WO2020127495A1 - Procédé de préstockage au moins d'un réactif dans un système microfluidique et système microfluidique - Google Patents

Procédé de préstockage au moins d'un réactif dans un système microfluidique et système microfluidique Download PDF

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Publication number
WO2020127495A1
WO2020127495A1 PCT/EP2019/085968 EP2019085968W WO2020127495A1 WO 2020127495 A1 WO2020127495 A1 WO 2020127495A1 EP 2019085968 W EP2019085968 W EP 2019085968W WO 2020127495 A1 WO2020127495 A1 WO 2020127495A1
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WO
WIPO (PCT)
Prior art keywords
reagent
protective layer
microfluidic system
drying
microfluidic
Prior art date
Application number
PCT/EP2019/085968
Other languages
German (de)
English (en)
Inventor
Stefanie Fischer
Original Assignee
Robert Bosch Gmbh
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 Robert Bosch Gmbh filed Critical Robert Bosch Gmbh
Publication of WO2020127495A1 publication Critical patent/WO2020127495A1/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
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/16—Reagents, handling or storing thereof

Definitions

  • the invention is based on a method for storing at least one reagent in a microfluidic system, in particular in a lab-on-chip system, and a microfluidic system according to the type of the independent claims.
  • EP 2 080 554 B1 discloses a method for storing a
  • Reagent in a microfluidic device Reagent in a microfluidic device.
  • the approach presented here presents a method for storing at least one reagent in a microfluidic system, in particular in a lab-on-chip system.
  • the method has a step of introducing the reagent into a cavity of the
  • microfluidic system as well as a step of applying one
  • Upstream of a reagent can be understood to mean prior introduction or filling of the cavity in the microfluidic system, such as, for example, a microfluidic channel, a microfluidic chamber or another microfluidic reaction volume prior to an actual, in particular subsequent, use of the reagent.
  • a reagent can be understood as a substance that shows a specific reaction when it comes into contact with certain other substances or materials. It can this is a synthesis reagent and in particular a detection reagent.
  • the protective layer protects the reagent from external influences at least temporarily. For example, undesired chemical reactions of the reagent with substances from its immediate environment can be prevented or at least delayed. Furthermore, the protective layer also offers protection against mechanical removal of the reagent by, for example, liquid streams and / or gas streams with which the microfluidic system, in particular the channels, cavities and / or reaction volumes therein, are flushed in order to, for example, air bubbles from the microfluidic system remove.
  • the protective layer at least partially covers the reagent, premature dissolution of the reagent by a fluid stream flushing the microfluidic system can advantageously be prevented or at least delayed. Furthermore, it can be achieved in this way that when the microfluidic system is rinsed with the fluid flow, parts of the reagent get into further channels, cavities and / or reaction volumes of the microfluidic system, thus contaminating them and disrupting processes taking place there.
  • the protective layer can also be applied completely to the reagent, to further cavities and / or to an entire surface of the microfluidic system.
  • a precisely metered amount of a material forming the later protective layer can be applied to the reagent in a simple manner. This can be done, for example
  • the protective layer has a polyether. Because polyethers can be adjusted in a simple manner with specifically adjustable chemical and
  • the melting point and / or a solubility of the polyether, for example in water, can be influenced in a targeted manner during the production of the polyether. It is particularly advantageous if the polyether is a polyethylene glycol (PEG).
  • PEG is a liquid or solid, water-soluble and non-toxic polymer, the properties of which can be adjusted by adjusting the chain length of the polymer.
  • the PEG can be the PEG 6000.
  • PEG 6000 has a number average mass of 6000 g per mol.
  • a protective layer can be applied to the reagent in a particularly simple and inexpensive manner.
  • the polyether is present in an aqueous solution. This allows it to be applied to the reagent as a protective layer in a simple manner. It is particularly advantageous if the polyether is present in aqueous solution with a concentration in a range from 2% to 40%, preferably in a range from 10% to 30% and particularly preferably in a range from 15% to 25%. This area is particularly suitable for applying the polyether to the reagent as a protective layer.
  • Protective layer a step of drying the protective layer takes place. This creates a protective layer that is particularly stable against external influences.
  • the drying of the protective layer can be understood to mean the escape of the water contained in the aqueous solution or in the protective layer. In particular, this can be understood to mean evaporation of the water. This can preferably be understood to mean evaporation of the water in air.
  • the drying step can comprise a lyophilizing or freeze-drying step.
  • the still liquid protective layer is first cooled down to such an extent that the water contained in it freezes, and then the frozen water is sublimated from the protective layer by lowering an ambient pressure.
  • the reagent can be applied in liquid form.
  • the reagent can be introduced into the cavity or into the microfluidic system in a particularly simple manner. It is advantageous if after the step of introducing the reagent in liquid form there is a step of drying the reagent present in liquid form. When the reagent dries, the escaping of the one present in the aqueous solution, in particular, can occur
  • Reagent contained water can be understood. In particular, this can be understood to mean evaporation of the water. This can preferably be understood to mean evaporation of the water in air.
  • the step of drying the reagent can comprise a step of heating the reagent so that evaporation or
  • Evaporation of the water is supported from the reagent present in particular in aqueous solution.
  • the protective layer can cover the reagent in a targeted and particularly effective manner.
  • the reagent and / or the protective layer are freeze-dried in the drying steps. Because this frees the reagent and / or the protective layer from the water contained therein in a particularly simple manner, so that a solid reagent and / or a solid protective layer is formed.
  • the microfluidic device comprises at least two cavities and in the step of introducing at least one reagent is introduced into each of the at least two cavities.
  • the microfluidic device can comprise a plurality of cavities and at least two or more cavities can each contain at least one reagent. It is therefore also possible for two or more reagents to be contained in one cavity, for example. It can optionally be provided that the reagents differ in the different cavities. This allows different reagents in different cavities Microfluidic device or the microfluidic system can be upstream, in order to use them later at given, determinable times.
  • the protective layer covers all reagents, preferably covers all cavities and particularly preferably covers the entire microfluidic system, the microfluidic system can be pre-rinsed with a liquid prior to its use in order to displace trapped air from the cavities of the system.
  • the step of introducing the reagent is repeated at least once.
  • the step of drying the reagent can also be repeated at least once. In this way, an amount of the reagent to be introduced or introduced into the cavity can advantageously be checked.
  • the step of applying the protective layer to the reagent is repeated at least once.
  • the thickness of the protective layer allows a mechanical resistance of the protective layer and thus the protective effect for the reagent underneath to be set at least in certain areas.
  • a duration can be set via the thickness of the protective layer, after which the upstream reagent is supplied for later use in the microfluidic system
  • introducing the reagent and applying the protective layer to the reagent are repeated at least once.
  • a plurality of reagents are each arranged in a stack-like manner one above the other by a protective layer.
  • the reagents each covered by a protective layer can be fed into their microfluidic system one after the other in time.
  • the aforementioned advantages also apply in a corresponding manner to a microfluidic system, in particular a lab-on-chip system, for storing at least one reagent.
  • the microfluidic system comprises at least one cavity for receiving the at least one reagent and is set up to carry out the steps of the method according to one of the previously described embodiments.
  • Figure 1 is a schematic representation of a microfluidic system for storing at least one reagent according to an embodiment
  • Figure 2 is a schematic representation of a cavity of the microfluidic system according to an embodiment
  • Figure 3 is a schematic representation of a cavity of the microfluidic system according to another embodiment. such as
  • FIG. 4 shows a flow chart of a method for storing at least one reagent in a microfluidic system according to one
  • Figure 1 is a schematic representation of a plan view of a
  • the microfluidic system has a plurality of cavities designed as microfluidic channels 2a, 2b, 2c, 2d and as a reaction chamber 3.
  • the channels 2a, 2b, 2c are in this case designed to hold a fluid and any substances dissolved therein, such as, for example.
  • the reaction chamber 3 is set up for a chemical, biochemical and / or biomolecular reaction between different fluids supplied from the channels 2a, 2b, 2c. Any reaction products or the previously supplied fluids can be removed from the reaction chamber 3 via the channel 2d.
  • the cavities Before using the microfluidic system 1, the cavities can be pre-rinsed with a liquid in order to displace trapped air from the channels 2a, 2b, 2c, 2d and / or the reaction chamber 3. This reduces the risk of entraining the air bubbles in a later process. It can be advantageous here to store reagents in the microfluidic channels 2a, 2b, 2c, 2d before this rinsing process.
  • the reagent is at least partially covered by a protective layer.
  • FIG. 2 shows a schematic illustration of a cross section of one of the channels 2a, 2b, 2c from FIG. 1.
  • a reagent 10 is introduced or upstream in the channel 2.
  • the reagent 10 is at least partially covered by a protective layer 20.
  • the reagent 10 can comprise, for example, oligonucleotides, in particular primers and / or probes.
  • the reagent 10 can, for example, be introduced into the channel 2 as a solid.
  • the reagent 10 can be introduced into the channel 2 in liquid form, in particular as an aqueous solution.
  • the reagent 10 is now at least partially, in particular completely, covered by a protective layer 20.
  • FIG. 3 shows a further development of the embodiment described in FIG. 2.
  • the reagent 10 introduced into the channel 2 is covered with a protective layer 20.
  • a further reagent 11 is introduced onto the protective layer 20, which in turn is covered by a further protective layer 21.
  • Both Reagents 10, 11 can be the same reagents. Alternatively, it can be provided that the reagents 10, 11 differ.
  • the reagents 10, 11 are arranged in a kind of stacked shape in the channel 2 and are encapsulated in themselves by the protective layers 20, 21.
  • Protective layer 20, 21 for the reagents 10, 11 can be set. This can ensure that the reagents 10, 11 are not prematurely detached and thereby washed out by a previous rinsing process of the microfluidic system, and that the reagents 10, 11 are supplied for their actual use at the desired or specific point in time.
  • FIG. 4 shows a flow chart of a method 100 for storing at least one reagent 10, 11 in a microfluidic system 1.
  • a reagent 10 is introduced into a channel 2 of a microfluidic system 1.
  • a protective layer 20 is applied to the reagent 10. The protective layer 20 is applied in such a way that the reagent 10 is at least partially covered by the protective layer 20.
  • the reagent 10 can be introduced into the channel 2 both in solid and in liquid form.
  • a drying step can subsequently take place in which the reagent 10 is dried.
  • the drying step can include heating and / or freeze-drying the reagent.
  • the protective layer can consist of a polyether, which is present in aqueous solution.
  • the protective layer 20 can be a
  • a step of drying the protective layer 20 can optionally take place analogously to the optional step of drying the reagent 10.
  • the method 100 can be carried out successively or simultaneously on different areas of the microfluidic system 1, in particular on different channels 2 of the Microfluidic system 1 take place, so that the method 100 can be easily integrated into a series production.
  • the reagent 10 in particular in liquid form, is introduced several times in succession into a channel 2 in order to increase the amount of the reagent 10 introduced overall.
  • the protective layer 20 is applied to the reagent 10 several times. As a result, a layer thickness of the protective layer 20 increases accordingly and thus the protective duration of the protective layer 20
  • the steps of introducing 110 the reagent 10 and applying 120 the protective layer 20 are carried out several times in succession. In this way, a stack of in each case encapsulated reagents 10, 11 is reached, which can be added to their respective processes one after the other in time.
  • Concentration of 10 mM are pipetted into channel 2 of the microfluidic system 1. This is followed by drying of the primer and / or
  • Probe solution for about two hours.
  • the drying of the primer and / or probe solution can be accelerated by increasing the temperature to approx. 60 degrees Celsius.
  • a pl of an aqueous PEG 6000 solution in a concentration of 20% (w / v) (weight / volume) is then applied to the dried reagent 10 as a protective layer 20.
  • the PEG 6000 solution is also dried for approx. One hour, so that the
  • the drying process can be accelerated by applying heat.
  • nl of the primer and / or probe solution are introduced into the microcavity in a concentration of one mM and dried. Due to the small amount of the primer and / or probe solution, drying takes place within a few seconds to minutes at room temperature. Below 50 nl of a PEG 6000 solution in a concentration of 5% (w / v) (weighl / volume) are applied to the primer and / or probe solution and also dried. As a result, the reagent 10 can also be protected in very small quantities with PEG 6000 as the protective layer 20.
  • Polyethylene glycols such as PEG 1000, PEG 2000, PEG 3000, PEG 4000,
  • PEG 5000 or polyethylene glycols with average molecular weights greater than 6000 g per mol can be used.

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Dispersion Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Physical Or Chemical Processes And Apparatus (AREA)

Abstract

L'invention concerne un procédé (100) de préstockage au moins d'un réactif (10, 11) dans un système microfluidique (1), notamment dans un système de laboratoire sur puce, comprenant les étapes consistant à introduire (110) le réactif (10, 11) dans une cavité (2a, 2b, 2c, 2d, 3) du système microfluidique (1) et appliquer (120) une couche protectrice (20, 21) sur le réactif (10, 11) de manière telle que le réactif (10, 11) soit recouvert au moins partiellement par la couche protectrice (20, 21).
PCT/EP2019/085968 2018-12-21 2019-12-18 Procédé de préstockage au moins d'un réactif dans un système microfluidique et système microfluidique WO2020127495A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102018222847.7 2018-12-21
DE102018222847.7A DE102018222847A1 (de) 2018-12-21 2018-12-21 Verfahren zum Vorlagern wenigstens eines Reagenzes in einem mikrofluidischen System und mikrofluidisches System

Publications (1)

Publication Number Publication Date
WO2020127495A1 true WO2020127495A1 (fr) 2020-06-25

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DE (1) DE102018222847A1 (fr)
WO (1) WO2020127495A1 (fr)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100216193A1 (en) * 2007-10-26 2010-08-26 Toppan Printing Co., Ltd. Reaction chip, reaction method, temperature controlling unit for gene treating apparatus and gene treating apparatus
EP2080554B1 (fr) 2008-01-16 2013-03-20 Samsung Electronics Co., Ltd. Procédé de stockage d'un réactif analytique dans un dispositif microfluidique
WO2018069056A1 (fr) * 2016-10-12 2018-04-19 Mycartis N.V. Cartouche pré-remplie

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101102532B1 (ko) * 2008-07-10 2012-01-03 삼성전자주식회사 시약 카트리지, 시약 카트리지를 구비하는 미세유동장치, 그 제조방법, 및 이를 이용한 시료분석방법

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100216193A1 (en) * 2007-10-26 2010-08-26 Toppan Printing Co., Ltd. Reaction chip, reaction method, temperature controlling unit for gene treating apparatus and gene treating apparatus
EP2080554B1 (fr) 2008-01-16 2013-03-20 Samsung Electronics Co., Ltd. Procédé de stockage d'un réactif analytique dans un dispositif microfluidique
WO2018069056A1 (fr) * 2016-10-12 2018-04-19 Mycartis N.V. Cartouche pré-remplie

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