WO2017186332A1 - Verfahren und vorrichtung zur temperaturüberwachung einer kryokonservierten biologischen probe - Google Patents
Verfahren und vorrichtung zur temperaturüberwachung einer kryokonservierten biologischen probe Download PDFInfo
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- WO2017186332A1 WO2017186332A1 PCT/EP2017/000405 EP2017000405W WO2017186332A1 WO 2017186332 A1 WO2017186332 A1 WO 2017186332A1 EP 2017000405 W EP2017000405 W EP 2017000405W WO 2017186332 A1 WO2017186332 A1 WO 2017186332A1
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- Prior art keywords
- indicator substance
- temperature
- chamber
- diol
- indicator
- Prior art date
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0236—Mechanical aspects
- A01N1/0263—Non-refrigerated containers specially adapted for transporting or storing living parts whilst preserving, e.g. cool boxes, blood bags or "straws" for cryopreservation
- A01N1/0268—Carriers for immersion in cryogenic fluid, both for slow-freezing and vitrification, e.g. open or closed "straws" for embryos, oocytes or semen
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0236—Mechanical aspects
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0236—Mechanical aspects
- A01N1/0242—Apparatuses, i.e. devices used in the process of preservation of living parts, such as pumps, refrigeration devices or any other devices featuring moving parts and/or temperature controlling components
- A01N1/0252—Temperature controlling refrigerating apparatus, i.e. devices used to actively control the temperature of a designated internal volume, e.g. refrigerators, freeze-drying apparatus or liquid nitrogen baths
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0236—Mechanical aspects
- A01N1/0263—Non-refrigerated containers specially adapted for transporting or storing living parts whilst preserving, e.g. cool boxes, blood bags or "straws" for cryopreservation
- A01N1/0273—Transport containers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01K—MEASURING TEMPERATURE; MEASURING QUANTITY OF HEAT; THERMALLY-SENSITIVE ELEMENTS NOT OTHERWISE PROVIDED FOR
- G01K11/00—Measuring temperature based upon physical or chemical changes not covered by groups G01K3/00, G01K5/00, G01K7/00 or G01K9/00
- G01K11/06—Measuring temperature based upon physical or chemical changes not covered by groups G01K3/00, G01K5/00, G01K7/00 or G01K9/00 using melting, freezing, or softening
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01K—MEASURING TEMPERATURE; MEASURING QUANTITY OF HEAT; THERMALLY-SENSITIVE ELEMENTS NOT OTHERWISE PROVIDED FOR
- G01K2203/00—Application of thermometers in cryogenics
Definitions
- the invention relates to a method for monitoring the temperature of a cryopreserved biological sample.
- the invention further relates to a device for temperature monitoring of a cryopreserved biological sample.
- Cryopreservation of cells has so far been the only way to reversibly halt vital processes at the cellular level in such a way that they can restart after being heated to physiological temperatures. Cryopreservation has become an indispensable element for clinics, pharmaceutical companies, species conservation, environmental protection and health care through large biobanks in recent decades.
- Biological material is stored in low-temperature compatible sample containers (cryobanks), eg. As tubes, straws and bags, different sizes. In cryopreservation, the stored biomaterials are frozen while maintaining the vitality of the sample material, mostly at temperatures below -80 ° C, for live collections below -140 ° C to the temperature of the liquid nitrogen.
- cryoprobe is also used below.
- cryopreservation For macroscopic samples, such. As blood or tissue, numerous techniques for sample storage at low temperatures have been developed. In modern medicine, genetic engineering and biology, there is a tendency to undergo increasingly small samples of cryopreservation. For example, small volumes of suspension (milliliter or below) are frozen with suspended cells or cell groups. Cryopreservation of cells from in vitro cultures predominantly takes place in a suspension. However, most of the biomedically relevant cells require theirs Propagation and orderly development of a substrate contact. Therefore, samples may be frozen after culturing in the substrate-bound state.
- the quality of the samples is crucial as they are used for cell therapies in clinics, the development of pharmaceuticals and biotechnology products, as national resources and much more.
- the storage time is several days to decades, with a tendency for long-term storage.
- the samples are stored in refrigerated containers, mostly in metal drawers and racks, with which they are subject to temperature fluctuations in new deposits or withdrawals.
- live deposits cells, cell suspensions and tissue parts
- the uninterrupted cold chain plays a decisive role, but also the avoidance of large temperature jumps in the frozen phase.
- a further object is to provide a device for monitoring the temperature of a cryopreserved biological sample, with which disadvantages of conventional techniques can be avoided.
- Another object is to provide a way to detect the simplest possible markers or mark, whether a cryoprobe has heated above a definable limit temperature, and even if only for a short time. The limit temperature must be determinable between -20 ° C and -140 ° C before freezing.
- the device or the method should be easy to handle, low temperature tolerant and adjustable. It may consume little or no energy and possibly cause only the least cost, since the storage of a bioprobe in a refrigerated state should cost only a few Euros in its total expenditure. This requirement must also meet the applicable materials.
- said objects are achieved by a method for monitoring the temperature of a cryopreserved biological sample.
- a device for monitoring the temperature of a cryopreserved biological sample is provided.
- the device for monitoring the temperature of a cryopreserved biological sample as an object per se should be disclosed and claimed.
- the statements relating to the device, in particular its advantageous embodiments, are thus to avoid repetition as disclosed purely as a device and apply as disclosed according to the method and claimable.
- the device for monitoring the temperature of a cryopreserved biological sample comprises a sample container having a receiving space (sample reservoir) for receiving a sample, in particular a biological sample.
- the receiving cavity may contain a cryopreserved biological sample.
- the device further comprises a chamber whose interior is not fluidly connected to the receiving space.
- the interior is also only partially filled with at least one indicator substance, the melting temperature at atmospheric pressure, ie at 1013.25 mbar, the indicator substance is in a range of -20 ° C to -140 ° C.
- An additional compartment is provided by the chamber of the device according to the invention, which can be used by the partial filling with the indicator substance as an indicator element or indicator device to indicate an undesirable exceeding of the limit temperature.
- the partially filled with indicator substance chamber is therefore hereinafter also referred to as indicator device.
- the chamber is arranged in the interior of the sample container, in particular in the receiving space and / or in the lid.
- the arrangement in the interior of the sample container offers the particular advantage that no additional space outside the sample container is required.
- the chamber can also be arranged on the outside of the sample container or integrated into a wall of the receiving space.
- the sample container is a container suitable for cryopreservation, for example a tube, a straw (also referred to as seminal tube), a bag for storing blood or stem cells, a box or another container suitable for cryopreservation.
- Such containers are also referred to as cryotubes, Kryostraw, cryobags, cryobox or generally as a cryocontainer.
- the sample container is a cryotube.
- Cryogenic tubes are also referred to as biobank or cryobank tubes.
- Cryotubes have a receiving space, which has an internal Forms cavity for receiving a biological sample.
- the cryovial usually also has a lid for closing the receiving space. The lid may have an engagement over which the lid can be rotated with a tool.
- the cryotube may also include a bottom member having an identifier, e.g. B. in the form of a machine-readable code.
- the method further comprises freezing the indicator substance, wherein the chamber for freezing the indicator substance is brought into a first position, so that the indicator substance in the liquid state flows into a first partial volume of the chamber and freezes there. Thereafter, in particular before and during the monitoring phase of the cryolayer, the chamber with the frozen indicator substance is brought into a second position, in which melting of the indicator substance by the influence of gravity leads to a configuration change of the chamber filling.
- the configuration change may be an at least partial displacement of the chamber fill and / or shape change of the chamber fill.
- the chamber fill may be formed by the indicator substance or by the indicator substance and by one or more solid bodies disposed in the chamber and having a higher density than the indicator substance.
- the indicator substance is frozen in such a geometry or position and the chamber in its position in the frozen state, for. B. at the storage temperature or at least below the specified limit temperature or melting temperature of the indicator substance, so that a melting of the indicator substance after the change in position lead to a visible displacement of the liquid or its boundary geometry or to a displacement of the fixed body arranged in the chamber, which are no longer frozen in this after melting the indicator substance.
- the device comprising the sample container with a cryopreserved sample and the at least one chamber with the frozen indicator substance, for cryopreservation, the at least one temperature monitoring chamber being arranged in the second position on the sample container ,
- a particular advantage of the invention is thus that a configuration change of the chamber filling, z.
- the indicator substance directly indicates whether a cryoprobe has heated above a definable limit temperature, and even if only for a short time. This can be quickly and easily detected by visual visual inspection or technically automated by means of a suitably equipped measuring device, without the sample must be removed or thawed from the sample container.
- the interior of the chamber may be subdivided into a plurality of separate subspaces, each of which is only partially filled with an indicator substance whose melting temperature is in a range of -20 ° C to -140 ° C, the indicator substances have different melting temperatures in the subspaces.
- each indicator substance is selected and / or their mixing ratio is adjusted so that their melting point corresponds to one of the monitored temperature limits.
- This embodiment offers the advantage that the achieved temperature intervals into which the sample has arrived can be narrowed down more precisely.
- the sample container and a chamber wall can be transparent or semi-transparent at least at one point, so that it is visible from the outside whether a configuration change, for. B. a change in position, the indicator substance is done.
- the entire wall of the sample container and the chamber is made transparent or semi-transparent.
- the indicator substance may contain an indicator additive which increases a detectability of a physical property of the indicator substance.
- the indicator additive can be, for example, a dye, so that the indicator substance colored or colored, d. H. is not transparent, and so their shape and / or location is better visually recognizable.
- any dye which satisfies at least the following conditions is suitable as the dye:
- the dye is selected from the group consisting of triphenylmethane dyes, rhodamine dyes, especially xanthenes, azo dyes and phenazine and phenothiazine dyes.
- the dye is selected from the group comprising Oil Red, Methyl Red, Brilliant Green, Rhodamine B, Neutral Red, Methylene Blue, or other dyes used to stain cells in cytology.
- the indicator additive may be particles, in particular nanoparticles, which increase a scattering effect and / or polarization effect of the indicator substance for electromagnetic radiation impinging on the indicator substance. As a result, a configuration change of the indicator substance can be detected more reliably by means of an optical transmission measurement, scattering measurement and / or polarization measurement.
- the indicator additive may be conductive particles. By adding conductive particles, the conductivity or impedance of the indicator substance can be influenced. In this way, a configuration change of the indicator substance can be detected by means of a conductivity measurement or impedance measurement.
- the device may comprise a measuring device which is configured to a configuration state of the chamber filling, for. As a location of the indicator substance in the chamber to detect.
- the measuring device may be an optical or optical-electrical measuring device to z. B. with an optical transmission, scattered light or reflection measurement to determine a configuration change of the indicator substance.
- the indicator substance it is possible to select a substance whose melting temperature corresponds to a predetermined limit temperature whose exceeding is to be monitored.
- the indicator substance is a liquid or a mixture of different liquids whose melting point corresponds to the desired limit temperature.
- a mixture of water (H 2 O) and ethanol (C 2 H 6 O), a mixture of water (H 2 O) and potassium hydroxide (KOH) or a mixture of water and an antifreeze may be selected as the indicator substance.
- the mixing ratio is adjusted according to the respective melting diagram, which indicates the course of the melting point as a function of the mixing ratio, so that the melting point of the liquid mixture has the desired value, namely the limit temperature to be monitored.
- the indicator substance comprises at least one alcohol selected from the group consisting of octan-1-ol, nonan-1-ol, propan-1, 2-diol, propane-1,3-diol, butane-1,2 -diol, butane-l, 3-diol, butan-2-ol, pentane-l, 5-diol, pentan-l-ol, cyclo- pentanol, benzyl alcohol is selected.
- the at least one alcohol is particularly preferably selected from propan-1, 3-diol, propan-1, 2-diol and butan-2-ol.
- the indicator substance comprises at least two different alcohol components:
- an alcohol selected from the group consisting of octan-1-ol, nonan-1-ol, propan-1, 2-diol, propan-1, 3-diol, butane-l, 2-diol, butane-l 3-diol, butan-2-ol, pentane-l, 5-diol, pentan-1-ol, cyclopentanol, benzyl alcohol;
- an alcohol selected from the group consisting of octan-1-ol, nonan-1-ol, propan-1, 2-diol, propane-1,3-diol, butane-1,2-diol, butane-1 , 3-diol, butan-2-ol, pentane-l, 5-diol, pentan-1-ol,
- the mixing ratio of the components a) and b) is set so that the melting temperature of the mixture within a temperature range of -20 ° C to -160 ° C, especially from -25 ° C to -160 ° C or -50 ° C to -150 ° C, lies.
- the indicator substance comprises one of the following combinations of components a) and b):
- this indicator mixture comprises, for example, propane-1,2-diol and butan-2-ol in a mixing ratio of 40 to 60% by volume (gives a melting temperature of about -90 ° C.), propane-1 , 2-diol and propane-1,3-diol in a mixing ratio of 30 to 70% by volume, or propane-1,3-diol and butan-2-ol in a mixing ratio of 30 to 70% by volume.
- the indicator substance preferably also comprises at least one dye as described above. Most preferably, this dye is selected from the group comprising Oil Red, Methyl Red, Brilliant Green and Rhodamine B.
- an even more specific embodiment is characterized in that the indicator substance two alcohols a) and b), which are selected from propan-l, 3-diol, propane-l, 2-diol and butan-2-ol, preferably in a mixing ratio as above, and a dye selected from the group consisting of Oil Red, Methyl Red, Brilliant Green and Rhodamine B.
- the concentration of the dye in the alcohol component can vary widely depending on the dye and alcohol.
- the concentration should be kept as low as possible during intensive staining, so that the color molecules do not change or increase the viscosity of the freezing and melting behavior of the alcohols in which they are dissolved.
- the dye In this case, the concentration is typically in a range of ⁇ 10% by volume, in particular ⁇ 1% or ⁇ 0.1%, ie in the percentile or per thousand or subpromute range.
- the limit temperature to be monitored does not correspond directly to the melting temperature of the indicator substance, but rather to that temperature above the melting temperature at which the viscosity of the molten substance has decreased so much that the required liquid transport can take place.
- This temperature is also referred to herein as a threshold temperature and is typically in a temperature range of 3-30 ° C or 5-30 ° C, for example 3-10 ° C, 3-20 ° C, 5-10 ° C or 5-20 ° C, above the nominal melting temperature.
- the indicator substance is therefore characterized in that the liquid mixture in a temperature range of 3-30 ° C or 5-30 ° C above the melting temperature has a viscosity in a range of 10 to 10 6 mPa * s, preferably 10 to 10 4 mPa * s.
- the sample container may have a lid, which is provided for closing the receiving space.
- the lid may have a shaft engaging with an upper end portion of the receiving space.
- the shank can be formed on a head part of the lid, so that in the mounted or screwed-on state of the lid, the head part sits on the receiving space, while the shank engages in an upper end region of the receiving space.
- the chamber in the lid, z. B. in the head part and / or the shaft to be integrated.
- the introduced indicator device can not contaminate a bioprobe stored in the receiving space, since it does not come into contact with it, but is enclosed in a cover which is to be used anyway.
- the chamber serving as an indicator device is stored and prepared spatially separated from the rest of the sample container together with the lid (eg freezing of the indicator substance in the first case). th situation).
- the shank of the lid has a cavity partially filled with indicator substance. Especially in the case of cryotubes, often only a lower partial volume of the receiving space is filled with the bioprobe, so that an upper partial volume can be used to arrange the indicator substance.
- the chamber is designed as a closed hollow body, which is arranged in the receiving space of the sample container below the lid.
- the closed hollow body may be loosely disposed on a cryopreserved biological sample present in the receiving space, e.g. B. be placed on this.
- the loose arrangement should mean that the at least one body is placed in the interior of the chamber and there, at least in the liquid state of aggregation of the indicator liquid, in principle, is free to move, unless he is frozen by the indicator substance.
- only a solid body may be arranged loosely in the chamber, whose volume is greater than that of the indicator substance.
- at least two solid bodies may be present in the interior of the chamber, one volume of the solid bodies being smaller than one volume of the indicator substance in each case.
- sample container refers in particular to a container designed for cryopreservation.
- the sample container is preferably produced using low-temperature-compatible plastic material for temperatures below -140.degree.
- the plastic material can tolerate repeated temperature changes without change and without damage.
- a plastic material is preferably used whose water absorption capacity is ⁇ 1% of the intrinsic mass, in particular ⁇ 0.1% of the intrinsic mass.
- Cryogenic storage elements according to the invention are based, for example, on polyurethane or polyethylene.
- biological sample refers to biological material such as cells, tissue, cell constituents, biological macromolecules, etc. which is subjected to cryopreservation in the sample container, possibly in a suspension and / or composite with a substrate material Substrate arranged for the adherence of biological cells, which are part of the biological sample, is arranged.
- FIG. 1-4 are schematic views of various embodiments of apparatus for monitoring the temperature of a cryopreserved biological sample
- FIG. 5 is a flowchart illustrating an embodiment of a method for monitoring the temperature of a cryopreserved biological sample
- FIG. FIGS. 6A, 6B, 7A are each a melting diagram of a liquid mixture
- FIG. 7B is a table with melting points of some pure liquids.
- FIG. Figure 8 shows a miscibility matrix of solvents.
- FIG. 1 The schematic sectional views A to E of FIG. 1 illustrate a first embodiment of the invention.
- FIG. 1A a cylindrical receiving part 1 of a cryotube is shown in section in FIG. 1A.
- the receiving cavity 2 formed by the cylindrical receiving part 1 has already been filled with the bioprobe 6 here.
- the bioprobe 6 can, for. B. be a cell suspension.
- FIG. 1B shows the cover 3, which can be screwed on via a thread 8, for the cryotube 1, 3, which is upside down and which closes the receiving part 1 and optionally has an engagement 4 above, via which the cover 3 can be fitted with a tool (FIG. not shown) in the case of automation can be rotated.
- the lid contains in the screw-in part, ie in the shaft 5, which engages in the receiving volume 2 in the screwed state, a chamber 11 which forms a hollow volume 12.
- an indicator substance 7 in the form of a liquid or a liquid mixture whose freezing point / melting point in the range of -20 ° C to -100 ° C is selected via the mixing ratio such that the melting point has the value of a temperature limit to be monitored. This will be explained in more detail below with reference to FIGS. 5 to 8.
- the cryovial 1 is frozen in the open state, as shown in Figure 1A, and the lid 3 is frozen to the storage temperature in the inverted position as shown in Figure 1B.
- the indicator substance 7 first collects in the liquid state under the influence of gravity in the partial volume 12b of the hollow volume 12 of the chamber 11 and freezes there during the cooling to the storage temperature or to a temperature which is at least below the melting temperature of the indicator substance 7.
- the screw-on cover 3 is rotated by 180 ° and, as in FIG. 1C and FIG. 1D, screwed in to close the cylindrical receiving part 1 shown in FIG. 1A.
- the indicator substance 7 frozen in the partial volume 12b remains in the upper partial volume 12b of the chamber.
- the lower partial volume 12 a is substantially free of indicator substance 7.
- the device 10 for temperature monitoring thus formed can now be cryothorged.
- the device 10 is stored in the cryogenic containers. If the bioprobe 6 is now brought above the melting point of the indicator substance 7 during any manipulation or accident situation in the storage tank, it becomes liquid, drips downwards, and the picture shown in FIG. IE results.
- the indicator substance continues to be located in the partial region 12a of the chamber 11 after a storage process. The state is shown in FIG. 1D. In this way, an inadmissible heating of the sample 6 is easily recognizable.
- the location of the indicator substance within the chamber 11 is optically detectable by sight. Through a horizontal detection, shown schematically by the dashed line 100, the position of the indicator substance 7 can also be determined in an automated optical way in the case of transparent or semi-transparent design of the cryotube 1, 3.
- Another advantage of the device 10 is the reusability of the lid 11 and the use of marker liquids used as an indicator substance 7 with a freely selectable freezing point.
- a melting point of -80 ° C is recommended, as there is a significant recrystallization of the ice in and around the cells, which leads to a reduction in the quality of the cryoprobe.
- a melting point of -30 ° C is recommended.
- FIG. 2 shows a further exemplary embodiment of a device 20 for monitoring the temperature of a cryopreserved biological sample.
- the device 20 in turn comprises a cryotube as a sample container and an indicator device 21 which can be arranged in the interior of the cryotube in the form of a hollow cylinder 21 or 21 a partially filled with indicator substance 7.
- a cryotube is shown, which may for example also be frozen.
- the cryovial again comprises a cylindrical receiving part 1, which forms a receiving cavity 2, which has already been filled with the bioprobe 6 here.
- the cryotube further comprises a cover 3 for the cryotube, which closes the receiving part 1.
- a closed hollow cylinder 21 is used, which is partially filled with an indicator substance 7 and can be used in this way as an indicator device for temperature monitoring.
- the indicator substance 7 is again selected so that its melting point is in the range of -20 ° C to -100 ° C and has the value of a temperature limit to be monitored.
- the hollow cylinder 21 is frozen in the position shown in Figure 1B outside of the cryotube.
- the indicator substance collects in the lower partial volume 22b and freezes there.
- the cryotube 1, 23 shown in Figure 1A is opened by the lid 23 is unscrewed, the hollow cylinder 21 is rotated by 180 ° on the already frozen bioprobe 6 and the cryotube 1, 23 closed again, so that in Figure IC shown image results.
- the hollow cylinder 21 thus lies loosely on the bioprobe, in an orientation in which the indicator substance is initially in the frozen state at the top in the cavity 22.
- the melting temperature of the indicator substance 7 has been reached, the liquid is again at the bottom of the cylinder 21, in the partial volume 21a, as can be seen that a critical limit temperature has been exceeded.
- FIG. 2D alternatively shows a four-chamber wood cylinder 21a which, instead of the hollow cylinder 21, can be used analogously and can be introduced into a cryotube 1, 23 in the same way.
- the interior of the hollow cylinder 21 is divided by partitions 25 into four fluidly isolated part cavities, which are each partially filled with an indicator substance.
- the indicator substances 7a, 7b, 7c, 7d in the partial cavities are different and have different melting points.
- the indicator substances 7a, 7b, 7c, 7d a staggering of the melting points of the indicator substances 7a, 7b, 7c, 7d, z.
- the frozen indicator substances 7a to 7c are still found in the upper region 22b of the hollow cylinder during an inspection, but the indicator substance 7d is found on the bottom (partial region 22a), a temperature of -110 ° C. was exceeded for the sample 6.
- the indicator substance 7c is also on the ground, when -80 ° C has been exceeded, all marker liquids 7a to 7d are on the ground and have even been exceeded at -20 ° C.
- the introduced into the receiving space 2 hollow cylinder 21 or 21a should be sterile on its surface or otherwise made germ and contamination free.
- FIG. 3 shows a further exemplary embodiment of a device 30 for temperature monitoring of a cryopreserved biological sample.
- the peculiarity of this device 30 in comparison with the device 10 shown in FIG. 1 is that the chamber 31 integrated in the lid 3 of the cryotube is not only partially covered with an indicium. torsubstanz 7 is filled, but also contains small solid body 33 which are loosely inserted into the interior or cavity 32 of the chamber 33.
- Figure 3A shows the receiving part 1 of a cryotube in section analogous to Figure 1A.
- the chamber which is integrated into the shaft 5 of the upside-down cover 3 is, as shown in FIG. 3B, filled with an indicator substance 7 only at the bottom.
- the bodies 33 may be, for example, metal balls.
- the receiving part 1 and the upside-down cover 3 are now brought to the storage temperature. As a result, the small bodies 33 freeze in the indicator liquid 7.
- FIG. 4 shows a further exemplary embodiment of a device 40 for monitoring the temperature of a cryopreserved biological sample.
- the peculiarity of this device 40 compared to the device 30 shown in Figure 3 is that in the in the lid 3 integrated chamber 41 in addition to the indicator substance, a comparatively large and heavy body 43 is instead of several small bodies.
- FIG. 4A the receiving part 1 of a cryovial for a bioprobe 6 is again shown in section.
- FIG. 4B again shows an upside-down lid 3 of the cryotube, in the interior of which again a chamber 41 is integrated, which forms a cavity 42 which is partially filled with an indicator substance 7. Furthermore, a large and heavy body 43 is loosely inserted into the cavity 42.
- the freezing of the indicator substance and storage of the device 40 is analogous to the device 30.
- the detachment of the large body 43 is even less susceptible to interference when exceeding the melting temperature of the indicator substance 7, at the same time he passes his sinking either an optically transparent route free or blocked.
- the optically transparent path is shown schematically by dashed line 100 in FIG. 4E.
- FIG. 5 illustrates, with reference to a flow chart, a method for monitoring the temperature of a cryopreserved biological sample.
- a device for temperature monitoring is provided, for example one of the devices 10, 20, 30, 30 or 40.
- a suitable liquid or liquid mixture is selected as the indicator substance 7.
- melting point can be set to a desired value, in particular in a range from -20 ° C to -140 ° C.
- FIG. 6A shows the curve of the melting point as a function of the mixing ratio of an alcohol and water, with which a temperature range between 0 ° C and -118 ° C can be covered with a moderate increase in viscosity with decreasing temperature.
- the ethanol content can be set to 93.5%.
- Melting points up to a value of just below -60 ° C can also be adjusted by admixing potassium hydroxide (KOH) to water, which is shown in FIG. 6B by means of a melting diagram is shown.
- KOH potassium hydroxide
- a mixture of water and cryoprotectant may also be used as the indicator substance, as illustrated by the melting diagram of Figure 7A.
- step S2 the indicator substance is then frozen in the chamber, wherein the chamber is brought into a first position during the freezing of the indicator substance.
- the first layer corresponds in each case to a position of the cover 3 in which it is turned upside down, as shown in FIG. 1B, 3B or 4B.
- step S3 the at least one chamber with the frozen indicator substance is brought into a second position and arranged in the interior of the sample container.
- the second layer is rotated by 180 ° to the first layer.
- the chamber is brought by screwing the lid 3 with the receiving part 1 in the second position.
- the hollow cylinder is rotated by 180 ° placed on the frozen bio sample 6 in the receiving space 2.
- the device can be stored with a cryoprobe in the receiving space of the sample container at a storage temperature below the melting temperature (step S4).
- step S5 it is checked whether an undesirable, albeit temporary, heating of the cryoprobe has taken place. For this purpose, it is checked whether an at least partial displacement and / or shape change of the chamber filling caused by a melting process has taken place, as has already been explained above in FIGS. 1 to 4. If this is the case, it can be concluded that the limit temperature (s) to be monitored have been exceeded.
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- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Physics & Mathematics (AREA)
- Hematology (AREA)
- General Physics & Mathematics (AREA)
- Thermal Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Sampling And Sample Adjustment (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Temperature Or Quantity Of Heat (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
Description
Claims
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US16/095,936 US20190133111A1 (en) | 2016-04-27 | 2017-03-31 | Method and device for monitoring the temperature of a cryopreserved biological sample |
CN201780025971.3A CN109152355A (zh) | 2016-04-27 | 2017-03-31 | 用于冷冻保存的生物样本的温度监测的方法和设备 |
JP2018556920A JP2019515286A (ja) | 2016-04-27 | 2017-03-31 | 凍結保存生体サンプルの温度監視用の方法及びデバイス |
EP17718477.7A EP3448152A1 (de) | 2016-04-27 | 2017-03-31 | Verfahren und vorrichtung zur temperaturüberwachung einer kryokonservierten biologischen probe |
KR1020187033936A KR20190004727A (ko) | 2016-04-27 | 2017-03-31 | 냉동보존된 생물학적 시료의 온도 모니터링 방법 및 기구 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
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DE102016005076.4 | 2016-04-27 | ||
DE102016005076.4A DE102016005076A1 (de) | 2016-04-27 | 2016-04-27 | Verfahren und Vorrichtung zur Temperaturüberwachung einer kryokonservierten biologischen Probe |
Publications (1)
Publication Number | Publication Date |
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WO2017186332A1 true WO2017186332A1 (de) | 2017-11-02 |
Family
ID=58579131
Family Applications (1)
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PCT/EP2017/000405 WO2017186332A1 (de) | 2016-04-27 | 2017-03-31 | Verfahren und vorrichtung zur temperaturüberwachung einer kryokonservierten biologischen probe |
Country Status (7)
Country | Link |
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US (1) | US20190133111A1 (de) |
EP (1) | EP3448152A1 (de) |
JP (1) | JP2019515286A (de) |
KR (1) | KR20190004727A (de) |
CN (1) | CN109152355A (de) |
DE (1) | DE102016005076A1 (de) |
WO (1) | WO2017186332A1 (de) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE102017003171A1 (de) | 2017-03-31 | 2018-10-04 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Tieftemperatur-indikatormischung, vorrichtung und verfahren zur überwachung eines temperaturübergangs bei tiefen temperaturen |
DE102018009805B4 (de) | 2018-12-18 | 2020-12-10 | Dräger Safety AG & Co. KGaA | Kühlelement, Kontrollsystem, Einfrierhalterung und Verfahren zur Kontrolle eines Kreislaufatemschutzgerätes |
CN110097819B (zh) * | 2019-05-09 | 2021-04-13 | 南京工程学院 | 一种基于液晶形变原理的冷链标签及其使用方法 |
Citations (7)
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US3701282A (en) * | 1970-06-01 | 1972-10-31 | Grace W R & Co | Cold storage temperature monitor |
DE3716972A1 (de) * | 1987-05-20 | 1988-12-01 | Holzer Walter | Kaelteindikator zur ueberwachung des kuehl- oder gefrierzustandes von lebensmitteln, arzneimitteln und dergleichen |
US5282684A (en) * | 1987-02-13 | 1994-02-01 | Walter Holzer | Device for monitoring the temperature of cooled or deep-frozen products |
WO2000047964A1 (en) * | 1999-02-10 | 2000-08-17 | Temp-Tell Limited | Thermal history indicators |
US20060078036A1 (en) * | 2004-10-12 | 2006-04-13 | Wieder Horst K | Method and Device for Indicating Temperature |
WO2007085385A1 (de) * | 2006-01-27 | 2007-08-02 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Probenträger und probenspeicher, insbesondere zur kryokonservierung biologischer proben |
FR3013836A1 (fr) * | 2013-11-28 | 2015-05-29 | Commissariat Energie Atomique | Dispositif de controle d'un parametre de temperature en vue de detecter une rupture dans une chaine de froid |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP4163665B2 (ja) * | 2004-08-03 | 2008-10-08 | 松下電器産業株式会社 | 温度変化検知器、温度変化検知装置、温度変化検知システム、食品容器、および包装材 |
US7387438B2 (en) * | 2005-11-02 | 2008-06-17 | Robert Parker | Reusable, resettable, critical temperature indicator |
DE102008028334B4 (de) * | 2008-06-13 | 2014-02-27 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Probenkammer-Adapter für die Kryokonservierung biologischer Proben |
DE102008031666B4 (de) * | 2008-07-04 | 2010-05-12 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Kryobehälter |
DE102012202565A1 (de) * | 2012-02-20 | 2013-08-22 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Sicherheitsetikett für gefriergut |
TWI467142B (zh) * | 2013-05-13 | 2015-01-01 | Ind Tech Res Inst | 分離式溫度指示裝置及其使用方法 |
DE102013108557B3 (de) * | 2013-08-08 | 2014-11-13 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Einrichtung zur irreversiblen Erfassung einer Überschreitung einer vorbestimmten Temperatur |
-
2016
- 2016-04-27 DE DE102016005076.4A patent/DE102016005076A1/de not_active Withdrawn
-
2017
- 2017-03-31 CN CN201780025971.3A patent/CN109152355A/zh active Pending
- 2017-03-31 US US16/095,936 patent/US20190133111A1/en not_active Abandoned
- 2017-03-31 WO PCT/EP2017/000405 patent/WO2017186332A1/de active Application Filing
- 2017-03-31 KR KR1020187033936A patent/KR20190004727A/ko unknown
- 2017-03-31 JP JP2018556920A patent/JP2019515286A/ja active Pending
- 2017-03-31 EP EP17718477.7A patent/EP3448152A1/de not_active Withdrawn
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3701282A (en) * | 1970-06-01 | 1972-10-31 | Grace W R & Co | Cold storage temperature monitor |
US5282684A (en) * | 1987-02-13 | 1994-02-01 | Walter Holzer | Device for monitoring the temperature of cooled or deep-frozen products |
DE3716972A1 (de) * | 1987-05-20 | 1988-12-01 | Holzer Walter | Kaelteindikator zur ueberwachung des kuehl- oder gefrierzustandes von lebensmitteln, arzneimitteln und dergleichen |
WO2000047964A1 (en) * | 1999-02-10 | 2000-08-17 | Temp-Tell Limited | Thermal history indicators |
US20060078036A1 (en) * | 2004-10-12 | 2006-04-13 | Wieder Horst K | Method and Device for Indicating Temperature |
WO2007085385A1 (de) * | 2006-01-27 | 2007-08-02 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Probenträger und probenspeicher, insbesondere zur kryokonservierung biologischer proben |
FR3013836A1 (fr) * | 2013-11-28 | 2015-05-29 | Commissariat Energie Atomique | Dispositif de controle d'un parametre de temperature en vue de detecter une rupture dans une chaine de froid |
Also Published As
Publication number | Publication date |
---|---|
DE102016005076A1 (de) | 2017-11-02 |
JP2019515286A (ja) | 2019-06-06 |
US20190133111A1 (en) | 2019-05-09 |
KR20190004727A (ko) | 2019-01-14 |
CN109152355A (zh) | 2019-01-04 |
EP3448152A1 (de) | 2019-03-06 |
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