EP3195933A1 - Device for cryo-asservation of tissue samples - Google Patents

Abstract

The invention relates to a device for the cryoasservation of tissue, in particular a sample carrier insert (10). The sample carrier insert (10) has a first insert section (11) which can be introduced into a sample carrier (1). The first insert portion (11) has a recess (12) for receiving a second insert portion (20) and for receiving at least one coolant. The second insert section (20) can be introduced into the recess (12) of the first insert section (11), preferably in a form-fitting manner, and the second insert section (20) furthermore has at least one receiving section (21, 22) for receiving at least one container (11). 31, 32).

Description

The present invention relates to a device for the cryoasservation of biological tissue samples, namely a sample carrier insert, a sample carrier with such a sample carrier insert and a sample carrier system.

For many diagnostic or scientific analyzes of surgically removed tissue samples, a freezing, in particular a blast freezing of the tissue samples, is necessary. This can be achieved in a simple way by immersing tissue samples directly in liquid nitrogen. However, this can lead to cryo-artefacts in the tissue, caused by the so-called Leydenfrost phenomenon. These cryo-artefacts appear in the tissue as microscopic holes or cracks. Such artifacts make the tissue almost useless for structural microscopic analysis.

The formation of cryogenic artifacts can be prevented by immersing the tissue in a suitable cold mediator, such as liquid isopentane, which is precooled. In particular, isopentane is inert to the tissue and therefore does not affect subsequent analyzes. Isopentane-mediated shock-freezing is the only common method that preserves biological tissue samples for both structural and molecular biological analyzes. However, since isopentane is a highly flammable and explosive substance, this method is often used only in laboratories. Because some substances in collected biological tissues disintegrate within minutes (such as ribonucleic acids, RNA), adequate preservation at the site of collection (eg in the operating room) is necessary. Therefore, surgically removed tissue samples in the operating room for special or scientific issues mostly shared. One part is then fixed in formalin solution, a standard preservation method for subsequent structural analyzes - unsuitable for RNA analysis, another part in a commercially available stabilizer solution (eg RNAlater ^® ) for subsequent RNA analysis, but does not reflect the structure of the Preserved tissue. This division into tissue sections with different preservation methods is problematic because some disease processes in the tissue may not be homogeneous but very discontinuous and focal. Thus, it may be that with the formalin-fixed portion of such a focus can be structurally detected, in the section for RNA analysis, however, is not present. On the other hand, clinical and scientific methods of analysis today use very small amounts of sample. Thus, many analysis procedures can be performed on or with single or few microscopic sections. Isopentane-mediated shock-frozen tissue is equally well-suited for all these analysis methods.
With the invention presented here, the safety risks in handling isopentane are minimized and the handling of the cryopreservation is facilitated.
For example, isopentane as a cold mediator is routinely used in muscle biopsies, typically with an open plastic cup containing isopentane placed in a nitrogen tank containing liquid nitrogen to thereby be cooled down to the temperature of the liquid nitrogen. The boiling point of nitrogen is -196 ° C. By contrast, isopentane, also referred to as 2-methylbutane, is only present in liquid form up to -159.7 ° C. Therefore, isopentane, when stored in the nitrogen tank, will freeze. After removal of the plastic container from the nitrogen container, therefore, the isopentane must first thaw again so far that at least one liquid layer forms in the isopentane cup. In this liquid phase, the tissue to be asserved can be dipped and cryopreserved.

The tissue so cryopreserved in this way can then be stored, for example, in a - 80 ° C freezer or in the gas or liquid phase of the liquid nitrogen until further analysis.

However, the use of isopentane is problematic since both the liquid and vapor phases of this substance are extremely flammable. Therefore, a use in particularly security-sensitive areas, such as operating theaters, as long as not special security concepts are implemented. However, this requires a considerable amount of time and money.

Accordingly, it has been complicated to quickly and safely freeze tissue samples in an operating room.

DE 31 49 624 A1 and CH 672 834 A5 relate to transportable devices that use a temperature control of a transport device using an active system in the form of a Peltier element. Sample containers or substances to be cooled are stored in a suitably cooled metal block.

US 4,262,494 relates to a transportable device for freezing, transporting and temporary storage of tissue or tissue samples using dry ice as the refrigerant.

The prior art has in common that the devices for freezing and short-term storage of tissue samples are indeed suitable. However, these devices are not capable of reducing tissue damage on freezing or simplifying the freezing process for a user.

It is therefore an object of the present invention to obviate at least one of the above-mentioned disadvantages and to provide a device which enables a simplified and higher safety requirements sufficient cryoasservation of tissue samples.

This object is achieved according to the invention by a sample carrier insert Claim 1, a sample carrier according to claim 13 and a sample carrier system according to claim 14. Incidentally, the object is achieved by a use according to claim 15 in a method for the cryoasservation of tissue samples.

According to a first aspect of the present invention, a sample tray is provided. This sample carrier insert has a first insert section, which is preferably reversibly insertable into a sample carrier (1), wherein the first sample carrier insert has a recess for receiving a second insert section of the sample carrier insert. For this purpose, the first insert section is designed as a reversibly insertable section. The first insert section can also have a fastening means which is designed to fasten the first insert section to a sample carrier, in particular to a sample carrier housing. Incidentally, the first insert portion for receiving at least one coolant is formed. The second insert section is designed such that it can be introduced into the recess of the first insert section, preferably in a form-fitting manner. Furthermore, the second insert section has at least one receiving section for receiving at least one container.

Such a sample carrier insert makes it possible to safely arrange and transport a container with refrigeration agent, preferably a sealable container, in a sample carrier. In this case, both the cold mediator and a tissue sample to be asserved by a stored in the sample holder insert coolant, preferably dry ice, frozen or stored frozen. The refrigerant can be stored at a low temperature necessary for cryopreservation and a step for defrosting the refrigerant can be avoided.

Thus, it is also possible according to the invention to enable tissue preparation, shock freezing with a cold promoter and transfer into a storage container in a device. This can simplify the cryopreservation process and increase the safety of the procedure. In addition, can on avoided in this way, that the tissue sample or the necessary materials for preservation between several devices must be transferred back and forth, which can reduce the risk of contamination of the tissue sample.

In some embodiments, the receiving portion of the second insert portion is formed with a predetermined shape, so that the receiving portion is formed in particular for receiving a container having a shape corresponding to the specific shape of the receiving portion.

This can in particular represent a safeguard against unintentional tilting, overturning or shifting of the container. Incidentally, in particular in a case in which the receiving portion and the container have a shape which is not purely rotationally symmetrical, twisting of the container in the receiving portion can also be prevented or at least reduced.

The coolant, which is provided in the recess of the first insert section, may in particular be dry ice or have dry ice. Alternatively, liquid nitrogen would also be conceivable as a coolant if care is taken to ensure that the sample carrier does not become colder than -160 ° C. by isolation or sufficient distance from the liquid nitrogen. The sample carrier insert, in particular the first insert section, is designed to receive the coolant, in particular to receive at least one dry ice plate. It is also conceivable that, in addition or in alternative embodiments, the second insert section is adapted to receive a coolant, such as a dry ice plate. For this purpose, or alternatively, the second insert portion may be formed with a recess for receiving coolant, preferably for receiving a dry ice plate. The first insert portion may also be formed integrally or integrally with a sample carrier in some embodiments. However, a multi-part construction can also allow a retrofitting of existing sample carriers or sample carrier systems.

By using dry ice as a coolant, the cold promoter can be cooled to a temperature of -78.64 ° C or stored at this temperature. Similarly, the first insert portion and / or the second insert portion can be cooled to this temperature or stored at this temperature. It goes without saying that the first and / or the second insert section can also be pre-cooled and stored in a freezer, for example a -80 ° C. freezer, before use in a sample carrier, so that the passive cooling, in particular by dry ice, only for temperature stabilization, but not for cooling down the insert sections, must be used. The first and / or second insert section advantageously have a highly conductive material, in particular metal, preferably aluminum. In particular embodiments, the first and / or the second insert section may have a corrosion-resistant, also seawater-proof, aluminum alloy (Al Mg and Al Mg Mn alloys), such as. B. AlMg3 (Euro standard designation: EN AW-5754). It is understood that in addition to aluminum, other metals with good corrosion resistance and thermal conductivity can be used without departing from the subject matter of the present invention.

The receiving portion of the sample carrier insert may have at least one projection and / or at least one discontinuously formed edge portion. In the context of the present invention, a "discontinuously formed" edge section is understood to mean a shape which has a non-continuous edge course, that is to say, for example, an edge or a side surface with a kink. In particular, such a discontinuous edge portion is a corner or edge of a recess in a surface. With such a shaped receiving portion and a container, which is shaped according to the shape of the receiving portion, therefore, a rotation of the container in the receiving portion can be reliably prevented when a form-fitting or almost form-fitting shaped container is introduced into the receiving portion.

Under the understanding of the term "discontinuous" should also fall such edge portions which are introduced into a surface, for example in the second insert portion, and having a plurality of edges which are interconnected via rounded corner or edge portions. For example, the edges between the faces of a box fall under the definition of a discontinuous section. As a consequence of such a shape, it is not possible for an object, which is positively inserted into such recess or recess, to be freely rotated about at least one of its axes of rotation.

Likewise, a protrusion formed, for example, at a bottom portion of the receiving portion, or a plurality of protrusions, may engage a correspondingly shaped bottom structure of a container, for example a cryopreserved tissue storage container or a container having a refrigerating agent with a correspondingly shaped bottom portion. The projection may also be formed as a tongue or lip, which protrudes from one of the surfaces of the recess, in particular from a bottom portion of the recess.

Preferably, in the receiving portion, an inner ring is introduced, for example, as an aluminum ring with a superior "crown", consisting of two opposite ring segments (approximately 60-85 ° arc) and a form-fitting depressed "bottom plate" is formed. Alternatively, it is possible that, according to some embodiments, in the receiving portion, the projection is formed as an internal thread. In this case, the receiving portion or the projection on the thread and / or one or more protruding lips for engaging in a provided on a container to be introduced external thread, which corresponds to the internal thread. In addition, it can be provided that the container has recesses for receiving or for latching at least one of the protruding lips. In this way, the container can be improved in the sample tray insert be stored and secured against accidental removal. Alternatively, the thread and the lips may also be formed in such a way that, for example, they have a sharp-edged design in such a way that they press or cut into a holding region of a container. Preferably, this is done in such a way that the container is impaired in any way in its function for the storage of tissue or refrigerant. The holding region of the vessel may be, for example, a bottom region or a correspondingly thick-walled section.

Incidentally, in the embodiments that a container is prevented from rotating in the recess by correspondingly forming the recess, it may be possible for the container or containers, which are introduced into the recess, to also be operated with one hand. The containers may in particular be closable containers, for example with a screw cap and / or screw cap. Since it may be particularly advantageous to open a vessel with only one hand, the internal thread in the recess may be formed in particular as a left-hand thread or have left-hand thread rings.

According to some embodiments, the second insert portion of the sample tray may include a plurality of receiving portions. The plurality of receiving portions may be provided in particular for receiving various types of containers.

"Various" should be understood in this context that containers of different shape, size and function in the or the receiving sections are receivable. Thus, for example, a receiving portion may be formed such that a container containing a refrigerant, such as isopentane, can be recorded against rotation therein. For example, a specific shape could be a quadrangular shape known from square headed screw-cap wide-mouth bottles. One or more recesses can be designed such that therein cryotubes of known dimensions, for example, those that a threaded / mating thread system with locking function, sometimes called TwistLock system, for example, the manufacturer FluidX Ltd. or Micronic, can be arranged.

In preferred embodiments, the second insert section is designed to accommodate a container that holds refrigeration agents, as well as to receive a plurality of, in particular five containers, which are provided for cryopreserving and storing the frozen tissue samples.

It is also conceivable that a recess, which is dimensioned such that a square-shaped vessel can be accommodated therein, has an additional recess with an internal thread corresponding to the thread of such a cryotube on a floor structure. Also, a threaded structure projecting from the floor structure can be formed in the receiving section.

In particular, the use of square screw cap wide-mouth bottles with a volume of 20 ml, which can be closed with a screw cap, may be advantageous, since the volumes of the cold mediator, in particular isopentane, are low. For example, it is sufficient, depending on the size of the tissue samples provided for cryopreservation, to use a cold mediator volume of only 5-10 ml. In this way, the amount of refrigerant and thus a security risk in dealing with the refrigerant can be reduced.

Incidentally, a plurality of such refrigerating agent containers can be kept in a sample carrier or in corresponding recesses of the sample holder insert. In this way, a separate refrigerant reservoir can be used for each tissue sample so that cross-contamination of tissue samples to be preserved can be reduced. This is also advantageous, for example, if tissue samples of several patients are to be taken in succession are. For this purpose, in some developments of the invention, for example, the first insert portion may be formed with one or more holders for receiving and storing refrigerating agent containers. In particular, the provision of two such holders provided on opposite sides of the second insert portion may allow containers to be stored in a holder which are still unconsumed and uncontaminated. In the other holder, however, those refrigerant reservoir can be stored, which have already been used and thereby possibly contaminated. In particular, the danger of cross-contamination of the tissue samples, in particular of the tissue samples of different types of tissue or of different tissue sources, can be reduced by such a storage system by means of one or more holders. In addition, this facilitates the handling of the system to successively asserve samples of several biological individuals or patients.

According to some embodiments of the present invention, a receiving portion for receiving a refrigerating agent container is provided in the second insert portion. In addition, a plurality, in particular five, receiving sections for receiving cryopreservation containers, also referred to as cryotubes, can be provided. The number of preservation container receiving sections may vary according to the methods used in practice.

The recess in the first insert section for receiving the second insert section can be formed centrally in the first insert section in a further development of the invention. "Central" in this context means that the recess is arranged centered in the sample carrier insert in the lateral direction, so that the recess when the sample carrier insert is introduced into a sample carrier is also arranged centered in the sample carrier in the lateral direction. In this way, the greatest possible distance to the walls of a sample carrier in each lateral direction, a lowest possible temperature or lowest possible temperature gradients are achieved at which the tissue samples or the refrigerants are stored.

Alternatively, the first recess of the first insert portion may be laterally offset in the lateral direction from the center of the first insert portion. In this way, a work surface can be provided or created on the surface of the first insert section. On this work surface, for example, the preparation of the tissue can be made.

In addition, the second insert section may have at least one color marking. This can in particular be formed adjacent to at least one receiving portion. Such a color marking serves to identify the recess with regard to at least one parameter. As a "characteristic" of the recess can be the size of the recess, for example, the diameter or lateral dimensions, the position of the recess in the second insert portion, the geometric depth of the recess, the specific shape of the recess, the type of projection, the snap is formed in a container, as far as it is provided, and / or the type of edge portion which is provided in the recess.

It is also conceivable that the color marking or the color markings on the second insert section correspond with color markings which are provided on the storage containers. On the containers, such markings can be produced, for example, either by production or by the user.

It is also possible that the color markers are interchangeable color markers, for example color plates or the like, which can be used as needed, replaced or removed. In addition, it is conceivable that, instead of or in addition to color markings and corresponding color marking elements for accommodating color markings, lids of containers have a predetermined color or are marked with a predetermined color. That way you can the lids themselves serve as a color marking, so that the assignment to the respective container is maintained when it is opened to fill it. Advantageously, color marking elements may be formed in the second insert section such that the color marking elements are suitable for receiving lids of the containers for storing the frozen tissue samples.

It is also conceivable that the preservation containers are used as sets, each set having a predefined color coding. Similar tissue samples can be given in each case in each case in similar containers, which can bring a routine user and thus an error reduction.

In addition, the first insert section and / or the second insert section may have a sample-cut template. This sample cut template may be provided as a two-dimensional pattern or as a three-dimensional pattern. For example, in the case of a two-dimensional pattern, it is conceivable that one or more squares of predetermined edge length may be engraved, milled, scored, or otherwise introduced into the surface on a work area or surface of the second insert section, or projecting from the surface is trained. The edge length of a square can be in particular 8.5 mm. It is understood that other dimensions, for example, between 6 mm and 10 mm can be provided. In general, it should be noted that the dimensions are based on the opening diameter of the container used for sample storage or of the cryotube.

Also, the sample cut template may be formed as a rectangle or otherwise. In general, it is conceivable that the sample cutting template predetermines a sectional shape and cut size which is useful for a subsequent processing of the tissue samples and therefore depends on the purpose of the further processing or analysis of the tissue sample. In this respect, even more complex shapes and / or sections may be predetermined by the sample section template.

Analogously, a three-dimensional sample section template, for example in the form of a cube with a predetermined edge length, for example 8.5 mm, can be arranged on the second insert section. It is understood that the statements regarding the two-dimensional sample-cut template apply equally to the shape and size of the three-dimensional sample-cut template.

Such a three-dimensional sample cutting template can be fastened to or on the surface of the insert section, for example, by means of a rod, preferably a metal rod. Alternatively, the rod may be inserted in the surface or in a hole provided in the surface.

The use of a sample slice template, particularly a sample slice template for cube-shaped tissue pieces with an edge length of no more than 8.5 mm, allows a precise definition of the volume of the tissue sample, in this case a volume of up to 0.6 ml in a refrigerating agent volume of about 5 ml, here isopentane, sufficiently fast to a temperature of below -40 ° C are cooled. In this way, the volume used by the refrigerant, in particular isopentane, can be reduced to a minimum, which can make the handling of the refrigerant during the cryo-aeration process safer due to the small volume. Furthermore, this ensures that a frozen piece of tissue / tissue sample can also be inserted into the respective opening diameter of the container or cryotube.

It should also be noted that the use of isopentane at a temperature of about -80 ° C, ie the temperature of the dry ice, in terms of safety-relevant issues is advantageous because at this temperature, the vapor pressure of isopentane by more than two powers of ten the room temperature is reduced.

According to a second aspect of the present invention, there is provided a sample carrier having a sample tray according to the first aspect of the invention. The sample holder insert can be integrally formed, integrated and / or reversibly introduced into the sample carrier, or arranged and fastened in it. This may, for example, also allow use of the invention in the form of a retrofit system for existing sample carriers.

In preferred embodiments of the present invention, all surfaces of the sample carrier, in particular the first insert section, the second insert section, a housing of the sample carrier as well as a handle, if provided, are made of or coated with a washable and disinfectable material. In particular, a housing of the sample carrier may be a plastic housing. A carrying handle of the sample carrier may advantageously comprise aluminum. Ensuring these hygiene criteria may allow use in an operating room.

According to a third aspect of the present invention, a sample carrier system is provided with a sample carrier according to the second aspect of the present invention, wherein the sample carrier system comprises at least one container, which is designed for receiving in at least one receiving portion of the sample carrier or a sample carrier insert. The at least one container is designed for the storage of a cold mediator, preferably isopentane. It is understood that it is also possible according to the various embodiments of the first aspect, that the sample carrier system further containers, for example, for the storage of cryo-tissue or the like, has.

The sample carrier or the sample carrier system also have a thermally insulated housing. The housing may be, for example, an insulating box commonly used for cooling or cooling storage of tissue samples. The housing may, for example, a cryostable plastic (ie stability to -80 ° C) have. Especially the housing can be made of polyethylene with an insulation made of polyurethane (PU full foam) arranged in the housing wall and a labyrinth seal. When using dry ice for cooling, in particular when using several, for example two to three dry ice plates, a temperature necessary for the cryoasservation and storage of tissue can thus be maintained in the sample carrier for longer periods of time, for example up to 24 hours and more, without having to top up with coolant. In addition, the cold mediator, here isopentane, immediately ready for use and does not first, as usual, heat or thawed to be present in usable, liquid form.

The sample carrier or the sample carrier system can be accommodated by the compact structure and the passive cooling system in an apparatus which may have a low weight, for example a weight of 6 kg or less. This makes the system easily portable. In addition, due to the passive cooling allows independence from a power supply. Thus, after preserving the tissue samples, slightly longer transports of the tissue samples can be carried out without the cold chain having to be interrupted for a short time after freezing the samples.

With the sample carrier or with the sample carrier system, it is possible to cryoassify a tissue sample taken from a tissue, for example muscle tissue. For this purpose, a cold agent is kept in a container. The container is stored in a recess in a first or second insert portion of the sample carrier. In addition, a coolant is stored in the first or second insert portion of the sample container, so that the temperature of the sample carrier or the sample carrier system at a predetermined temperature, in particular a temperature between about -40 ° C and -80 ° C, can be maintained or Temperature can be lowered to a value in this range.

The tissue to be preserved is then in the recess located in the Container introduced. The fabric is shock-frozen by the refrigerant in the container and can be placed in a storage container. The storage container may also be arranged in a recess of the first or second insert portion.

Due to the special positive reception of the containers and the possible formation of projections, discontinuous sections and / or threads in the recesses of the first and / or second insert section, it may be possible for a user to open and / or close the containers with one hand can, so that the use of the sample carrier insert according to the invention, sample carrier and / or sample carrier system can simplify a method for cryoasservieren. This is particularly advantageous in isopentane-filled containers, since the isopentane-filled container sits against rotation in the recess and therefore does not have to be opened to open it. Thus, the container can not fall out of the hand and fall unintentionally.

It is conceivable for the first and / or the second insert section of the sample carrier to be stored in a freezer prior to cryoasservation in order to be cooled to at least the predetermined temperature or below before being introduced into the sample carrier housing.

In addition, it is conceivable that prior to the introduction of the preserved tissue into the storage container, ie before or after the shock-freezing of the tissue, the tissue is cut into a suitable shape. For this purpose, the fabric can be stored and cut on a work surface created on one of the insert sections. In particular, a sample cutting template provided on the first and / or second insert portion may serve as a template for cutting the tissue sample to a predetermined extent.

According to the invention, a method may comprise, for example, steps listed below.

First, in some implementations, the first and / or the second insert section may be pre-cooled in a freezer, for example at a temperature of about -80 ° C. The optionally pre-cooled insert sections are then inserted into the sample carrier.

It is understood that in some embodiments, in addition to or as an alternative to cooling with the passive coolant, such as dry ice, the sample carrier may also be a sample carrier having an active cooling device. In this case, a cooling, for example, by means of a Peltier element, a refrigerant circuit or otherwise done.

Before using the device according to the invention, a plurality of containers, in particular square tubes, can be held in a holder of the sample holder insert, which containers are filled with a predetermined volume of cold agent. The filling of the cold mediator can be carried out under sterile conditions to reduce or eliminate cross-contamination of the tissue samples during their further processing. Such filled and unused containers may be provided with a seal for identification.

During tissue preservation, the sample carrier is opened, and it is checked whether a container is already provided in the receiving section provided for this purpose. If this is the case, it is checked before use whether this container is still unused, ie whether the seal is already damaged or not. If the container is already in use, an unused container can be removed from the reserved supply in the first insert section and arranged in the second insert section.

If the held containers have not already been stored refrigerated, the container can be pre-cooled in the second insert section for a predetermined time, for example 10 minutes. This can reduce the vapor pressure of the refrigerant and make handling accordingly safer. In addition, a freezing process can be accelerated in this way and it can be ensured that a shock-like freezing of the tissue by a sufficiently low temperature is possible.

In a further step, containers, in particular a container set, can be arranged for cryopreservation in the remaining receiving sections of the second or also of the first insert section. The container set is preferably again arranged in the receiving sections for a predetermined time, for example 5 min, prior to use, ie filling with a tissue sample, so that the containers can be pre-cooled. This prevents shock-frozen tissue from thawing or thawing in the containers or adhering to a warm inner wall of the container and freezing. Preferably, a container set is used whose container lids are marked with different colors. The containers can then be opened and the colored or color-coded lids can be inserted into the color marking elements. In this way, an easy allocation of the container can be ensured even when the container is open.

For tissue preservation, tissue that has been removed is then placed on a sterile gauze compress in a dish and rinsed, preferably with NaCl. The tissue is then fixed on the gauze compress, for example with tweezers, and cut into small pieces with a scalpel. When cutting, a sample section template can be used for uniform preparation. In this way, samples with at least approximately the same size can be generated.

The chopped pieces of tissue are then successively placed in the precooled refrigerant, preferably liquid isopentane, in the prepared container. It should be noted that the tissue does not stick to the container wall above the liquid level, which can lead to the formation of cryo-artifacts on redissociation to damage the fabric or by the unmediated (without liquid refrigerant).

After a sufficient freezing time, for example after 10-15 seconds, the tissue is removed from the Isopentanbad and distributed in the prepared container, also referred to as tissue tube or cryocontainer distributed.

The isopent container used in these steps is then screwed on and placed in a receptacle of the first insert section.

The filled tissue tubes are screwed on and stored in the sample carrier until further use or transport.

If several samples are frozen and prepared, the cryocontainers can be removed from the second insert section and either in further receiving sections, which may be formed, for example, in the first insert section, or in another, similarly constructed transport container, in particular with a pre-cooled, passively cooled and / or or good thermally conductive insert section.

The sample holder can be wiped externally with a disinfectant cloth after use.

Figur 1

einen Probenträger mit einem ersten Einsatzabschnitt eines Probenträgereinsatzes gemäß einer Ausführungsform der vorliegenden Erfindung;

Figur 2

den Probenträger gemäß Figur 1 mit dem ersten Einsatzabschnitt und mit einem zweiten Einsatzabschnitt des Probeträgereinsatzes;

Figur 3

einen vergrößerten Ausschnitt des zweiten Einsatzabschnitts des Probeträgereinsatzes; und

Figur 4

einen weiteren vergrößerten Ausschnitt des zweiten Einsatzabschnitts;

Figur 5

einen zweiten Einsatzabschnitt eines Probenträgereinsatzes gemäß einer weiteren Ausführungsform der vorliegenden Erfindung;

Figur 6

einen Probenträger mit dem ersten Einsatzabschnitt eines Probenträgereinsatzes gemäß Figur 5 gemäß der weiteren Ausführungsform der vorliegenden Erfindung.

Details, further details and further exemplary embodiments of the present invention are explained below with reference to the figures. Showing:

FIG. 1

a sample carrier having a first insert portion of a sample tray according to an embodiment of the present invention;

FIG. 2

according to the sample carrier FIG. 1 with the first insert section and with a second insert section of the sample carrier insert;

FIG. 3

an enlarged section of the second insert portion of the sample carrier insert; and

FIG. 4

another enlarged section of the second insert section;

FIG. 5

a second insert portion of a sample tray according to another embodiment of the present invention;

FIG. 6

a sample carrier with the first insert portion of a sample carrier insert according to FIG. 5 according to the further embodiment of the present invention.

FIG. 1 shows a sample carrier 1 with a sample carrier insert 10. The sample carrier insert 10 has a housing 2, which in the illustrated embodiment has a substantially rectangular shape and enclosing a cavity. On an upper surface an opening is provided in the sample carrier 1, which is closable by means of a lid 3. The lid 3 has for this purpose a sealing means 4, which in the in FIG. 1 shown embodiment is designed as a circumferential seal that seals the opening of the housing 2 against the lid 3 in the closed state of the lid 3.

It is also conceivable that in embodiments of the invention, the lid is provided with an additional insulation. The insulation can be arranged on the inside, that is, a side facing the cavity of the housing 2, of the lid 3. Such insulation can increase the insulation performance and thus reduce consumption of coolant, especially dry ice. In addition to the cover 3, the inside of the housing 2 can be provided with an additional insulation. The isolation can be formed, for example by a Armaflex ^® insulating mat, which is covered by a stainless steel sheet.

Furthermore, a carrying device is provided on the sample carrier 1, which is designed here as a hinged carrying handle 5. The carrying handle 5 may also comprise aluminum or consist of aluminum. The sample carrier 1 also has a closure 6, which in the closed state urges the lid 3 against the housing 2 and thus increases a sealing effect of the sealing means 4. The closure may have zugelastische loops. In this way, for example, in the case of a caused by the volatile or sublimating coolant pressure in the sealed housing, damage or destruction of the housing can be avoided by the pressure. Thus, with increasing pressure in the housing, the cover would open against the tensile force of the elastic loops until a sufficient pressure equalization has taken place. In addition, the closure may for example comprise at least one hook.

The housing 2 of the sample holder 1 surrounds a cavity. In this cavity of the sample carrier 1, a sample carrier insert 10 can be introduced. The sample carrier insert 10 shown here consists of two components, a first insert section 11 and a second insert section 20. FIG. 1 shows only the first insert portion 11 of the sample carrier insert 10. The outer contours of the first insert portion 11 correspond to the inner contours of the housing 2, so that the first insert portion 11 may be formed positively arranged in a lateral direction in the sample carrier 1. Preferably, the first insert portion 11 is reversibly introduced into the sample carrier 1.

The first insert portion 11 has a recess 12. The recess 12 is in the embodiment shown here a substantially cuboidal recess formed in or around the center, in the lateral plane, of the insert portion 11. The shape and the dimensions of the recess 12 preferably correspond to the shape or the outer dimensions of cooling elements, in particular dry ice plates. Of course, it is also conceivable that as a coolant also dry ice pellets can be used. It is understood that the first insert section 11 may be a solid block, in particular a metal block. Preferably, this metal is aluminum. However, other materials are possible analogously. Preferably, the first insert portion 11 comprises a material having a high thermal conductivity.

In alternative embodiments, the first insert portion may also be formed from a bent metal or sheet metal. This can significantly reduce the weight of the sample carrier. In addition, a larger volume of coolant can be stored in the sample carrier in this way. Preferably, the second insert portion is formed in this case as a stainless steel pan. It is also possible that an additional insulation, for example in the form of Armaflex ^® mats, is provided on the inside of the housing 2 such that the insulation fills a space between the first insert portion 11 and the housing 2. In this way, a thermal insulation of the sample carrier 1 can be improved.

As in FIG. 2 can be seen, the first insert portion 11 also has a fastening means 13, which is designed for attachment of the first insert portion 11 to the sample carrier 1. The fastening means 13 may comprise, for example, a screw connection. By reversibly provided arranging the first insert portion 11 in the sample carrier 1, the first insert portion can be precooled before use, for example in a freezer, so that the necessary cooling capacity can be reduced by the coolant and thus allows use of the sample carrier 1 over a longer period can be.

In an alternative embodiment, however, the first insert portion may also be integrally or inseparably connected to the housing 2 of the sample carrier 1.

As in further FIG. 2 can be seen, a second insert portion 20 can be inserted into the recess 12 of the first insert portion 11 and can be stored therein. In this case, preferably before introducing the second insert section 20, coolant, in particular dry ice or dry ice plates, is arranged in the recess 12. The second insert portion 20 is therefore preferably stored on the coolant. In this way, a direct contact between the coolant and the second insert portion 20 can be achieved, so that an improved cold bridge can be formed. In this way, a cooling of the second insert portion 20 can be achieved.

Advantageously, the second insert section 20 comprises a material with high thermal conductivity, in particular aluminum or other materials. In this way, heat can be easily dissipated through the body of the second insert portion 20, and cooling can be enabled or further improved.

The lateral dimensions of the second insert portion 20 are shown in FIG FIG. 2 designated embodiment smaller than the lateral dimensions of the recess 12. Therefore, the second insert portion 20 can be easily stored in the recess 12. In other embodiments, it is conceivable that the lateral dimensions of the second insert section 20 correspond to those of the recess 12 of the first insert section 11. In this case, the second insert portion 20 can be positively inserted into the first insert portion 11. Thus, shifting or blurring of the second insert portion 20 in the recess 12 can be reduced or prevented. This can be particularly advantageous if dry ice pellets or another volatile coolant is used as the coolant. Further, on the edge of the recess 12 extending in the insertion direction guide grooves or guide ribs may be provided which cooperate with corresponding guide ribs or guide grooves radially outward on the second insert portion 20 to an optimal alignment of the second insert portion 20 in the recess 12 to facilitate and prevent tilting of the second insert portion 20.

The second insert portion 20 has in the illustrated embodiment also has a receiving portion 21 which is suitable from its dimensions and its shape for receiving a container 31. In alternative embodiments, two or more such receiving sections 21 may be formed in the second insert section 20, but additionally or alternatively also in the first insert section 11. In addition, the second insert portion 20 has a plurality of additional receiving portions 22, which may differ in their dimensions and the shape of the aforementioned receiving portion 21. The recesses 22 are dimensioned and shaped so that they can preferably accommodate a different type of containers 32 with appropriate shape and size.

Incidentally, shows FIG. 2 two color markings 24, which are provided for marking an associated receiving portion in the second insert portion 20. The color markings 24 can here be reversibly arranged on or in the second insert section 20. In the in FIG. 2 For example, five receiving portions 22 are provided, wherein the second and fourth receiving portion is covered with a container 32. Each of the receiving sections 22 is assigned a color marking element 25, here in the form of a depression in the second insert section 20, wherein in the embodiment shown only the corresponding second and fourth color marking element 25 is actually provided with a color marking 24. Thus, the ink marks 24 may mark the respective containers 32 in the receiving portions 22 and allow differentiation between the various containers.

The color marking elements 25 can also be designed and dimensioned such that they can receive a lid of a container 32. In this way, a safe storage of the lid of the container during the preservation procedure be ensured. It is particularly conceivable that the lid of the container 32 itself have a predetermined color or are marked with a predetermined color.

It is understood that, alternatively, a plurality of receiving sections 21 for receiving containers 31 may be provided, for example for the provision of a refrigerating agent.

It is also conceivable in alternative embodiments that corresponding receiving portions are provided in the first insert portion 11. In particular, in such a case can then be dispensed with the provision of a receiving portion in the second insert portion 20, if it should be desired, for example, to increase the working surface on the second insert portion 20 or to provide generally there.

The second insert portion 20 is also provided with a hinged carrying handle 26. The carrying handle 26 allows for easier insertion and removal of the second insert section 20 into and out of the recess 12 of the first insert section 11. The carrying handle 26 can likewise comprise aluminum or consist of aluminum. In addition, it is conceivable that the carrying handle to facilitate handling with a sheath is provided, on the one hand has a low thermal conductivity and hygienic (as good disinfectable) and on the other hand is resistant to cold, such as polytetrafluoroethylene (Teflon ^® ).

FIG. 2 , as well as the enlarged partial view in FIG. 3 also shows a cube 34 which can be used as a sample cutting template. The cube 34 preferably has an edge length of 8.5 mm. However, the cube can also have other edge lengths, depending on the size of the tissue samples that are to be preserved.

As in FIG. 3 clearly visible, the cube 34 is connected by a rod 35 with the second insert portion 20. Alternatively, the sample cut template may also have a different shape or be formed directly on the surface of the first and / or second insert portion without a fastener or as a two-dimensional pattern. FIG. 3 also shows a container 31 for receiving a cold mediator and / or for receiving a tissue sample, on the surface of the second insert portion 20 adjacent to a receiving portion 21.

FIG. 4 shows a plan view of a receiving portion 22 for receiving the other container 32, in particular cryotubes or Kryoasservierungsbehälter, which usually serve to store the prepared tissue samples. As can be seen, the receiving portion 22 has a thread 23 along its inner wall. In the receiving section but instead of the thread and an aluminum ring with a superior "crown" consisting of two opposite ring segments (about 60 - 85 ° arc) and a form-pressed depressed "bottom plate" may be introduced. In addition, the receiving portion 22 in the in FIG. 4 also shown on a bottom portion a protruding tongue 23 'on. The internal thread 23 as well as the tongue 23 'are used for fastening and anti-rotation of containers 32, which are formed with an engaging in the thread 32 and a projection to the tongue 32' complementary engagement form. Preferably, the thread 32 is a left-hand thread. In this way, a Kryoasservierungsbehälter be stored for storage of tissue samples in the receiving portion 22 and stored securely therein. Incidentally, the special design of the receiving portions 21, 22 allow a one-handed opening or closing of the container 31, 32.

It is further in the embodiment of the FIG. 4 to recognize that at the bottom portion of the receiving portion 22, an opening 28 is provided. The opening 28 is formed centrally in the bottom portion. The opening 28 can be closed by means of a pressed-in plug of metal, in particular aluminum be. However, in alternative embodiments, the bottom portion of the receiving portion 22 may be formed entirely without an opening. By eliminating holes, it may be possible to prevent dry ice structures, for example in the form of pins, from forming in the openings and in the receiving sections 22. Such dry ice structures could in extreme cases even push the containers 23 out of the receiving sections 22. This can hinder the workflow and lead to confusion of the tissue samples. This can be counteracted advantageously. This can be equally applied to the receiving portions 21 and container 31.

The figures shown are merely examples in this respect, which are intended to provide a better understanding of the present invention.

FIG. 5 shows a second insert portion 20 according to an embodiment of the invention. In this case, a receiving portion 21 for receiving a container 31 in a central region of the second insert portion 20 is cuboid. Around three of the four edges of the receiving portion 21 are further receiving portions 22, here five receiving portions 22, formed which are adapted to receive a respective container 32. Adjacent to the receiving portions 22 color marking elements 25 are formed. In the embodiment shown according to FIG. 5 The color marking elements 25 are recesses in the surface of the second insert section 20. The position of the color marking elements 25 is selected so that each color marking element 25 is in each case designed as a clear nearest neighbor to one of the receiving section 22. In addition, the recesses of the color marking elements 25 are dimensioned such that they can receive a lid of the container 32 provided in the receiving sections 22. By colored provision of the container lid they can serve as a color mark 24, so that a clear identification of the samples is always possible in the containers, even if the insert section moves and the position of the container is changed thereby.

FIG. 6 shows a sample carrier system with a sample carrier 1 according to an embodiment of the present invention. In this case, on the surface of the first insert portion 10 in the lateral direction on opposite sides in a region which faces a surface of the housing 2 of the sample carrier 1, a holder 40 or 40 'for receiving and holding a plurality of containers 31 is provided.

In the illustrated embodiment, the brackets 40, 40 'extend between a front surface and a rear surface respectively parallel to one in plan view of FIG FIG. 6 left and right side surface of the housing 2. Each of these brackets 40, 40 'is for receiving four containers 31, which is particularly suitable for the provision of a cold mediator such as isopentane, or 2-methylbutane. The brackets 40, 40 'are designed as rails or clamping rails, which rest on the surface of the second insert section. The rails or clamping rails make it possible to clamp up to 4 containers 31 between the container inner wall and the rail.

The brackets 40, 40 'are reversibly fastened to the first insert section 10 by fastening means 41. The attachment can also be irreversible, for example by welding, soldering or gluing. However, it is advantageous to ensure that the fastening is on the one hand kryofest and effective at low temperatures, and on the other hand, that the respective mounting against water, detergents and disinfectant is resistant.

In alternative embodiments, the brackets 40, 40 'may also be integrally formed with the first insert portion 10. In addition, it is possible that at least one of the holders 40, 40 'is formed as a recess in the second receiving portion.

Provision of two holders 40, 40 ', in particular on opposite sides of the sample carrier 1, may be advantageous. In a first holder 40 can in this case, as in FIG. 6 shown holding container 31 A, which were used unused and ideally under sterile conditions. These containers 31 A may be provided with a seal 33, to indicate that these containers 31 A are unused. After use of the containers 31A, and thus after breakage of the seal 33, the containers are marked as used containers 31B due to the broken seal and can be stored in the second holder 40 'until disposal.

Consequently, all containers can always be closed and stored, so that the volumes of refrigerants, in particular isopentane, which are accessible at a particular time, are reduced to a minimum. This can facilitate the handling of the refrigerant and make it safer.

LIST OF REFERENCE NUMBERS

1

sample carrier

2

casing

3

cover

4

sealing

5

handle

6

shutter

10

Sample carrier insert

11

first insert section

12

recess

13

fastener

20

second insert section

21

receiving portion

22

receiving portion

23

thread

23 '

tongue

24

color coding

25

Color marker

26

handle

31

container

31 a

sealed container

31 B

used container

32

container

33

seal

34

Sample cutting template / Cube

35

pole

40, 40 '

brackets

41

fastener

Claims (15)

A sample carrier insert (10), wherein the sample carrier insert (10) has a first insert section (11) which is preferably reversibly insertable into a sample carrier (1), wherein the first insert section (11) has a recess (12) for receiving a second insert section (20 ) of the sample carrier insert (10) and for receiving at least one coolant, and
wherein the second insert section (20) can be introduced into the recess (12) of the first insert section (11), preferably positively, and the second insert section (20) further comprises at least one receiving section (21, 22) for receiving at least one container (11). 31, 32). A sample holder insert (10) according to claim 1, wherein the receiving portion (21, 22) is formed in a predetermined shape for receiving a container (31, 32) having a shape corresponding to the shape of the receiving portion (21, 22). Sample holder insert (10) according to one of claims 1 or 2, wherein the sample holder insert (10), in particular the first insert portion (11), for receiving the coolant, preferably of dry ice, in particular for receiving at least one dry ice plate is formed. Sample holder insert (10) according to one of the preceding claims, wherein the receiving portion (21, 22) has at least one projection (23, 23 ') and / or at least one discontinuously formed edge portion. Sample holder insert (10) according to claim 4, wherein the projection as an internal thread (23) and / or as a pressed-inner ring in the receiving portion (22) is formed. A sample tray (10) according to any one of the preceding claims, wherein the second insert portion (20) has a plurality of receiving portions (21, 22), preferably for receiving a variety of containers (31, 32). Sample holder insert (10) according to one of the preceding claims, wherein the recess (12) of the first insert portion (11) is formed centrally in the first insert portion (11). A sample holder insert (10) according to any one of claims 1 to 6, wherein the recess (12) of the first insert portion (11) is laterally offset from a center of the first insert portion (11). Sample holder insert (10) according to one of the preceding claims, wherein the second insert section (20) has at least one color marking (24, 25) adjacent to at least one receiving section (21, 22) for identifying the recess with regard to at least one parameter: size of the recess , Position of the recess, depth of the recess, shape of the recess, type of projection (23, 23 ') and / or type of edge portion. Sample holder insert (10) according to claim 9, wherein the color marking (24, 25) is designed as an exchangeable color marking. The sample carrier insert (10) according to one of the preceding claims, wherein the first insert section (11) and / or the second insert section (20) has a sample section template (34), wherein the sample section template (34) is preferably provided as a two-dimensional pattern or three-dimensional pattern. Sample holder insert (10) according to one of the preceding claims, wherein the first insert portion (11) and / or the second insert portion (20) comprises a material with high thermal conductivity, preferably aluminum and / or stainless steel. Sample carrier (1), in particular for the cryopreservation of biological samples, comprising: - A sample carrier housing (2) and - A sample carrier insert (10) according to any one of claims 1 to 12, wherein the sample carrier insert (10) integrally, integrally and / or reversibly introduced into the sample carrier housing (2) is arranged and attachable therein. Sample carrier system with a sample carrier according to claim 13 and at least one container (31, 32) for receiving in at least one receiving portion (21, 22) of the sample carrier (1), wherein at least one of the containers (31, 32) for storing a cold mediator, preferably isopentane , is trained. Use of a sample holder insert (10) according to one of claims 1 to 12 or of a sample holder (1) according to claim 13 in a method for cryoasservating tissue.

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