WO2017129541A1 - Tube à structure microfluidique - Google Patents

Tube à structure microfluidique Download PDF

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Publication number
WO2017129541A1
WO2017129541A1 PCT/EP2017/051372 EP2017051372W WO2017129541A1 WO 2017129541 A1 WO2017129541 A1 WO 2017129541A1 EP 2017051372 W EP2017051372 W EP 2017051372W WO 2017129541 A1 WO2017129541 A1 WO 2017129541A1
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WO
WIPO (PCT)
Prior art keywords
tube
microfluidic structure
reaction
sample material
bioanalytical
Prior art date
Application number
PCT/EP2017/051372
Other languages
German (de)
English (en)
Inventor
Frank Daniel Scherag
Jürgen Rühe
Thomas Brandstetter
Original Assignee
Albert-Ludwigs-Universität Freiburg
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 Albert-Ludwigs-Universität Freiburg filed Critical Albert-Ludwigs-Universität Freiburg
Priority to EP17701136.8A priority Critical patent/EP3408025B1/fr
Publication of WO2017129541A1 publication Critical patent/WO2017129541A1/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
    • 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 having a microfluidic structure, in particular to a tube in the form of a pipette in whose tip the microfluidic structure is arranged. Further embodiments relate to performing a test in a pipette (pipette tip), for example, to carry out a multi-stage bioanalytical reaction in a conical (pipette) tube by integration of a microfluidic device which generates at least two compartments. Modifications of pipette tips and corresponding functional forms are in part widespread [1 -3]. The known approaches merely describe a simplification of individual analysis steps, in particular sample preparation and sample processing.
  • Microfluidic systems that are able to perform several coherent analysis steps are particularly advantageous when the smallest sample volumes have to be processed and analyzed in a closed sequence [4].
  • the analysis steps are almost always discontinuous, ie in principle always several discrete reaction spaces required or at least several start-stop phases necessary to receive an analyte to prepare for the detection reaction and finally perform the analyzing step.
  • the goal is usually that automation and a closed sequence of all necessary steps should reduce possible manual errors, especially for very small sample volumes.
  • the LAB-ON-A pipette system [7, 8] combines a special "micro aspiration tip" tip with a micro-fluidic analyzing unit to optimize especially genetic single-cell analyzes In this system All reagents necessary for the detection reaction are also pre-stored.
  • Lab-on-a-pipette system is a special advantageous addition of common lab-on-chip systems (dt.
  • Examples include the wetting of surfaces and, associated therewith, the filling of compartments without air bubbles [9], the formation of gas bubbles [10] as well as the pressure compensation [1 1, 12]. Due to the vertical limitation of the flat structure and the horizontal position of the fluid management, no natural separation of gas and aqueous phase can take place. Furthermore, in order to move liquid volumes and to ensure a smooth automated process, both special and large and complex instrument platforms have to be used [13, 14]. Both factors mean that current solutions are considered practicable but hardly practicable. An extremely complex fluidics is usually expensive for a single use in production and also carries a higher risk of susceptibility. Large and complex instrument platforms usually provide solutions for individual fluidics to which they have been tailored.
  • microfluidic platforms are the interface between sample and fluidics, ie the loading / filling of the microfluidic unit. With a few exceptions [7, 8], lab-on-chip systems do not have adequate possibilities to directly record and process a sample.
  • US Pat. No. 8,394,625 B2 describes a lab-on-a-pipette system which is a combination of a lab-on-chip system and a microtip.
  • the microtip is an extension of a chip platform, wherein the fluidics and analytics is done in the lab-on-chip system.
  • 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, wherein the pipe section defines 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 a gas movement in the operating room.
  • EP 1 381 468 B1 describes a device for taking an aliquot of a sample from a sealed container.
  • WO 201 1/032228 A1 describes an experimental unit which 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, with the tip serving only for sample collection.
  • WO 2006/029387 A1 describes a handy and portable microfluidic device which comprises a cleaning chip and a syringe-like device for sample taking and movement. Only a preparation or purification of the sample takes place.
  • the present invention is therefore based on the object to provide a concept which on the one hand requires a simpler structure and on the other hand allows a more reliable examination of the sample.
  • Embodiments of the present invention provide a tube having a portion 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, wherein the microfluidic structure is configured to perform a multi-step bioanalytical reaction with the sample material.
  • a microfluidic structure is used to analyze a sample by a multistage bioanalytical reaction, located directly in a tapered region of a tube, such as a tip of a pipette tip. This allows a reliable and cost-effective performance of complex, multi-step reactions directly within the tapered region of the tube.
  • Some embodiments make it possible to perform bioanalytical detection reactions, which take place in several spatially or temporally separate steps, in the vessels that directly serve for the collection and withdrawal of sample material (e.g., pipette tubes).
  • sample material e.g., pipette tubes
  • This is made possible by a subdivision of the vessel volume by means of a movable fluidics. Its incorporation into the vessel volume thereby forms at least two separate reaction spaces / compartments.
  • tapered tubes are considered in which a tubular fluidity rests on the inner cone.
  • Corresponding functional designs solve common production-related and microfluidic problems of chip-based structures that have more than one reaction space.
  • Other embodiments include performing a sequential, multi-stage bioassay in the volume of a tapered tube (in the form and function of a pipette tip), characterized in that an integrated microfluidic device produces at least two compartments preceded by analytical reagents and with a sample liquid in time sequence (successive / consecutive) can be reacted.
  • the peculiarity of the integrated microfluidics is that a nested tubular channel system loosely rests on the inner wall of the cone, thereby forming compartments and sealing them functionally.
  • the compartments formed are separated by (annular) gaps / openings of hydrophobic surfaces.
  • Some embodiments allow, in a very simple and inexpensive manner, the implementation of complex, multi-stage and cohesive reactions within the volume of a pipette tube, as is commonly used for the collection and removal of sample material.
  • FIG. 1 is a schematic cross-sectional view of a tube having an area tapering toward one end of the tube and a microfluidic structure disposed in the tapered area of the tube, according to FIG.
  • 2a-2b are schematic cross-sectional views of a tube with one to one end of
  • FIG. 3 is a schematic cross-sectional view of a tube having an area tapering toward one end of the tube and a microfluidic structure at least partially disposed in the tapered area of the tube according to an embodiment of the present invention
  • FIGS. 4a-4b show a photograph and a schematic cross-sectional view of a tube with an area tapering towards one end of the tube and a microfluidic structure disposed in at least partially in the tapered area of the tube according to an embodiment of the present invention
  • Figures 5a-5e show schematic cross-sectional views of the tube shown in Figure 2 during different steps of a method of assaying a sample material with the tube, according to an embodiment of the present invention
  • FIGS. 6a-6d show schematic cross-sectional views of the tube shown in FIG. 2 during and after different steps of a method for removing a sample material after performing the multistage bioanalytical
  • FIGS. 7a-7e are schematic cross-sectional views of the tube shown in FIG. 2 at different stages of a method of inspecting a sample material with the tube, according to one embodiment of the present invention; a photograph of a pipette tip through a filter disc;
  • FIG. 9 shows a photograph of a gel electrophoresis of two PCR products.
  • FIG. 10 is a flowchart of a method according to an embodiment of the present invention.
  • FIG. 1 shows a schematic cross-sectional view of a tube 100 having a region 104 tapering toward an end 102 of the tube 100 and a microfluidic structure 106 disposed in the tapered region 104 of the tube 100 according to one 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, wherein the microfluidic structure 106 is configured to perform a multi-stage bioanalytical reaction with the sample material.
  • the tapered portion 104 of the tube 100 may be tapered (tapered-shaped) toward the end 102 of the tube 100.
  • the tapered portion 104 of the tube 100 may thus be in the shape of a tip or pointed.
  • the tube 100 can only consist of the tapered region 104.
  • the tube 100 can also optionally have a cylindrical (hollow-cylindrical) region 110.
  • the tube may be both a pipette tip and a pipette having a tip with the microfluidic structure 106 disposed within the tip 104.
  • the microfluidic structure 106 may be disposed only within the tapered portion 104 of the tube 100. However, it is also possible that the microfluidic structure 106 extends over the tapered region out to the conical portion 1 10 extends. In this case, the microfluidic structure is thus (only) at least partially disposed within the tapered region 104 of the tube.
  • 2 a and 2 b show schematic cross-sectional views of a tube 100 with a region 104 tapering towards an end 102 of the tube 100 and a microfluidic structure 106 which is at least partially disposed in the tapered region 104 of the tube 100, according to an embodiment of FIGS present invention. In comparison with the exemplary embodiment shown in FIG. 1, details of the microfluidic structure 106 are shown in FIGS. 2 a and 2 b.
  • the microfluidic structure 106 may have a movable, tubular component 14.
  • the movable, tubular component 14 may be connected to the opening 108 of the tube.
  • the microfluidic structure 106 may be designed to transport a sample material received via the opening 108 of the tube via the movable, tubular component 14 in a first direction (in FIGS. 2a and 2b "upwards").
  • the microfluidic structure 106 may have at least two reaction chambers 1 2_1 to 12_3 (two reaction chambers 1 12_ 1 and 1 12_ 2 in FIG. 2 a, three reaction chambers 1 12_ 1 to 1 12_ 3 in FIG. 2 b) for carrying out the multi-stage bioanalytical reaction.
  • the (at least) two reaction chambers 1 12_1 to 1 12_3 can be separated from each other.
  • the microfluidic structure 106 can be designed to be the sample material (previously transported by the movable, tubular component 14 in the first direction (upwards) for carrying out the multistage bioanalytical reaction through the at least two reaction chambers 1 12_ 1 to 1 12_ 3 in FIG a second direction (in Fig. 2 to "down") to transport.
  • microfluidic structure 106 shown in FIG. 2 it is thus possible to carry out the multistage bioanalytical reaction sequentially or in time-separated steps.
  • an analytical reagent for the multi-stage bioanalytical reaction can be arranged upstream of at least one of the at least two reaction chambers 1 12 1 to 12 12 for this purpose.
  • analytical Reagents may be upstream, wherein the fluidic structure 106 may be formed to the analytical reagents with to react the sample material in chronological order to perform the multistep bioanalytical reaction.
  • the microfluidic structure 106 may have at least one annular component 16 1 and 16_ 2 which bears against an inner wall of the tube.
  • the movable, tubular component 1 14 is arranged so that it rests in the tapering portion 104 of the tube on an inner wall of the tube and at least partially on the at least one annular component 1 16_1 and 1 16_2.
  • the movable, tubular component 1 14 can in this case abut the annular component 16 so that the formed at least two reaction chambers 11 1 to 12 12 are separated from one another by openings of hydrophobic surfaces or hydrophobic capillary openings.
  • the tube 100 may further include a filter or O-ring 120.
  • the sample material can be absorbed via the O-ring 120.
  • FIGS. 2 a and 2 b show schematic representations of tip-shaped tubes 100 for taking in and taking up sample material with integrated tubular fluidics, which subdivides the volume of tube 100 into two or three separate reaction regions 1 12_ 1 to 1 12_ 3 and the passage multi-stage detection reactions allows.
  • Sample liquids may be picked up very precisely externally via the tapered portion of the tube (tip) 104 and may enter the first compartment 112_1 in the proximal (widened) area of the first compartment 112 via a central tubular channel 14 loosely fitting the inner conical portion 104 Get pipette tube 100.
  • the liquid transport is characterized in that it runs primarily in the direction of the cylinder / cone axis, ie vertically by means of pressure differences and centrifugal forces.
  • the liquid transport preferably takes place via centrifugal forces and in the direction of the cone tip, so that the liquid of a proximal compartment is forced via the hydrophobic capillary opening into the nearest distal compartment (in the direction of the cone tip).
  • the capillary opening length, width, surface area
  • the applied centrifugal force the transport from compartment to compartment can be controlled. This allows the local and temporal separation of reaction processes.
  • FIG. 3 shows a schematic view of a tube 100 having a region 104 tapering towards an end 102 of the tube 100 and a microfluidic structure 106 at least partially disposed in the tapered region 104 of the tube 100, according to one embodiment of the present invention Invention.
  • the microfluidic structure 106 comprises four reaction chambers 1 12_ 1 to 1 12_ 4 that are formed via three annular components 1 16_ 1 to 1 16_ 3 and the movable, tubular component 14.
  • FIG. 3 shows a schematic representation of a microfluidic device 100 with a total of 4 compartments 1 12_1 to 112_4, which are separated by differently broad and long capillary gaps / openings.
  • the two middle compartments (2 and 3) 1 12_2 and 1 12_3 each have an annular side pocket 1 18_1 and 1 18_2 in which liquid can be retained, e.g. for separate reactions or to skim off superfluous reaction solutions.
  • compartment 4 (1 12_4) result from the arrangement shown here, three reaction chambers with dead end function. If the tube-shaped channels 104 are moved counter to the taper, the liquid can be emptied towards the tip.
  • the microfluidic device 106 integrated into the conical pipette tube 100 can also be designed such that individual compartments 1 12_ 2 and 12_ 3 form dead-end pockets 1 18_ 1 and 1 18_ 2.
  • reaction solutions can be divided and reduced in volume prior to transport to the next compartment.
  • the last (distal) compartment 1 12_4 closest to the tip may be characterized by being sealed from the integrated tubular channel system in the direction of the cone.
  • the centrifugal forces for liquid transport simultaneously favor the self-sealing construction.
  • the hydrophobic surfaces of the capillary gaps and the structure of the tubular channel system can prevent evaporation of the liquid, so that even reactions in the range of the boiling point of the reaction solution (eg PCR (Polymerase Chain Reaction) with temperatures of up to 95 ° C) over a long and practicable period are possible.
  • the targeted release of the reaction solution from the distal (tip-facing) compartment can be initiated by movement of the tubular channel system 14 against the taper.
  • reagents preferably freeze-dried
  • pre-stored By pre-storing all reagents necessary for a reaction, only the liquid uptake of the sample to be examined is necessary.
  • An example of a three-stage system would be eg 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 in a temporal and spatial separation for further reactions. Another example would be the storage of reagents in the individual compartments, which allow a start-stop function for the reaction.
  • Embodiments provide several advantages, as described below.
  • One advantage is that the structure and functional principle of the microfluidics integrated into the pipette tube do not require any elaborate processes and methods 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.
  • the arrangement of the liquid transport pressure equalization and gas bubble formation, as well as the rapid bubble-free filling of the compartments pose no problems.
  • the liquid intake can be done manually and without electrical pumps in a very precise manner (eg also on standard laboratory pipettes). Furthermore, at all stages of the analysis, the liquid remains in the pipette tube (single tube solution).
  • a transfer of the sample into the analyzing microfluidics is not necessary. Further, the self-sealing construction of the fluidics within the pipette tube allows both centrifugation and heating of the reaction solution without loss of fluid. Further, for detection (optional), the reaction solution can be released quickly and easily. Ultimately, the shape and absence of electrical interfaces enable easy parallelization and cost-effective integration into standard laboratory equipment. The following describes a structure with which at least two separate reactions, such as cell lysis / DNA extraction and PCR, can be performed in a pipette tip.
  • a microfluidic structure which comprises (1) a filter tip (eg ep Dualfilter TIPS®), (2) a silicone tube “long, thin” (eg inner diameter 0.5 mm, outer diameter 1, 3 mm, length 25 mm) , (3) a silicone tube “thick, short” (eg inside diameter 1, 58 mm, outside diameter 3, 18 mm, length 4 mm), (4) instead of the dual filter of the filter tip also a short piece of an additional silicone tube can be used (eg Inner diameter 0.5 mm, outer diameter 3.7 mm, length 3 mm), and (5) a spiral wire (eg twisted flat wire made of stainless steel 0, 1 x 0.5 mm).
  • a filter tip eg ep Dualfilter TIPS®
  • a silicone tube “long, thin” eg inner diameter 0.5 mm, outer diameter 1, 3 mm, length 25 mm
  • a silicone tube “thick, short” eg inside diameter 1, 58 mm, outside diameter 3, 18 mm, length 4 mm
  • a commercially available pipette eg Eppendorf Research®
  • a bench top centrifuge eg peqlab®, PerfectSpin® mini centrifuge with rotor insert for 0.5 ml reaction vessels
  • a PCR device for capillaries or Tubes eg Roche LightCycler 1 .5 ®
  • FIG. 4 a shows a photo and FIG. 4 b shows a schematic cross-sectional view of a tube 100 with a region 104 tapering towards an end 102 of the tube 100 and a microfluidic structure 106 arranged in at least partially in the tapered region 104 of the tube 100 is, according to an embodiment of the present invention.
  • the microfluidic structure 106 comprises two reaction chambers 1 12_1 and 1 12_2, which are formed via an annular component 1 16_1 and a movable, tubular component 1 14, as has already been described in detail with reference to FIG. 2 a.
  • an O-ring or filter 120 can be seen in FIGS. 4a and 4b.
  • Fig. 4a shows a photograph and Fig.
  • 4b is a schematic drawing showing the structure of the fluidics 106 and the resulting compartments 1 12_1 and 1 12_2 in the pipette tip 104. The photo was taken for better visibility of the hoses blue (first hatching) and for the rubber ring green-yellow striped (second hatching).
  • FIGS. 5a-5e show schematic cross-sectional views of the tube 100 shown in FIG. 2 during different steps of a method of inspecting a sample material with the tube 100, according to one embodiment of the present invention.
  • Fig. 5a shows a schematic cross-sectional view of the tube 100 prior to a step of soaking up the sample material 130.
  • Fig. 5a can an inner side of the movable, tubular component 1 14 lysis reagents are 122 upstream, while in the second reaction chamber 1 12_2 PCR reagents 124 can be upstream
  • Fig. 5b shows a schematic cross-sectional view of the tube 100 after a step of sucking the sample material 130, whereby the Sample material 130 was transported through the movable, tubular component 1 14 in the first reaction chamber 1 12_1.
  • Fig. 5c shows a schematic cross-sectional view of the tube 100 in a step of centrifuging, whereby the sample material from the first reaction chamber 1 12_1 enters the second reaction chamber 1 12_2.
  • FIG. 5d shows a schematic cross-sectional view of the tube 100 after the centrifuging step, whereby the sample material 130 has completely passed from the first reaction chamber 12_1 into the second reaction chamber 12_2, where the sample material contacts the PCR reagents 124 to a second bioanalytical reaction with the sample material 130 comes.
  • Fig. 5e shows a schematic cross-sectional view of the tube 100 during the second bioanalytical reaction.
  • Figures 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) the choice of suitable lysing 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 by fluorescent dyes (turquoise (fourth hatching)).
  • 6a to 6d show schematic cross-sectional views of the tube 100 shown in FIG. 2 during different steps of a method for taking a sample material after performing the multi-stage bioanalytical reaction, according to an exemplary embodiment of the present invention.
  • Fig. 6a shows a schematic cross-sectional view of the tube 100 after performing the multistage bioanalytical reaction.
  • the sample material 130 (together with any reagents) is located in the second reaction chamber 12_2.
  • Fig. 6b shows a schematic cross-sectional view of the tube 100 after a step of applying
  • Fig. 6c shows a schematic cross-sectional view of the tube 100 in a step of displacing the movable, tubular component 1 14 by applying the removal device 132 in the movable, Tubular Component 1 14.
  • FIG. 6d shows a schematic cross-sectional view of the tube 100 after the step of displacing the moveable tubular component 14, allowing the sample material 130 to escape from the tube 100.
  • FIGS. 6a to 6d show schematic representations of how, after the end of the reaction, by subsequently applying a spiral-wound wire 132, the reaction liquid 130 can be released for further examinations.
  • the wire 132 may be inserted from the pipette side (top) or, in the case of the filter, from the tip (bottom) into the pipette tip. In both cases, with the wire 132, the long, thin tube 1 14 is moved upward, so that the liquid 130 can escape through the tip by operating the pipette.
  • FIGS. 7a-7e show schematic cross-sectional views of the tube 100 shown in FIG. 2 during different steps of a method of inspecting a sample material with the tube 100, according to one embodiment of the present invention.
  • Figure 7a shows a schematic cross-sectional view of the tube 100 prior to a step of soaking up the sample material 130.
  • the sampling device 132 eg, a spiral wire
  • the lysis reagents 122 may be upstream of the sampling device 132.
  • the second reaction chamber 1 12_2 PCR reagents 124 may be upstream.
  • 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 14 into the first reaction chamber 112.
  • FIG. 7c shows a schematic cross-sectional view of the tube 100 in a step of centrifuging, whereby the sample material from the first reaction chamber 1 12_1 enters the second reaction chamber 1 12_2.
  • FIG. 7 d shows a schematic cross-sectional view of the tube 100 the centrifuging step, whereby the sample material 130 has completely passed from the first reaction chamber 1 12_ 1 into the second reaction chamber 1 12_ 2 where the sample material contacts the PCR reagents 124, resulting in a second bioanalytical reaction with the sample material 130.
  • FIG. 5 e shows a schematic cross-sectional view of the tube 100 after the second bioanalytical reaction and displacement of the moveable tubular component, allowing the sample material to escape from the tube 100.
  • FIGS. 7a to 7e show schematic representations of an application similar to Figs. 5a to 5e.
  • the wire 132 is already part of the fluidics at the beginning.
  • the lysis reagents 124 are in this case even pre-deposited on the surface of the wire 132 and washed off during the absorption of the sample 130 of this.
  • 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 as described in FIGS. 5b to 5e, but in addition the function of FIG. 6a to 6d described already integrated.
  • FIG 8 shows a photograph of a pipette tip (lab in a pipette tip, German laboratory in a pipette tip) through a filter disk.
  • a pipette tip is a PCR product with the dye SYBR Green I which can be excited in the wavelength range of fluorescein with blue light ( ⁇ 495nm) and emits green light (-520 nm).
  • FIG. 9 shows a photograph of a gel electrophoresis (1.5% agarose, 70 V, 45 min.) Of two PCR products.
  • the positive control (Pos.) was tempered in a plastic capillary.
  • the pipette tip was filled with 10 ⁇ of the PCR solution.
  • the temperature was controlled in a Roche LightCycler 1.5 ®.
  • the method 200 includes a step 202 of receiving a sample material through an opening of a tube, the tube having a tapered portion toward an end of the tube forming the opening. Further, the method 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 a supplement / extension, this represents one embodiment essential part of the analyzing system and the fluidics.
  • aspects have been described in the context of a device, it will be understood that these aspects also constitute a description of the corresponding method such that a block or device of a device is also to be understood as a corresponding method step or feature of a method step , Similarly, aspects 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 may be performed by a hardware device (or using a hardware device). commodity), such as a microprocessor, a programmable computer, or an electronic circuit. In some embodiments, some or more of the most important method steps may be performed by such an apparatus.
  • the embodiments described above are merely illustrative of the principles of the present invention.
  • Exner, T., pipette tip for enzyme-linked immunosorbent assay includes tracer coated on surface (s) from inside and outside surfaces and proximal fluid receiving and delivery end of pipette tip, and for mixing with liquid sample drawn into pipette tip, HAEMATEX RES PTY LTD (HAEM-Non-standard). p. 16th
  • Gjerde, D.T. Purifying / concentrating analyte, e.g. protein from sample solution involves passing it into pipette tip column having packed bed of extraction medium; passing wash solution / desorption solvent; and passing through second pipette tip column, GJERDE D T (GJER-Individual) PHYNEXUS INC. (PHYN-Non-standard).
  • GJERDE D T GJER-Individual PHYNEXUS INC. (PHYN-Non-standard).
  • Gaitas, A. and A. Basu Lab-on-a-pipette apparatus useful for analyzing small analyte volumes in a genetic diagnosis comprising a pipette and a microfluidic component, GAITAS A (GAIT-Individual) BASU A (BASU-Individual) BASU A (BASU-individual).
  • GAITAS A GAIT-Individual
  • BASU A BASU-Individual
  • BASU-individual BASU-individual

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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)

Abstract

Dans des modes de réalisation cités à titre d'exemple, l'invention concerne un tube présentant une zone se rétrécissant en direction d'une de ses extrémités, une ouverture à l'extrémité du tube, permettant l'introduction d'une substance à analyser; et une structure microfluidique située à l'intérieur du tube dans la zone se rétrécissant, et reliée à l'ouverture, la structure microfluidique étant conçue pour la conduite d'une réaction bioanalytique en plusieurs étapes avec la substance à analyser.
PCT/EP2017/051372 2016-01-27 2017-01-24 Tube à structure microfluidique WO2017129541A1 (fr)

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DE102016201181.2A DE102016201181B3 (de) 2016-01-27 2016-01-27 Röhre mit einer mikrofluidischen Struktur
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Publication number Priority date Publication date Assignee Title
WO2020214224A1 (fr) * 2019-04-18 2020-10-22 Siemens Healthcare Diagnostics Inc. Dispositif microfluidique intégré avec adaptation de pipette

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