WO2015051993A1 - Procédé pour mélanger des liquides et système microfluidique basé sur la force centrifuge - Google Patents

Procédé pour mélanger des liquides et système microfluidique basé sur la force centrifuge Download PDF

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Publication number
WO2015051993A1
WO2015051993A1 PCT/EP2014/070288 EP2014070288W WO2015051993A1 WO 2015051993 A1 WO2015051993 A1 WO 2015051993A1 EP 2014070288 W EP2014070288 W EP 2014070288W WO 2015051993 A1 WO2015051993 A1 WO 2015051993A1
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WIPO (PCT)
Prior art keywords
mixing chamber
mixing
centrifugal system
liquid
centrifugal
Prior art date
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PCT/EP2014/070288
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German (de)
English (en)
Inventor
Melanie HOEHL
Arne Kloke
Juergen Steigert
Jens Liebeskind
Felix Von Stetten
Nils Paust
Original Assignee
Robert Bosch Gmbh
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Publication of WO2015051993A1 publication Critical patent/WO2015051993A1/fr

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/50273—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the means or forces applied to move the fluids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F25/00—Flow mixers; Mixers for falling materials, e.g. solid particles
    • B01F25/50—Circulation mixers, e.g. wherein at least part of the mixture is discharged from and reintroduced into a receptacle
    • B01F25/51—Circulation mixers, e.g. wherein at least part of the mixture is discharged from and reintroduced into a receptacle in which the mixture is circulated through a set of tubes, e.g. with gradual introduction of a component into the circulating flow
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F29/00—Mixers with rotating receptacles
    • B01F29/30—Mixing the contents of individual packages or containers, e.g. by rotating tins or bottles
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F33/00—Other mixers; Mixing plants; Combinations of mixers
    • B01F33/30—Micromixers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F33/00—Other mixers; Mixing plants; Combinations of mixers
    • B01F33/40—Mixers using gas or liquid agitation, e.g. with air supply tubes
    • B01F33/403—Mixers using gas or liquid agitation, e.g. with air supply tubes for mixing liquids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F33/00—Other mixers; Mixing plants; Combinations of mixers
    • B01F33/40—Mixers using gas or liquid agitation, e.g. with air supply tubes
    • B01F33/408—Controlling
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F33/00—Other mixers; Mixing plants; Combinations of mixers
    • B01F33/40—Mixers using gas or liquid agitation, e.g. with air supply tubes
    • B01F33/409—Parts, e.g. diffusion elements; Accessories
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J7/00—Apparatus for generating gases
    • B01J7/02—Apparatus for generating gases by wet methods
    • 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
    • B01L2300/0803—Disc shape
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0409—Moving fluids with specific forces or mechanical means specific forces centrifugal forces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/046—Chemical or electrochemical formation of bubbles

Definitions

  • the present invention relates to a method for mixing liquids in a mixing chamber in a microfluidic centrifugal system and to a microfluidic centrifugal system suitable for this purpose.
  • the processing and processing of biological, biochemical or chemical samples is based essentially on the handling of liquids.
  • various aids in particular pipettes and various reaction vessels, are used in order to be able to carry out the various processes during manual handling with the aid of various laboratory equipment.
  • Lab-on-a-Chip systems are microfluidic systems that combine the full functionality of a macroscopic laboratory on a plastic card-sized plastic substrate.
  • German published patent application DE 10 2010 003 223 A1 describes a centrifugal system for the automated performance of biochemical, biological or chemical processes.
  • the system comprises a cartridge-based centrifugation unit comprising two or more turret-type bodies arranged axially one above the other. These bodies, the different cavities Contain (reaction spaces) are rotated by means of an integrated ballpoint pen mechanism in response to the acting centrifugal force against each other, so that a controlled fluid guidance through the individual reaction spaces of the device is achieved. Liquids are in this case transported along the force vector of the centrifugal force from radially inner points to radially outer points. An acceleration change of the centrifuge activates the integrated ballpoint pen mechanism, whereby the revolvers can be rotated against each other and the cavities can be switched to each other. With such a system can be different
  • Lysis buffer come into contact and can be digested. Mixing in centrifugal systems poses a particular challenge since, as such, centrifuging generally causes separation of the phases and thus counteracts mixing.
  • Bubble column reactors are known in the field of process engineering, with ascending gas bubbles being used for the mixing of liquids. Due to the flow rate, different flow states can be produced, so that, especially at high flow rates, a strong turbulence and thus a marked mixing effect can be achieved (eg shenchez Mirön, A. et al., Mixing in bubble column and airlift reactors, Chemical Engineering Research and Design 82 (2004) 1367).
  • the use of gas bubbles is also known in general microfluidics. For example, Garstecki, P. et al.
  • centrifugable ampoule is used.
  • the trapped gas bubble in the ampule is reciprocated by changes in the acceleration of the liquid within the ampule, causing turbulence.
  • multiple accelerator changes of the rotor are required, which is a critical material load for the centrifuge and the
  • Mixing chamber provided in a microfluidic centrifugal system.
  • gas bubbles are generated in a chamber of the centrifugal system.
  • These gas bubbles are introduced from below into the mixing chamber via a structure which is provided for a substantially liquid-free gas bubble transfer.
  • the liquids or phases to be mixed are present, for example two liquid phases.
  • the introduced gas bubbles displace the liquid in the mixing chamber during the ascent and thus create a hydrodynamic fluidization within the liquid, which leads to the mixing of the phases. With this method, it is possible to realize a mixing process in micro-fluidic centrifugal systems which requires very little technical effort.
  • This method can therefore be used with great advantage, for example, also for disposable cartridges, which are provided for the automated processing of, for example, biochemical processes or assays.
  • the method according to the invention is particularly suitable, for example, for stacked microfluidic centrifugal systems, as known from the already mentioned German published patent application DE 10 2010 003 223 A1 are.
  • the method according to the invention can also be used for other centrifugal systems, in particular for cartridge-based systems.
  • the method of the invention utilizes gas bubbles for the purpose of mixing phases in a microfluidic centrifugal system, with gas bubble generation integrated into the centrifugal system.
  • the gas bubbles are transferred substantially liquid-free into the mixing chamber.
  • substantially refers to the general advantage of having only gas bubbles and no other liquids or reagents from the gas bubble generating chamber enter the mixing chamber, which in turn increases the efficiency of the mixing process Depending on the application, it may be harmless if such entrainment occurs, in which case the gas bubble transfer does not have to be completely free of liquid
  • the required components can be very easily pre-stored and completely integrated into the centrifugal system.
  • the particular advantage of the method according to the invention is that the mixing in the centrifugal system is carried out without an acceleration change, so that no further material loading takes place in the centrifuge or the centrifugal system itself.
  • Components for mixing according to the invention require no moving parts. Another advantage is that the mixing according to the invention in the microfluidic centrifugal system can be operated in a wide centrifugal acceleration range. The mixing according to the invention can easily be integrated as a unit operation in a total protocol and with standard
  • the structure which is provided for a substantially liquid-free gas bubble transfer can in particular be a valve or a valve-like structure.
  • a tortuous channel in particular a U-shaped channel, can be used.
  • a siphon which acts as a passive valve.
  • the gas bubbles within the centrifugal system are generated.
  • the gas bubbles may be generated, for example, by being released from a pressure chamber in which the gas is stored upstream within the centrifugal system, if necessary, and the structure provided for a substantially liquid-free gas bubble transfer, for example via a siphon, into the mixing chamber reach. It is particularly preferred if the gas itself is first generated directly in the centrifugal system if necessary.
  • the gas and the gas bubbles can be generated by a chemical reaction.
  • the generation of the gas can in this case be carried out, for example, such that one or more reagents and optionally at least one catalyst are kept separate from one another in the centrifugal system.
  • the reagents or, for example, a reagent and a catalyst are brought together so that gas is generated in the course of the triggered chemical reaction.
  • the formed gas enters the structure in the form of bubbles, which is intended for a substantially liquid-free gas bubble transfer, for example into the siphon.
  • This overpressure expresses possibly existing liquid out of the structure.
  • the structure connects the gas generating chamber and the mixing chamber with each other. Above a critical overpressure Ap crit.
  • a gas bubble is ejected into the mixing chamber.
  • the overpressure in the structure suddenly drops off.
  • the pressure by the gas generation increases again until Ap crit is reached again and the next bubble is ejected.
  • the bubble production is thus repeated continuously.
  • the gas bubbles are introduced continuously into the mixing chamber and lead by their upward movement to a mixing of the liquids or phases.
  • the mixing effect can be further enhanced by bursting of the gas bubbles in the mixing chamber. Such bursting occurs especially at elevated centrifugal accelerations.
  • the mixing chamber is open at the top, so that the generated gas after the mixing process in the surrounding vessel, in particular in the surrounding cartridge, escape and finally escape into the interior of the centrifuge, so that a vent is ensured.
  • the generation of the gas bubbles on the basis of a chemical reaction has the particular advantage that in this way high gas production rates can be realized.
  • gas production can be activated at a defined time.
  • the corresponding reagents and / or catalysts can readily be stored over a relatively long period of time, for example one year or more, so that appropriately equipped centrifugal systems can be stored without further ado.
  • V end gas volume
  • V pre pre- storage volume
  • gaseous oxygen (O 2 ) from hydrogen peroxide (H 2 O 2 ) is particularly suitable, since hydrogen peroxide is si- is more likely to be stored and assigned to a lower security level.
  • Hydrogen peroxide may advantageously be provided as a stabilized solution, for example as a 30% hydrogen peroxide solution.
  • This solution is contacted with a catalyst, such as manganese oxide, which triggers gas production.
  • This chemical reaction is readily integrated into a centrifugal system with little effort.
  • the Morrisrung and activation of the liquid reagents can be realized for example via a Dornstech mechanism in a stacked microfluidic system with mutually rotatable revolvers.
  • micro-packaging units that can be opened, for example, via centrifugally-sensitive fractures.
  • Such micro-packaging units are known from the publication by van Oordt, T. et al. (Miniature stick-packaging - an industrial technology for pre-storage and release of reagents in lab-on-a-chip Systems, Lab on a Chip 13 (2013) 288) known per se.
  • the liquid is discharged from the mixing chamber via a siphon structure or a siphon.
  • the siphon can be controlled in particular via the filling level in the mixing chamber.
  • the invention further comprises a microfluidic centrifugal system, which is provided in particular for the automated processing of liquids.
  • the centrifugal system comprises at least one gas bubble generating chamber, at least one structure which is provided for a substantially liquid-free gas bubble transfer, in particular a siphon, and furthermore at least one mixing chamber.
  • the liquids or liquid are included, which is to be mixed according to the invention.
  • the structure which is intended for a substantially liquid-free gas bubble transfer, connects the gas bubble generating chamber to the mixing chamber, so that the gas bubbles generated in the gas bubble generating chamber can be introduced into the mixing chamber, in particular from below.
  • the microfluidic centrifugal system is particularly suitable for carrying out the method according to the invention for mixing liquids in the mixing chamber as already described.
  • the microfluidic centrifugal system is preferably a cartridge-based system which preferably has at least two bodies arranged axially one above the other.
  • the bodies may be rotatable or displaceable relative to each other in response to a centrifugal force or equivalent force.
  • Within the body one or more cavities are located as reaction spaces.
  • the individual cavities or reaction spaces can be interconnected so that a predeterminable fluid flow can be generated.
  • the rotation or displacement of the bodies relative to one another can be achieved, for example, by means of an integrated ballpoint pen technology which is activated by an acceleration change of the centrifuge.
  • a system is known in principle from the German patent application DE 10 2010 003 223 A1 and can be used for the automated implementation of various biochemical, chemical and / or biological processes.
  • the inventive design of such a centrifugal system enables a very simple and effective mixing of different phases, in particular of liquid phases, in the system, so that the functionalities and the effectiveness of such a centrifugal system can be considerably improved by the design according to the invention.
  • the structure intended for substantially liquid-free gas bubble transfer is a structure which acts as a valve or which is a valve. It is essential here that the structure is suitable for the forwarding of gas, but at the same time largely precludes the passage of liquids or reagents from the gas bubble production chamber into the mixing chamber. This structure may continue, for example, a convoluted
  • Channel in particular a U-shaped channel, be.
  • a siphon which acts as a passive valve.
  • this structure which is provided for introducing the gas bubbles into the mixing chamber, is equipped with at least one diessigabscheide Modell.
  • the integrated te liquid separation structure prevents any entrained liquids in the form of aerosols or particles that arise in the course of chemical gas generation, are introduced into the mixing chamber and can lead to contamination there.
  • the remplissigabscheide Modell for example, in an expansion of the structure, in particular the siphon exist. In this expansion, the gas flow is slowed down so that entrained particles escape from the flow and settle. Aerosols can condense to droplets by the longer residence time in the remplissigabscheide Modell and also settle. In this way, the liquid-free gas transfer through the siphon can be further improved.
  • a flow obstruction can be provided in the mixing chamber. This obstacle may be arranged so that the gas bubbles are introduced on only one side of the obstacle, so that the Mammutpumpen effect enhances the mixture.
  • the mammoth pump effect can be explained as follows: If the resulting gas is directed to one side of an obstacle, then the liquid on this side has a lower average density pi ⁇ p2 than the other side since gravity acts equally on both liquid columns on the side of the less dense liquid, a higher level h 1 > h 2 -
  • a pumping around the liquid can be achieved, wherein the upper portion of the obstacle forms an overflow.
  • this upper region of the obstacle is provided with means for separating the liquid flow, for example with one or more projections, so that the liquid is divided into several paths when overflowing. This measure can also increase the mixing effect by breaking up the sequence of superimposed liquid layers.
  • the flow obstacle may further comprise at least one transverse channel, with which a so-called Venturi nozzle effect is achieved. Due to the Venturi effect, the mixing can be further enhanced by the introduced gas bubbles. This effect is based on the lowest static pressure at the narrowest point of the rising stream. At this narrowest point is the transverse channel. At the junction of the transverse channel with the rising stream creates a suction that draws liquid into the rising stream from the transverse channel. As a result of this embodiment, liquids from different regions of the mixing chamber are mixed with one another, so that an overall increased mixing takes place.
  • the discharge of the mixed liquid (s) or of the mixed phases from the mixing chamber can take place in different ways.
  • an outflow may be provided, for example, at the bottom of the mixing chamber, which is opened once by a mandrel or the like when the mixing process is completed.
  • the discharge can be realized by a pierceable foil.
  • a siphon system and in particular a discharge siphon can be used for the outflow.
  • the siphon can be controlled via the filling level in the mixing chamber. As the fill level in the mixing chamber rises above the apex of the siphon, the liquid may flow to a lower level compared to the lowest point of the mixing chamber. As the leaking fluid creates a suction, the mixing chamber is deflated in this way.
  • FIG. 1A schematic representation of the components for carrying out the method according to the invention
  • FIG. 1B schematic representation of the pressure profile in the siphon during the gas bubble generation
  • Figure 2 is a schematic representation of the components for the mixing method according to the invention for illustrating the chemical gas generation.
  • 3 is a schematic representation of the components for the mixing process according to the invention with liquid separation structure in the siphon;
  • FIG. 4 schematic representation of the mixing chamber with flow obstacle
  • FIG. 5 schematic representation of the mixing chamber with flow obstacle and venturi structure
  • Fig. 8A / B views of a turret half as part of a centrifugal system having structures for carrying out the method according to the invention
  • Figure 9 shows a turret half as part of a centrifugal system according to the invention with remplissigabscheide Modellen in the supply siphon area.
  • FIG. 10 is a plan view of a revolver as part of a centrifugal system illustrating the various fluid paths.
  • FIG. 1A-F schematic views of a turret half as part of a
  • Centrifugal system having structures for carrying out the method according to the invention for illustrating the processes of a biochemical purification protocol
  • FIG. 12 shows a graphic representation of a centrifugation time protocol for carrying out a DNA extraction with mixing according to the invention in a centrifugal system.
  • Fig. 1A shows the components in a microfluidic centrifugal system required for carrying out the mixing process according to the invention.
  • gas bubbles 1 1 are generated, which are introduced via a siphon 12 into a mixing chamber 13 from below.
  • the movement in the liquid induced by the rising bubbles 11 in the liquid within the mixing chamber 13 effects the mixing effect.
  • the components shown are integrated in a centrifugal system (not shown in detail).
  • the arrow 14 indicates the centrifugal force acting on this system from radially inward to radially outward.
  • the generation of gas within the gas generating chamber 10 creates an overpressure in the siphon. This is illustrated by the schematic representation of the pressure curve in FIG. 1B.
  • the overpressure pushes any mixing liquids possibly contained in the siphon 12 in the direction of the mixing chamber 13 or keeps liquid away from the siphon 12.
  • a critical overpressure Ap crit An ejection of a gas bubble 1 1 takes place in the mixing chamber 13.
  • the pressure in the siphon 12 drops abruptly. Subsequently, the pressure in the siphon 12 rises through the
  • This mixing system can be integrated with little effort into a fluidic overall system, in particular in a microfluidic centrifugal system, wherein an activation of the gas generation, the supply lines to the mixing chamber and a forwarding of the mixture from the mixing chamber is added to the basic system.
  • Fig. 2 illustrates the chemical gas generation in the gas generating chamber 20.
  • catalyst particles 25 which can be pre-stored as solid particles.
  • a solution with one or more suitable reagents 26 is introduced into the gas generating chamber 20.
  • gas is formed, which essentially flows via the siphon 22 liquid-free in the mixing chamber 23 in the form of gas bubbles 21 is introduced.
  • the catalyst particles 25 may be, for example, manganese oxide.
  • the reagent solution may be, for example, a 30% hydrogen peroxide solution. The hydrogen peroxide is converted by the catalytic action of manganese oxide 25 to water and gaseous oxygen.
  • a remplissigabscheide Cook 35 is shown within the siphon 32.
  • the chemical reactions that occur within the gas generating chamber 30 can entrain liquids in the form of aerosols or particles. This could lead to contamination within the mixing chamber 33. Due to the sossigabscheide Design 35, in which the gas flow is slowed by expansion of the gas stream, the entrained particles get out of the flow and settle. Aerosols can be used in the
  • Fig. 4 illustrates the so-called mammoth pump effect for enhancing the mixing effect.
  • a flow obstruction 45 is arranged, wherein the gas bubbles 41, which pass through the siphon 42 into the mixing chamber 43, on one side of the obstacle 45, in this illustration on the left side, are introduced into the mixing chamber 43 from below.
  • the liquid On the left side of the flow obstruction 45, the liquid has a lower average density (pi ⁇ p 2 ).
  • equilibrium is established as a result of the acting gravitation, so that the level on the side of the less dense liquid increases (h> h 2 ).
  • Transverse channel 56 may be referred to as a "venturi tube.”
  • the ascending fluid stream may be referred to as a "main tube.”
  • this embodiment forms in a certain way a "Venturi nozzle", which is also shown in enlarged detail in Fig. 5.
  • FIG. 6 illustrates a first possibility for subsequent discharge of the mixed liquid (s) from the mixing chamber 63.
  • an outflow 65 in the form of an initially closed opening 65 is provided in the lower region of the mixing chamber 63. It may, for example, be an opening provided with a pierceable foil.
  • FIG. 7 Another possibility for forwarding the mixed liquid (s) from the mixing chamber 73 is illustrated in FIG. 7.
  • a siphon system 75 is used.
  • the siphon 75 is controlled by the filling level. As soon as the filling level in the mixing chamber 73 rises above the uppermost point (arrow) of the siphon 75, the liquid can flow off to a lower level compared with the lowest point of the mixing chamber 73. To reach the uppermost point, the level in the mixing chamber 73 can be increased. The leaking fluid creates a suction so that the mixing chamber 73 deflates.
  • FIGS. 8 to 12 illustrate the implementation of the method according to the invention and of the centrifugal system according to the invention for a cartridge-based system with a plurality of mutually rotatable revolvers, as is known from German Offenlegungsschrift DE 10 2013 003 223 A1.
  • the centrifugal system can be used, for example, for a DNA
  • FIGS. 8A and 8B show the same servo half 800 in various ways of representation. The dimensions of such a, approximately cylindrical turret, which is assembled from two halves, for example, 38 mm in height and 23 mm in diameter.
  • the microfluidic channel structures may, for example, have a width of 600 ⁇ m and a depth of, for example, 1 mm.
  • the channel structures are located in the bonding plane of the two turret halves. In this embodiment, only small aspect ratios occur in the manufacture of the microchannels, so that a precise production in injection molding is easily possible.
  • the mixing chamber 83 At the center of the turret half 800 is the mixing chamber 83.
  • catalyst particles 85 Within the underlying gas generating chamber 80 are catalyst particles 85 (FIG. 8B), for example manganese oxide particles.
  • Revolver 800 in the region of the gas generating chamber 80 an opening 84 (Fig. 8A) is provided, via which the gas generating chamber with the catalyst particles 85 can be charged.
  • a small chamber 86 via which the or the required reagents for gas generation are introduced into the gas generating chamber 80 via the channel 88.
  • the reagent solution for example a hydrogen peroxide solution
  • the chemical reaction in the gas generating chamber 80 can be started so that the formed gas bubbles are introduced into the mixing chamber 83 via the siphon 82.
  • FIG. 9 shows a further embodiment of a revolver of the centrifugal system, again shown here as a turret half 900. This embodiment is comparable in many parts with the turret half 800.
  • two separation chambers 95 and 96 are provided in the region of the inlet siphon 92.
  • the deposition chambers 95 and 96 form liquid separation structures of the siphon 92, thereby avoiding any carryover of liquids or contaminants into the mixing chamber 93 in general.
  • the first deposition chamber 95 captures particles or foam residues which, for example, due to foam formation in the gas generation chamber 90, come close to the gas outlet and can thereby be dragged off. As a result of the centrifugal force acting these particles or
  • Foam residues within the separation chamber 95 directed downwards.
  • the flow velocity is reduced by the deposition chambers 95 and 96, whereby the entrainment of droplets or aerosols is difficult. Due to the longer residence time of the aerosols is within the Abscheide Modellen 95 and
  • Fig. 10 shows a plan view of a revolver, which is composed of two turret halves.
  • the lower turret half is formed for example by the turret half 800 or 900, which have been explained in more detail in Fig. 8 and 9 respectively.
  • the dashed line indicates the bonding plane with the second (upper) turret half 100.
  • the turret half 100 has a cavity 101 (column cavity), which is provided for an extraction matrix.
  • the piercing spikes 87, 89, 102 and 103 On the compound turret are the piercing spikes 87, 89, 102 and 103.
  • mandrel 87 By means of the mandrel 87 can be released from an overlying turret reagent solution, such as a hydrogen peroxide solution and in the gas generating chamber, located in the lower part of the turret 800/900 is initiated. With the mandrel 103 above lying liquid reservoirs can be pierced, the liquids pass directly into the column cavity 101. With the other thorns 102 and 89 is a
  • Liquid transfer causes the underlying mixing chamber.
  • a typical protocol for extraction of human, viral or bacterial DNA can be performed.
  • a protocol for extraction of human, viral or bacterial DNA includes the following steps:
  • FIGS 11A-F illustrate the various steps A-F of this
  • step A 200 ⁇ lysis buffer is introduced into the mixing chamber 83 by an overlying reservoir is pierced by means of the mandrel 89.
  • step B the sample (200 ⁇ ) is transferred into the mixing chamber 83.
  • a 30% hydrogen peroxide solution (200 to 500 ⁇ ) is passed into the gas generating chamber 80.
  • an overlying reservoir is pierced with the mandrel 87.
  • the chamber volume of the gas generating chamber 80 is for example about 600 ⁇ .
  • Oxygen gas is generated by the contact of the hydrogen peroxide with the catalyst 85, which precedes the gas generating chamber 80, for example, 5 to 500 mg of MnO 2 . Due to the resulting overpressure, any existing liquid will leak out the siphon 82 (gas transfer channel) displaced. The generated gas comes in the form of bubbles in the mixing chamber 83 (step C). The rising bubbles cause a turbulence of the liquids, so that it comes to a mixing effect within the mixing chamber 83.
  • the reaction of, for example, 500 .mu.l 30% H 2 0 2 solution to 500 mg Mn0 2 particles allows at a
  • step D 200 ⁇ binding buffer is introduced into the mixing chamber 83. Mixing of the binding buffer with the lysate is carried out by the ascending bubbles.
  • additional liquid is added in step E which does not biochemically interfere with the batch, for example 300 ⁇ of a mixture of lysis and binding buffer or
  • the level in the mixing chamber 83 increases over the vertex of the discharge siphon 81.
  • centrifugation for example, with an acceleration of 300 x g, the projecting over the vertex liquid on the other side is pressed down. In this way, a suction is created, which pulls the other liquid behind it, so that the outflow via the connection 91 to the extraction column takes place (step F).
  • the liquid phase in the siphon should be uninterrupted. For this reason, a gas-tightness of the component and the bonding is generally beneficial. Tearing off the liquid flow or the entry of gas bubbles would hinder the suction effect.
  • Outlet the mixture can be transferred in the direction of the radially outer extraction matrix and centrifuged from there through the extraction matrix.
  • the subsequent washing and elution steps are carried out via a separate fluidic path, in which the piercing mandrel 103 (FIG. 10) is inserted. In this case, the liquids are transferred directly into the column cavity 101.
  • FIG. 12 shows schematically a centrifugation time protocol which can be used for the conversion of a DNA extraction.
  • the accelerations a S wi and a S w2 set the cutoff frequencies for the switching operation of
  • Cartridge mechanism which induces the rotation of the processing turrets. ed. For a rotation you have to accelerate once over a S wi. The rotation or the switching process is then completed by a deceleration to a S w2. Depending on the design of the cartridge mechanism, the release can take place either during acceleration or when decelerating.
  • the maximum acceleration a max is used to force the flow through the extraction matrix.
  • the acceleration a mix at which the mixing is carried out, is freely selectable between a S w2 and a max .
  • the first two acceleration pulses on a S wi cause transfer of lysis buffer and the sample into the mixing chamber.
  • the hydrogen peroxide is added to the catalyst in the gas bubble generating chamber simultaneously with the sample, thereby activating gas bubble generation. This is followed by a hold time at a mix , during which the lysis takes place.
  • the binding buffer is transferred into the mixing chamber and there is a further hold time at a mix , during which now the binding mixture is incubated.
  • the discharge siphon is activated by adding further liquid.
  • the mixture is passed via the discharge siphon to the extraction matrix.
  • the mixture is then centrifuged at a max through the extraction matrix.
  • the buffers are added directly to the extraction matrix and centrifuged at a max .
  • a typical centrifugal acceleration for a max is between about 4000 to 6000 x g.
  • a S wi may be at 200 to 600 xg
  • a S w2 may typically be between 0 and 200 xg.

Abstract

L'invention concerne un procédé consistant à mélanger des liquides dans une chambre de mélange (13) d'un système microfluidique basé sur la force centrifuge afin de produire des bulles de gaz (11) dans une chambre (10) du système basé sur la force centrifuge. Les bulles de gaz sont introduites dans la chambre de mélange (13) au moyen d'une structure permettant le transfert (12) des bulles de gaz réalisé sensiblement sans liquide.
PCT/EP2014/070288 2013-10-08 2014-09-24 Procédé pour mélanger des liquides et système microfluidique basé sur la force centrifuge WO2015051993A1 (fr)

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DE102013220264.4A DE102013220264A1 (de) 2013-10-08 2013-10-08 Verfahren zum Mischen von Flüssigkeiten und mikrofluidisches Zentrifugalsystem
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GB2562762B (en) * 2017-05-24 2022-07-13 Univ Heriot Watt Microfluidic mixing
CN109490058B (zh) * 2018-11-16 2021-04-20 南京爱思唯志生物科技有限公司 一种适于液体混合的微流控系统及方法
CN112481123B (zh) * 2020-11-16 2022-02-15 大连理工大学 一种研究剪切力和生化因子梯度调控细胞划痕修复的微流控系统及方法
DE102022203875B3 (de) 2022-04-20 2023-06-15 Hahn-Schickard-Gesellschaft für angewandte Forschung e.V. Handhabung zweier flüssigkeitsvolumina

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005016532A2 (fr) * 2003-06-13 2005-02-24 Corning Incorporated Systeme automatise de chambre de reaction pour essais biologiques
US20050272144A1 (en) * 2004-06-08 2005-12-08 Konica Minolta Medical & Graphic, Inc. Micro-reactor for improving efficiency of liquid mixing and reaction
EP1970565A1 (fr) * 2007-03-14 2008-09-17 Samsung Electronics Co., Ltd. Unité de pompe et système microfluide centrifuge l'incorporant
US20090075801A1 (en) * 2007-09-19 2009-03-19 Dalibor Hodko Counter-centrifugal force device
US20110103174A1 (en) * 2009-10-29 2011-05-05 Samsung Electronics Co., Ltd. Microfluidic device comprising gas providing unit, and methods of mixing liquids and forming emulsion using the same
WO2012131598A1 (fr) * 2011-03-28 2012-10-04 Biosurfit, S.A. Changement, dosage et pompage de liquide

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5511880A (en) 1994-09-27 1996-04-30 Spacelabs Medical, Inc. Method and apparatus for storing and mixing a plurality of fluids and body fluid sampling cartridge using same
DE102010003223B4 (de) 2010-03-24 2014-09-18 Albert-Ludwigs-Universität Freiburg Vorrichtung zum Einsetzen in einen Rotor einer Zentrifuge, Zentrifuge und Verfahren zum fluidischen Koppeln von Kavitäten
DE102013003223A1 (de) 2013-02-27 2014-08-28 Haver & Boecker Ohg Packmaschine und Verfahren zum Füllen von Offensäcken

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005016532A2 (fr) * 2003-06-13 2005-02-24 Corning Incorporated Systeme automatise de chambre de reaction pour essais biologiques
US20050272144A1 (en) * 2004-06-08 2005-12-08 Konica Minolta Medical & Graphic, Inc. Micro-reactor for improving efficiency of liquid mixing and reaction
EP1970565A1 (fr) * 2007-03-14 2008-09-17 Samsung Electronics Co., Ltd. Unité de pompe et système microfluide centrifuge l'incorporant
US20090075801A1 (en) * 2007-09-19 2009-03-19 Dalibor Hodko Counter-centrifugal force device
US20110103174A1 (en) * 2009-10-29 2011-05-05 Samsung Electronics Co., Ltd. Microfluidic device comprising gas providing unit, and methods of mixing liquids and forming emulsion using the same
WO2012131598A1 (fr) * 2011-03-28 2012-10-04 Biosurfit, S.A. Changement, dosage et pompage de liquide

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
CHOI Y H ET AL: "A micropump operating with chemically produced oxygen gas", SENSORS AND ACTUATORS A, ELSEVIER SEQUOIA S.A., LAUSANNE, CH, vol. 111, no. 1, 1 March 2004 (2004-03-01), pages 8 - 13, XP004489315, ISSN: 0924-4247, DOI: 10.1016/J.SNA.2003.10.005 *

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