Device for obtaining sample material

Abstract

The invention describes a device for obtaining sample material (1) from a sample (2). A control device (10) controls a cutting tool (3) such that the cutting tool (3) acts mechanically on the sample (2) within a boundary (4) of the sample material (1) to be obtained, in a mechanically oscillating state. The control device (10) also controls a removal device (9) to receive the sample material (1).

Description

[0001] The invention relates to a device for obtaining sample material from a sample. The sample is, for example, a biological sample and is preferably a thin tissue section.

[0002] Obtaining sample material from thin tissue sections is an important step in many areas of medical and biological research, as well as diagnostics. This is usually performed manually by specially trained personnel. However, when very small tissue sections or even individual cells or cell components are to be removed, manual preparation reaches its limits. The high time consumption also hampers high-throughput applications.

[0003] Laser microdissection systems have become established for the preparation of particularly small tissue fragments from thin sections. These systems are based on the following principle: Using a laser focused on the thin section, the sample material is severed at defined locations. A suitable removal mechanism transfers the excised tissue into a designated sample container. The removed tissue can then be further processed.

[0004] DE 10 2011 001 474 A1 describes a device for laser microdissection in which a user specifies a cutting line which is then automatically followed.

[0005] Many laser dissection systems use UV lasers with a wavelength of approximately 355 nm. The wavelength is selected to minimize energy input into the tissue, preventing significant heating or associated tissue damage. Furthermore, the selected wavelength of light is intended to prevent nucleic acid alterations. However, it has been shown that the yield and quality of tissue cut with a laser are lower compared to mechanical preparations.

[0006] An alternative device for tissue microdissection is disclosed in DE 199 32 032 C2. In this device, a needle is set into vibration, which facilitates tissue transection. It is also proposed that an acoustic sensor register whether the needle tip touches the sample carrier. The needle movement is controlled by skilled personnel with electronic assistance. The tissue fragment is then removed using a micropipette.

[0007] It is also known to cut tissue samples with a milling cutter (see the discussion of the prior art in EP 4 177 662 A1). A rinsing device is known for this purpose, which flushes the treated area with a special solution and collects the liquid together with the comminuted tissue fragments. The size of the milling cutter represents a limiting factor for the smallest diameter (approximately 200 µm) of the areas to be cut.

[0008] DE 101 47 950 A1 describes a device for holding samples. An adhesive medium is cooled on a pipette until an adhesive force is exerted on the sample. The pipette is moved relative to the sample, or the sample is moved relative to the pipette.

[0009] WO 98 / 36261 A1 discloses a transfer film made of a material that absorbs light in the infrared range and melts in the process. If the film is used as part of a stamp, the cut tissue fragment can stick and be removed from the thin section. Using an IR laser to fix the cut sample material to a support increases the system's cost and complexity, making it more susceptible to failure. Furthermore, the stamp system carries the risk that tissue fragments will not be completely captured if the tissue does not adhere completely to the stamp due to its material properties. Removal using this type of contact method also increases the risk of contamination.Heating of the polymer stamp by the IR laser can also prove problematic for certain sample types, as it can lead to nucleic acid changes and thus reduce the significance of certain testing methods.

[0010] It is also known to apply the sample material to the underside of a support. Once the support and sample are cut to size, the aforementioned stamp can be used to hold the film and sample section.

[0011] In WO 97 / 29355 A1, a laser beam catapults the tissue sample into a vessel without contact. The UV laser is focused below the previously cut area, triggering a shock wave. The tissue to be removed is exposed to laser radiation during both cutting and removal, which increases the likelihood of nucleic acid alterations. Furthermore, there is a risk that the tissue fragment cannot be transferred correctly if the laser is unable to completely remove the fragment or if the tissue material remains (partially) stuck together due to its composition.

[0012] In DE 100 43 506 C1, the excised tissue sample is drawn into the vessel by gravity. The thin sections are applied to PEN (polyethylene naphthalate) membrane slides.

[0013] The object underlying the invention is to improve the preparation of tissue samples in terms of reliability and at the same time to enable the highest possible throughput.

[0014] The invention solves the problem by a device for obtaining sample material from a sample, wherein the device has at least one separating tool, a removal device and a control device, wherein the control device controls the separating tool in such a way that the separating tool, in a mechanically oscillating state, mechanically acts on the sample at least in a region of a boundary of the sample material to be obtained, and wherein the control device controls the removal device in such a way that the removal device receives the sample material.

[0015] The device according to the invention allows, in particular, microdissection of, for example, tissue fragments or cells from samples, which may be, for example, thin tissue sections. To obtain the sample material, a cutting tool mechanically acts on the sample within a defined boundary. The action preferably involves cutting, severing, or perforating the sample. In particular, no laser is used, so that laser-induced heating of the sample or changes in nucleic acids cannot occur. The sample material is essentially subjected to minimal stress and therefore provides an excellent basis for further investigations.

[0016] To separate the sample material from a defined area of ​​the sample, the cutting tool is moved relative to the sample by the control device, traversing the boundary—i.e., the cutting edge—once or several times, if necessary. In one embodiment, a complete separation of the sample material from the rest of the sample is achieved.

[0017] Control is dependent on data regarding the boundaries of the sample material. This data can be, for example, coordinates or coordinates in conjunction with size specifications relating to the area to be removed. For the data input – e.g., by the operating personnel – an overview image of the sample is taken, which serves as the basis for plotting cutting edges and parameters. In one embodiment, the control device automatically determines and implements data on the cutting movements and, if necessary, further work steps from the cutting edges thus specified. Alternatively or additionally, parameters for the extraction of other sample materials can be adopted or adapted.

[0018] To control the movement, one design involves the cutting tool being fixed in space, and the sample being moved—e.g., using a motorized linear stage. Alternatively, the sample is fixed in space, and the cutting tool is moved. The two variants can also be combined.

[0019] The cutting tool performs mechanical vibrations during the action. Depending on the design, these vibration modes are transverse and / or axial. The vibration form can be a pure sinusoidal vibration or a superposition of sinusoidal vibrations of different frequencies, phases, and / or amplitudes. In one design, the frequency of the fundamental vibration at which the cutting tool oscillates depends on the type of sample and is specified accordingly. For example, the frequency is in the ultrasonic frequency range and is therefore greater than 20 kHz. The control device controls the relative movement between the cutting tool and the sample. Automating the movement increases accuracy and simplifies processing. Furthermore, greater throughput is possible. Furthermore, the invention enables remote control and thus also use in telemedicine.The selected cutting technique also allows very small areas to be prepared, especially those with a diameter of less than 20 µm.

[0020] The control device also controls the sampling device, which collects the sample material from the sample. Thus, the sampling of the sample material from the sample is also automated. This increases the overall speed of sample processing and sample extraction.

[0021] Overall, the device according to the invention allows the components involved in microdissection to be controlled automatically, significantly increasing accuracy and throughput. The user is only required to define the sample areas to be extracted and to equip them with consumables such as pipette tips or sample containers, depending on the respective design.

[0022] To facilitate the removal of the sample material, e.g. from a slide, or to be able to collect the sample material, e.g. using a pipette as part of the sampling device, the sample material is divided into smaller fragments by the separation tool in one embodiment.

[0023] One embodiment provides for the cutting tool to be designed as a micro-tool. In another embodiment, it is a needle tool, a micro-blade, or a micro-chisel.

[0024] One embodiment includes the device further comprising an actuator, the actuator being configured to excite the cutting tool to mechanical vibrations, and the control device controlling the actuator. In this embodiment, the control device additionally controls the vibrations of the cutting tool. In one embodiment, the actuator comprises at least one piezoelectric element. Alternatively, it is a voice coil actuator.

[0025] One embodiment of the device provides that the device further comprises a sensor device, and that the sensor device is configured to determine and / or monitor a position between the cutting tool and a sample carrier, and that the control device receives data from the sensor device. In this embodiment, a sample carrier is present, on which the sample is applied or fixed for the application of the device. The position of the cutting tool relative to the sample carrier is coupled to the relative position between the cutting tool and the sample (and even specifically to the boundary of the sample material). Therefore, in this embodiment, the data from the sensor device supports the control of the movement of the cutting tool relative to the sample.

[0026] In one embodiment, an image recording device is provided that creates an image of the sample. The image of the sample—in particular, a stained or unstained thin section—is captured, for example, by a CMOS camera, which in one embodiment is connected to a microscope. In one embodiment, the recording device is part of the sensor device for determining the position of the cutting tool or is the sensor device itself. In one embodiment, the sensor device also includes an illumination device.

[0027] In one embodiment, the position of the cutting tool is at least a distance between the cutting tool and the sample carrier. In this variant, it is thus monitored in particular whether the cutting tool touches the sample carrier. It is preferably ensured that the cutting tool and a sample carrier on which the sample is located do not come into contact with each other. Care is therefore taken to ensure that the cutting tool maintains a necessary distance. In particular, damage to the cutting tool is prevented by placing it on the sample carrier, which is made of glass, for example.

[0028] One embodiment provides for the sensor device to be designed as an optical measuring device. The sensor device has a suitable imaging optics that observes the contact point of the cutting tool on the sample from a suitable direction.

[0029] Alternatively or additionally, the sensor device is designed as a capacitive measuring device or as a conductivity measuring device.

[0030] Alternatively or additionally, the sensor device is part of the actuator system.

[0031] According to a further embodiment, the sensor devices evaluate a transmission of the vibrations of the cutting tool to the sample carrier as a measure of possible contact.

[0032] One embodiment of the device includes a dispenser device, the dispenser device being configured to dispense an auxiliary medium for taking the sample material, and the control device controlling the dispenser device. The dispenser device is, for example, a dosing system that dispenses an auxiliary medium. In particular, the control device controls the position from which or to which the auxiliary medium is dispensed. Thus, either the dispenser device is moved relative to the sample carrier and / or the sample carrier is moved relative to the dispenser device. Alternatively or additionally, the quantity of the auxiliary medium is controlled. The auxiliary medium is preferably a liquid.

[0033] The dispenser device draws the auxiliary medium, for example, from a reservoir included in the device. In one embodiment, the auxiliary medium is applied in the form of—preferably small—droplets onto the area of ​​the sample to be prepared. This can be done before the sample material is cut out, as an intermediate step, or after it has been cut out.

[0034] One embodiment provides for the sampling device to collect the sample material with the auxiliary medium. In this embodiment, the auxiliary medium carries the sample material, so that the sample material is also collected when the auxiliary medium is removed.

[0035] One embodiment involves the dispenser device also serving as a withdrawal device. In this version, a common device performs the functions of both the dispenser device and the withdrawal device. These functions include, for example, two different operating modes. In one embodiment, the common dispenser / withdrawal device has two different function heads, one for dispensing the auxiliary medium and the other for collecting the sample material.

[0036] One embodiment provides for the dispenser device to be designed as a pipetting device. For this purpose, the dispenser device preferably has a suitable holder for pipette tips. In one variant, the holder is equipped with an ejection system so that different pipette tips can be picked up and ejected automatically. This allows the removal of various tissue fragments without the risk of cross-contamination. The possible use of standard consumables instead of highly specialized consumables enables cost-effective and rapid processing of thin tissue sections.

[0037] The invention is explained in more detail with reference to the following figures. Fig. 1 shows a purely schematic first embodiment of the device and Fig. 2 shows a second embodiment of the device.

[0038] In the Fig. 1 shows how a smaller area than sample material 1 can be cut out of a sample 2, which is, for example, a thin section.

[0039] The sensor device 11, which is embodied here as a camera, captures the sample 1, allowing a user to define the boundary 4 of the sample material 1 to be separated. This is done, for example, by the user tracing the boundary 4 with an input device on a recording of the sample 2. The corresponding data is transmitted to the control device 10, which automatically executes the subsequent steps.

[0040] The actual removal of the sample material 1 is performed by the cutting tool 3, which is a micro-tool shaped like a needle, blade, or chisel. Thus, a mechanical force is applied to the sample 2.

[0041] The cutting tool 3 is excited to mechanical vibrations by an actuator 12. The generation of vibrations and the guidance of the cutting tool 3 along the boundary 4 are controlled by the control device 10. The control device 10 accesses the image data from the sensor device 11.

[0042] To prevent the cutting tool 3 from being damaged by being placed on the sample carrier 7, which is made of glass, for example, a further sensor device 6 is provided, which monitors the distance between the cutting tool 3 and the sample carrier 7 and thus also between the cutting tool 3 and the sample 2. The sensor device 6 is part of the actuator 12, which registers whether the cutting tool 3, as an oscillating unit, changes its oscillation properties due to contact with the sample carrier 7.

[0043] Once the separation tool 3 has sufficiently separated the sample material 4 from the surrounding sample 2, the control device 10 allows the dispenser device 8 to release an auxiliary medium 5, preferably in the form of a liquid, onto the sample material 1.

[0044] The auxiliary medium 5 consists, for example, at least partially of a tris(hydroxymethyl)aminomethane (also called TRIS, THAM, or TRIS buffer) or of a TE buffer comprising TRIS and EDTA (ethylenediaminetetraacetic acid). The auxiliary medium 5 is preferably obtained by adding, for example, one of the aforementioned substances with a small amount of a detergent, which is, for example, a non-ionic surfactant from the group of octylphenol ethoxylates (well-known examples include Triton X-100 or Nonidet P40).

[0045] The aid 5 simplifies the task of the sampling device 9 to collect the sample material 1 and to place it in a collecting vessel 16 - not shown here - as a transport container (cf. Fig. 2) to be transferred.

[0046] Even during the removal of the sample material 1, the control device 10 monitors the process via the sensor device 11 designed as a camera to determine whether the sample material 1 separates well from the remaining sample 2.

[0047] The Fig. Figure 2 shows a more concrete design of the device for obtaining the sample material 1.

[0048] The sample 2 is located on the sample carrier 7, which rests on a work platform 13. In the illustrated embodiment, the work platform 13 accommodates pipette tips 14, a reservoir 15 for the auxiliary medium 5, and a collecting vessel 16 for the sample material 1.

[0049] Located below the work platform 13 is the sensor device 6, which optically detects from below whether the cutting tool 3 is touching the sample carrier 7. The sensor device 6 is designed, in particular, as an image recording device. In addition, the microscope, which is arranged inverted in the illustrated embodiment, can be assigned an object illumination device (not shown here), which can preferably be operated with various illumination modes. These modes include, for example, incident light / dark field with a preferably limited, contrast-enhancing wavelength range adapted to the application.

[0050] The actuator 12 is designed such that it excites the cutting tool 3 to oscillate - here along the longitudinal axis of the cutting tool 3 - and that it can move the cutting tool 3.

[0051] The dispensing device 8 and the removal device 9 are formed here by a component which uses different pipette tips 14 for dispensing the auxiliary medium 5 and for taking up the sample material 1. List of reference symbols 1 Sample material 2 Sample 3 Cutting tool 4 Boundary 5 Auxiliary medium 6 Sensor device 7 sample carriers 8 Encoder device 9 Removal device 10 Control device 11 Sensor device 12 Actuator 13 Work platform 14 Pipette tip 15 Reservoir for the auxiliary medium 16 collecting vessel QUOTES CONTAINED IN THE DESCRIPTION

[0000] This list of documents submitted by the applicant was generated automatically and is included solely for the convenience of the reader. This list is not part of the German patent or utility model application. The DPMA assumes no liability for any errors or omissions. Cited patent literature

[0000] DE 10 2011 001 474 A1

[0004] DE 199 32 032 C2

[0006] EP 4 177 662 A1

[0007] DE 101 47 950 A1

[0008] WO 98 / 36261 A1

[0009] WO 97 / 29355 A1

[0011] DE 100 43 506 C1

[0012]

Claims

[1] Device for obtaining sample material (1) from a sample (2), wherein the device has at least one separating tool (3), a removal device (9) and a control device (10), wherein the control device (10) controls the separating tool (3) in such a way that the separating tool (3) in a mechanically oscillating state acts mechanically on the sample (2) at least in a region of a boundary (4) of the sample material (1) to be obtained, and wherein the control device (10) controls the removal device (9) in such a way that the removal device (9) receives the sample material (1). [2] Device according to claim 1, wherein the separating tool (3) is designed as a micro-tool - preferably as a needle tool, micro-blade or micro-chisel. [3] Device according to claim 1 or 2, wherein the device further comprises an actuator (12), wherein the actuator (12) is designed to excite the separating tool (3) to mechanical vibrations, and wherein the control device (10) controls the actuator (12). [4] Device according to one of claims 1 to 3, wherein the device further comprises a sensor device (6), wherein the sensor device (6) is designed to determine and / or monitor a position - in particular a distance - between the separating tool (3) and a sample carrier (7), and wherein the control device (10) receives data from the sensor device (6). [5] Device according to claim 4, wherein the sensor device (6) is designed as an optical measuring device, and / or wherein the sensor device (6) is designed as a capacitive or conductivity measuring device, and / or wherein the sensor device (6) is part of the actuator system (12). [6] Device according to one of claims 1 to 5, wherein the device further comprises a dispenser device (8), wherein the dispenser device (8) is designed to dispense an auxiliary medium (5) for the removal of the sample material (1), and wherein the control device (10) controls the transmitter device (8). [7] Device according to claim 6, wherein the sampling device (9) receives the sample material (1) with the auxiliary medium (5). [8] Device according to claim 6 or 7, wherein the donor device (8) also serves as a removal device (9). [9] Device according to one of claims 6 to 8, wherein the donor device (8) is designed as a pipetting device.

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