EP2095876A1 - Cover for a sample carrier - Google Patents

Abstract

The cover device (101) has a sticky surface area for covering a sample carrier (5) in liquid-tight fashion after connection of the cover device to the sample carrier in the sticky surface area. The sticky surface area comprising a thermoplastic or poly dimethyl siloxane (PDMS) sylgard 184 is formed by applying an adhesive layer is applied on carrier material. The thickness of the adhesive layer is 1-1000 mu m. An independent claim is included for method for producing cover device.

Description

The invention relates to a covering device for a sample carrier and to a method for producing a covering device or a sample carrier.

Particularly in the fields of biology, biochemistry or medicine, sample carriers are used to examine molecules, for example DNA, or cells. The substances to be investigated are generally added with a liquid into or onto a sample carrier and can then be examined, for example, by methods of microscopy (transmitted-light microscopy, fluorescence microscopy, confocal microscopy, etc.).

From the DE 101 48 210 For example, a sample carrier in the form of a flow chamber is known. This flow chamber has in a base plate at least one channel which is connected on both sides to a liquid reservoir.

In order to provide suitable sample chambers or reservoirs, various components are often interconnected to obtain the desired sample chamber structures in this way. Different methods are known for connecting these various components. One possible method is in the EP 1 579 982 described in which a plastic body is exposed to a swelling agent-containing vapor and then pressed at room temperature with another plastic body.

In this known method, it is necessary that a swelling agent suitable for the respective components used is provided, with which the surface of the plastic body is then dissolved. It is therefore the object of the present invention to provide a covering device for a sample carrier, which can be easily connected to the sample carrier.

This object is achieved by a covering device according to claim 1.

According to the invention, a covering device for a sample carrier is provided, comprising a sticky surface area for connection to the sample carrier, wherein the sticky surface area is designed such that the sample carrier is covered in a liquid-tight manner in the sticky surface area after connecting the covering device to the sample carrier.

In particular, because of the sticky surface area, such a covering device has the advantage that it can be connected to the sample carrier at substantially any time. On the other hand, such a covering device can be connected to sample carriers made of a variety of different materials in a simple manner.

The sample carrier itself can be present in a wide variety of shapes or geometries. For example, it may merely be a coverslip or a foil. Alternatively, however, the sample carrier may already have structures for sample intake, for example a sample chamber or a reservoir.

"Sticky" in this context means that at room temperature, in particular from 20 to 25 ° C, and 30 - 40% relative humidity stickiness exists. The stickiness should consist in particular when connecting covering device and sample carrier. After bonding, the cover should be liquid-tight even in the presence of liquid or medium or 100% humidity.

In particular, the sticky surface area may be permanently tacky and / or non-hardening. This makes it possible, at least on dry surfaces, to connect the cover device to a sample carrier multiple times.

The covering device may comprise a carrier material to which an adhesive layer for forming the sticky surface area is applied.

This makes possible the structures desired for this production for the covering device, in particular from a non-sticky material. In particular, the covering device may comprise a blank which comprises or consists of the carrier material. In this blank, the adhesive layer is then applied to predetermined areas.

The adhesive may in particular be a pressure-sensitive adhesive. Such a pressure-sensitive adhesive remains highly viscous and permanently tacky after application to the carrier material. The viscosity may be in particular 50-5000 mPa s. Such a pressure-sensitive adhesive does not harden. The adhesive may be a two-component adhesive comprising a base material and a crosslinker. In particular, base material and crosslinker may be mixed in an asymmetrical ratio.

The sticky surface area may in particular be structured perpendicular to the surface. The sticky surface area may thus have elevations or otherwise be irregular or non-planar.

For example, the adhesive layer may be applied by placing the surface to be tackified in an e.g. dipped freshly mixed pressure sensitive adhesive, and then pressed against a smooth or textured surface for a predetermined period of time. With a structured (stamped) surface, one can thus obtain a structured adhesive layer. When used as a pressure-sensitive adhesive, e.g. PDMS Sylgard 184 in a mixing ratio of 35: 1 and plastic structured as a molding surface, it is possible to obtain e.g. Create microstructures such as channels. Such structures may e.g. be applied to initially dry cell growth surfaces. Thus, directed growth of neuronal axons or chemotactic effects of cells can be investigated.

The adhesive layer may have a thickness of 1 .mu.m to 1000 .mu.m, in particular 10 .mu.m to 100 .mu.m. This makes it possible to achieve precise connection. The adhesive layer may in particular be elastic.

In the cover devices described above, the tacky surface area may comprise a thermoplastic or a silicone, in particular PDMS. With these materials, such a covering device can be produced in a suitable manner. For example, the surface area of a blank for the capping device in the surface area may be coated with an adhesive comprising or consisting of one of said materials. For example, a coating with asymmetrically mixed PDMS can be provided.

The sticky surface area can be designed in such a way that the covering device can be detached again from the latter without destruction and / or residue after it has been connected to the sample carrier.

This allows multiple times to connect a cover device to one or more sample carriers.

The tacky surface area may be such that the tensile strength after bonding to a coverslip is at least equal to the tensile strength of the reference material PDMS Sylgard 184, available from Dow Corning, having a blend ratio of 20: 1 of base material to crosslinker is. Such a minimum tensile strength, determined according to the protocol described below, is thus at least four times higher than the tensile strength of Sylgard 184 with a mixing ratio of 10: 1. In particular, the tensile strength may be at least twice and / or at most equal to twenty times the tensile strength of the reference material.

To determine the tensile strength of the sticky surface of a capping device, a test cylinder is made of the material to be tested, with the circular base area of the cylinder used for the measurement having an RMS roughness between 0.1 nm and 10 nm. For example, if the tensile strength of a cover device made of a sticky material is determined, the test cylinder is made of this material. When the covering device comprises a blank provided with an adhesive layer, the test cylinder is made from the material of the blank having the above roughness and the adhesive layer (in the thickness used for the blank) is applied.

The round base of the test cylinder has an area of 20 mm ² , the corresponding diameter is 5.4 mm. The cylinder is suspended vertically with the axis of rotation symmetrical so that the normal vector points downwards on the surface used for the test. From the bottom, a conventional Menzel cover glass (25.5 mm wide and 75.5 mm long) is glued to the base with the cover glass weighted. The data refer to a test procedure with a gravitational acceleration of 9.81 m / s ² . To glue the cover glass to the base, the cover glass is pressed against the cylinder for 10 seconds at a pressure of 3.0 × 10 ⁴ Pa.

The tensile strength results from the total mass (including the mass of the cover glass), which sticks to the base surface for at least 10 seconds before it comes loose and falls off. The determination is carried out at room temperature and a relative humidity between 30 and 40%.

For comparison, a reference test cylinder from the reference material Sylgard 184, Dow Corning, with the dimensions mentioned above is always produced. For the reference test cylinder, the mixing ratio of base material to crosslinker is 20: 1. The reference test cylinder thus has a sticky surface on which at least four times the mass, compared with a corresponding test cylinder of Sylgard 184 with a mixing ratio of 10: 1, while remaining stuck for at least 10 seconds.

For each total mass (coverslip with weight) the process is carried out at least ten times. Each time the residence time is measured until the cover glass is peeled off. A material to be tested has at least a tensile strength corresponding to a certain total mass, if in the process for the particular total mass an average residence time (at least 10 times) of at least 10 seconds has been obtained. This should apply to both the reference material and the material to be tested.

The cover device and / or the sample carrier may comprise a plastic or glass. In particular, they may consist of this material. However, the capping devices and the sample carrier need not comprise the same material or type of material; this is just a possibility. The covering device may in particular consist of a sticky carrier material. However, predetermined areas of the covering device can be treated such that the surface in these areas is no longer sticky. In principle, the entire covering device, a part of the covering device or only the sticky surface region can comprise or consist of a thermoplastic or a silicone. The covering device may consist of one piece.

The covering device may in particular be an injection-molded part or comprise an injection-molded blank. A surface area of the injection-molded part or the injection-molded blank can then be treated in such a way that a sticky surface area is provided.

The covering device may comprise or consist of an elastic material. This may be, for example, an elastic thermoplastic. The elastic material may in particular have a Young's modulus of 1 kPa to 1 MPa, in particular 1 kPa to 300 kPa. An elastic material allows a suitable adaptation of the covering device to the sample carrier during the connection.

In the case of the covering devices described above, only part of the surface, in particular only part of the contact surface with the sample carrier, can be made sticky. In this way it can be achieved that surface areas which are required for handling, for example, are not sticky. Below the contact surface with the sample carrier the surface of the covering device is meant, which is designed or intended for connection to the sample carrier or rests after the connection to the sample carrier. This Teilklebrigkeit can be achieved, for example, by only certain parts of a blank are provided with an adhesive layer or by a blank of a sticky material in a region that are not part of the contact surface with the sample carrier in particular, treated (passivated or plasma treated, for example) in that the surface in these areas is no longer sticky.

In particular, the invention also provides a covering device for a sample carrier, comprising at least one covering element for covering a surface region of the sample carrier, wherein the covering device is designed such that the surface region of the sample carrier is covered in a liquid-tight manner after connecting the covering device to the sample carrier. The at least one cover element thus has a contact surface with the sample carrier. In particular, the covering devices described above may comprise at least one such cover element.

At least part of the sticky surface area of the cover device may be arranged on at least one cover element. This represents a possibility to connect the cover liquid-tight with the sample carrier. In this case, the sticky surface area is preferably arranged on the contact surface of the cover element with the sample carrier. The sticky surface area may, but need not, be present or arranged on further surface areas of the cover device.

The possible properties of at least one cover element described here and below can apply to a cover element, a part of the cover elements or to each cover element.

As an alternative to the variant described above, at least one cover element can not have a sticky surface area. In particular, then the contact surface of the cover can not be provided with a sticky surface. In this case, the cover device has a sticky surface area in another area such that the cover element achieves a liquid-tight cover.

At least one cover member may comprise an elastic material. In this way you can in particular by pressing the cover to the sample carrier reach a liquid-tight covering during and after connection to the cover device. The elastic material may in particular have a Young's modulus between 1 kPa and 1 MPa, in particular 1 kPa to 300 kPa. It may, for example, be a thermoplastic or a silicone, in particular PDMS.

The elastic material may further be a sticky material. The sticky material may in particular be non-hardening and / or permanently tacky.

The covering device can have a substrate with a planar surface, on which at least one covering element is arranged.

At least one cover element may in particular be designed in such a way that the cover device does not have a contact surface in a region surrounding the at least one cover element (partially or completely), ie it does not abut against it in this surrounding region after the connection to the sample support. In particular, the at least one cover element may have a cylindrical shape. The base of the cylinder can be basically arbitrary. For example, the footprint may be a circle or a polygon. By means of such cover elements in column form can cover precisely certain surface areas. A base of the cylinder can form a contact surface.

Alternatively or additionally, at least one cover element may have a web shape. When connected to a sample carrier, a cover element in the form of a web allows a boundary wall to be formed for a sample chamber or a reservoir. The web shape can be straight or curved.

At least one cover element may have a height of 10 μm to 30 mm, in particular 50 μm to 10 mm, preferably 100 μm to 3 mm. A cylindrical cover element can have a base area of 0.01-500 mm ² , in particular of 0.1 to 50 mm ² . A cover element in web form may have a width of 50 μm to 30 mm, in particular 100 μm to 10 mm, and / or a length of more than 1 mm, in particular more than 10 mm.

At least one cover element may be designed such that it is pressed together after connecting the cover device with the sample carrier. Due to the compressed Condition after the connection, a liquid-tight cover can be achieved in an advantageous manner.

In this case, the at least one cover element can be designed such that its height after connection is reduced by 1 to 20%, in particular by 5 to 15%, compared with the height before joining. This makes it possible to achieve a suitable contact pressure for producing a liquid-tight cover.

The cover devices described above may comprise a plurality of cover elements. The cover elements can be designed differently or the same. For example, the cover elements may have a same shape with different dimensions. In particular, the covering device can have a plurality of covering elements in the form of cylinders, which have different basic surface dimensions. The height of the cover elements can be the same.

In principle, the sample carrier to which the covering device is connected can be structured or unstructured, in particular structured and / or unstructured surface regions. In the unstructured case, it may have a flat surface or flat surface areas.

In particular, the sample carrier may comprise a sample chamber or a reservoir, and the covering device may be designed to cover a surface area in the interior of the sample chamber. In particular, the covering device may have at least one covering element for covering a surface area in the interior of the sample chamber. For this purpose, in particular the sample chamber can have an opening to the environment. In particular, at least one cover element may be formed to cover a surface area at the bottom of the sample chamber.

The covering device can be designed such that it can be arranged completely inside the sample chamber. The covering device is thus designed as an insert for a sample chamber. Furthermore, the covering device and / or at least one covering element can be designed such that a sample chamber and / or a cavity is formed by the covering device and / or the at least one covering element after connection to the sample carrier. This sample chamber (reservoir) or this cavity can in particular in the interior of a (existing) sample chamber be arranged of the sample carrier. The covering device can furthermore be designed in such a way that a channel is formed after connection to the sample carrier.

A part of the tacky surface area may be arranged to be located outside the interior of the sample chamber after connecting the capping device to the sample carrier. In particular, the entire sticky surface area can be arranged in such a way. This can be avoided, for example, that remain in the interior of a sample chamber adhesive residue after removal of the cover.

For connecting the covering device and the sample carrier, the sticky surface may alternatively or additionally also be arranged on the sample carrier. The invention thus also provides a sample carrier comprising a sticky surface area for connection to a cover device, the sticky surface area being sticky, so that the sample carrier is covered in a liquid-tight manner in the sticky surface area after connecting the cover device to the sample carrier.

Such a sample carrier may, but need not, be used in conjunction with the previously described capping devices. The sample carrier may in particular also have the features described above.

The invention further provides a sample carrier system comprising a sample carrier and a cover device associated therewith, wherein the cover device and / or the sample carrier are formed as described above.

The sample carrier may comprise or consist of a foil. The film may have a thickness of 1 to 250 μm, in particular 100 to 200 μm. The sample carrier may comprise a substrate having a planar surface upon which one or more sample chambers or reservoirs are disposed. Alternatively, one or more sample chambers may be formed in the substrate. In particular, for forming a sample chamber, the substrate may have a hole, for example a blind hole or a through hole. In the case of a blind hole, the bottom of the blind hole forms the bottom of the sample chamber. The sample carrier may further comprise a bottom plate which is connected to the substrate, for example to form a bottom for a through hole. The bottom plate may have a thickness of 1 μm - 1.5 mm; it may be in the form of a film, in particular as described above.

The sample carrier may have one or more, for example formed in the substrate, channels. The channels are preferably aligned at least partially parallel to the base of the sample carrier or the substrate. A substrate may have a planar surface in which at least one groove is formed. This surface is preferably opposite the surface on which one or more sample chambers are formed. By covering the at least one groove with a bottom plate, a channel is formed.

The cover device, the sample carrier, the substrate and / or the bottom plate can be made of plastic or a silicone, in particular with a predetermined intrinsic fluorescence, in particular less than or equal to the self-fluorescence of COC or COP or a conventional cover glass (for example, pure white glass of the hydrolytic class 1 (such Menzel cover glass, in particular with the strength No. 1.5), and / or with a predetermined refractive index, in particular> 1.2 and / or <1.7, can be formed with such a high-quality optical material can be in an advantageous manner However, the substrate and / or the bottom plate may also be made of glass, and the cover device, the sample carrier, the substrate and / or the bottom plate may be flexibly formed All these elements may be injection molded.

Possible plastics are, for example, COC (cyclo-olefin copolymer), COP (cyclo-olefin polymer), PE (polyethylene), PS (polystyrene), PC (polycarbonate) or PMMA (polymethyl methacrylate). For example, Sylgard 184, available from Dow Corning, or other commercial, cell-compatible, single or multi-component silicones may be used as the silicone.

The capping device and / or the sample carrier may have the width and length of a conventional microscope slide (having a width of 25.5 mm and a length of 75.5 mm) or a multi-titre plate (85.5 mm × 127.5 mm). The cover device and / or the sample carrier may have a height of 0.1 to 15 mm. In particular, the covering device can have a height of 1 to 3 mm. A substrate of a sample chamber, in particular in a plane region, can have a height of 0.1 to 5 mm, in particular of 1 to 2 mm.

A sample chamber or a channel of the sample carrier can have a void volume of 0.1 μl to 3 ml, in particular 20 μl to 2.5 ml. In a substrate of a sample chamber Trained sample chambers or channels may in particular have a void volume of 0.1 to 200 .mu.l, in particular of 20 .mu.l - 150 .mu.l. The height of such sample chambers or channels may be 5 μm to 1 mm, in particular 0.1 mm-0.5 mm, the width may be 10 μm to 5 mm, in particular 0.5 mm to 2 mm. In the case of sample chambers formed on a substrate, their height may be 1 mm to 15 mm, in particular 5 mm to 10 mm.

• Spritzgießen eines Rohlings aus einem Silikon oder einem Thermoplasten,
• Beschichten eines vorherbestimmten Oberflächenbereichs mit einer Klebstoffschicht.

The invention further provides a method of manufacturing a capping device or a sample carrier, comprising the steps of:

• Injection molding of a blank made of a silicone or a thermoplastic,
• Coating a predetermined surface area with an adhesive layer.

In this case, the blank may not be sticky. The adhesive may in particular be a pressure-sensitive adhesive. The adhesive may be a two-component adhesive comprising a base material and a crosslinker. In particular, base material and crosslinker can be mixed in an asymmetrical ratio.

• Bereitstellen eines Rohlings aus einem klebrigen Material,
• Behandeln eines vorherbestimmten Oberflächenbereichs, um diesen nicht mehr klebrig zu machen.

As an alternative, the invention provides a method of manufacturing a capping device or a sample carrier, comprising the steps of:

• Providing a blank of a sticky material,
• Treat a predetermined surface area to make it no longer tacky.

Again, the blank can be injection molded. In the treatment of a sticky surface area in order to make it no longer tacky, it may be, for example, a passivation, in particular by plasma treatment.

The covering devices or sample carriers produced by means of these methods can have the properties and features described above.

Further features and advantages will be described below with reference to the exemplary figures.

Fig. 1

ein Beispiel einer Abdeckvorrichtung und eines Probenträgers,

Fig. 2

ein weiteres Beispiel einer Abdeckvorrichtung,

Fig. 3.

ein weiteres Beispiel einer Abdeckvorrichtung und eines Probenträger,

Fig. 4

ein weiteres Beispiel einer Abdeckvorrichtung und eines Probenträger, und

Fig. 5

ein weiteres Beispiel einer Abdeckvorrichtung und eines Probenträgers.

Showing:

Fig. 1

an example of a cover device and a sample carrier,

Fig. 2

another example of a covering device,

Fig. 3.

another example of a cover device and a sample carrier,

Fig. 4

another example of a capping device and a sample carrier, and

Fig. 5

another example of a cover device and a sample carrier.

At the in Fig. 1 Shown embodiment, a cover 101 is provided. The cover 101 has a recess 102 having a predetermined height. As a result, a circumferential edge is produced, through which a contact surface 103 is formed.

In the recess 102, a plurality of cover members 104 is arranged. These cover elements have the shape of circular cylinders of the same height. However, the cover elements have different base dimensions. For example, the diameters of the base can be 0.1 mm, 0.2 mm, 0.3 mm, 0.4 mm, 0.5 mm, 0.6 mm and 0.7 mm. From each circular cylinder are two copies available, the arrangement of the cylindrical columns is designed in opposite directions in two rows to average out any fluidic effects in the investigations.

The covering device 101 is connected to a sample carrier 105 in a liquid-tight manner, as can be seen in the lower part of the figure. The sample carrier 105 is designed in the form of a Petri dish and thus has a sample chamber or a reservoir. The cover device 101 is connected to the bottom 107 of the sample carrier and is thus arranged completely within the sample chamber.

After joining, both the border 103 of the covering device and the covering elements 104 cover corresponding surface areas at the bottom 107 of the sample carrier 105 in a liquid-tight manner.

At the in Fig. 1 Shown embodiment, two through holes 106 are further provided in the recess 102, with which a filling and venting of the resulting cavity can be performed after connecting the cover device with the sample carrier.

In principle, the covering device should be designed such that it rests in a liquid-tight manner on a corresponding surface of the sample carrier. The corresponding surfaces of the sample carrier can be smooth, rough or porous. A smooth surface has a roughness between 0.1 nm to 10 microns. A rough surface has a roughness of more than 10 μm. A porous surface may have pores with diameters of 0.01 μm to 50 μm. Porous surfaces may in particular comprise filter membranes, as used, for example, for filtering microorganisms or for carrying out transmembrane migration assays.

For this purpose, the covering device is preferably made of a silicone or a thermoplastic. For example, the cover device may comprise a silicone injection molding blank. The injection molding blank, for example, from PDMS, z. Sylgard 184. This two-component silicone gives complete crosslinking when the base material and the crosslinker are mixed in a ratio of 10: 1, which is referred to as symmetrical with respect to the crosslinking active groups in the base material and crosslinker. In this case, the resulting material has a Young's modulus of 57 kPa. At such a mixing ratio, the material is not sticky.

In principle, the desired connection between covering device and sample carrier can be achieved if the entire contact surface of the covering device or of the sample carrier is coated with a pressure-sensitive adhesive. On the part of the cover device, both the border and the cylinder base surfaces of the cover elements 4 could be provided with the adhesive for this purpose.

Alternatively, however, the adhesive is only applied to the border. The cover elements thus do not remain sticky. In order to achieve a liquid-tight cover, however, the cover elements are then made slightly higher than the depth of the recess 2 of the cover.

For example, the total thickness of the cover device may be 2 mm. The recess 2 may have a depth of 1 mm. However, the height of the columns 4 arranged in the recess 2 is 1.1 mm, so that the columns protrude by 0.1 mm beyond the recess. The contact surface of the border is provided with a 10 μm thick adhesive layer.

If the covering device and the sample carrier are pressed against one another, an adhesive bond is produced. The cover elements are pressed against the sample carrier and deform, resulting in a liquid-tight cover. During deformation, the height of the cover elements is reduced. In the example shown, the reduction is at most 10%, which, however, depends on whether the areas of the capping device on which the columns are arranged are also deformed.

If the entire covering device is made of a homogeneous, elastic material, the ceiling (or the bottom) of the recess 102 will also deform. Alternatively, however, the covering device may have an inhomogeneous material distribution, wherein the covering elements would have a higher elasticity than the rear side of the covering device.

A separate coating with an adhesive can be avoided if the material of the covering device or the corresponding blank is sticky at least at certain points of the surface. For example, asymmetric mixtures of PDMS can be used for this purpose. For mechanically stable, but strongly sticky surfaces, mixing ratios of 20: 1 to 60: 1 (base material: crosslinker) can be used. When using Sylgard 184, a residue-free removal of the adhesive from the slide after removal of the cover at mixing ratios of 10: 1 to 35: 1 can be achieved.

In principle, the covering device can also have a variable mixing ratio, for example a mixing ratio gradient, across the thickness. The distribution of the mixing ratio may be selected so that the side facing the sample carrier has sticky surfaces and is made more elastic. In the direction of the side facing away from the sample carrier, the stickiness and the elasticity decrease. Thus, the side facing away from the sample carrier has an increased hardness in order to give the insert stability, which also facilitates the removal of the covering device.

According to an alternative method of manufacture, an injection molding blank made of an elastic thermoplastic having a Young's modulus of 1 kPa to 1 MPa is produced. This blank can be made sticky, for example, by coating with sticky silicone or by means of plasma deposition. According to another possibility, in one second injection molding step, overmoulding predetermined areas of the blank with a tacky thermoplastic layer.

If a blank for the cover device is made of a substantially homogeneous, sticky material, certain surface areas can be made non-sticky by a passivation. A corresponding, spatially structured passivation can be carried out, for example, by a masked plasma treatment with the aid of mask technology.

In particular, the in Fig. 1 The embodiment shown makes it possible to cover liquid-tight circular surfaces with precisely predetermined diameters. In principle, however, the cover elements can also have other geometries. Thus, the base surfaces of the cylinder may have a polygonal shape. Furthermore, the cover can be in the form of straight or curved webs having a width of a few microns to a few mm. By such webs can be formed separate sample chambers or reservoirs. The length of straight webs can be up to a few cm as needed.

If such a covering device is connected to a sample carrier and then cells are seeded in or on the sample carrier, then these cells come into contact only with regions of the sample carrier which are not covered by the covering device. If the cells are those which can adhere to the bottom of a sample chamber, a confluent cell lawn can form, which is delimited by the cover elements. If the covering devices are removed, the cells can also grow in the previously covered areas of the sample carrier. By analyzing such ingrowth processes, statements about the behavior of the cells can be made. This is particularly true when cell ingrowth is analyzed as a function of the respective sample support surfaces (physical, chemical or biological properties, such as those achieved by certain treatments) or as a function of substances in the cell suspension.

In this way, for example, so-called wound-healing assays can be carried out. In this case, covering devices can be used by which circles with different diameters (or other geometric figures) are covered in one or more sample chambers. A corresponding cover device is in Fig. 1 shown.

After removal of the covering device from the sample carrier, in whose sample chamber or sample chambers a confluent cell lawn is formed, quantitative statements about cell growth (proliferation) can be made on the basis of the different circular diameters. This has the particular advantage that the cell growth does not have to be permanently monitored (for example by means of video microscopy), but can be measured at a specific time on the basis of the diameter of the just overgrown circle.

Corresponding investigations can be carried out in particular when multititer plates are used as sample carriers, in which case the covering device then has cover elements for the corresponding sample chambers. A covering device for a multi-animal plate may consist of one piece.

The described covering devices can furthermore be used for a structured coating of surfaces. After connecting the capping device to a sample carrier, the same or different coating substances may be filled into the one or more sample chambers of the sample carrier to produce corresponding surface functionalizations. After removal of the covering device, one thus has sample chambers in which certain areas are surface-functionalized and others are not. When using cell-adhesive coatings can thus grow a cell lawn only in certain areas. In this way, the nutrient requirements of a cell culture in a sample chamber can be reduced, so that it can then manage for longer periods of time without a change of media, or can be kept in smaller vessels or in materials with lower gas permeability.

Another area of application are so-called invasion assays. In this case, a cover device is provided on a homogeneously functionalized cell culture surface, for example plasma-treated polystyrene, which are provided with webs as cover elements such that after the connection two reservoirs separated by a partition wall are obtained. Thereafter, a monolayer of fibroblasts is cultured in one of the reservoirs and a monolayer of cancer cells is cultured in the other reservoir. After removal of the cover there are two cell spots separated by a cell-free strip whose width is given by the thickness of the previously existing partition. Since the surface of the sample carrier is functionalized in this area for cell adhesion, the cells can then grow in the strip and interact with each other. Thus, for example, the invasive behavior or metastasis behavior of Tumor cells are examined in connective tissue, for example, specific cell markers are used in the form of fluorescent dyes to distinguish the tumor cells clearly from the fibroblasts.

The sample carrier may consist of one piece, for example in the form of a cover slip, a foil or a Petri dish. Alternatively, the sample carrier may comprise a substrate and a bottom plate connected thereto, wherein in the substrate, for example, a groove is provided, so that a channel is formed by the bottom plate. The substrate may alternatively or additionally have through-holes, so that corresponding sample chambers or reservoirs are formed by the bottom plate.

The sample carrier may be made of a similar or the same material as the covering device, or be wetted with a corresponding material. This allows an advantageous connection of the two elements.

The bottom plate can have a thickness of up to 2 mm. Thicknesses of up to 200 μm are particularly advantageous for use with high-resolution microscopy. Larger thicknesses are used for low resolution microscopy, such as those used in wound healing assays.

Base plates having a thickness in the range of 1 μm can be used in combination with the described covering devices, in particular for laser microdissection of cell monolayers or pieces of tissue which can be fixed by means of the sticky surfaces of the covering devices. Depending on the design, the areas that do not rest on a piece of fabric can also serve for sealing and / or fixing.

Another example of a cover device 201 is shown in FIG FIG. 2 shown. Here, the cover device is intended as an insert for a sample chamber. The cover device 201 has a plurality of webs, through which, after connection to a sample carrier, two sample chambers or reservoirs 202 and 203 are formed, which are separated by a partition wall 205. The bottom surface 204 of the capping device is tacky while the sidewalls and top surface are non-sticky to facilitate handling of the insert, for example, with tweezers or other gripping tools.

FIG. 3 shows a sample carrier system comprising the cover 201 and a sample carrier 301. The cover is connected via its sticky bottom liquid-tight to the bottom 302 of the sample carrier. The two reservoirs 202 and 203 allow cells to be initially separated and cultivated after removing the cover device in the same volume. In the sample carrier shown, the bottom 302 is preferably made of optically high-quality material for high-resolution microscopy.

FIG. 4 illustrates another example of a sample carrier system with a sample carrier 401, this time with a plurality of capping devices 201. The sample carrier has a substrate 402 with a planar surface on which a plurality of sample chambers (reservoirs) 403 is arranged. An insert 201 can be introduced into the interior of each of the sample chambers 403 in order to produce two (partial) reservoirs in this way. Each insert is liquid-tightly connected at its bottom to the bottom of the respective sample chamber 403. In the resulting sub-sample chambers then appropriate investigations can be carried out as described above.

In FIG. 5 Another example of a cover device 501 is shown. The cover device 501 has a substrate with a planar surface 508, on which a cover element 506 is arranged. A sample carrier 504 has a substrate with a planar surface, on which two sample chambers are arranged, which are connected to one another via a channel 503 arranged in the substrate. In turn, a sample chamber is formed in the substrate through the channel. The channel 503 can be formed, for example, by a groove which is introduced into the underside of the substrate and covered with a film.

In the substrate of the sample carrier, an opening 502 is provided, which leads into the channel 503. The side wall of the opening 502 may be vertical or conical. The cover member 506 is formed as needed so that it fills the opening partially or completely. In particular, the sides 507 of the cover 506 partially or completely abut against the side wall. The planar surface 508 of the substrate of the cover device is tacky in order to connect the cover device in this area in a liquid-tight manner to the sample carrier. Thus, the opening 502 can be closed.

According to an alternative, the substrate of the covering device FIG. 5 also be omitted. In this case, for example, the sides 507 would be sticky in order to connect the cover device liquid-tight with the sample carrier.

It is understood that the embodiments described above are only to be understood as examples, and the individual features can also be combined with each other in a different manner.

Claims (20)

Covering device for a sample carrier, comprising a sticky surface area for connection to the sample carrier, wherein the sticky surface area is formed such that the sample carrier is covered liquid-tight after connecting the covering device to the sample carrier in the sticky surface area. Covering device according to claim 1, wherein the covering device comprises a carrier material, on which an adhesive layer for forming the sticky surface area is applied. Covering device according to claim 2, wherein the adhesive layer has a thickness of 1 .mu.m to 1000 .mu.m. Covering device according to one of the preceding claims, wherein the tacky surface area comprises a thermoplastic or a silicone, in particular PDMS. Covering device according to one of the preceding claims, wherein the tacky surface area is formed such that the covering device after connecting to the sample carrier of this again nondestructive and / or residue-solvable. Covering device according to one of the preceding claims, wherein the tacky surface area is formed such that the tensile strength after bonding with a coverslip is at least equal to the tensile strength of the reference material Sylgard 184 with a mixing ratio of 20: 1 of base material to crosslinker. Covering device according to one of the preceding claims, wherein the covering device and / or the sample carrier comprises a plastic, in particular an elastic plastic, or glass. Covering device according to one of the preceding claims, wherein the covering device consists of a sticky carrier material. Covering device according to one of the preceding claims, comprising at least one cover member for covering a surface region of the sample carrier, wherein the covering device is designed such that the surface region of the sample carrier is covered liquid-tight after connecting the covering device with the sample carrier. Covering device according to claim 9, wherein at least a part of the sticky surface area is arranged on at least one cover element. Covering device according to claim 9, wherein at least one cover member has no sticky surface area. Covering device according to one of claims 9 - 11, wherein at least one cover member comprises an elastic material. Covering device according to one of claims 9 - 12, wherein at least one cover element has a cylindrical shape or a web shape. Covering device according to one of claims 9 - 13, wherein at least one cover element is designed such that it is compressed after connecting the cover device with the sample carrier. Covering device according to one of claims 9 - 14, wherein the sample carrier comprises a sample chamber, and at least one cover element for covering a surface area is formed in the interior of the sample chamber. Covering device according to claim 15, wherein a part of the sticky surface area is arranged such that it is arranged after connecting the covering device with the sample carrier outside of the interior of the sample chamber. Sample carrier comprising a sticky surface area for connection to a cover device, wherein the sticky surface area is sticky, so that the sample carrier is covered liquid-tight after connecting the cover device with the sample carrier in the sticky surface area. Sample carrier system comprising a sample carrier and a cover device connected thereto, wherein the covering device according to one of claims 1 - 16 and / or the sample carrier is formed according to claim 17. Method for producing a covering device or a sample carrier with the steps: Injection molding a blank of silicone or a thermoplastic, Coating a predetermined surface area with an adhesive layer. Method for producing a covering device or a sample carrier with the steps: Providing a blank of a sticky material, Treat a predetermined surface area to make it no longer tacky.

Images