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WO2025181542A1 - Cellule pour un système d'analyse microfluidique - Google Patents

Cellule pour un système d'analyse microfluidique

Info

Publication number
WO2025181542A1
WO2025181542A1 PCT/IB2024/063217 IB2024063217W WO2025181542A1 WO 2025181542 A1 WO2025181542 A1 WO 2025181542A1 IB 2024063217 W IB2024063217 W IB 2024063217W WO 2025181542 A1 WO2025181542 A1 WO 2025181542A1
Authority
WO
WIPO (PCT)
Prior art keywords
cell
reagent
well
port
cells
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/IB2024/063217
Other languages
English (en)
Inventor
Bartolomeo Della Ventura
Raffaele Velotta
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Elicadea Srl
Materias SRL
Original Assignee
Elicadea Srl
Materias SRL
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 Elicadea Srl, Materias SRL filed Critical Elicadea Srl
Publication of WO2025181542A1 publication Critical patent/WO2025181542A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

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
    • B01L3/50—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
    • B01L3/502715—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 interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
    • 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
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
    • 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
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
    • B01L2200/027—Fluid interfacing between devices or objects, e.g. connectors, inlet details for microfluidic devices
    • 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
    • B01L2200/0689—Sealing
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0829—Multi-well plates; Microtitration plates
    • 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
    • B01L2300/0877—Flow chambers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/16—Microfluidic devices; Capillary tubes

Definitions

  • the present invention is generally placed, in general, in the field of microfluidic systems; in particular, the invention refers to a cell for a microfluidic analysis system.
  • Microfluidics is an interdisciplinary field that deals with the manipulation and control of liquids at micro and nanometric scales. It has had a significant impact on biosensor technology and in the analysis of biological samples through the development of miniaturized devices with applications in different fields.
  • Microfluidic technologies have made automation possible and improved analytical capacity, allowing numerous experiments to be performed quickly in parallel. This allows the creation of portable and low-cost devices, suitable for use in the field or in healthcare facilities with limited resources. Such devices can significantly improve access to medical care in remote or disadvantaged areas and provide rapid results for timely and accurate diagnosis.
  • a single microfluidic unit, or cell which can be connected in series with other units, offering greater control over flows in the culture medium and more uniform reagent distribution.
  • the possibility of connecting multiple units in series allows the use of a single culture medium, reducing the use of reagents.
  • microfluidics in PDMS used in the known art presented while having excellent qualities in terms of flexibility, does not always allow good sealing in the wells due to its mechanical characteristics. These limitations of this material may lead to liquid leaks in the case of high pressures, a necessary condition for feeding multiple cells with a single fluid flow.
  • the cell for the microfluidic system comprises within a vertical inlet duct of reagent fluids and a vertical outlet duct of reagent fluids.
  • the cell also includes a well formation for channelling the fluid introduced by the vertical inlet duct of reagent fluids towards the vertical outlet duct.
  • the claimed system comprising a multi-well plate that allows a high degree of customization, is efficient for various numbers of cells and allows the type of culture table to be changed according to the specific requirements.
  • figure 1 is a perspective view of a cell for a microfluidic system according to an embodiment of the present invention
  • figure 2 is a top view of a cell for a microfluidic system according to an embodiment of the present invention
  • figure 3 is a bottom view of a cell for a microfluidic system according to an embodiment of the present invention
  • figure 4 shows two perspective views of a multi-well plate for a microfluidic system according to an embodiment of the present invention.
  • figure 5 is a bar graph reporting the experimental results of the absorption peak of cells arranged in series according to an embodiment of the present invention
  • figure 6 shows the resulting data from theoretical simulations on the pressure of cells arranged in series according to an embodiment of the present invention.
  • FIG. 1 a perspective view of a cell 10 for a microfluidic analysis system according to an embodiment of the present invention is shown.
  • the cell 10 is designed to be traversed by a reagent flow to be supplied to a culture table 80 where biological samples, for example, may be present.
  • the cell 10 comprises a hollow main container 20 with an external side wall 30 that defines an internal chamber 40, a lower base 70 having a flow port 50, and a well formation 60 that extends vertically from the base 70 into the internal chamber 40, circumscribing the flow port 50.
  • This well formation 60 when the cell 10 is placed vertically on the culture table 80, delimits an analysis volume 90 having as its base a subsurface of the culture table 80.
  • FIG 2 a bottom view of the cell 10 is shown, making the flow port 50 visible.
  • the flow port in a preferred embodiment, is connected to a lower outlet port 91 of a vertical inlet duct 90 of reagent fluids to introduce the reagent fluids into the analysis volume 90.
  • the well formation 60 which has a height less than the height of the external side wall 30, is arranged to channel the fluid, introduced by the lower outlet port 91 of the vertical inlet duct 90 for introducing reagent fluids connected to the flow port 50, from the analysis volume 90 to the internal chamber 40 of the hollow container 20.
  • the well formation 60 is coaxially internal to the external side wall 30 of the hollow container 20 and defines with it a cylindrical annular interspace.
  • the cell 10 also includes a vertical outlet duct 100 for discharging reagent fluids from the cell 10 with a lower inlet 101 arranged in the internal chamber 40 adjacent to an upper edge 61 of the well formation 60.
  • the cell 10 also includes an upper part 110 of the hollow main container 20 comprising an upper inlet port 92 of the vertical inlet duct 90 and an upper outlet port 102 of the vertical outlet duct 100 of the reagent fluids.
  • FIG. 3 a top view of the cell 10 is shown in an embodiment of the present invention that shows the upper inlet port 92 of the vertical inlet duct 90 and the upper outlet port 102 of the vertical outlet duct 100.
  • the cell 10 can be, for example, 4 cm high with the upper inlet port 92 having a diameter of 0.9 mm and the upper outlet port 102, instead, a diameter of 0.6 mm.
  • the external side wall 30 has one or more projections 31 that are designed to be housed in corresponding locking seats 121 present in a plurality of wells 120 of a multi-well plate 130 associated with the culture table 80.
  • the locking seats 121 are configured to cooperate with one of the projections 31 to lock and support a cell 10 in a vertical position.
  • FIG. 3 a perspective view of a multi-well plate 130 with the corresponding wells 120 according to an embodiment of the present invention is shown.
  • the locking seats 121 of the well 120 are shaped to cooperate with the projections 31 of the external side wall 30 locking the cell 10 in a detachable manner according to a bayonet configuration.
  • the base 70 also includes a sealing element 140 in a closed-ring shape that surrounds a lower part of the well formation 60 so that, when the cell 10 is locked through the locking seats 121 as described above, the analysis volume 90 is sealed.
  • the sealing element 140 can be, for example, a toroidal O-ring.
  • the sealing element 140 may have an internal diameter of 6.2 mm and an external diameter of 9.6 mm.
  • the present invention also relates to a microfluidic analysis system for implementing the flow of reagent fluids to a culture table 80, which will now be illustrated.
  • the system includes a multi-well plate 130 with a plurality of wells 120 configured for the temporary support and locking of a corresponding plurality of cells 10 as described in the previous embodiments.
  • the cells 10 are arranged to be fluidically connected and traversed in series by a reagent fluid flow through a plurality of connecting channels 150 that connect each upper outlet port 102 of the vertical outlet duct 100 of a preceding cell 10 with the upper inlet port 92 of the vertical inlet duct 90 of a subsequent cell 10.
  • a reagent fluid flow through a plurality of connecting channels 150 that connect each upper outlet port 102 of the vertical outlet duct 100 of a preceding cell 10 with the upper inlet port 92 of the vertical inlet duct 90 of a subsequent cell 10.
  • the flow management of the system claimed here can be carried out, for example, by a peristaltic pump or a continuous pump.
  • the microfluidic analysis system can be made through an additive manufacturing process such as 3D printing.
  • this additive manufacturing process may be carried out using an acrylonitrile butadiene styrene filament.
  • the locking seats 121 of each well 120 are arranged in such positions that, in a locked condition in which the projections 31 of the cells 10 are engaged in the well seats 121, the respective closed-ring sealing elements 140 are vertically compressed and tightly surround the bases of the analysis volumes 90.
  • the uniform and highly efficient coating of a nanostructured surface with gold nanoparticles was demonstrated.
  • the nanostructure was used to exploit a physical phenomenon called "localized surface plasmon resonance" to monitor the coating of the surface through spectrophotometric measurements in a simple and fast way.
  • the shift of the absorption peak was observed.
  • the greater the measured peak shift the greater the surface coverage by the biological element (mouse-produced standard IgG antibody at a concentration of 25 pg/mL).
  • the concentration and volumes of the antibody solution used for this test were excessive, with an amount of material that in principle would have been enough to saturate approximately 200 wells.
  • the result of the simulation confirms the uniformity of the wells of the various cells positioned in series.

Landscapes

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

Abstract

La présente invention concerne une cellule pour un système d'analyse microfluidique, la cellule étant adaptée pour être traversée par un flux de fluide réactif et comportant un contenant principal creux doté d'une paroi latérale extérieure délimitant une chambre intérieure, une base inférieure comportant un orifice et une formation de puits, un orifice de sortie inférieur pour l'entrée des fluides réactifs, un orifice de sortie vertical pour l'évacuation des fluides réactifs de la cellule avec un orifice d'entrée inférieur agencé dans la chambre intérieure adjacente à un bord supérieur de la formation de puits et une partie supérieure contenant un orifice d'admission supérieur et un orifice d'évacuation supérieur.
PCT/IB2024/063217 2024-02-26 2024-12-27 Cellule pour un système d'analyse microfluidique Pending WO2025181542A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
IT202400004036 2024-02-26
IT102024000004036 2024-02-26

Publications (1)

Publication Number Publication Date
WO2025181542A1 true WO2025181542A1 (fr) 2025-09-04

Family

ID=91072981

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IB2024/063217 Pending WO2025181542A1 (fr) 2024-02-26 2024-12-27 Cellule pour un système d'analyse microfluidique

Country Status (1)

Country Link
WO (1) WO2025181542A1 (fr)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011137058A1 (fr) * 2010-04-30 2011-11-03 Merck Sharp & Dohme Corp. Réacteur avec plaque à puits multiples et système s'y rapportant
US10538726B2 (en) * 2010-10-07 2020-01-21 Vanderbilt University System and method for microdialysis imaging and regional fluidic delivery and control and applications of same
WO2020081740A1 (fr) * 2018-10-17 2020-04-23 Northwestern University Plateforme de culture tissulaire ayant de multiples chambres de puits en raccordement fluidique par l'intermédiaire de canaux microfluidiques et de vannes de sélection
US10633624B2 (en) * 2017-01-19 2020-04-28 Essen Instruments, Inc. Methods and apparatus for perfusion and environment control of microplate labware

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011137058A1 (fr) * 2010-04-30 2011-11-03 Merck Sharp & Dohme Corp. Réacteur avec plaque à puits multiples et système s'y rapportant
US10538726B2 (en) * 2010-10-07 2020-01-21 Vanderbilt University System and method for microdialysis imaging and regional fluidic delivery and control and applications of same
US10633624B2 (en) * 2017-01-19 2020-04-28 Essen Instruments, Inc. Methods and apparatus for perfusion and environment control of microplate labware
WO2020081740A1 (fr) * 2018-10-17 2020-04-23 Northwestern University Plateforme de culture tissulaire ayant de multiples chambres de puits en raccordement fluidique par l'intermédiaire de canaux microfluidiques et de vannes de sélection

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
RAUTI ROSSANA ET AL: "Transforming a well into a chip: A modular 3D-printed microfluidic chip", APL BIOENGINEERING, AMERICAN INSTITUTE OF PHYSICS, 2 HUNTINGTON QUADRANGLE, MELVILLE, NY 11747, vol. 5, no. 2, 28 April 2021 (2021-04-28), XP012256008, DOI: 10.1063/5.0039366 *

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