EP1805495A2

EP1805495A2 - Receiving unit for biological objects and method and device for treating biological objects placed in the receiving unit

Info

Publication number: EP1805495A2
Application number: EP05774733A
Authority: EP
Inventors: Karin SCHÜTZE; Yilmaz Niyaz; Raimund SCHÜTZE
Original assignee: PALM Microlaser Technologies GmbH
Current assignee: PALM Microlaser Technologies GmbH
Priority date: 2004-08-30
Filing date: 2005-08-16
Publication date: 2007-07-11
Also published as: WO2006024392A2; WO2006024392A3; DE102004041941B4; DE102004041941A1

Abstract

The invention relates to a receiving unit for biological objects, which comprises a plate-shaped element (1) provided with a plurality of openings (2). Said openings are arranged, in particular, in a manner such that they correspond to the openings (10a) of a microtiter plate or multiwell plate (10). As a result, biological objects (5) can be introduced in a simple manner, for example, by means of laser-catapulting, into the receiving element and from there into the microtiter plate or multiwell plate. The invention also relates to a device for thermally treating biological objects which are arranged in the receiving element.

Description

Recording device for biological objects and method and apparatus for processing in the recording unit recorded biological objects

The present invention relates to a recording unit for biological objects, which can serve in particular for receiving biological objects obtained by means of laser microdissection with subsequent laser catapulting. Furthermore, the present invention relates to a device for processing recorded in the receiving unit biological objects, in particular for performing a cyclic heat treatment, for example, for a polymerase chain reaction ("Polymerase Chain Reaction", PCR) and a method for transferring the biological objects from the recording unit in an associated multi-container element such as a microtiter plate.

The so-called "Laser Pressure Catapulting" (LPC) described in the Applicant's WO 97/29355 A is an efficient method for sorting and obtaining individual biological objects which are arranged on a planar support, in particular a selected biological object separated from a surrounding further biological mass by a laser beam and then catapulted the thus free-prepped biological object then by means of a laser pulse from the carrier to a collecting device.

To obtain a larger number of biological objects, a device is proposed in WO 01/73397 A1 of the Applicant, in which biological objects are automatically catapulted into a plurality of containers, for example a so-called microtiter plate. Such a device is shown in simplified form in FIG.

In the arrangement shown in FIG. 8, a biological mass 35, for example a cell culture, is arranged on a light-transmitting slide 34. Above the slide 34, a microtiter plate 30 (shown as a detail in cross-section) is mounted such that its openings point to the slide 34. As indicated by arrows B, the microtiter plate 30 can be moved parallel to the slide 34. Furthermore, the device comprises a laser 32, for example a nitrogen laser, whose laser beam 33 can be directed and focused onto the biological mass 35 via a microscope optical system (not shown). The microscope optics serves at the same time for optical control and for selecting selected sections of the biological mass 35.

The selected sections are cut out with the laser beam 33 and then catapulted out of the biological mass 35 by a targeted laser pulse. This catapult can be targeted in such a way that the biological object 36 thus released lands in a selected depression of the microtiter plate 30. Since the distance over which targeted catapulting of the biological objects 36 is possible (optimally up to 5 mm) is limited, it is necessary to fill the microtiter plate to a corresponding height with a recording medium 31 on which the biological objects 36 stick to it. The occupied by the recording medium 31 volume is not readily available for subsequent processes or the recording medium must first be resolved for this purpose. In addition, different filling levels of the recording medium 31 are necessary for different geometries of the microtiter plate 30 or other recording units having a multiplicity of depressions, such as so-called multiwell plates. Furthermore, the volume necessary for further processing or analysis of the biological objects 36 depends on the subsequent processing method. Therefore, microtiter plates 30 need to be individually filled with the recording medium 31 according to dimensions and intended use of the biological objects, which is expensive.

As an alternative, the biological objects 36 can be collected in individual collection containers such as, for example, reaction vessels and from there, for example by means of manual pipetting, be transferred into corresponding microtiter plates for further sample processing or analysis. Such manual pipetting is time consuming and involves the risk of contamination of the sample.

It is therefore an object of the present invention to provide a recording unit, which allows an optimal distance for the described laser catapulting and a simple further processing of the correspondingly obtained biological objects in commercially available multicontainer elements such as microtiter and microwell plates. Accordingly, it is a further object of the present invention to provide a corresponding method for transferring the biological objects from the receiving unit into such a multi-container element. In addition, it is an object of the present invention to provide an apparatus for easily and efficiently processing or analyzing the biological objects accommodated in such a receiving unit. In particular, a device for the particular cyclic heating of biological objects, which are received in the receiving unit, are provided. Such cyclic heating is needed, for example, to carry out the polymerase chain reaction (PCR), a process for amplifying DNA strands.

These objects are achieved by a receiving unit according to claim 1, claim 4, claim 11 or claim 18, an apparatus according to claim 30 and a method according to claim 41, respectively. The dependent claims each define preferred or advantageous embodiments.

According to the invention, a biological object receiving unit is proposed which comprises a plate-shaped element having a plurality of openings for receiving the biological objects passing from a first side of the plate-shaped element to a second side of the plate-shaped element. According to a first aspect of the invention, the receiving unit is adapted to cooperate with an associated multi-tank element which comprises a plurality of containers, wherein the openings of the receiving unit are arranged corresponding openings of the container of the container element. Such multi-container elements may be, for example, commercially available microtiter or multiwell plates.

According to an advantageous and preferred second aspect of the present invention, the openings contain a biological object receiving medium, for example a hydrogel or a liquid. The recording medium can contain means for further processing or analysis of a recorded biological object, for example means for carrying out a PCR method.

According to an advantageous and preferred third aspect of the present invention, a film is stretched on one side of the plate, which is preferably designed such that biological objects remain adhered to the side of the film facing the openings. If necessary, the film can be pretreated chemically or physically. For the film, a material is preferably used whose strength can be reduced by a treatment, for example by irradiation with ultraviolet radiation. The However, film can also be chosen so thin that it can be torn off without such a treatment, for example by means of a punch.

Of course, the above aspects of the present invention are freely combinable with each other.

According to a fourth aspect of the present invention, there is provided a biological object accommodating unit comprising a plate-shaped member having a plurality of apertures opened toward a first side of the plate-shaped member, which are closed on a second side of the plate-shaped member with a bottom. These openings are designed according to the above-described first aspect of the invention for cooperation with an associated multi-container element, wherein the openings of the receiving unit are arranged corresponding to the openings of the container of the container element.

The bottom of the receiving unit according to the fourth aspect of the present invention is preferably thinner than 0.5 mm and is optically transparent, i. for light radiation used for analysis purposes, in particular laser radiation, permeable.

Also, the openings of the receiving unit according to the fourth aspect may include a biological object receiving medium according to the second aspect of the present invention.

Generally, the plate-shaped element is preferably between 1 and 3 mm thick and made of a plastic such as polycarbonate.

The side walls of the plate-shaped element are preferably inclined to the vertical, for example by an angle of about 9 °, which facilitates the handling of the plate-shaped element.

Such a receiving unit with a plate-shaped element has the advantage that biological objects can be conveyed in the openings in a simple manner, for example by means of laser pressure catapulting. The receiving unit can then be placed on a corresponding multi-container element, and the biological objects can - for example, by centrifuging or by treatment with a punch, which is adapted to the openings of the plate-shaped element - are transported into containers of the multi-container element for further processing there. However, it is also possible to carry out at least a first processing of the biological objects in the plate-shaped element itself. It is particularly advantageous if the plate-shaped element according to the third aspect of the invention is covered with a foil or according to the fourth aspect has a bottom, since so small containers are formed.

A preferred application here is the implementation of the polymerase chain reaction. For this purpose, the required substances are either added to the openings, or they are already present in an existing according to the second aspect of the invention recording medium. Preferably, the plate-shaped element is then covered on the other side with a film to prevent evaporation. This film may in particular be a heating foil for carrying out thermal cycles or a heat-permeable foil. In the second case, the receiving unit is then placed in a device according to the invention with a flat heating element in order to carry out necessary heat cycles for carrying out the polymerase chain reaction. The field of use of the device is not limited to receiving units according to the invention, but the device can in principle be used together with any flat receiving units.

The invention will be explained in more detail below with reference to the accompanying drawings with reference to preferred embodiments. Show it:

1 is a plan view of a first embodiment of an inventive recording unit,

FIG. 2 is a cross-sectional view taken along line A-A of FIG. 1; FIG.

3A is a cross-sectional view of a second embodiment of a receiving unit according to the invention corresponding to the cross-sectional view of Fig. 2,

3B is a cross-sectional view of a third embodiment of a receiving unit according to the invention corresponding to the cross-sectional view of Fig. 2,

4A to 4C process steps for obtaining biological objects using a recording unit according to the invention,

5A and 5B process steps for processing biological objects located in the recording unit according to the invention, 6 shows schematically a device for covering a receiving device according to the invention with a film,

7 shows an exemplary embodiment of a device according to the invention for cyclically heating the receiving unit according to the invention and biological objects located therein, and

8 shows a device for obtaining biological objects according to the prior art.

In Fig. 1 is a plan view of a first embodiment of a receiving unit according to the invention for biological objects is shown. The receiving unit consists essentially of a plate 1, which may be made of a plastic such as polycarbonate and preferably has a thickness between 1 and 3 mm. In principle, however, other materials and thicknesses of the plate 1 are conceivable. The plate 1 has a plurality of openings 2, which in the present embodiment are arranged in a rectangular matrix and pass from an upper side of the plate to an underside of the plate. The openings 2 are selected in size, number and shape such that they fit to container openings of a multi-container element such as a microtiter plate or a multiwell plate. In the case of a commercially available 96-well microtiter plate with round container openings arranged in eight rows of 12 containers, the plate 1 would also comprise 96 openings 2 in eight rows of 12 openings, with size, shape and position of the openings of size, position and shape of the openings would correspond to the container of the microtiter plate. The same applies to so-called multiwell plates or other multi-container elements. The plate may additionally have a positioning aid or marking, for example in the form of a chamfer 22. By means of such a positioning marking, the recording unit according to the invention can be aligned with a multi-tank unit assigned to it. In addition, such a positioning mark is helpful to avoid confusion between openings, which are located in corresponding positions.

The openings 2 each contain a recording medium 3. As can be seen in particular in the cross-sectional view of FIG. 2 along a line AA of FIG. 1, the recording medium 3 can in particular fill the entire area of the openings 2, which can be seen in FIG. But it is also conceivable that the openings 2 only partially through the recording medium 3 are filled, or that the recording medium 3 completely fills the openings 2.

As a recording medium 3, for example, a hydrogel is suitable, wherein hydrogel in this context any substandard is to be understood, which can form colloidal dispersions with water and solidify, for example by gelling. An example of this is agarose. If the openings 2 are not too large, a liquid is conceivable as a recording medium, which adheres to the edges of the openings 2 by adhesion forces. As the recording medium 3, for example, any chemically inert substance having adhesiveness can be used.

Since, as will be described later, the recording medium 3 serves to record biological objects, which are subsequently processed further, it is preferable that the recording medium 3 is inert with respect to the planned further processing of the biological object, that is, not influenced by subsequent analysis reactions and the like. Alternatively, the recording medium may also contain substances which serve the subsequent processing. For example, buffers or pre-batches can be incorporated in a hydrogel as the recording medium.

It should also be noted that the recording medium 3 is filled depending on the durability already in the production of the plates 1 or shortly before the use of the receiving unit in the openings 2.

FIG. 3A shows a cross-sectional view of a second exemplary embodiment of a receiving unit according to the invention, the view corresponding to that of FIG. 2. The second embodiment of FIG. 3A is constructed substantially like the receiving unit according to the first embodiment, that is, it comprises a plate 1 with openings 2 according to the first embodiment. In contrast to this, the openings of the disk 1 according to the second embodiment do not contain a recording medium. In the embodiment of Fig. 3A is further on one side (in Fig. 3A of the underside) of the plate 1, an optionally pretreated film 4 stretched over the openings 2 in order to close them. In this case, the film 4 is preferably adhesive on the side 2 facing the openings, so that biological objects conveyed into the openings 2 adhere to the film 4. The term "film" is to be interpreted broadly in this context, it may be a single or double membrane or even a material with a thickness in the millimeter range.Of course, the first and the second embodiment are also combinable, that is, both the film 4 and the recording medium 3 may be provided. In this case, the film 4 need not have any special adhesive properties.

The film 4 may for example be laminated to the plate 1.

In Fig. 3B is a cross-sectional view of a third embodiment of a receiving unit according to the invention is shown. The view again corresponds to that of Figures 2 and 3A. The third exemplary embodiment illustrated in FIG. 3B differs from the second exemplary embodiment illustrated in FIG. 3A in that, instead of the film 4 from FIG. 3A, a bottom 37 is now provided, which is preferably integrally or integrally formed with the plate 1. The bottom has a thickness of preferably ≦ 0.5 mm and is optically transparent. As a result, for example, fluorescence measurements as described later with reference to FIG. 7 are possible. In addition, with a permeability to the laser radiation used for the laser pressure catapulting described above, it is possible to carry out further catapult processes or pulse-like transport processes from the recording unit according to the invention.

For carrying out the above-mentioned fluorescence measurements, it is advantageous if the recording unit according to the invention is made of a plastic material which itself has no fluorescence.

Also in the embodiment of FIG. 3B, a recording medium may be disposed in the openings 2 as in the first embodiments, or the floor 37 may be provided with an adhesive layer on the side of the openings 2 so that biological objects adhere to the floor 37.

In general, the receiving units according to the invention of FIGS. 1-3B should be made of a material which does not impair the biological objects and, if appropriate, any reactions carried out with them in the receiving unit. Generally, it is advantageous in the receiving units shown in Figures 1-3B, when the side walls of the plate 1 with respect to the vertical slightly, for example between 5 ° and 15 °, preferably 9 °, beveled, as this is the handling of the receiving unit when inserted into a Holder in a microscope arrangement or even when coupling with a multi-container element such as a microtiter plate as described below. The openings 2 can also open slightly to one side, preferably to the side from which the receiving unit is later filled with biological objects. For this purpose, the side walls of the openings 2 can be inclined at an angle of 2 ° to 7 °, preferably 4 °, with respect to the vertical.

Furthermore, it is advantageous if the individual openings of the receiving unit are labeled so as to facilitate, for example, logging which biological object is introduced into which opening. The inscription of the opening can correspond to that of the already mentioned assigned multi-container element. Such a label may for example denote the rows of openings 2 in Fig. 1 with letters A, B, etc. and the columns of openings with numbers 1, 2, 3, etc.

In the following, it will now be described with reference to FIGS. 4A to 4C how the recording unit according to the invention can be used for obtaining and subsequently processing biological objects.

FIG. 4A shows a first step of a method according to the invention for obtaining biological objects. For this purpose, a device corresponding to that already described in the introduction to the description with reference to FIG. 8 is used, in which a biological mass 6 is arranged on a transparent slide 7. By means of a laser beam 9 generated by a laser 8 parts of the biological mass 6 can be cut out and then catapulted with a targeted laser pulse in a filled with the recording medium 3 opening 2 of the receiving unit of FIGS. 1 and 2. The recording unit according to the invention can be moved according to arrows B on the slide 7 to select an opening 2, in which a respective biological object 5 is to be catapulted. The method of laser pressure catapulting itself is explained in detail in the documents cited in the introduction to the description and will therefore not be described in detail here.

Compared to the method described in the introduction to the description with reference to FIG. 8, the use of the receiving unit according to the invention offers the advantage of an optimal distance for the catapulting operation. In addition, a visual control of the biological objects 5 located in the openings 2 is possible in a simple manner, since the biological objects 5 are close enough to an objective also used to focus the laser beam 9 (not shown) in order to be able to focus on them and They can thus be observed through the lens. In general, however, the biological objects can also be introduced in other ways into the openings 2 of the receiving unit, for example by pipetting.

In a next step, which is shown in Fig. 4B, the receiving unit is then placed on an associated multi-container unit, for example a microtiter plate 10 with a plurality of containers or recesses 10A. In this case, the receiving unit is positioned such that in each case an opening 2 is arranged above a recess 10A. The placement of the receiving unit can in principle both be such that the biological objects 5 are remote from the microtiter plate 10, as well as such that they are facing her. In principle, it is also possible to use the combination of receiving unit and microtiter plate 10 shown in FIG. 4B in an already connected manner in a device for laser pressure catapulting, such as that shown in FIG. 4A. In the receiving unit according to the third embodiment of FIG. 3B, however, it is preferable to position the bottom 37 on the side facing away from the microtiter plate 10, since otherwise the bottom 37 would have to be broken for the transfer of the biological objects in the microtiter plate 10 described below ,

Subsequently, the arrangement shown in Fig. 4B is inserted into a conventional microtiter centrifuge. During centrifugation, the biological objects 5 are centrifuged together with the respective recording medium 3 into the respectively assigned containers 10A, which leads to the result shown in FIG. 4C. Since the recording medium 3 has only a small volume, essentially the entire volume of the containers 10A is available for subsequent processing steps of the biological objects 5 in the microtiter plate 10. Since, as already mentioned, the recording medium is preferably inert, it does not interfere with the following processing. The plate 1 can then either - in the case of a disposable plate - be disposed of or refilled with a recording medium after appropriate cleaning to be reused.

As an alternative to centrifuging, it is also conceivable to dissolve the recording medium 3, for example by heating, so that the biological objects 5 fall under the action of gravity into the containers 10A. Another alternative is the use of a punch, which presses the biological objects 5 together with the respective recording medium 3 in the associated container 10A. Such a punch may for example be in the form of a roll with corresponding nubs which are adapted to the openings 2, wherein the roll is rolled over the plate 1. If not, as shown in FIGS. 4A to 4C, the first embodiment of FIGS. 1 and 2, but the second embodiment of the collecting unit of FIG. 3A is used, it is possible with a correspondingly thin-selected film 4, this also by Centrifuging together with the respective biological object 5 into the recesses 10A transfer. Optionally, the film 4 tears only partially and thus remains hanging on the plate 1. It is also conceivable to use films which, for example, can be changed by irradiation with ultraviolet radiation in such a way that they become brittle in order to facilitate the subsequent centrifuging. Finally, the use of a punch as described above is also possible with foils.

In the third embodiment of Fig. 3B, as already mentioned, the bottom 37 is preferably positioned on the side facing away from the microtiter plate 10, so that z. B. the centrifugation described above is readily possible.

It should be noted that instead of the microtiter plate 10, a single container whose container opening covers a plurality of openings 2 can be used if several biological objects 5 are to be further processed in a single container.

In the case of the embodiment of FIG. 3A or 3B, it is possible to carry out at least part of the reactions necessary for processing the biological objects directly in the recording unit according to the invention. An example of this is shown schematically in FIGS. 5A and 5B. FIG. 5A shows a section of the recording unit according to the invention of FIG. 3A after being loaded with biological objects 5. A digestive solution 11 is now added to the individual openings, which results in the biological objects 5 being "digested", which means that, for example, individual DNA strands 12 are present in the solution 11. These DNA strands 12 can then be combined are transferred into a microtiter plate with the solution 11 as described above.

It is also possible to carry out a polymerase chain reaction directly or additionally to the "digesting" of biological objects shown in FIGS. 5A and 5B directly in the receiving device according to the invention provided.

In this case, the receiving unit according to the invention from FIG. 2 on both sides or the receiving unit according to the invention from FIG. 3A or 3B is advantageously also on the upper side covered with a film 4 to avoid evaporation effects during heating. A suitable device is shown schematically in FIG. Here, a film 4 is unwound from a roll 13 and pressed onto the plate 1 of the receiving unit. The film may either comprise adhesive so that it will adhere to the plate 1 automatically, or an additional adhesive may be used. The covering with the film 4 can be done automatically in a corresponding device or manually.

Instead of a flexible film which can be unwound from the roll 13, a thicker, rigid film is also conceivable, which is placed on the plate 1 and attached to it e.g. is fixed by lamination.

In Fig. 7, an embodiment of a device according to the invention for cyclically heating a receiving unit according to the invention is shown. The device shown in Fig. 7 comprises a flat heating element 16 with openings 17 which are arranged according to the openings 2 of the plate 1 of the receiving unit according to the invention. The planar heating element 16 may be constructed, for example, from Peltier heating elements. But it is also conceivable to use a heating foil. If this is transparent, the openings 17 are dispensable. A control unit 20 controls or regulates a temperature of the planar heating element 16. For this purpose, for example, a desired temperature profile can be predetermined. For example, for the polymerase chain reaction typically a plurality of heating cycles with temperatures between about 96 ° C for the separation of double-stranded DNA, 37 ° C to 65 ° C for the attachment of so-called primers and DNA polymerase to the individual DNA strands and 65 ° C to 80 ⁰ C for so-called elongation, in which the respectively missing second strand of DNA is generated, go through.

The preferably covered with two sheets 4 as evaporation protection plate 1 is placed directly on the flat heating element 16. In the openings 2 of the plate 1 are biological objects 5 - optionally after a "digestion" as shown in Fig. 5A and 5B -, the collecting means 3 and / or additionally added substances 21. If a suitable hydrogel is used as the recording medium liquefies It is particularly advantageous if the substances required for the reaction to be carried out, in the case of the polymerase chain reaction, in particular the DNA polymerase, are incorporated in the hydrogel, in which case these substances become liquid during the liquefaction of the hydrogel This saves an additional step in which the substances 21 would have to be introduced into the openings 2, which reduces the risk of contamination. additionally or alternatively required substances 21 in a separate step before pipetting shown in Fig. 6 with foil in the openings 2.

To control the running in the openings 2 of the plate 1 reaction, a lighting device with a light source 14 and an optical system 15 is provided, which preferably illuminates the plate 1 over its entire width. Furthermore, a detection device 18 with an evaluation unit 19 is provided.

In the case of the polymerase chain reaction, for example, a fluorescent dye is added to the openings (either incorporated into the recording medium or separately), for example SYBR-Green, which is incorporated into the resulting DNA during the reaction in order to monitor the detection in real time. This dye is then excited by the illumination device 14, 15 to fluoresce, which are detected by the openings 17 in the planar heating element 16 of the detection device 18. The intensity of fluorescence is a measure of the number of DNA strands generated.

Various embodiments of the detection device 18 are conceivable. On the one hand, the detection device 18 can be designed as a single planar detector, which measures the total fluorescence that arises in the openings 2. In this case, the evaluation unit 19 can determine the corresponding reaction sequence for each of the openings 2. However, it is also conceivable that the detection device 18 comprises a plurality of detection fields, each detection field being associated with an opening 2 of the plate 1. Hereby, an individual detection of the course of the reaction in the individual openings 2 is possible. For example, a CCD camera may be used as the detector. In this variant, it is also conceivable that the fluorescence signal is conducted by means of glass fibers to corresponding detectors. Finally, it is also conceivable that the detection device 18 is designed as a scanner, which the individual openings 2, e.g. scanned sequentially by means of a lens. Also in this case, the evaluation unit 19 can determine the course of the reaction for each chamber separately.

Instead of the transmitted light measurement of the fluorescence, a reflection fluorescence measurement is also conceivable. In this case, the illumination device 14, 15 would be arranged on the same side of the plate 1 as the detection device 18.

The illustrated device for cyclic heating has the particular advantage that it very compact and in particular by the use of the recording unit according to the invention can be carried out flat, with a desired reaction in a plurality of openings 2 can take place simultaneously, which means a considerable time savings.

If a heating foil 16 is used instead of Peltier elements as a flat heating element 16, it can also be stretched directly on the plate 1 as a replacement for one of the foils 4.

In order to ensure the best possible distribution of the heat generated by the flat heating element 16 to the openings 2, it is advantageous that the plate 1 and the films 4 are made of a sufficiently thermally conductive material. For the fluorescence measurements, the films 4 should also be translucent.

It is to be noted that for use in the cyclic heating apparatus of Fig. 7, the openings 2 of the receiving unit may be loaded with biological objects 5 other than by the described laser catapulting, for example by pipetting.

Due to the space-saving design, it is possible to integrate the device of FIG. 7 in a microscope assembly for performing a laser microdissection or a laser catapulting, so that both the extraction and the further processing of the biological objects in a compact device can be carried out. For this purpose, for example, the planar heating element 16 may be integrated in a holder for the receiving unit according to the invention.

In addition, the device may comprise the device shown in Fig. 6 schematically for covering the plate 1 with the film 4.

After carrying out the reaction in the device from FIG. 7, the reaction product, in the case of the polymerase chain reaction also referred to as amplificate, can be transferred into a microtitre or multiwell plate according to the method shown in FIGS. 4A to 4C and further processed there. to be analyzed.

Claims

PAT EN TA NSPR O CHE

A biological object holding unit (5) comprising a plate-shaped member (1) having a plurality of openings (2) for receiving the biological matter from a first side of the plate-shaped member (1) to a second side of the plate-shaped member (1) Objects, wherein the openings (2) corresponding to container openings of containers (10A) of an associated multi-tank element (10) are arranged.

2. Recording unit according to claim 1, characterized in that a shape of a cross section of the openings (2) and / or a number of openings (2) corresponds to a shape or a number of container openings of the associated multi-container element (10).

3. Recording unit according to claim 1 or claim 2, characterized in that the associated multi-container element is a microtiter plate or a multiwell plate.

A biological object holding unit comprising a plate-shaped member (1) having a plurality of openings (2) passing from a first side of the plate-shaped member (1) to a second side of the plate-shaped member (1), and having one in the openings (2) arranged recording medium (3) for receiving the biological objects.

5. Recording unit according to claim 4, characterized in that the recording medium (3) comprises an adhesive, in particular a hydrogel.

6. Recording unit according to claim 4 or 5, characterized in that the recording medium (3) comprises a liquid.

7. Recording unit according to one of claims 4 to 6, characterized in that the recording medium (3) completely fills a cross-sectional area of the openings (2).

8. Recording unit according to one of claims 4 to 7, characterized in that the recording medium (3) comprises means for processing the recorded biological objects (5).

9. Recording unit according to claim 8, characterized in that the means for processing the biological objects (5) comprise means for carrying out a polymerase chain reaction.

10. Recording unit according to one of claims 4 to 9, characterized in that the receiving unit is designed according to one of claims 1 to 3.

A biological object holding unit comprising: a plate-shaped member (1) having a plurality of openings (2) for receiving the biological objects passing through from a first side of the plate-shaped member (1) to a second side of the plate-shaped member (1); with cover means (4) applied to the first side of the plate-shaped element (1), the covering means (4) being applied so as to cover the openings (2).

12. A receiving unit according to claim 11, characterized in that the cover means (4) on one of the second side of the plate-shaped member (1) facing side comprises an adhesive.

13. Recording unit according to claim 11 or 12, characterized in that the covering means (4) is designed such that the strength of the covering means (4) can be reduced by irradiation.

14. Recording unit according to one of claims 11 to 13, characterized in that the covering means (4) is translucent.

15. Recording unit according to one of claims 11 to 14, characterized in that the covering means (4) comprises heating means.

16. Recording unit according to one of claims 11 to 15, characterized in that the covering means comprises a preferably chemically or physically treated film (4).

17. Recording unit according to one of claims 11 to 16, characterized in that the receiving unit is designed according to one of claims 1 to 10.

18. A biological object receiving unit (5), comprising a plate-shaped element (1) having a plurality of openings (2) formed on a first side of the plate-shaped element (1) and extending on a second side of the plate-shaped element (1) Floor (37) are completed, wherein the openings (2) corresponding to container openings of containers (10 A) of an associated multi-tank element (10) are arranged.

19. Recording unit according to claim 18, characterized in that the plate-shaped element (1) and the bottom (37) are integrally formed.

20. Recording unit according to claim 18 or 19, characterized in that the bottom (37) is thinner than 0.5 mm.

21. Recording unit according to one of claims 18 to 20, characterized in that the bottom (37) is translucent.

22. Recording unit according to one of claims 18 to 21, characterized in that the openings (2) comprise a recording medium.

23. Recording unit according to claim 22, characterized in that the recording medium is designed according to claim 5 or 6.

24. Recording unit according to one of claims 18 to 23, characterized in that a shape of a cross section of the openings (2) and / or a number of openings (2) corresponds to a shape or a number of container openings of the associated multi-container element (10).

25. Recording unit according to one of claims 18 to 24, characterized in that the associated multi-container element is a microtiter plate or a multiwell plate.

26. Recording unit according to one of claims 1 to 25, characterized in that the plate-shaped element (1) has a thickness between 1 and 3 mm.

27. Recording unit according to one of claims 1 to 26, characterized in that the plate-shaped element (1) is made of a plastic material.

28. receiving unit according to claim 27, characterized in that the plastic material comprises polycarbonate.

29. Recording unit according to one of claims 1 to 28, characterized in that the receiving unit positioning means (22) for positioning the receiving unit on an associated multi-container element (10).

30. Apparatus for the thermal treatment of biological objects (5) located in openings of a receiving unit according to one of claims 1 to 29, with heating means (16) having a heating surface which is at least the area of the first

Side or the second side of the receiving unit, wherein the receiving unit with its first and second sides flat on the

Heizoberfiäche can be applied, and with control means (20) for controlling a predetermined temperature profile of

Heating means (16).

31. The device according to claim 30, characterized in that the heating means (16) comprise Peltier elements.

32. Apparatus according to claim 30 or 31, characterized in that the heating means (16) comprise a heating foil.

33. The apparatus of claim 32, characterized in that the heating foil is configured such that it can be fastened to cover the openings (2) of the receiving unit to the receiving unit.

34. Device according to one of claims 30 to 33, characterized in that the heating means (16) at least at locations at which openings in the surface of the receiving unit to the heating means (2) of the receiving unit are positioned, is translucent.

35. Apparatus according to claim 34, characterized in that the heating means (16) are transparent.

36. Device according to one of claims 30 to 35, characterized in that the device comprises illumination means (14, 15) for illuminating the biological objects (5) in the receiving unit.

37. Device according to one of claims 30 to 36, characterized in that the device comprises detection means (18) for detecting a light emission in the recording unit.

38. Apparatus according to claim 37, characterized in that the detection means comprise a plurality of detection units, wherein in each case a detection unit of an opening (2) of the receiving unit is associated.

39. Device according to one of claims 30 to 38, characterized in that the device comprises means (13) for covering openings (2) of the receiving unit with a covering means (4).

40. Apparatus according to claim 39, characterized in that the cover center! a foil (4).

41. A method for obtaining biological objects (5), wherein a biological object (5) in an opening (2) of a receiving unit according to one of claims 1 to 29 is introduced, wherein the receiving unit is coupled to a container element such that the biological object (5) containing opening (2) of the receiving unit with a

Container opening of the container element (10) corresponds, and wherein the biological object (5) is transferred from the opening (2) in the container element.

42. The method according to claim 41, characterized in that the container element (10) is a multi-container element with a plurality of containers

(10A) with respective container openings, each opening (2) of the receiving unit corresponding to a respective container opening.

43. The method according to claim 41 or 42, characterized in that the biological object (5) is transferred by centrifuging into the container element (10).

44. The method according to claim 41 or 42, characterized in that the biological object (5) is transferred by mechanical action into the container element (10).

45. The method according to claim 44, characterized in that the mechanical action is exerted by a punch.

46. The method according to any one of claims 41 to 45, characterized in that the biological object (5) is introduced by a targeted laser pulse in the opening (2).

47. Method according to one of claims 41 to 46, characterized in that before the coupling of the receiving unit with the container element (10), a reaction is carried out involving the at least one biological object (5) in the receiving unit.

48. The method according to claim 47, characterized in that the reaction comprises a digestive reaction.

49. The method of claim 47 or 48, characterized in that the reaction comprises a polymerase chain reaction.

50. The method according to claim 49, characterized in that the polymerase chain reaction is carried out in a device according to one of claims 30 to 40.

51. The method according to any one of claims 41 to 50, characterized in that the container element (10) is a microtiter plate or a multiwell plate.

52. The method according to any one of claims 41 to 46, characterized in that the method is carried out simultaneously with a plurality of located in a plurality of openings (2) of the receiving unit biological objects (5).