EP3408025B1 - Tube à structure microfluidique et méthode d'utilisation associée - Google Patents

Tube à structure microfluidique et méthode d'utilisation associée Download PDF

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Publication number
EP3408025B1
EP3408025B1 EP17701136.8A EP17701136A EP3408025B1 EP 3408025 B1 EP3408025 B1 EP 3408025B1 EP 17701136 A EP17701136 A EP 17701136A EP 3408025 B1 EP3408025 B1 EP 3408025B1
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EP
European Patent Office
Prior art keywords
tube
reaction
microfluidic structure
sample material
bioanalytical
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EP17701136.8A
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German (de)
English (en)
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EP3408025A1 (fr
Inventor
Frank Daniel Scherag
Jürgen Rühe
Thomas Brandstetter
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Albert Ludwigs Universitaet Freiburg
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Albert Ludwigs Universitaet Freiburg
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • 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
    • B01L3/02—Burettes; Pipettes
    • B01L3/0275—Interchangeable or disposable dispensing tips
    • 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
    • B01L3/502707—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 manufacture of the container or its components
    • 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
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
    • 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
    • B01L2200/16—Reagents, handling or storing thereof
    • 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
    • B01L2300/0832—Geometry, shape and general structure cylindrical, tube shaped
    • 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
    • B01L2300/0832—Geometry, shape and general structure cylindrical, tube shaped
    • B01L2300/0838—Capillaries
    • 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
    • B01L2300/087—Multiple sequential chambers
    • 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
    • B01L2300/16—Surface properties and coatings
    • B01L2300/161—Control and use of surface tension forces, e.g. hydrophobic, hydrophilic
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0409—Moving fluids with specific forces or mechanical means specific forces centrifugal forces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
    • B01L2400/049—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics vacuum
    • 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
    • B01L2400/08—Regulating or influencing the flow resistance
    • B01L2400/084—Passive control of flow resistance
    • B01L2400/088—Passive control of flow resistance by specific surface properties

Definitions

  • Embodiments of the present invention relate to a tube with a microfluidic structure, in particular to a tube in the form of a pipette in the tip of which the microfluidic structure is arranged. Further exemplary embodiments relate to carrying out an investigation in a pipette (pipette tip), for example carrying out a multi-stage bioanalytical reaction in a conical (pipette) tube by integrating a microfluidic device which generates at least two compartments.
  • the lab-on-a-pipette system is a special, advantageous addition to common lab-on-chip systems (laboratory-on-a-chip systems) with an upstream microtip, which is only used for targeted and exact Sample collection is used.
  • the analyzing lab-on-chip unit (fluidics and temperature control) was integrated into a specially developed pipette. The description of the production of corresponding microfluidic structures also corresponds to that of classic chip-based production processes.
  • the US 2007/0059215 A1 shows a micropipette in combination with a MEMS flow sensor.
  • the micropipette comprises a main section, an operating section and a pipe section, the pipe section defining an operating space.
  • the detection device is arranged at a suitable position in the operating space of the pipe section.
  • the MEMS flow sensor detects gas movement in the operating room.
  • the EP 1 381 468 B1 describes a device for taking an aliquot of a sample from a sealed container.
  • WO 2011/032228 A1 describes an experimental unit that has two interfaces - an interface for a pipette and an interface for a tip (sample holder). The experimental unit is thus connected in series between the tip and the pipette, the tip being used only for sample collection.
  • the WO 2006/029387 A1 describes a handy and portable microfluidic device that includes a cleaning chip and a syringe-like device for sample collection and movement. The sample is only prepared or purified.
  • the EP 0 803 064 B1 discloses a portable device for taking a fluid sample.
  • the device comprises a syringe, a housing having a first and second end, the first end being attached to the syringe, a receiving chamber contained in the housing, a filter device which has at least one hollow fiber membrane which is sealed by a plug of hardened adhesive in the A housing is contained between the receiving chamber and the second end, a piercing device which is attached to the second end of the housing, the housing being in fluid communication with the syringe and the piercing device, and a sensing device which is in the housing between the receiving chamber and the first end of the housing is included.
  • the object of the present invention is therefore to create a concept which on the one hand requires a simpler structure and on the other hand enables a more reliable examination of the sample.
  • Embodiments of the present invention provide a tube having an area tapering toward one end of the tube; an opening at the end of the tube for receiving a sample material; and a microfluidic structure disposed within the tube in the tapered region of the tube and connected to the opening, the microfluidic structure configured to perform a multi-stage bioanalytical reaction with the sample material; wherein the microfluidic structure has at least two reaction chambers for carrying out the multi-stage bioanalytical reaction; wherein the microfluidic structure has at least one ring-shaped component which bears against an inner wall of the tube; and wherein the microfluidic structure includes a moveable tubular component that abuts the inner wall of the tube in the tapered region of the tube and at least partially against the at least one annular component to form the at least two reaction chambers, the moveable tubular component being included the opening is connected; and wherein the tube is a tip of a pipette or a pipette.
  • a microfluidic structure which is arranged directly in a tapering region of a tube, such as, for example, a tip of a pipette or pipette tip, is used for analyzing or examining a sample by means of a multi-stage bioanalytical reaction.
  • a tube such as, for example, a tip of a pipette or pipette tip
  • Some exemplary embodiments make it possible to carry out bioanalytical detection reactions, which take place in several spatially or temporally separate steps, in the vessels which are used directly for the uptake and removal of sample material (e.g. pipette tubes).
  • sample material e.g. pipette tubes
  • This is made possible by subdivision of the vessel volume with the help of a movable fluid system. Its incorporation into the vessel volume forms at least two separate reaction spaces / compartments.
  • Tapered tubes in which a tubular fluid system sits on the inner cone are regarded as particularly suitable and advantageous designs. Appropriate functional designs solve common production-related and microfluidic problems of chip-based structures that have more than one reaction space.
  • FIG. 1 For exemplary embodiments, include the implementation of a sequential, multi-stage bioanalytical method (bioassay) in the volume of a conically tapering tube (in the form and function of a pipette tip), which is characterized in that an integrated microfluidic device generates at least two compartments in which analytical reagents are stored and can be reacted with a sample liquid in chronological order (successively / consecutively).
  • the peculiarity of the integrated microfluidic is that a nested tubular channel system sits loosely on the inner wall of the cone, thereby forming compartments and sealing them functionally.
  • the compartments formed are separated from one another by (ring-shaped) gaps / openings in hydrophobic surfaces.
  • Some exemplary embodiments make it possible to carry out complex, multi-stage and coherent reactions within the volume of a pipette tube in a very simple and inexpensive manner, as is usually used for taking up and taking out sample material.
  • FIG. 10 shows a schematic cross-sectional view of a tube 100 with a region 104 tapering towards one end 102 of the tube 100 and a microfluidic structure 106, which is arranged in the tapered region 104 of the tube 100, according to an exemplary embodiment of the present invention.
  • the microfluidic structure 106 is connected to an opening 108 at the end 102 of the tube 100 for receiving a sample material, the microfluidic structure 106 being designed to carry out a multi-stage bioanalytical reaction with the sample material.
  • the tapering region 104 of the tube 100 can be tapered towards the end 102 of the tube 100 or be conical (hollow-conical).
  • the tapered region 104 of the tube 100 can thus have the shape of a tip or be tip-shaped.
  • the tube 100 can only consist of the tapered region 104.
  • the tube 100 can optionally also have a cylindrical (hollow cylindrical) region 110.
  • the tube can thus be both a pipette tip and a pipette with a tip, the microfluidic structure 106 being arranged within the tip 104.
  • the microfluidic structure 106 can be arranged only or exclusively within the tapered region 104 of the tube 100. However, it is also possible that the microfluidic structure 106 extends over the tapered region extends into the conical region 110. In this case, the microfluidic structure is (only) at least partially arranged within the tapered region 104 of the tube.
  • FIG. 14 shows schematic cross-sectional views of a tube 100 with a region 104 tapering towards one end 102 of the tube 100 and a microfluidic structure 106, which is arranged at least partially in the tapered region 104 of the tube 100, according to an exemplary embodiment of the present invention. Compared to that in Fig. 1 embodiment shown in 2a and 2b Details of the microfluidic structure 106 are shown.
  • the microfluidic structure 106 can have a movable, tubular component 114.
  • the moveable tubular component 114 may be connected to the opening 108 of the tube.
  • the microfluidic structure 106 can be designed to transfer a sample material received via the opening 108 of the tube via the movable, tubular component 114 in a first direction (in 2a and 2b to "up") to transport.
  • the microfluidic structure 106 can have at least two reaction chambers 112_1 to 112_3 (two reaction chambers 112_1 and 112_2 in Fig. 2a ; three reaction chambers 112_1 to 112_3 in Fig. 2b ) to carry out the multi-stage bioanalytical reaction.
  • the (at least) two reaction chambers 112_1 to 112_3 can be separated from one another.
  • the microfluidic structure 106 can be designed to move the sample material (previously transported in the first direction (upward) by the movable tubular component 114) in order to carry out the multi-stage bioanalytical reaction through the at least two reaction chambers 112_1 to 112_3 in a second direction (in Fig. 2 to "down") to transport.
  • an analytical reagent for the multi-stage bioanalytical reaction can be upstream in at least one of the at least two reaction chambers 112_1 to 112_3.
  • an analytical reagent for the multi-stage bioanalytical reaction can be upstream in at least one of the at least two reaction chambers 112_1 to 112_3.
  • analytical reagents upstream wherein the fluidic structure 106 may be configured to with the analytical reagents bring the sample material to the reaction in time order to carry out the multi-stage bioanalytical reaction.
  • the microfluidic structure 106 can have at least one ring-shaped component 116_1 and 116_2, which abuts an inner wall of the tube.
  • the movable, tubular component 114 is arranged such that it rests in the tapering region 104 of the tube on an inner wall of the tube and at least partially on the at least one annular component 116_1 and 116_2.
  • the movable, tubular component 114 can rest against the annular component 116 such that the at least two reaction chambers 112_1 to 112_3 formed are separated from one another by openings of hydrophobic surfaces or hydrophobic capillary openings.
  • Tube 100 may further include a filter or O-ring 120.
  • the sample material can, for example, be sucked up via the O-ring 120.
  • 2a and 2b show schematic representations of tip-shaped tubes 100 for taking up and taking out sample material with integrated tubular fluidics, which subdivides the volume of the tube 100 into two or three separate reaction areas 112_1 to 112_3 and enables multi-stage detection reactions to be carried out.
  • Sample liquids can be taken up very precisely from the outside via the conically tapering part of the tube (tip) 104 and can enter the first compartment 112_1 in the proximal (widened) area of the pipette via a central tubular channel 114 which loosely rests on the inner conical area 104 Tube 100.
  • the liquid transport is characterized in that it runs primarily in the direction of the cylinder / cone axis, ie vertically with the aid of pressure differences and centrifugal forces.
  • compartments 112_1 to 112_3 the liquid is preferably transported via centrifugal forces and in the direction of the cone tip, so that the liquid of a proximal compartment is forced into the closest distal compartment (in the direction of the cone tip) via the hydrophobic capillary opening.
  • the transport from compartment to compartment can be controlled, for example, by varying the capillary opening (length, width, surface) and the centrifugal force applied. This enables the spatial and temporal separation of reaction processes.
  • FIG. 10 shows a schematic view of a tube 100 with a region 104 tapering towards one end 102 of the tube 100 and a microfluidic structure 106, which is arranged at least partially in the tapered region 104 of the tube 100, according to an exemplary embodiment of the present invention.
  • the microfluidic structure 106 comprises four reaction chambers 112_1 to 112_4, which are formed by three ring-shaped components 116_1 to 116_3 and the movable, tubular component 114.
  • FIG. 3 shows a schematic representation of a microfluidic device 100 with a total of 4 compartments 112_1 to 112_4, which are separated from one another by capillary gaps / openings of different widths and lengths.
  • the two middle compartments (2 and 3) 112_2 and 112_3 each have an annular side pocket 118_1 and 118_2 in which liquid can be retained, for example for separate reactions or for skimming off unnecessary reaction solutions.
  • compartment 4 (112_4) the arrangement shown here results in three reaction spaces with a dead end function. If the tubular channels 104 are moved against the taper, the liquid can be emptied in the direction of the tip.
  • the microfluidic 106 integrated into the conical pipette tube 100 can also be designed such that individual compartments 112_2 and 112_3 form dead end-like pockets 118_1 and 118_2.
  • This allows reaction solutions to be divided up and reduced in volume before being transported to the next compartment.
  • the last (distal) compartment 112_4 closest to the tip can be characterized in that it is sealed in the direction of the cone by the integrated tubular channel system.
  • the centrifugal forces for liquid transport simultaneously promote the self-sealing structure.
  • the hydrophobic surfaces of the capillary gaps and the structure of the tubular channel system prevent evaporation of the liquid, so that even reactions in the boiling point of the reaction solution (e.g.
  • PCR polymerase chain reaction
  • the targeted release of the reaction solution from the distal (facing the tip) compartment can be initiated by moving the tubular channel system 114 against the taper.
  • Reagents preferably freeze-dried
  • By storing all the reagents necessary for a reaction only the liquid absorption of the sample to be examined is necessary. This means that liquid samples from only a few microliters can be processed in the tapered tube (pipette tube) after completion of the detection reaction until the detection reaction is complete.
  • the detection can then take place externally in tube 100 or after the reaction solution has been released.
  • Devices and objects for generating pressure differences can be used for operation and for carrying out the assay.
  • possible application examples are all multi-stage bioanalytical methods of sample preparation and preparation with detection reaction.
  • a preferred example of this would be nucleic acid analysis.
  • a simple two-compartment system would be, for example, the combination and implementation of a so-called liquid DNA extraction with subsequent PCR.
  • An example of a three-stage system would be an RT-PCR consisting of cell disruption, reverse transcription and PCR in spatial separation.
  • Further application examples are enzymatic reactions in which the products can be used for further reactions in a temporal and spatial separation.
  • Another example would be the storage of reagents in the individual compartments, which enable a start-stop function for the reaction.
  • Embodiments create several advantages, as described below.
  • One advantage is that the structure and the functional principle of the microfluidics integrated in the pipette tube do not require any complex processes and procedures as are usually used in the microstructuring of chip structures.
  • the vertical arrangement of the compartments and the vertical liquid transport solve common problems of chip-based microfluidics.
  • pressure equalization and gas bubble formation, as well as the rapid filling of the compartments without air bubbles pose no problems.
  • the liquid can be taken in manually and without electrical pumps in a very precise manner (for example, using standard laboratory pipettes).
  • the liquid remains in the pipette tube (single-tube solution) during all steps of the analysis.
  • the self-sealing structure of the fluid system within the pipette tube enables both the centrifugation and the heating of the reaction solution without loss of liquid. Furthermore, the reaction solution can be released quickly and easily for detection (optional). Ultimately, the form and the lack of electrical interfaces enable simple parallelization as well as cost-effective integration into the standard laboratory equipment.
  • a microfluidic structure can be built up, which (1) a filter tip (e.g. ep Dualfilter TIPS®), (2) a silicone tube “long, thin” (e.g. inside diameter 0.5 mm; outside diameter 1.3 mm; length 25 mm) , (3) a silicone hose "thick, short” (e.g. inner diameter 1.58 mm; outer diameter 3.18 mm; length 4 mm), (4) instead of the dual filter of the filter tip, a short piece of an additional silicone hose can also be used (e.g.
  • the control elements can be (1) a commercially available pipette (e.g. Eppendorf Research ®), (2) a table centrifuge (e.g. peqlab ®, PerfectSpin ® Mini centrifuge with rotor insert for 0.5 ml reaction tubes) and (3) a PCR device for capillaries or Tubes (e.g. Roche LightCycler 1.5 ®) are used.
  • Fig. 4a shows a photo
  • Fig. 4b 10 shows a schematic cross-sectional view of a tube 100 with a region 104 tapering towards one end 102 of the tube 100 and a microfluidic structure 106, which is arranged in at least partially in the tapered region 104 of the tube 100, according to an exemplary embodiment of the present invention.
  • the microfluidic structure 106 comprises two reaction chambers 112_1 and 112_2, which are formed via an annular component 116_1 and a movable, tubular component 114, as already described in relation to FIG Fig. 2a has been described in detail.
  • 4a and 4b to recognize an O-ring or filter 120.
  • Fig. 4a shows a photo shoot
  • Fig. 4b a schematic drawing showing the structure of the fluidics 106 and the resulting compartments 112_1 and 112_2 in the pipette tip 104.
  • blue hoses first hatching
  • green-yellow stripes second hatching
  • FIG. 5a-5e show schematic cross-sectional views of the in Fig. 2 Tube 100 shown in or after different steps of a method for examining a sample material with tube 100, according to an embodiment of the present invention.
  • Fig. 5a 10 is a schematic cross-sectional view of tube 100 prior to a step of soaking up sample material 130.
  • Lysis reagents 122 can be stored in front of an inside of the movable, tubular component 114, while PCR reagents 124 can be stored in the second reaction chamber 112_2.
  • Fig. 5b shows a schematic cross-sectional view of the tube 100 after a step of sucking up the sample material 130, whereby the sample material 130 has been transported through the movable, tubular component 114 into the first reaction chamber 112_1.
  • Fig. 5c shows a schematic cross-sectional view of the tube 100 during a centrifugation step, whereby the sample material passes from the first reaction chamber 112_1 into the second reaction chamber 112_2.
  • FIG. 5d shows a schematic cross-sectional view of tube 100 after the centrifugation step, whereby sample material 130 has completely passed from first reaction chamber 112_1 to second reaction chamber 112_2, where the sample material contacts PCR reagents 124, causing a second bioanalytical reaction comes with the sample material 130.
  • FIG. 10 shows a schematic cross-sectional view of tube 100 during the second bioanalytical reaction.
  • 5a to 5e show an exemplary schematic sequence in the case of upstream reagents (eg freeze-dried) for liquid DNA extraction (blue (first hatching), eg lysis of cell suspensions) with subsequent PCR (yellow (second hatching).
  • upstream reagents eg freeze-dried
  • lysis reagents 122, cell lysate, or DNA extract can be mixed directly with the reagents 124 of the PCR (green (third hatching))
  • the course of the reaction can be visualized using fluorescent dyes (turquoise (fourth hatching)).
  • FIG. 6a to 6d show schematic cross-sectional views of the in Fig. 2 Tube 100 shown in or after different steps of a method for taking a sample material after carrying out the multi-stage bioanalytical reaction, according to an embodiment of the present invention.
  • FIG. 6a is a schematic cross-sectional view of tube 100 after performing the multi-stage bioanalytical reaction.
  • the sample material 130 (together with any reagents) is located in the second reaction chamber 112_2.
  • Figure 10 shows a schematic cross-sectional view of tube 100 after an application step a removal device 132 (eg a spiral wire) into the movable tubular component 114.
  • Fig. 6c shows a schematic cross-sectional view of the tube 100 in a step of shifting the movable tubular component 114 by applying the removal device 132 into the movable tubular component 114.
  • FIG. 14 shows a schematic cross-sectional view of tube 100 after the step of displacing movable tubular component 114, allowing sample material 130 to escape from tube 100.
  • 6a to 6d show schematic representations of how the reaction liquid 130 can be released for further investigations after the end of the reaction by subsequent application of a spiral wire 132.
  • the wire 132 can be inserted into the pipette tip in the case of the O-ring from the pipette side (top) or in the case of the filter from the tip (bottom). In both cases, the wire 132 is used to move the long, thin tube 114 upward so that the liquid 130 can escape through the tip by actuating the pipette.
  • FIG. 7a to 7e show schematic cross-sectional views of the in Fig. 2 Tube 100 shown in or after different steps of a method for examining a sample material with tube 100, according to an embodiment of the present invention.
  • Fig. 7a 10 is a schematic cross-sectional view of tube 100 prior to a step of soaking up sample material 130.
  • the removal device 132 for example a spiraled wire
  • the lysis reagents 122 can be arranged upstream on the removal device 132.
  • PCR reagents 124 can be arranged upstream in the second reaction chamber 112_2.
  • Fig. 7b shows a schematic cross-sectional view of the tube 100 after a step of sucking up the sample material 130, whereby the sample material 130 has been transported through the movable, tubular component 114 into the first reaction chamber 112_1.
  • Fig. 7c shows a schematic cross-sectional view of the tube 100 during a centrifugation step, whereby the sample material passes from the first reaction chamber 112_1 into the second reaction chamber 112_2.
  • FIG. 7d 14 shows a schematic cross-sectional view of tube 100 of FIG the step of centrifuging, whereby the sample material 130 has completely passed from the first reaction chamber 112_1 into the second reaction chamber 112_2, where the sample material comes into contact with the PCR reagents 124, resulting in a second bioanalytical reaction with the sample material 130.
  • Fig. 5e shows a schematic cross-sectional view of the tube 100 after the second bioanalytical reaction and the displacement of the movable tubular component, whereby the sample material can escape from the tube 100.
  • 7a to 7e show schematic representations of an application similar to that 5a to 5e .
  • the wire 132 is part of the fluidics right from the start.
  • the lysis reagents 124 are even stored on the surface of the wire 132 and washed off the sample 130 when it is sucked up.
  • An advantage here is a better mixing of the lysis reagents 122 with the sample 130 and a simpler introduction of the lysis reagents 122.
  • the further course is the same as in FIG 5b to 5e described, in addition, however, the function of as below 6a to 6d already integrated.
  • Fig. 8 shows a photo of a pipette tip (lab in a pipette tip) through a filter disc.
  • the pipette tip there is a PCR product with the dye SYBR Green I which can be excited with blue light ( ⁇ 495nm) in the wavelength range of fluorescein and emits green light ( ⁇ 520 nm).
  • Fig. 9 shows a photo of a gel electrophoresis (1.5% agarose; 70 V; 45 min.) of two PCR products.
  • the positive control (item) was tempered in a plastic capillary.
  • PCR in tip the pipette tip was filled with 10 ⁇ l of the PCR solution.
  • the temperature was controlled in a Roche LightCycler 1.5 ®.
  • FIG. 12 shows a flow diagram of a method 200 according to an embodiment of the present invention.
  • the method 200 includes a step 202 of receiving a sample material through an opening of a tube, the tube having a tapered area toward an end of the tube that forms the opening.
  • the method further includes a step 204 of performing a multi-stage bioanalytical reaction with a microfluidic structure disposed within the tube in the tapered region of the tube and connected to the opening.
  • the pipette tip In contrast to conventional concepts, in which the pipette tip is only seen as an addition / extension, this represents one in exemplary embodiments an essential part of the analyzing system and the fluidics.
  • aspects have been described in connection with a device, it goes without saying that these aspects also represent a description of the corresponding method, so that a block or a component of a device is also to be understood as a corresponding method step or as a feature of a method step. Analogously, aspects that have been described in connection with or as a method step also represent a description of a corresponding block or detail or feature of a corresponding device.
  • Some or all of the method steps can be carried out by a hardware apparatus (or using a hardware device). Apparatus), such as a microprocessor, a programmable computer or an electronic circuit. In some embodiments, some or more of the most important process steps can be performed by such an apparatus.

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

Claims (13)

  1. Tube (100), aux caractéristiques suivantes:
    une zone (104) effilée vers une extrémité (102) du tube (100);
    une ouverture (108) à l'extrémité (102) du tube (100) destinée à recevoir un échantillon de matière; et
    une structure micro-fluidique (106) qui est disposée à l'intérieur du tube (100) dans la zone effilée (104) du tube (100) et qui est connectée à l'ouverture (108), la structure micro-fluidique (106) étant conçue pour réaliser une réaction bio-analytique en plusieurs étapes avec l'échantillon de matière;
    dans lequel la structure micro-fluidique (106) présente au moins deux chambres de réaction (112_1; 112_3) pour réaliser la réaction bio-analytique en plusieurs étapes;
    dans lequel la structure micro-fluidique (106) présente au moins un composant annulaire (116_1; 116_2) qui s'appuie contre une paroi intérieure du tube (100); et
    dans lequel la structure micro-fluidique (106) présente un composant tubulaire mobile (114) qui s'appuie, dans la région effilée (104) du tube (100), sur la paroi intérieure du tube (100) et au moins partiellement sur l'au moins un composant annulaire (116_1; 116_2) pour former les au moins deux chambres de réaction (112_1; 112_3), le composant tubulaire mobile (114) étant connecté à l'ouverture (108); et
    dans lequel le tube (100) est une pointe d'une pipette ou une pipette.
  2. Tube (100) selon la revendication 1, dans lequel la structure micro-fluidique (106) est conçue pour réaliser la réaction bio-analytique en plusieurs étapes de manière séquentielle.
  3. Tube (100) selon l'une des revendications 1 à 2, dans lequel la structure micro-fluidique (106) est conçue pour réaliser la réaction bio-analytique en plusieurs étapes en étapes séparées dans le temps.
  4. Tube (100) selon l'une des revendications 1 à 3, dans lequel les au moins deux chambres de réaction (112_1; 112_3) sont séparées l'une de l'autre.
  5. Tube (100) selon l'une des revendications 1 à 4, dans lequel dans au moins l'une des au moins deux chambres de réaction (112_1; 112_3) est entreposé en amont un réactif analytique (122; 124) pour la réaction bio-analytique en plusieurs étapes.
  6. Tube (100) selon l'une des revendications 1 à 5, dans lequel dans les au moins deux chambres de réaction (112_1; 112_3) sont entreposés en amont les réactifs analytiques (122; 124);
    dans lequel la structure fluidique (106) est conçue pour faire réagir les réactifs analytiques (122; 124) avec l'échantillon de matière dans l'ordre chronologique pour réaliser la réaction bio-analytique en plusieurs étapes.
  7. Tube (100) selon l'une des revendications 1 à 6, dans lequel le composant tubulaire mobile (114) s'appuie sur l'au moins un composant annulaire (116_1; 116_2) de sorte que les au moins deux chambres de réaction (112_1; 112_3) formées soient séparées l'une de l'autre par des ouvertures de surfaces hydrophobes ou des ouvertures capillaires hydrophobes.
  8. Tube (100) selon l'une des revendications 1 à 7, dans lequel le composant tubulaire mobile (114) est amovible ou déplaçable pour retirer l'échantillon de matière du tube (100) après avoir effectué la réaction bio-analytique en plusieurs étapes.
  9. Tube (100) selon l'une des revendications 1 à 8, dans lequel la structure micro-fluidique (106), ou au moins un composant de la structure micro-fluidique (106), est conçue de manière mobile ou déplaçable.
  10. Procédé (200) pour effectuer une réaction bio-analytique en plusieurs étapes par un tube (100) selon l'une des revendications 1 à 9, dans lequel le procédé présente le fait de:
    recevoir (202) un échantillon de matière à travers l'ouverture du tube (100); et
    réaliser (204) la réaction bio-analytique en plusieurs étapes par la structure micro-fluidique du tube (100).
  11. Procédé (200) selon la revendication 10, dans lequel le procédé (200) présente par ailleurs le fait de:
    transporter l'échantillon de matière à la réception de l'échantillon de matière à travers la structure micro-fluidique (106) dans une première direction; et
    transporter l'échantillon de matière pour effectuer la réaction bio-analytique en plusieurs étapes à travers la structure micro-fluidique (106) dans une deuxième direction;
    dans lequel la première direction et la deuxième direction sont opposées.
  12. Procédé (200) selon l'une des revendications 10 à 11, dans lequel le procédé (200) présente par ailleurs le fait de:
    transporter l'échantillon de matière à travers la structure micro-fluidique (106) par des différences de pression et/ou des forces centrifuges.
  13. Procédé (200) selon la revendication 12, dans lequel le composant tubulaire mobile (114) s'appuie sur l'au moins un composant annulaire (116_1; 116_2) de sorte que les au moins deux chambres de réaction (112_1; 112_3) formées soient séparées l'une de l'autre par des ouvertures de surfaces hydrophobes ou des ouvertures capillaires hydrophobes, dans lequel le procédé (200) présente par ailleurs le fait de:
    commander le transport de l'échantillon de matière entre les au moins deux chambres de réaction (112_1:112_2) par variation de l'ouverture capillaire et/ou de la force centrifuge appliquée.
EP17701136.8A 2016-01-27 2017-01-24 Tube à structure microfluidique et méthode d'utilisation associée Active EP3408025B1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102016201181.2A DE102016201181B3 (de) 2016-01-27 2016-01-27 Röhre mit einer mikrofluidischen Struktur
PCT/EP2017/051372 WO2017129541A1 (fr) 2016-01-27 2017-01-24 Tube à structure microfluidique

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EP3408025A1 EP3408025A1 (fr) 2018-12-05
EP3408025B1 true EP3408025B1 (fr) 2020-03-18

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Publication number Priority date Publication date Assignee Title
US20220193668A1 (en) * 2019-04-18 2022-06-23 Siemens Healthcare Diagnostics Inc. Integrated microfluidic device with pipette adaptation
EP4382204A1 (fr) * 2022-12-05 2024-06-12 Amiprox Pte. Ltd. Dispositif de traitement integre d'echantillons biologiques

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US5171537A (en) * 1991-05-06 1992-12-15 Richard E. MacDonald Activated immunodiagnostic pipette tips
GB9426251D0 (en) * 1994-12-24 1995-02-22 Fsm Technologies Ltd Device
US6817256B2 (en) 2001-02-27 2004-11-16 Alfa Wassermann, Inc. Pipette sampling system
JP2008512128A (ja) 2004-09-09 2008-04-24 マイクロフルイディク システムズ インコーポレイテッド 抽出装置及び試料準備方法
US20060105391A1 (en) * 2004-11-12 2006-05-18 Promega Corporation Device and method for separating molecules
WO2006057225A1 (fr) * 2004-11-25 2006-06-01 Matsushita Electric Industrial Co., Ltd. Capteur
US20070059215A1 (en) 2005-09-14 2007-03-15 Pacific Image Electronics Co., Ltd. Micro pipette sensing device
ES2624930T3 (es) * 2007-09-29 2017-07-18 El Spectra, Llc Punta de pipeta instrumentada
AU2010295250A1 (en) 2009-09-18 2012-04-05 Minifab (Australia) Pty Ltd Instrumented pipette
US8394625B2 (en) 2010-05-02 2013-03-12 Angelo Gaitas Lab-on-a-pipette
WO2014035986A1 (fr) * 2012-08-28 2014-03-06 Akonni Biosystems Inc. Procédé et kit de purification d'acides nucléiques

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EP3408025A1 (fr) 2018-12-05
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