WO2019025205A1 - Cellule capillaire et utilisation de la cellule capillaire - Google Patents

Cellule capillaire et utilisation de la cellule capillaire Download PDF

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Publication number
WO2019025205A1
WO2019025205A1 PCT/EP2018/069696 EP2018069696W WO2019025205A1 WO 2019025205 A1 WO2019025205 A1 WO 2019025205A1 EP 2018069696 W EP2018069696 W EP 2018069696W WO 2019025205 A1 WO2019025205 A1 WO 2019025205A1
Authority
WO
WIPO (PCT)
Prior art keywords
capillary
capillary cell
support device
cell
sample
Prior art date
Application number
PCT/EP2018/069696
Other languages
German (de)
English (en)
Inventor
Thomas Fritzsche
Ulf-Dietrich Braumann
Original Assignee
Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V.
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 Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. filed Critical Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V.
Publication of WO2019025205A1 publication Critical patent/WO2019025205A1/fr

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Classifications

    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/34—Microscope slides, e.g. mounting specimens on microscope slides
    • 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
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5082—Test tubes per se
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N21/6456—Spatial resolved fluorescence measurements; Imaging
    • G01N21/6458—Fluorescence microscopy
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/16—Microscopes adapted for ultraviolet illumination ; Fluorescence microscopes
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
    • G02B21/365—Control or image processing arrangements for digital or video microscopes
    • G02B21/367—Control or image processing arrangements for digital or video microscopes providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • G01N21/03—Cuvette constructions
    • G01N2021/0346—Capillary cells; Microcells

Definitions

  • the present application is a capillary cell according to the preamble of claim 1 and a use of a capillary cell in a microscopy method, such as a Lichtblattmikroskopie compiler or a selective piain Illumination microscopy (SPIM) method.
  • a microscopy method such as a Lichtblattmikroskopie compiler or a selective piain Illumination microscopy (SPIM) method.
  • the illumination and detection axes are orthogonal to each other.
  • a capillary cell in which the sample to be examined is introduced.
  • the capillary cell or the sample contained therein can be moved plane-wise along the detection axis so as to see different levels illuminated by the light sheet.
  • An example of such a capillary cell is shown in DE 10 2012 108 158 AI.
  • the capillary cell described therein comprises, in addition to an envelope with a limiting side wall and an upper and a lower lid, a closure and an inflow, by means of which the capillary cell can be flushed through with a solution.
  • the drain is in the first one
  • Document CA 2 684 221 A1 shows a device or a method in which a fluid, in which particles to be examined are located, flows through a capillary. The particles moving with the fluid are measured as they flow through a measuring range. Via porous structures, substances can be added to the fluid inside the capillary, such as, for example, growth-inhibiting substances, poisons or cell-damaging chemicals.
  • the documents DE 10 2014 004 851 AI and US 2016/0 001 285 AI show a vertical reaction vessel in the form of an insert.
  • the insert is adapted to form a receptacle, with at least one top opening located at the top, and a second opening located at the top for pressure equalization and an overflow.
  • the reaction vessel consists of a dimensionally stable base body, wherein the dimensionally stable base body forms at least one non-capillary reaction space as the inner volume with at least one semipermeable membrane as the side wall.
  • the document EP 0 515 883 A2 shows a device for the safe removal of blood from a storage vessel with an aperture holder, in which a
  • Aperture exhibiting part is held and to which a capillary is attached.
  • the storage vessel can be closed by a cover part.
  • the lid part has an area through which the capillary can be inserted into the storage vessel when the lid part is closed.
  • the document US 5 126 238 A shows a hollow fiber bioreactor system and a method for cell proliferation.
  • the use of a hollow fiber membrane or a bioreactor comprising a hollow fiber membrane is suggested as well as a circuit connected to a first inflow and a first outflow and having means for introducing fluids into the circulation, the reagents and Contain nutrient-containing basal medium.
  • the capillary cell comprises an outer shell of a light-permeable material in the visual spectrum (380 nm-800 nm) or near infrared spectrum (800 nm-1500 nm), preferably in the visual spectrum-transmissive region.
  • suitable materials are glass, in particular borosilicate glass, quartz glass, optical glass or special optical glass, and plastics such as fluorinated ethylene propylene (FEP), polymethyl methacrylate (PMMA), polystyrene (PS), cellulose acetate butyrate (CBS).
  • FEP fluorinated ethylene propylene
  • PMMA polymethyl methacrylate
  • PS polystyrene
  • CBS cellulose acetate butyrate
  • the shell comprises at least a side wall and a lower lid.
  • the envelope also has an inflow and an outflow by means of which a nutrient solution can be flushed through the inflow into the envelope and drained off through the outflow.
  • the capillary cell has a support device for a sample to be examined, which is connected to the shell.
  • the support device divides the sheath into a lower chamber extending between the lower lid and the receptacle and an upper part extending between an upper boundary of the sheath and the support device.
  • the support device is designed such that it comprises a permeable structure.
  • the support device is preferably attached to the side wall and allows no further gap between the side wall and the support device. In this way, for example, in the lower one Chamber filled nutrient solution, which is circulated for example by the outflow and the inflow, only pass through the permeable structure of the support device in the upper part of the capillary cell.
  • the support device is suitable for carrying biological samples on top, i. the upper part of the Kapillarzelle facing side of the support device to be arranged.
  • the sample is in a fixed location and can thus be made available for long-term observations.
  • nutrient solution flushed into the lower chamber can be absorbed by the permeable structure of the sample to be examined.
  • the sample to be examined can pull the nutrient solution upwards solely due to the acting capillary forces through the permeable structure.
  • a support device with a permeable structure for example, a filter, for example, a ceramic filter with a pore size of 2 ⁇ .
  • a nutriprimed lower chamber of the capillary cell wets the lower side of the support and the permeable structure, which is so moistened with the nutrient solution and the nutrient solution is sucked through the filter to the top of the support due to osmotic forces. Since the sample to be examined is placed on the upper side of the support device, it absorbs the nutrient solution and is thus either kept alive for long-term observation or ensures the biological functionality of the sample to be examined.
  • the lower cap of the capillary cell may for example be integrally connected to the side wall.
  • the capillary cell has an upper lid, which thus closes the upper part of the capillary cell to an upper chamber.
  • this cover may be integrally connected to the side wall.
  • the top lid is detachably connected to the sample.
  • the sample to be examined in a simple manner can be stored on the support device.
  • the support device is mechanically stable enough to carry the weight of the sample to be examined. Further embodiments can be found in the subordinate claims.
  • the inflow and outflow are directed into the lower chamber so that the lower part of the sheath is a flow chamber.
  • the entire lower chamber may be flushed with a nutrient solution, such as agarose.
  • a nutrient solution such as agarose.
  • openings in the lower lid or openings in the side wall, which limits the lower chamber are introduced.
  • the outflow and the inflow can be equipped with hoses, which enable a simpler connection of the micropump to the inflow and outflow.
  • Such hoses may for example consist of plastics such as polyvinyl chloride (PVC), polyurethanes (PUR), fluorinated ethylene propylene (FEP) or silicone or glass.
  • PVC polyvinyl chloride
  • PUR polyurethanes
  • FEP fluorinated ethylene propylene
  • silicone or glass plastics such as polyvinyl chloride (PVC), polyurethanes (PUR), fluorinated ethylene propylene (FEP) or silicone or glass.
  • PVC polyvinyl chloride
  • PUR polyurethanes
  • FEP fluorinated ethylene propylene
  • silicone or glass such as silicone or glass.
  • the inflow is passed through the lower lid and connected to another capillary. That capillary is inside the capillary cell shell or inside the sidewall and carries the nutrient solution.
  • the capillary preferably protrudes through the support device and extends to the upper limit of the shell or up to an upper lid on which the drain is arranged
  • the further capillary can be interrupted so that a nutrient solution passing through the further capillary can moisten the support device or the permeable structure or the permeable structure can absorb the nutrient solution.
  • the further capillary is formed with a semipermeable membrane, wherein the semipermeable membrane adjoins the lower chamber, so that a fluid exchange between the lower chamber and the further capillary can take place.
  • the further capillary also projects into the upper part and also includes here a semipermeable membrane.
  • the semipermeable membrane of the further capillary is located exclusively in the lower chamber or in the lower chamber and in the region of the support device.
  • the envelope comprises an upper lid which forms the upper boundary (even if this lid were, for example, still surmounted by the side wall).
  • This upper lid may have a gas inlet and a gas outlet system so that the upper chamber bounded by the upper lid and the support means is flushable with a gas.
  • the gas inlet system may have only a common inlet and outlet or have two separate inlet and outlet openings, which are respectively in the upper lid or in the side wall which defines the upper chamber, are introduced.
  • the permeable structure is a filter, in particular a ceramic filter or a microfilter. It is also possible to design the permeable structure in the form of a semipermeable membrane, for example a proton exchange membrane.
  • a typical pore size can be from ⁇ , ⁇ to ⁇ .
  • the capillary cell presented here can be used in various sizes. For example, the height of the capillary cell can be between 3 and 12 cm.
  • the diameter of a round capillary with a substantially circular side wall may be in the range between 500 ⁇ and 4 cm, preferably between 2 mm and 15 mm. For other geometries, an interval of the possible cross-sectional area of the capillary cell can be calculated from the above diameters.
  • the volume of the capillary cell may be, for example, 7 mm 3 to 20 cm 3 in the lower chamber.
  • the upper part can hold a similar volume.
  • the volume of the capillary cell, the lower chamber and the upper part depends inter alia on the diameter of the capillary cell.
  • the support device may have a volume of typically 0.1 mm 3 to 5 cm 3 , and / or the support device may have an area of typically 2 mm 2 to 5 cm 2 .
  • the capillary cell presented here is particularly suitable for use in a light sheet microscopy method.
  • FIG. 1A schematic representation of a capillary cell
  • FIG. 1B Use of such a capillary cell in a light-sheet microscope
  • FIG. 2 shows further embodiments of a capillary cell, which is shown in longitudinal section;
  • FIG. and Fig. 3 further embodiment of a capillary cell, which is also shown in longitudinal section.
  • FIG. 1A shows a capillary cell 1 with an outer shell 3, which is defined by a side wall 5 and a lower cover 7. Furthermore, a support device 9 is shown within the shell 3, which is designed as a permeable structure. Between the lower cover 7 and the supporting device 9 there is a lower chamber 11, wherein in the side wall 5, which delimits the lower chamber 11, an inflow 13 and an outflow 15 are present to supply the lower chamber 11 with a nutrient solution, For example, from agarose, to flow through and so to create a constant flow.
  • the ceramic filter has a disc with a diameter of 2.5 cm.
  • the ceramic filter is glued to the side wall.
  • the nutrient solution is sucked through the support device 9 from the lower side 17 to the upper side 19, and thus can be a sample to be examined located on the upper side 19 of the support device 9 provide with the nutrient solution.
  • the capillary cell 1 shown here is suitable, for example, to deposit a rat hippocampus on the surface 19 and to supply it with nutrients by means of a nutrient solution flushed into the lower chamber 11.
  • the hippocampus prepared in this way can be introduced together with the capillary cell 1 into a light-sheet microscope assembly and examined over a period of days or weeks in order to investigate the activity of the hippocampus in a long-term experiment. It will be explained with reference to FIG. 1B how an examination of the sample in the capillary cell can be carried out by means of light-sheet microscopy.
  • FIG. 1B shows the capillary cell 1 with a sample 30 arranged therein, which is arranged on the support device 9.
  • the light-sheet microscope 40 has a plurality of parts, only a few of which are explicitly shown.
  • a detection objective 42 can be seen and a matrix screen 44, on which the light waves (shown here schematically as reference symbol 46) impinge and, for example, are assembled into images by a computer unit.
  • the sample 30 is transilluminated out of the plane of the drawing with a light sheet 48.
  • This light sheet or the capillary with sample can be pushed back and forth along the direction z, so as to illuminate different levels of the sample 30 to be examined. In this way, different levels can be displayed on the matrix screen 44 and assembled into an overall image or a three-dimensional model of the sample to be examined.
  • the capillary cell 1 is located within a sample chamber 50, which is intended to prevent the objective 42 or the objective for the irradiation of the
  • Light sheets 48 are touched by the capillary cell 1 and thus could be damaged.
  • it can be connected, for example, at its upper end to a holder 60, so that a discharge into the sample chamber 50 is simply possible. If the sample to be examined or the lower chamber of the capillary cell is now flushed with a nutrient medium, the sample to be examined 30 can remain in the experimental setup for several days or even weeks, and images of different levels can be taken continuously.
  • a capillary cell 70 is shown in longitudinal section. Similar to the capillary cell 1 shown in FIG. 1A, the capillary cell of FIG. 2 may have a round, oval, quadrangular or rectangular cross-section.
  • the capillary cell has a sheath 72 having a sidewall 74 and a bottom lid 76 integral with the sidewall connected, and an upper lid 78 which is designed to be removable, so that the sample 80 can be placed on the support device 82.
  • the lower chamber 86 disposed between the lid 76 and the lower surface 84 of the support 82 has an inlet 88 and a drain 90.
  • the inlet 88 projects into the lower chamber 86 as a small capillary 92 that does not protrude over the entire length of the lower chamber 86 and supplies it with a nutrient medium needed for the sample 80.
  • This nutrient medium may be carried out through the drain 90, which also includes a small capillary 94. In this way, a flow chamber is created in the lower chamber 86.
  • the support device 82 is at the edge
  • the upper chamber 100 is connected to a gas purging system 104 via an inlet 102.
  • the gas of the gas purging system 104 is, for example, nitrogen or carbon dioxide or another gas or gas mixture, which requires numerous samples to be examined in order to be permanently functional or suitable for the experiments.
  • a separate outlet may be present in addition to the inlet.
  • the capillary cell 70 In order to access the capillary cell 70 for the light sheet microscopy or SPIM process, it is made of a borosilicate glass.
  • the support device with the solid region 96 and the semipermeable membrane 98 is manufactured in advance and subsequently introduced into the capillary cell 70. In this case, it is glued to the side wall 74. Thus, it is only possible via the semi-permeable membrane 98 to exchange media between the upper and lower chambers.
  • the capillary cell 110 has a shell 112 with a side wall 114 and a bottom cover 116 and an upper cover 118.
  • the upper cover 118 is designed to be detachable again, see FIG that a sample to be examined 120 can be stored on the support device 122.
  • the support device 122 shown here is for example a filter.
  • the jig 122 and the lower lid 116 define a lower chamber 124.
  • the upper lid 118 and the support surface 122 define an upper cell 126.
  • the capillary cell 110 also has a rinsing system for supplying the sample 120 to be examined with a nutrient solution.
  • the nutrient solution is passed through a capillary 128, whose inflow 130 is arranged on the lower lid 116 and whose outflow 132 is on the upper lid 118.
  • the further capillary 128 thus passes over the entire height of the capillary cell.
  • the further capillary 128 is also guided by the bearing surface 122. If a nutrient solution is now flushed into the further capillary 128 by the inflow 130, the nutrient solution can enter the lower chamber 124, since a delimiting wall 134 of the further capillary 128 is a semi-permeable membrane through which the nutrient solution passes, and thus gradually can fill lower chamber.
  • the semi-permeable membrane is located exclusively in the area of the support 122, ie, only along the small portion 136.
  • the further capillary 128 has solid, ie impermeable, walls in the upper chamber 126 which do not allow any nutrient fluid to escape into the upper chamber 126.
  • the top cover 118 also contains a gas inlet 102, which can be connected to a gas flow system 104. In this way, the sample 120 can also be rinsed with the gas.

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  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Optics & Photonics (AREA)
  • General Health & Medical Sciences (AREA)
  • Multimedia (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Computer Vision & Pattern Recognition (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

L'invention concerne une cellule capillaire (1, 70, 110), notamment destinée à une utilisation dans un microscope optique, notamment dans un microscope à feuille de lumière (40), la cellule capillaire comprenant une enveloppe externe (3, 72, 112) en un matériau transparent dans le spectre visible. L'enveloppe possède également une paroi latérale et un couvercle. L'enveloppe renferme un dispositif formant platine (9, 82, 122) qui comporte entre autre une structure transparente.
PCT/EP2018/069696 2017-07-31 2018-07-19 Cellule capillaire et utilisation de la cellule capillaire WO2019025205A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102017213138.1 2017-07-31
DE102017213138.1A DE102017213138B9 (de) 2017-07-31 2017-07-31 Kapillarzelle und Verwendung einer Kapillarzelle

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WO2019025205A1 true WO2019025205A1 (fr) 2019-02-07

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WO (1) WO2019025205A1 (fr)

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5126238A (en) 1990-02-15 1992-06-30 Unisyn Fibertec Corporation Hollow fiber cell propagation system and method
EP0515883A2 (fr) 1991-05-28 1992-12-02 Dade International Inc. Appareil pour prélever sans risque le sang d'un récipient
CA2684221A1 (fr) 2007-04-12 2008-10-23 Regents Of The University Of Minnesota Systemes et procedes d'analyse d'une particule
WO2009049740A1 (fr) * 2007-10-09 2009-04-23 Carl Zeiss Microlmaging Gmbh Procédés de positionnement d'échantillons biologiques dans un dispositif à microscope
EP2174715A1 (fr) * 2008-10-13 2010-04-14 Roche Diagnostics GmbH Pointe de pipette dotée d'un matériau de séparation
DE102012108158A1 (de) 2012-09-03 2014-03-06 Johann Wolfgang Goethe-Universität Kapillarzelle, Anordnung und Verfahren zur Aufnahme, zur Positionierung und zur Untersuchung einer mikroskopischen Probe
DE102014004851A1 (de) 2014-03-31 2015-10-01 Scienova Gmbh Vertikales funktionelles Reaktionsgefäß
US20160000128A1 (en) 2013-02-15 2016-01-07 Nestec S.A. Food composition and its use

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5126238A (en) 1990-02-15 1992-06-30 Unisyn Fibertec Corporation Hollow fiber cell propagation system and method
EP0515883A2 (fr) 1991-05-28 1992-12-02 Dade International Inc. Appareil pour prélever sans risque le sang d'un récipient
CA2684221A1 (fr) 2007-04-12 2008-10-23 Regents Of The University Of Minnesota Systemes et procedes d'analyse d'une particule
WO2009049740A1 (fr) * 2007-10-09 2009-04-23 Carl Zeiss Microlmaging Gmbh Procédés de positionnement d'échantillons biologiques dans un dispositif à microscope
EP2174715A1 (fr) * 2008-10-13 2010-04-14 Roche Diagnostics GmbH Pointe de pipette dotée d'un matériau de séparation
DE102012108158A1 (de) 2012-09-03 2014-03-06 Johann Wolfgang Goethe-Universität Kapillarzelle, Anordnung und Verfahren zur Aufnahme, zur Positionierung und zur Untersuchung einer mikroskopischen Probe
US20160000128A1 (en) 2013-02-15 2016-01-07 Nestec S.A. Food composition and its use
DE102014004851A1 (de) 2014-03-31 2015-10-01 Scienova Gmbh Vertikales funktionelles Reaktionsgefäß

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
ANNA KAUFMANN ET AL: "Multilayer mounting enables long-term imaging of zebrafish development in a light sheet microscope", DEVELOPMENT, 1 September 2012 (2012-09-01), pages 3242 - 3247, XP055509449, Retrieved from the Internet <URL:http://www.mbl.edu/zebrafish/files/2013/03/MultilayerMountingZebrafish_2012.pdf> [retrieved on 20180925], DOI: 10.1242/dev.082586 *

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DE102017213138B9 (de) 2019-06-27

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