WO2003029817A2 - Procede et dispositif pour manier et/ou manipuler de la matiere biologique, eventuellement pour extraire de la matiere cellulaire d'un prelevement tissulaire - Google Patents

Procede et dispositif pour manier et/ou manipuler de la matiere biologique, eventuellement pour extraire de la matiere cellulaire d'un prelevement tissulaire Download PDF

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Publication number
WO2003029817A2
WO2003029817A2 PCT/EP2002/010883 EP0210883W WO03029817A2 WO 2003029817 A2 WO2003029817 A2 WO 2003029817A2 EP 0210883 W EP0210883 W EP 0210883W WO 03029817 A2 WO03029817 A2 WO 03029817A2
Authority
WO
WIPO (PCT)
Prior art keywords
instrument
biological material
tissue sample
partial area
cell
Prior art date
Application number
PCT/EP2002/010883
Other languages
German (de)
English (en)
Other versions
WO2003029817A3 (fr
Inventor
Klaus Schaller
Original Assignee
Olympus Biosystems Gmbh
Schaller, Ines
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 Olympus Biosystems Gmbh, Schaller, Ines filed Critical Olympus Biosystems Gmbh
Priority to AU2002362528A priority Critical patent/AU2002362528A1/en
Publication of WO2003029817A2 publication Critical patent/WO2003029817A2/fr
Publication of WO2003029817A3 publication Critical patent/WO2003029817A3/fr

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/42—Low-temperature sample treatment, e.g. cryofixation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
    • G01N2001/2833—Collecting samples on a sticky, tacky, adhesive surface
    • G01N2001/284—Collecting samples on a sticky, tacky, adhesive surface using local activation of adhesive, i.e. Laser Capture Microdissection

Definitions

  • the invention relates generally to a method and a device for handling or / and manipulating biological material, in particular at least one biological cell and / or a cell assembly or / and parts of cells.
  • the invention also relates specifically to a method and an apparatus for extracting cell material from a tissue sample in order to extract the extracted cell material e.g. to be placed in receptacles and to be subjected to further tests there.
  • individual biological objects for example cells or cell assemblies
  • individual biological objects can be handled and manipulated and, if desired, extracted from a tissue sample, or the individual cells or cell assemblies or cell parts can be selected and manipulated, handled and, if desired, sorted out from a large number of similar biological objects.
  • the biological objects are generally arranged on a flat support, usually a glass plate, in large numbers, for example 10 9 , and they can also be present on the support in a solution or in a particularly liquid surrounding medium. If, for example, individual cells or cell assemblies are to be isolated from a tissue sample, it is first necessary to separate the selected areas from the surrounding tissue sample.
  • this is done mechanically, for example by cutting out or cutting off the cells from the surrounding tissue sample, for example with a scalpel-like device.
  • the mechanical separation can also take place in a punching process.
  • a high-energy laser beam focused on the plane of the tissue sample can be used to separate the desired cells, the beam guidance of which is directed with the aid of lenses or mirrors in such a way that the desired cells are cut out.
  • Automatic image acquisition and processing systems are used to select the desired area, which contains the cells or cell assemblies or cell parts to be examined and possibly extracted, with which it is possible to select the areas to be examined automatically.
  • the image processing systems are connected to the microscope on which the carrier with the tissue sample is arranged, so that an enlarged image of the tissue sample is fed to the image processing system.
  • EP 0 879 408 B1 discloses a method for sorting and obtaining biological objects, in which the biological objects are freely prepared with the aid of a laser beam, and then the biological object to be examined with a further laser shot from the carrier a collecting device, for example a collecting vessel, is catapulted away.
  • WO 97/1 3838 Another method for isolating cell material is known from WO 97/1 3838, in which the cell material can be brought into contact with a selectively activatable surface, for example a sample probe. After the cell material has been separated from the surrounding tissue sample, a holding force is exerted on the cell material by activating the above-mentioned surface, so that it adheres to the activatable surface and can be spatially transported, for example, to a collecting vessel.
  • activatable is understood to mean, inter alia, that the cells are due to chemical adhesive properties adhere to the activatable surface due to electrostatic or electromagnetic interactions or due to the action of heat on the activatable surface.
  • this object is achieved with reference to claim 1 by a method for handling or / and manipulating biological material, in particular at least one biological cell and / or a cell assembly or / and at least one cell part, with the steps: providing biological material, for example a tissue sample, on a support; indirect or direct contacting of the biological material or a sub-area thereof with a section of a movable hand or manipulation instrument; Activation of the instrument in order to generate a holding force between the holding section on the one hand and the biological material or partial area on the other hand; Moving the instrument to manipulate and / or manipulate the biological material or the portion thereof; it is provided according to the invention that the instrument is activated by active cooling of at least the holding section.
  • biological material for example a tissue sample
  • the invention provides an apparatus for solving the problem, in particular for carrying out the method according to the invention or its further developments, comprising at least one, in particular by means of, movable relative to a carrier At least one positioning arrangement that can be moved relative to the carrier handling and / or manipulation instrument with a holding section that can be brought into direct or indirect contact with biological material arranged on the carrier, in particular at least one biological cell and / or a cell assembly and / or at least one cell part , whereby a holding force acting on the biological material or a partial area thereof can be generated on the holding section, whereby it is provided according to the invention that at least the holding section of the instrument can be cooled by means of at least one cooling device assigned or associated with the instrument in order to prevent the biological material or of the partial area thereof and / or to generate a holding force based on cooling of an adhesive medium acting between the holding section and the biological material or the partial area thereof.
  • Tissue sample comprising the steps of: providing a tissue sample on a support; Selection of an area of a tissue sample from the
  • Sampling device is carried out by cooling, as well as a device for extracting cell material from a tissue sample, with a sampling device which can be brought into contact with a selected area of the tissue sample and activated in order to To generate holding force between the sampling device and the selected area, wherein the sampling device is movable, in particular spatially movable to remove the selected area, which is characterized in that the sampling device is provided with at least one device or devices for cooling.
  • the main idea of the invention is that a device known per se for handling or manipulating or for extracting individual cells from a tissue sample is used, the instrument, hereinafter referred to as a sampling device without restriction of generality, which detaches the separated or free prepares the cells to be examined, for example, takes them to collecting vessels for further examination, is provided with a device for cooling.
  • a sampling device which is preferably designed as a pipette
  • an adhesive force is exerted on the cell material.
  • the sampling device is cooled until an adhesive medium changes its physical state from liquid to solid, ie freezes.
  • adhesive medium is understood to mean either the cell material itself, i. H .
  • the cooling takes place from usually room temperature below the freezing point or solidification point or other relevant state change point of the adhesive medium, in the case of a water-based adhesive medium, approximately one aqueous solution, so under O 'C. For example, by cooling down to a sb - 1 0 * C, if necessary. even on sth a - 20 * C.
  • the sampling device is cooled until the cells or cell parts or the solution in which the cells are arranged on the carrier freeze to the sampling device. This can either be done by slowly cooling the sampling device, for example the tip of the pipette, or preferably in a shock-like manner, in order to freeze in particular live cells of a tissue sample in a shock-like manner and not damage them as a result.
  • a liquid drop in particular an aqueous solution suitable for cell physiology, is provided at the tip of the sampling device or on the holding section of the instrument, which - after being brought into contact with the surface of the cell or cell group to be extracted - by cooling the sampling device or the instrument or at least the holding section of the same is frozen in order to be able to handle, if necessary, extract the desired cells or cell parts.
  • a micropipette known per se can be used, in which, for example, the aqueous solution can be sucked in or ejected with the aid of a pump device.
  • the pumping device pumps until the desired drop forms at the tip of the pipette.
  • a Peltier element In order to cool the sampling device, and in particular its tip or the holding section, which is brought into contact with the cells, it is proposed that a Peltier element be used.
  • the Peltier element is characterized in that, depending on the direction of current flow, it either generates heat or a cooling effect entry.
  • the Peltier element can be arranged, for example, in the area of the tip of a metallic sample probe.
  • For cooling current flows through the element in one direction, and after the cells have been brought into the desired position, the current direction is reversed in order to accelerate the thawing of, for example, the drop of liquid, for example in order to deposit the cells in collecting vessels.
  • a controller is preferably provided, by means of which the cooling / thawing can be controlled, in order not to damage the cells to be examined by heat input, in particular when thawing.
  • a cooling fluid for example liquid nitrogen
  • a cooling fluid for example liquid nitrogen
  • the sampling device or the instrument such as a probe or pipette, is made of metal.
  • the sampling device can be provided with a through-channel arranged in its interior, through which the cooling fluid flows, in order to cool the sampling device.
  • individual cells or cell groups arranged in cavities are not to be examined next to one another, e.g. B. be selected with an automatic image processing system, but a single tissue sample, it is necessary to prepare the cells to be extracted with a separator from the surrounding tissue sample. This is preferably done with a cutting laser that can be focused and controlled on the tissue sample.
  • both the carrier and the sampling device are equipped with a three-dimensional positioning system, on the one hand to move the carrier in all three spatial directions into the desired position, for example in the focus of a laser and on the other hand to move the tip of the sampling device to a selected position on the carrier.
  • a positioning system can be carried out in a simple manner with slide-like bearings known to the person skilled in the art.
  • the control of the positioning systems is preferably coupled to the electronic image data processing in order to automatically select and extract the cells or cell groups to be examined.
  • Either an upright microscope or an inverse microscope can be used as the microscope, with an inverse microscope the actual microscope being arranged below the carrier and the sampling device being arranged above the carrier on which the tissue sample is provided.
  • a simple construction of the entire device can be achieved with an inverted microscope.
  • the device can also, for. B. in the manner of a turret interchangeable head, be provided with further devices for exerting a holding force on the cell material to be extracted. These can be either electrostatic or electromagnetic holding forces, with another sampling device and e.g. B. the carrier a potential difference or an electromagnetic field is generated. Furthermore, a pipette can be used, which is provided with a pump device in order to suck the cells to be extracted into the pipette and to bring them to the collecting vessel.
  • Fig. 1 shows an example of a device according to the invention in a schematic representation.
  • Fig. 1 shown embodiment of the invention is a microscope 1, on which a carrier 2, preferably a glass plate, is arranged.
  • the microscope has an optical focusing element 3 with which light beams can be focused on the object 4 arranged on the carrier 2.
  • an optical combining / separating element 5 preferably a dichroic mirror, with which, for example, the rays coming from the cutting laser 6 are deflected and directed through the focusing element 3 onto the object 4.
  • an image processing system 7 arranged below the union / separating element 5 is provided, which automatically examines the object 4 or the individual cells arranged on the carrier 2 and the desired ones to be examined. Selects cells or cell groups.
  • the carrier 2 is mounted displaceably in all three spatial directions X, Y, Z with a positioning system known to the person skilled in the art, which is not shown here to simplify the drawing.
  • the cells or the cell material to be examined are freed from the surrounding tissue sample if individual cells or cell groups from a tissue sample are to be examined, here either the beam guidance of the laser 6 being influenced as desired or the carrier 2 being displaced. A cutting line 8 of the desired shape is thereby obtained.
  • a sampling device here in the form of a pipette 9, which can also be referred to as an instrument, for collecting vessels 10
  • a drop 11 of an aqueous solution is provided at the tip of the pipette 9, for example with the aid of a pump device connected to the pipette 9, and the cut-out cell material adheres to the surface of the solution by means of adhesive forces.
  • a cooling element 12 is also arranged on the pipette 9, with the aid of which the tip of the pipette 9 and the drop 11 arranged thereon can be frozen with the cell material.
  • the cooling device 1 2 can advantageously have a Peltier element as a cooling element, so that by reversing the direction of current flow, the cooling element 1 2 can also be used as a heating element for accelerating thawing of the drop at the location of the collecting vessel 10.
  • the cooling element 1 2 can, however, also be a tube through which liquid nitrogen flows, which rotates around the pipette 9 or is arranged therein.
  • the instrument 9, here the pipette 9 - as indicated here by the arrows - can be moved in all spatial directions X, Y, Z with a positioning system 1 3, preferably via a slide bearing, around the pipette 9 to one of the collecting vessels 10 to spend where the cell material can be deposited by thawing the frozen drop.

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  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Sampling And Sample Adjustment (AREA)
  • Microscoopes, Condenser (AREA)

Abstract

L'invention concerne un procédé et un dispositif permettant d'extraire de la matière cellulaire d'un prélèvement tissulaire, la matière cellulaire extraite étant ensuite placée p. ex. dans des récipients collecteurs en vue d'analyses ultérieures. Selon l'invention, la matière à extraire est mise en contact avec une sonde de prélèvement et une force d'adhérence est exercée sur la matière cellulaire par refroidissement de cette sonde de prélèvement.
PCT/EP2002/010883 2001-09-28 2002-09-27 Procede et dispositif pour manier et/ou manipuler de la matiere biologique, eventuellement pour extraire de la matiere cellulaire d'un prelevement tissulaire WO2003029817A2 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2002362528A AU2002362528A1 (en) 2001-09-28 2002-09-27 Method and device for handling or/and manipulation of biological material possibly for the extraction of cell material from a tissue sample

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE10147950.6 2001-09-28
DE2001147950 DE10147950C2 (de) 2001-09-28 2001-09-28 Verfahren und Vorrichtung zur Extraktion von Zellmaterial aus einer Gewebeprobe

Publications (2)

Publication Number Publication Date
WO2003029817A2 true WO2003029817A2 (fr) 2003-04-10
WO2003029817A3 WO2003029817A3 (fr) 2003-12-24

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PCT/EP2002/010883 WO2003029817A2 (fr) 2001-09-28 2002-09-27 Procede et dispositif pour manier et/ou manipuler de la matiere biologique, eventuellement pour extraire de la matiere cellulaire d'un prelevement tissulaire

Country Status (3)

Country Link
AU (1) AU2002362528A1 (fr)
DE (1) DE10147950C2 (fr)
WO (1) WO2003029817A2 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005093391A1 (fr) * 2004-03-29 2005-10-06 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Procede pour exciter des molecules d'un premier etat a un second etat avec un signal optique
WO2006131260A3 (fr) * 2005-06-08 2007-04-19 Palm Microlaser Tech Gmbh Procede et dispositif de manipulation d'objets
US20130109047A1 (en) * 2010-07-08 2013-05-02 Biomerieux S.A. Method of sampling and/or depositing a sample of biological matter and device implementing such a method

Citations (7)

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Publication number Priority date Publication date Assignee Title
DE19616216A1 (de) * 1996-04-23 1997-10-30 P A L M Gmbh Verfahren und Vorrichtung zur Gewinnung von laserdissektierten Partikeln wie biologische Zellen bzw. Zellorganellen, Chromosomenteilchen etc.
WO1998003628A1 (fr) * 1996-07-19 1998-01-29 Bayer Aktiengesellschaft Dispositif pour separer des micro-objets
DE19804800A1 (de) * 1998-02-08 1999-08-12 Malte Dr Med Boehm Verfahren zur automatisierten Bergung planar ausgebrachter Objekte vom Objekttisch und zu deren Transfer in nachgeordnete Reaktonsträger
WO2000034757A1 (fr) * 1998-12-10 2000-06-15 The Government Of The Unites States Of America Represented By The Secretary, Department Of Health And Human Services Conceptions de microdissection par piegeage laser sans contact
EP1067374A2 (fr) * 1999-07-09 2001-01-10 Eppendorf Ag Dispositif pour la microdissection d'un tissu
US6251467B1 (en) * 1994-03-01 2001-06-26 The United States Of America As Represented By The Department Of Health And Human Services Isolation of cellular material under microscopic visualization
DE19964054A1 (de) * 1999-12-30 2001-07-05 Trace Biotech Ag Vorrichtung zum Übertragen kleiner Flüssigkeitsmengen

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5843657A (en) * 1994-03-01 1998-12-01 The United States Of America As Represented By The Department Of Health And Human Services Isolation of cellular material under microscopic visualization
WO1997029354A1 (fr) * 1996-02-05 1997-08-14 Bayer Aktiengesellschaft Procede et dispositif pour trier et recuperer des objets biologiques deposes sur un support plat, tels que des cellules biologiques ou des organites cellulaires, des coupes histologiques, des particules de chromosomes etc., au moyen de faisceaux laser
US6103518A (en) * 1999-03-05 2000-08-15 Beecher Instruments Instrument for constructing tissue arrays

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6251467B1 (en) * 1994-03-01 2001-06-26 The United States Of America As Represented By The Department Of Health And Human Services Isolation of cellular material under microscopic visualization
DE19616216A1 (de) * 1996-04-23 1997-10-30 P A L M Gmbh Verfahren und Vorrichtung zur Gewinnung von laserdissektierten Partikeln wie biologische Zellen bzw. Zellorganellen, Chromosomenteilchen etc.
WO1998003628A1 (fr) * 1996-07-19 1998-01-29 Bayer Aktiengesellschaft Dispositif pour separer des micro-objets
DE19804800A1 (de) * 1998-02-08 1999-08-12 Malte Dr Med Boehm Verfahren zur automatisierten Bergung planar ausgebrachter Objekte vom Objekttisch und zu deren Transfer in nachgeordnete Reaktonsträger
WO2000034757A1 (fr) * 1998-12-10 2000-06-15 The Government Of The Unites States Of America Represented By The Secretary, Department Of Health And Human Services Conceptions de microdissection par piegeage laser sans contact
EP1067374A2 (fr) * 1999-07-09 2001-01-10 Eppendorf Ag Dispositif pour la microdissection d'un tissu
DE19964054A1 (de) * 1999-12-30 2001-07-05 Trace Biotech Ag Vorrichtung zum Übertragen kleiner Flüssigkeitsmengen

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005093391A1 (fr) * 2004-03-29 2005-10-06 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Procede pour exciter des molecules d'un premier etat a un second etat avec un signal optique
US7224452B2 (en) 2004-03-29 2007-05-29 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Method of exciting molecules out of a first state into a second states using an optical signal
WO2006131260A3 (fr) * 2005-06-08 2007-04-19 Palm Microlaser Tech Gmbh Procede et dispositif de manipulation d'objets
US7923679B2 (en) 2005-06-08 2011-04-12 Carl Zeiss Microimaging Gmbh Method and device for handling objects
US20130109047A1 (en) * 2010-07-08 2013-05-02 Biomerieux S.A. Method of sampling and/or depositing a sample of biological matter and device implementing such a method
JP2013531505A (ja) * 2010-07-08 2013-08-08 ビオメリュー 生物学的物質のサンプルのサンプリング及び／又は沈着方法及び該方法を実施する装置
US9650662B2 (en) * 2010-07-08 2017-05-16 bio Meriéux, S.A. Method of sampling and/or depositing a sample of biological matter and device implementing such a method
US10144948B2 (en) 2010-07-08 2018-12-04 Biomerieux Method of sampling and/or depositing a sample of biological matter and device implementing such method

Also Published As

Publication number Publication date
WO2003029817A3 (fr) 2003-12-24
DE10147950C2 (de) 2003-12-04
AU2002362528A1 (en) 2003-04-14
DE10147950A1 (de) 2003-04-24

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