EP1768782B1 - Dispositif de refroidissement pour echantillons biologiques - Google Patents

Dispositif de refroidissement pour echantillons biologiques Download PDF

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Publication number
EP1768782B1
EP1768782B1 EP05716364A EP05716364A EP1768782B1 EP 1768782 B1 EP1768782 B1 EP 1768782B1 EP 05716364 A EP05716364 A EP 05716364A EP 05716364 A EP05716364 A EP 05716364A EP 1768782 B1 EP1768782 B1 EP 1768782B1
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EP
European Patent Office
Prior art keywords
coolant
cooling device
cooling
wall
chamber
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.)
Active
Application number
EP05716364A
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German (de)
English (en)
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EP1768782A1 (fr
Inventor
Young-Joo Oh
Günter FUHR
Heiko c/o IBMT ZIMMERMANN
Uwe Schön
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Fraunhofer Gesellschaft zur Forderung der Angewandten Forschung eV
Original Assignee
Fraunhofer Gesellschaft zur Forderung der Angewandten Forschung eV
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Priority to PL05716364T priority Critical patent/PL1768782T3/pl
Publication of EP1768782A1 publication Critical patent/EP1768782A1/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
    • B01L1/00—Enclosures; Chambers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/02—Water baths; Sand baths; Air baths
    • F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
    • F25—REFRIGERATION OR COOLING; COMBINED HEATING AND REFRIGERATION SYSTEMS; HEAT PUMP SYSTEMS; MANUFACTURE OR STORAGE OF ICE; LIQUEFACTION SOLIDIFICATION OF GASES
    • F25D—REFRIGERATORS; COLD ROOMS; ICE-BOXES; COOLING OR FREEZING APPARATUS NOT OTHERWISE PROVIDED FOR
    • F25D29/00—Arrangement or mounting of control or safety devices
    • F25D29/001—Arrangement or mounting of control or safety devices for cryogenic fluid systems
    • F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
    • F25—REFRIGERATION OR COOLING; COMBINED HEATING AND REFRIGERATION SYSTEMS; HEAT PUMP SYSTEMS; MANUFACTURE OR STORAGE OF ICE; LIQUEFACTION SOLIDIFICATION OF GASES
    • F25D—REFRIGERATORS; COLD ROOMS; ICE-BOXES; COOLING OR FREEZING APPARATUS NOT OTHERWISE PROVIDED FOR
    • F25D3/00—Devices using other cold materials; Devices using cold-storage bodies
    • F25D3/10—Devices using other cold materials; Devices using cold-storage bodies using liquefied gases, e.g. liquid air
    • F25D3/102—Stationary cabinets
    • 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
    • B01L2200/14—Process control and prevention of errors
    • B01L2200/143—Quality control, feedback systems
    • B01L2200/147—Employing temperature sensors
    • 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
    • B01L2300/18—Means for temperature control
    • B01L2300/1838—Means for temperature control using fluid heat transfer medium
    • 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
    • B01L2300/18—Means for temperature control
    • B01L2300/1838—Means for temperature control using fluid heat transfer medium
    • B01L2300/1844—Means for temperature control using fluid heat transfer medium using fans
    • 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
    • B01L2300/18—Means for temperature control
    • B01L2300/1894—Cooling means; Cryo cooling

Definitions

  • the invention relates to a cooling device for biological samples, in particular for the investigation, manipulation and processing of cryoprobes, according to the preamble of claim 1.
  • cryoprobes wherein the vitality-preserving storage of such cryoprobes takes place in so-called cryotanks, in which liquid nitrogen is.
  • a cooling device which may consist for example of a tub at the bottom of which liquid nitrogen is, which is also referred to as nitrogen and slowly evaporates, so that in the tub located cryoprobe is still sufficiently cooled.
  • a transparent protective bell can be placed on the tub, which can be in the wall of the protective glove glove sleeves, via which an operator can manipulate the cryoprobe located in the tub.
  • a disadvantage of the above-described known cooling device for manipulation, processing or examination of Cryo sampling is the unsatisfactory temperature stability and distribution within the tub.
  • a cooling device comprising a cooling space having an outer wall and a metal-lattice inner wall, wherein liquid nitrogen is passed as a cooling medium in the space between the outer wall and the inner wall to cool samples.
  • a comprehensive temperature control is disclosed, inter alia, with a heater with fans and passages for gas circulation.
  • the invention is therefore based on the object to improve the initially described cooling device accordingly.
  • a further aim may be to improve the temperature stability in the cooling device, to allow the adjustment of the temperature, to optimize the temperature distribution within the cooling device, to avoid fogging of the protective bell and to minimize the moisture in the cooling space.
  • the invention is based on the technical knowledge that the unsatisfactory temperature stability and distribution within the cooling trough in the known cooling device is caused by the fact that the outgassing of the nitrogen from nitrogen within the cooling diffuses undefined, so that the desired working temperature is difficult to achieve and can hardly be regulated.
  • the uncontrolled outgassing of nitrogen from the nitrogen lake also leads to the fogging of the attached protective bell.
  • the invention therefore comprises the general technical teaching to avoid a nitrogen lake in the cooling device and instead to introduce gaseous nitrogen in a controlled manner into the cooling space.
  • the cooling device has for receiving refrigerated goods to a cooling space which is bounded by an inner wall and an outer wall, wherein there is a gap between the inner wall and the outer wall, in which a coolant supply line opens.
  • the coolant eg liquid nitrogen
  • the coolant is thus not introduced directly into the cooling space, but into the space between the inner wall and the outer wall of the cooling space, wherein the inner wall is permeable to the coolant, so that the coolant from the space between the outer wall and the inner wall enters through the inner wall into the cooling space.
  • a buffer material is arranged in the intermediate space between the inner wall and the outer wall of the cooling space, which temporarily absorbs the coolant introduced into the intermediate space and discharges it continuously through the inner wall into the cooling space.
  • the buffer material is therefore porous, for example, to be able to buffer liquid nitrogen.
  • the outer wall of the cooling space is preferably impermeable to the coolant, in contrast to the inner wall of the cooling space, in order to prevent leakage of the coolant outwards into the environment.
  • the outer wall preferably thermally insulating, in order to avoid a cooling of the environment or a heating of the cooling device.
  • the inner wall of the cooling space preferably consists of a thermally conductive material, such as metal, in order to improve the heat transfer from the inner cooling space to the coolant in the clearance.
  • the material of the inner wall not only has a good thermal conductivity, but also has a high specific heat capacity, so that the inner wall with its heat capacity as a thermal buffer counteracts undesirable temperature fluctuations.
  • the inner wall is substantially grid-shaped, so that the coolant located in the intermediate space can outgas largely unhindered in the refrigerator.
  • the cooling chamber is trough-shaped and has a peripheral edge on its upper side
  • the coolant supply line preferably has a coolant distributor which extends along the peripheral edge of the cooling space and distributes the coolant over its length into the intermediate space between the cooling space Interior wall and the outer wall of the refrigerator introduces.
  • the coolant is thus introduced evenly into the intermediate space between the inner wall and the outer wall of the cooling space, which advantageously leads to a uniform temperature distribution in the cooling space, since the cooling space is uniformly cooled from all sides.
  • a heating element is arranged in the cooling space in order to heat the cooling space or to thaw the refrigerated goods located in the cooling space.
  • this heating element is arranged under or in a heating plate, wherein the heating plate preferably has a plurality of passages, which allow gas circulation.
  • the cooling device according to the invention it is also possible with the cooling device according to the invention to place a removable protective bell on the cooling space in order to avoid the penetration of moisture into the cooling space.
  • this protective bell is at least partially transparent in order to allow a visual inspection of the refrigerated goods located in the refrigerator.
  • the protective bell has a sample lock, through which the goods to be cooled can be introduced into the cooling space or removed from the cooling space, wherein the sample lock largely prevents heat exchange with the environment.
  • a cold gas outlet can be arranged on the underside of the protective bell and / or on the upper side of the cooling space, via which coolant or cold gas can escape from the cooling space.
  • This cold gas outlet causes a large temperature gradient in the height of the cold gas outlet, wherein the temperature above the cold gas outlet is substantially higher than below the cold gas outlet. In this way, a fogging of the protective bell is advantageously prevented.
  • the cooling device preferably has a in the Refrigerator arranged temperature sensor to measure the temperature in the cold room or to regulate.
  • a controllable coolant valve is provided which adjusts the amount of coolant supplied or the coolant flow.
  • the actual temperature control is then performed by a temperature controller, which is connected on the input side to the temperature sensor and the output side controls the coolant valve according to a predetermined temperature setpoint.
  • the control of the coolant valve by the temperature controller can in this case take place via a clock generator which alternately opens and closes the coolant valve, wherein the opening and closing times of the coolant valve are predetermined by the clock and set by the temperature controller.
  • the coolant is thus supplied discontinuously by the coolant valve opens and closes alternately.
  • the temperature sensor for detecting the temperature in the cooling space is arranged here at the processing position of the cooling space in order to measure or regulate the optimum processing temperature in the cooling space.
  • the temperature controller therefore preferably regulates the temperature in the cold room so that no coolant lake forms at the bottom of the cold room.
  • the coolant is preferably liquid nitrogen, but the invention is not limited to nitrogen as a coolant, but can also be realized with other liquid or gaseous coolants which flow into the space between the inner wall and the outer wall of the refrigerator can be initiated.
  • the cooling device according to the invention can be used for various temperature ranges, such as at temperatures of about -150 ° C, -130 ° C, -80 ° C, -40 ° C, + 4 ° C or + 37 ° C, the above-mentioned Temperature ranges may include, for example, a range of ⁇ 10 ° C, ⁇ 5 ° C or ⁇ 2 ° C.
  • a temperature of 37 ° C is advantageous because the growth temperature of biological cells is then optimal.
  • a temperature of + 4 ° C offers the advantage that the physiological processes in the cells are slowed down.
  • cell damage is less (e.g., with tropsia and DMSO).
  • the invention encompasses not only the above-described cooling device as a device, but also the use of such a cooling device for the examination, processing and / or manipulation of a cryoprobe.
  • cooling device 1 is used for temperature control of a cold room for receiving cryoprobes in an investigation, manipulation and / or processing.
  • the cooling device 1 a cryogenic sump 2 with a trough-shaped, open-topped cooling chamber 3, wherein the cryogenic sump 2, a removable protective cover 4 is placed, which prevents the ingress of moisture from the environment into the refrigerator and in detail FIG. 2 is shown.
  • the protective bell 4 has to introduce the cryoprobes into the cooling chamber 3 and to remove the cryoprobes from the cooling chamber 3 to a sample lock 5, which is attached to the side of the protective bell 4 and during insertion of the cryoprobes or when removing the cryoprobes heat exchange with the Environment largely prevented and minimizes the moisture in the refrigerator 3.
  • the protective bell 4 has on its upper side a lamp 6 in order to illuminate the cooling space 3 and thereby facilitate the manipulation of the cryoprobes located in the cooling space 3.
  • the protective bell 4 itself consists of a transparent material, which allows a simple visual inspection by an operator.
  • cryotube 2 At the top of the cryotube 2 is located at the front on a control and display panel 10, where the temperature in the refrigerator 3 can be displayed and adjusted.
  • the cooling of the cooling space 3 takes place here by liquid nitrogen, which is supplied from a nitrogen tank (eg an Apollo tank) via a nitrogen line 11, wherein the nitrogen line 11 does not open directly into the cooling chamber 3 to the formation of a nitrogen lake at the bottom of the refrigerator 3 to avoid.
  • the nitrogen line 11 opens via an electrically controllable coolant valve 12 in a coolant supply line 13, wherein the coolant supply line 13 along the peripheral edge of the trough-shaped Refrigerator 3 extends and releases the liquid nitrogen distributed over the length.
  • the cooling space 3 is in this case bounded by a lattice-shaped inner wall 14 made of metal, which is enclosed by an outer wall 15, the inner wall 14 and the outer wall 15 enclosing a gap in which a buffer material 16 is arranged.
  • the coolant supply line 13 is arranged in the lateral direction between the inner wall 14 and the outer wall 15 above the buffer material 16 and has downwardly directed outlet openings, is discharged through the liquid nitrogen from the interior of the coolant supply line 13 into the buffer material 16.
  • the buffer material 16 absorbs the liquid nitrogen and releases it continuously through the grid-shaped inner wall 14 into the cooling space 3.
  • the coolant valve 12 operates in this case discontinuously by the coolant valve 12 either closes or opens.
  • the control of the coolant valve 12 takes place here by a clock generator 17, wherein the opening time T OPEN and the closing time T ZU for the coolant valve 12 are set by a controller 18 to meter the coolant.
  • the control takes place here as a function of the temperature in the cooling space 3, which is measured by a temperature sensor 19, wherein the temperature sensor 19 is arranged at the processing position of the cooling space 3.
  • the temperature sensor 19 measures a temperature T IST and supplies this to a subtractor 20 which, as a further input variable, sets a target value T SOLL for the temperature in receives the cooling chamber 3 and calculates a target-actual deviation .DELTA.T.
  • the controller 18 sets the opening time T AUF and the closing time T ZU for the coolant valve 12 such that the desired temperature (for example -130 ° C.) prevails in the cooling space 3 without forming a nitrogen lake at the bottom of the cooling space 3.
  • the desired temperature for example -130 ° C.
  • a heating plate 21 is arranged on the bottom of the cooling space 3, which enables heating of the cryoprobe and the cooling space 3.

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  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Combustion & Propulsion (AREA)
  • Physics & Mathematics (AREA)
  • Mechanical Engineering (AREA)
  • Thermal Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Sampling And Sample Adjustment (AREA)

Claims (17)

  1. Dispositif de refroidissement (1) servant à refroidir un produit à refroidir, en particulier pour l'analyse, la manipulation et/ou le traitement d'échantillons cryogéniques, comprenant :
    - un espace de refroidissement (3) destiné à recevoir le produit à refroidir,
    - une paroi intérieure (14) délimitant l'espace de refroidissement (3), la paroi intérieure (14) étant perméable au fluide de refroidissement,
    - une paroi extérieure (15),
    - un espace intermédiaire entre la paroi extérieure (15) et la paroi intérieure (14) ainsi
    - qu'une conduite d'amenée de fluide de refroidissement (11, 13) servant à introduire un fluide de refroidissement, la conduite d'amenée de fluide de refroidissement (11, 13) débouchant dans l'espace intermédiaire entre la paroi intérieure (14) et la paroi extérieure (15), et alimentant le fluide de refroidissement dans l'espace intermédiaire,
    caractérisé en ce que
    - dans l'espace intermédiaire est disposé un matériau tampon (16), lequel absorbe temporairement le fluide de refroidissement alimenté dans l'espace intermédiaire, et le transfère continuellement dans l'espace de refroidissement (3) en le faisant passer à travers la paroi intérieure (14) et en ce que
    - le matériau tampon (16) est poreux.
  2. Dispositif de refroidissement (1) selon la revendication 1, caractérisé en ce que la paroi intérieure (14) a sensiblement la forme d'une grille.
  3. Dispositif de refroidissement (1) selon l'une quelconque des revendications précédentes, caractérisé en ce que la paroi intérieure (14) se compose d'un matériau thermiquement conducteur.
  4. Dispositif de refroidissement (1) selon la revendication 3, caractérisé en ce que la paroi intérieure (14) est essentiellement en métal.
  5. Dispositif de refroidissement (1) selon l'une quelconque des revendications précédentes, caractérisé en ce que l'espace de refroidissement (3) a la forme d'une cuvette et comporte sur sa face supérieure un bord périphérique.
  6. Dispositif de refroidissement (1) selon la revendication 5, caractérisé en ce que la conduite d'amenée de fluide de refroidissement (11, 13) comprend un distributeur de fluide de refroidissement (13), lequel s'étend le long du bord périphérique de l'espace de refroidissement (3) et introduit le fluide de refroidissement, réparti sur sa longueur, dans l'espace intermédiaire.
  7. Dispositif de refroidissement (1) selon l'une quelconque des revendications précédentes, caractérisé en ce qu'un élément chauffant (21) est disposé dans l'espace de refroidissement (3).
  8. Dispositif de refroidissement (1) selon la revendication 7, caractérisé en ce que l'élément chauffant est placé sous une plaque chauffante (21), la plaque chauffante (21) comportant plusieurs passages (22) qui permettent une circulation de gaz.
  9. Dispositif de refroidissement (1) selon l'une quelconque des revendications précédentes, caractérisé en ce qu'une cloche de protection amovible (4) est placée sur l'espace de refroidissement (3).
  10. Dispositif de refroidissement (1) selon la revendication 9, caractérisé en ce que la cloche de protection (4) est au moins partiellement transparente.
  11. Dispositif de refroidissement (1) selon la revendication 9 ou 10, caractérisé en ce que la cloche de protection (4) comprend un sas à échantillons (5).
  12. Dispositif de refroidissement (1) selon l'une quelconque des revendications précédentes, caractérisé en ce que sur la face inférieure de la cloche de protection (4) et/ou sur la face supérieure de l'espace de refroidissement (3) est agencée une sortie de gaz froid (9), par l'intermédiaire de laquelle le fluide de refroidissement et/ou le gaz froid peut s'échapper de l'espace de refroidissement (3).
  13. Dispositif de refroidissement (1) selon l'une quelconque des revendications précédentes, caractérisé par
    - un capteur de température (19) disposé dans l'espace de refroidissement (3) afin de mesurer la température dans l'espace de refroidissement (3),
    - une soupape de fluide de refroidissement (12) pouvant être commandée, destinée au réglage de la quantité de fluide de refroidissement alimenté,
    - un régulateur de température (18) servant à réguler la température dans l'espace de refroidissement (3), le régulateur de température (18) étant relié côté entrée au capteur de température (19) et côté sortie à la soupape de fluide de refroidissement (12).
  14. Dispositif de refroidissement (1) selon la revendication 13, caractérisé en ce que le régulateur de température (18) est relié à la soupe de fluide de refroidissement (12) par l'intermédiaire d'un générateur de rythme (17), le générateur de rythme (17) ouvrant et fermant alternativement la soupape de fluide de refroidissement (12).
  15. Dispositif de refroidissement (1) selon la revendication 13 ou 14, caractérisé en ce que le capteur de température (19) est disposé sur une position de traitement dans l'espace de refroidissement (3).
  16. Dispositif de refroidissement (1) selon l'une quelconque des revendications précédentes, caractérisé en ce que le fluide de refroidissement est de l'azote liquide.
  17. Utilisation d'un dispositif de refroidissement (1) selon l'une quelconque des revendications précédentes en vue d'analyser, de traiter et/ou de manipuler un échantillon cryogénique.
EP05716364A 2004-07-19 2005-03-24 Dispositif de refroidissement pour echantillons biologiques Active EP1768782B1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PL05716364T PL1768782T3 (pl) 2004-07-19 2005-03-24 Urządzenie chłodnicze dla próbek biologicznych

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102004034827A DE102004034827A1 (de) 2004-07-19 2004-07-19 Kühleinrichtung für biologische Proben
PCT/EP2005/003162 WO2006007884A1 (fr) 2004-07-19 2005-03-24 Dispositif de refroidissement pour echantillons biologiques

Publications (2)

Publication Number Publication Date
EP1768782A1 EP1768782A1 (fr) 2007-04-04
EP1768782B1 true EP1768782B1 (fr) 2008-07-02

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Application Number Title Priority Date Filing Date
EP05716364A Active EP1768782B1 (fr) 2004-07-19 2005-03-24 Dispositif de refroidissement pour echantillons biologiques

Country Status (9)

Country Link
US (1) US20070169488A1 (fr)
EP (1) EP1768782B1 (fr)
AT (1) ATE399593T1 (fr)
DE (2) DE102004034827A1 (fr)
DK (1) DK1768782T3 (fr)
ES (1) ES2308455T3 (fr)
PL (1) PL1768782T3 (fr)
PT (1) PT1768782E (fr)
WO (1) WO2006007884A1 (fr)

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KR102009505B1 (ko) * 2019-01-17 2019-08-12 주식회사 엘지화학 유전자 증폭 모듈
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WO2005010499A2 (fr) * 2003-07-18 2005-02-03 Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. Dispositif et procede de manipulation d'un echantillon

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ATE399593T1 (de) 2008-07-15
ES2308455T3 (es) 2008-12-01
PT1768782E (pt) 2008-09-19
DK1768782T3 (da) 2008-09-29
DE102004034827A1 (de) 2006-03-16
WO2006007884A1 (fr) 2006-01-26
PL1768782T3 (pl) 2008-12-31
US20070169488A1 (en) 2007-07-26
EP1768782A1 (fr) 2007-04-04
DE502005004582D1 (de) 2008-08-14

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