments actifs (7), lesquels sont disposés de telle sorte qu'ils comportent ensemble au moins une zone de chevauchement (5) et forment ensemble une chambre de réaction (6), dans lequel une membrane (7e) active est disposée entre les premier et second canaux (3, 4) dans la zone de chevauchement (5) des premier et second canaux (3, 4), la chambre de réaction (6) étant divisée en un premier espace de réservoir (9) et en un second espace de réservoir (10).
  2. Système micromécanique microfluidique selon la revendication 1, caractérisé en ce que d'autres canaux (16, 18, 21) sont fournis, dans lequel des membranes (7e) sont disposées entre les espaces de réservoir (9,10,19) appartenant aux canaux (16,18,21) dans les zones de chevauchement (5) de plus de deux canaux (16, 18, 21), lesquels espaces de réservoir formant la chambre de réaction (6).
  3. Système micromécanique microfluidique selon l'une des revendications 1 ou 2, caractérisé en ce que les membranes (7e) entre les premier, second et éventuellement autres espaces de réservoir (9, 10, 19) sont constituées d'un matériau soluble dans un liquide.
  4. Système micromécanique microfluidique selon l'une quelconque des revendications précédentes, caractérisé en ce qu'il comporte une chambre de réaction (6) comprenant au moins un premier espace de réservoir (9) et un second espace de réservoir fonctionnant comme un dépôt (11), dans lequel un élément actif (7f) est disposé dans le dépôt (11).
  5. Système micromécanique microfluidique selon la revendication 4, caractérisé en ce que l'élément actif (7f) est conçu sous la forme de système de distribution de substances actives et/ou d'autres substances dans la zone de base du dépôt (11).
  6. Système micromécanique microfluidique selon l'une quelconque des revendications précédentes, caractérisé en ce que les éléments actifs (7) sont conçus pour être activés par la présence de liquide en tant que grandeur d'environnement.
  7. Système micromécanique microfluidique selon l'une quelconque des revendications précédentes, caractérisé en ce que les éléments actifs (7) sont conçus pour être définitifs par une sélection appropriée de matériaux ou par leur dimensionnement ou par une combinaison appropriée de la séquence temporelle précitée ainsi que du comportement dans le temps du mélange de différents liquides (13, 14).
  8. Système micromécanique microfluidique selon l'une quelconque des revendications précédentes, caractérisé en ce que les éléments actifs (7) sont constitués d'hydrogels réticulés chimiquement et/ou physiquement réticulables.
  9. Système micromécanique microfluidique selon la revendication 8, caractérisé en ce que les éléments actifs (7) sont constitués d'hydrogels choisis dans un groupe de polymères réticulés, de préférence de polyacrylamides, d'alcools polyvinyliques, de polyacrylates, d'hydroxycellulose, de polyvinylpyridines ou de polyglycols tels que le polyéthylène glycol, le polypropylène glycol et leurs dérivés.
  10. Système micromécanique microfluidique selon l'une quelconque des revendications 1 à 7, caractérisé en ce que les éléments actifs (7) sont conçus à base de polymères non réticulés, de sels ou de substances organiques naturelles telles que les saccharides.
  11. Procédé de commande de processus microfluidique dans un système micromécanique microfluidique selon l'une quelconque des revendications 1 à 10, comprenant les étapes suivantes :
    - l'introduction d'un premier liquide (13) dans un premier canal (3) et dans un premier espace de réservoir (9),
    - l'introduction d'un second liquide (14) dans un second canal (4) et dans un second espace de réservoir (10),
    - la fermeture et la séparation des espaces de réservoir (9, 10) par des éléments actifs (7a), lesquels sont conçus sous la forme de barrières d'agent gonflant, et, ainsi liée, de quantification des volumes de liquide (13, 14) dans les espaces de réservoir (9, 10)
    - l'interconnexion des espaces de réservoir (9, 10) à la chambre de réaction (6) par ouverture de l'élément actif (7e) dans la zone de chevauchement (5) des premier et second canaux (3, 4), ensuite suivie du mélange d'un premier et d'un second liquide (13, 14) dans la chambre de réaction (6), laquelle est formée dans la zone de chevauchement (5) des premier et second canaux (3, 5), dans lequel la séquence temporelle du mélange des premier et second liquides (13, 14) dans la chambre de réaction (6) est déterminée par les propriétés de l'élément actif (7e).
  12. Procédé selon la revendication 11, caractérisé en ce que la séquence temporelle du mélange des premier et second liquides (13, 14) dans la chambre de réaction (6) est déterminée par une sélection appropriée de matériaux ou par le dimensionnement ou par une combinaison appropriée de sélection de matériaux et de dimensionnement des éléments actifs (7), lesquels sont conçus comme solubles dans un liquide ou sous la forme de barrière d'agent gonflant.
  13. Procédé selon la revendication 11, comprenant en outre :
    - la dissolution d'une membrane (7e) soluble dans un liquide, laquelle divise la chambre de réaction (6) en un premier espace de réservoir (9) et en un second espace de réservoir (10), par les premier et second liquides (13, 14) avant le mélange des premier et second liquides (13, 14).
  14. Utilisation d'un système micromécanique microfluidique selon l'une quelconque des revendications 1 à 10 et d'un procédé selon l'une quelconque des revendications 11 à 13, destinée à la mise en œuvre des processus basés sur des réactions antigène-anticorps, la mise en œuvre des processus basés sur la méthode de culture, la commande et/ou la détection des processus basés sur une réaction en chaîne par polymérase et la détection de l'activité enzymatique dans un processus biochimique.
EP13717014.8A 2012-04-13 2013-04-11 Procédé et dispositif de commande de processus ciblée dans un processeur microfluidique doté d'éléments actifs intégrés Active EP2836302B8 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102012206042A DE102012206042B4 (de) 2012-04-13 2012-04-13 Verfahren und Vorrichtung zur gezielten Prozessführung in einem Mikrofluidik-Prozessor mit integrierten aktiven Elementen
PCT/EP2013/057631 WO2013153181A1 (fr) 2012-04-13 2013-04-11 Procédé et dispositif de commande de processus ciblée dans un processeur microfluidique doté d'éléments actifs intégrés

Publications (3)

Publication Number Publication Date
EP2836302A1 EP2836302A1 (fr) 2015-02-18
EP2836302B1 true EP2836302B1 (fr) 2021-12-08
EP2836302B8 EP2836302B8 (fr) 2022-01-12

Family

ID=48139927

Family Applications (1)

Application Number Title Priority Date Filing Date
EP13717014.8A Active EP2836302B8 (fr) 2012-04-13 2013-04-11 Procédé et dispositif de commande de processus ciblée dans un processeur microfluidique doté d'éléments actifs intégrés

Country Status (4)

Country Link
US (1) US9272281B2 (fr)
EP (1) EP2836302B8 (fr)
DE (1) DE102012206042B4 (fr)
WO (1) WO2013153181A1 (fr)

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3461559A1 (fr) * 2015-06-11 2019-04-03 Neofluidics LLC Plaque de puits entraînée par pipette manuelle ou électronique pour le stockage de nano-litres de gouttelettes et procédés d'utilisation associés
DE102016124059B4 (de) 2016-07-25 2019-12-19 Leibniz-Institut Für Festkörper- Und Werkstoffforschung Dresden E.V. Vorrichtung für die mikrofluidik
WO2018200896A1 (fr) 2017-04-28 2018-11-01 Neofluidics, Llc Dispositifs fluidiques à puits de réaction, et utilisations associés
EP3665262A4 (fr) 2017-08-09 2021-09-01 Neofluidics, LLC Dispositifs et procédés pour essai biologique
CA3082074A1 (fr) 2017-11-10 2019-05-16 Neofluidics, Llc Circuit fluidique integre et dispositif de manipulation de gouttelettes et procedes associes
US11998885B2 (en) 2018-10-26 2024-06-04 Unchained Labs Fluidic devices with reaction wells and constriction channels and uses thereof
DE102022125010A1 (de) 2022-09-28 2024-03-28 Leibniz-Institut für Festkörper- und Werkstoffforschung Dresden e.V. (IFW Dresden e.V.) Mikrofluidisches Bauteil

Family Cites Families (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE110142T1 (de) 1989-06-14 1994-09-15 Westonbridge Int Ltd Mikropumpe.
US5798215A (en) * 1993-02-18 1998-08-25 Biocircuits Corporation Device for use in analyte detection assays
US5659171A (en) 1993-09-22 1997-08-19 Northrop Grumman Corporation Micro-miniature diaphragm pump for the low pressure pumping of gases
DE69533554T2 (de) * 1994-11-10 2005-01-27 Orchid Biosciences, Inc. Flüssigkeitsverteilungssystem
US6030580A (en) * 1997-10-31 2000-02-29 Enerfab, Inc. Method of aseptically transporting bulk quantities of sterile products
US6283718B1 (en) 1999-01-28 2001-09-04 John Hopkins University Bubble based micropump
JP3548858B2 (ja) 2001-01-22 2004-07-28 独立行政法人産業技術総合研究所 流量の制御方法及びそれに用いるマイクロバルブ
DE10157317A1 (de) 2001-11-23 2003-06-05 Gesim Ges Fuer Silizium Mikros Grundelement eines Mikrofluidik-Prozessors
DE10202996A1 (de) 2002-01-26 2003-08-14 Eppendorf Ag Piezoelektrisch steuerbare Mikrofluidaktorik
EP1746158A4 (fr) * 2004-05-07 2009-11-25 Konica Minolta Med & Graphic Microreacteur pour essais, appareil pour essais genetiques et procede pour essais genetiques
US20060090800A1 (en) 2004-10-18 2006-05-04 Applera Corporation Fluid processing device including size-changing barrier
KR20060088780A (ko) 2005-02-02 2006-08-07 삼성전자주식회사 마이크로 벨브 장치 및 이를 응용한 장치
US20080069729A1 (en) 2005-02-16 2008-03-20 Mcneely Michael R Liquid Valving Using Reactive or Responsive Materials
US20100240022A1 (en) * 2006-06-23 2010-09-23 Mcneely Michael R Reagent preparation and valving design for liquid testing
DE102006051535A1 (de) 2006-10-27 2008-12-18 Andreas Dr. Richter Automatischer Mikrofluidik-Prozessor
WO2011157735A2 (fr) * 2010-06-15 2011-12-22 Carlsberg Breweries A/S Récipient à refroidissement automatique et dispositif de refroidissement

Also Published As

Publication number Publication date
DE102012206042B4 (de) 2013-11-07
EP2836302A1 (fr) 2015-02-18
US20150044688A1 (en) 2015-02-12
WO2013153181A1 (fr) 2013-10-17
US9272281B2 (en) 2016-03-01
EP2836302B8 (fr) 2022-01-12
DE102012206042A1 (de) 2013-10-31

Similar Documents

Publication Publication Date Title
EP2836302B1 (fr) Procédé et dispositif de commande de processus ciblée dans un processeur microfluidique doté d'éléments actifs intégrés
EP1846160B1 (fr) Nouveaux porte-echantillons microfluidiques
EP1807208B1 (fr) Dispositif d'analyse de proteine et d'adn integree et automatisee dans une cartouche a usage unique, procede de production de cette cartouche et procede de fonctionnement de l'analyse de proteine et d'adn a l'aide de cette cartouche
DE60029582T2 (de) Patrone zum durchführen einer chemischen reaktion
DE112007003160B4 (de) Automatischer Mikrofluidik-Prozessor
DE102007044889B4 (de) Diagnosetestsystem
DE102005054923B3 (de) Vorrichtung und Verfahren zur Aufbereitung einer Probe
EP1566215A2 (fr) Plateforme microstructurée et procédé de manipulation d'un liquide
EP3143119B1 (fr) Procédé et dispositif pour traiter un échantillon de matière biologique et systeme d'analyse d'un échantillon de matière biologique
EP1198294A1 (fr) Support de reaction microfluide presentant trois niveaux d'ecoulement et une couche protectrice transparente
EP1522343B1 (fr) Dispositif de test analytique comprenant une matrice hydrophile pour former un canal capillaire, son utilisation et procédé pour déterminer un analyte dans un liquide.
DE19910392B4 (de) Mikrosäulenreaktor
DE102005049976A1 (de) Anordnung zur integrierten und automatisierten DNA- oder Protein-Analyse in einer einmal verwendbaren Cartridge, Herstellungsverfahren für eine solche Cartridge und Betriebsverfahren der DNA- oder Protein-Analyse unter Verwendung einer solchen Cartridge
DE102006020716B4 (de) Mikrofluidik-Prozessor
EP2624954B1 (fr) Procédé de nettoyage d'une cavité microfluidique
EP2169383A1 (fr) Organe pour l'écoulement à travers des cuvettes et son utilisation
EP2525225B1 (fr) Installation et procédé d'analyse de la différentiation de cellules
WO2010040758A1 (fr) Dispositif et procédé permettant d'effectuer plusieurs réactions de pcr en parallèle selon le procédé par écoulement continu
EP2745936B1 (fr) Système fluidique avec absorbent et gel de polymere reversible
EP2894456A1 (fr) Système microfluidique et procédé de préparation et d'analyse d'un échantillon contenant des cellules d'un matériau biologique
DE102007009806A1 (de) Flüssigkeitshandhabungssystem
EP1510254A2 (fr) dispositif et procédé de détection d'un analyte dans un fluide
DE102016124059B4 (de) Vorrichtung für die mikrofluidik
DE202018006869U1 (de) Mikrofluidisches System für die digitale Polymerasekettenreaktion (dPCR) einer biologischen Probe
WO2021013595A1 (fr) Système de laboratoire sur puce comportant au moins une partie fonctionnalisée

Legal Events

Date Code Title Description
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

17P Request for examination filed

Effective date: 20141103

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

RIN1 Information on inventor provided before grant (corrected)

Inventor name: RICHTER, ANDREAS, PROF. DR.

Inventor name: ALLERDISSEN, MERLE

Inventor name: GREINER, RINALDO

DAX Request for extension of the european patent (deleted)
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: EXAMINATION IS IN PROGRESS

17Q First examination report despatched

Effective date: 20170510

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

Free format text: STATUS: EXAMINATION IS IN PROGRESS

GRAP Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOSNIGR1

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

Free format text: STATUS: GRANT OF PATENT IS INTENDED

INTG Intention to grant announced

Effective date: 20210706

GRAS Grant fee paid

Free format text: ORIGINAL CODE: EPIDOSNIGR3

GRAA (expected) grant

Free format text: ORIGINAL CODE: 0009210

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

Free format text: STATUS: THE PATENT HAS BEEN GRANTED

REG Reference to a national code

Ref country code: DE

Ref legal event code: R108

Ref document number: 502013016008

Country of ref document: DE

AK Designated contracting states

Kind code of ref document: B1

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

REG Reference to a national code

Ref country code: GB

Ref legal event code: FG4D

Free format text: NOT ENGLISH

REG Reference to a national code

Ref country code: AT

Ref legal event code: REF

Ref document number: 1453338

Country of ref document: AT

Kind code of ref document: T

Effective date: 20211215

Ref country code: CH

Ref legal event code: PK

Free format text: BERICHTIGUNG B8

Ref country code: CH

Ref legal event code: EP

RBV Designated contracting states (corrected)

Designated state(s): AL AT BE BG CH CY CZ 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

REG Reference to a national code

Ref country code: IE

Ref legal event code: FG4D

Free format text: LANGUAGE OF EP DOCUMENT: GERMAN

REG Reference to a national code

Ref country code: DE

Ref legal event code: R107

Ref document number: 502013016008

Country of ref document: DE

REG Reference to a national code

Ref country code: NL

Ref legal event code: FP

REG Reference to a national code

Ref country code: LT

Ref legal event code: MG9D

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: RS

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

Ref country code: LT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

Ref country code: FI

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

Ref country code: BG

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20220308

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: SK

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

Ref country code: SE

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

Ref country code: NO

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20220308

Ref country code: LV

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

Ref country code: HR

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

Ref country code: GR

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20220309

Ref country code: ES

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: SM

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

Ref country code: RO

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

Ref country code: PT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20220408

Ref country code: EE

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

Ref country code: CZ

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: PL

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: IS

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20220408

PLBE No opposition filed within time limit

Free format text: ORIGINAL CODE: 0009261

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

Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: DK

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

Ref country code: AL

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

26N No opposition filed

Effective date: 20220909

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: SI

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

REG Reference to a national code

Ref country code: BE

Ref legal event code: MM

Effective date: 20220430

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: MC

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

Ref country code: LU

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20220411

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: BE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20220430

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: IE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20220411

REG Reference to a national code

Ref country code: AT

Ref legal event code: MM01

Ref document number: 1453338

Country of ref document: AT

Kind code of ref document: T

Effective date: 20220411

P01 Opt-out of the competence of the unified patent court (upc) registered

Effective date: 20230524

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: NL

Payment date: 20230417

Year of fee payment: 11

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: AT

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20220411

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: IT

Payment date: 20230428

Year of fee payment: 11

Ref country code: FR

Payment date: 20230417

Year of fee payment: 11

Ref country code: CH

Payment date: 20230502

Year of fee payment: 11

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: GB

Payment date: 20230413

Year of fee payment: 11

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: HU

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT; INVALID AB INITIO

Effective date: 20130411

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: MK

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208

Ref country code: CY

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20211208