DE102006032637A1 - Device for the information of biological samples e.g. cells/tissues, comprises sound producing mechanism, which produces sound wave in the sample to be treated, a reception for a container, mixer with a retainer for mix element, and drive - Google Patents

Abstract

The device for the information of biological samples e.g. cells/tissues, comprises a sound producing mechanism, which produces a sound wave in the sample to be treated, a reception (2) for a container (1), which receives the sample, a mixer with a retainer (3) for a mix element (4), and a drive (5) for the movement of the reception and/or the mix element. The mix element is brought into the container, when the container is arranged in the reception or in a further reception. The reception encompasses the container in cheek like. The drive is arranged for the oscillating- or horizontal movement of the reception. The device for the information of biological samples e.g. cells/tissues, comprises a sound producing mechanism, which produces a sound wave in the sample to be treated, a reception (2) for a container (1), which receives the sample, a mixer with a retainer (3) for a mix element (4), and a drive (5) for the movement of the reception and/or the mix element. The mix element is brought into the container, when the container is arranged in the reception or in a further reception. The reception encompasses the container in cheek like. The drive is arranged for the oscillating- or horizontal movement of the reception. The mix element is held in the retainer that is moved translationally or rotatively by the drive. A flexible shaft is arranged between the drive and the mixer. A sound horn, whose sound exit opening is arranged in such a manner that sound horn lies at the container wall of the container held in the reception or a further reception. The sound producing mechanism consists of an elastic element, with which the sound exit opening of the sound horn is pressed against the container wall of the container. A container receives the sample for the reception. Loose particles are present in the container for the support of the information of the biological material. An adapter is applicable in the reception.

Description

The The invention relates to a device for digestion (lysis) of biological Samples, having a sound generating device, which in the aufzuschließenden Sample, which is preferably in the form of an aqueous suspension, a sound wave, preferably an ultrasonic wave can generate.

From the US 6,100,084 is known in the field of lysis of cells the use of ultrasonic waves. The cells to be lysed are held in an ultrasound wave transmitting medium in a container. This is closed by a membrane, on which an electrode and a layer of piezoelectric material are formed. When the membrane is excited by excitation of the piezoelectric material, the ultrasound wave transmitting liquid and, above all, the cells are excited with the ultrasonic wave. This leads to the lysis of the cells.

According to US 6,100,084 For example, loose particles may be added to the solution to assist the so-called sonication (disruption of biological materials by the use of ultrasound). A comparable device for lysis of biological materials with a sound generating device, which can generate an ultrasonic wave in the suspension, is also out US 2005/0064598 A1 known. It is also described there that glass beads are added to the suspension.

In front In this background, the invention is based on the object, the to improve known devices.

• – eine Aufnahme für einen Behälter, der die Probe aufnehmen kann,
• – einen Mixer mit einer Halterung für ein Mixelement, das in den Behälter eingebracht werden kann, wobei der Behälter in der Aufnahme oder in einer anderen Aufnahme angeordnet ist und
• – einen Antrieb zur Bewegung der Aufnahme und/oder des Mixelements (4) aufweist.

This object is achieved with a device for the digestion of biological samples with a sound generating device ( 8th ), which can generate a sound wave in the sample to be treated, dissolved, the

• A receptacle for a container capable of holding the sample,
• A mixer with a holder for a mixing element, which can be introduced into the container, wherein the container is arranged in the receptacle or in another receptacle, and
• A drive for moving the receptacle and / or the mixel element ( 4 ) having.

advantageous Embodiments are specified in the subclaims.

The Invention is based on the idea in the lysis device furthermore to provide a mixer in addition to the sound generating device, with which the biological sample to be disrupted can be treated. Especially preferred is additionally intended to the sound generating device, a mixer of the type so-called Homogenisators added become.

The The device has a receptacle for a container that holds the treatment to be treated can absorb biological material. Suitable containers especially laboratory standard Sample containers such as. 50 ml Falcon tube. Furthermore, a mixer is provided, which has a holder for a mixing element, that in the container can be introduced. The container is held in place by a receptacle. A drive is provided to move the recording and / or the mixing element.

The inventive device allows it for the digestion of biological materials such as cells and tissues three different treatment methods to perform with only one device, namely the so-called "Sonication", the so-called. Homogenization and in a preferred embodiment the so-called "Bead Mill Disruption "(the Treatment of the sample with the help of loose particles, the sample to be digested enclosed and moved to treat the sample). Such particles are preferably spherical and serve the digestion of the Mechanically support sample material.

By doing with the invention, a device is provided, which is the implementation of all These three treatment methods will allow the activity of such devices using user significantly simplified. He can spontaneously for decide the best treatment method of the material to be broken up. Further deleted the need for Each treatment method in the laboratory a separate device vorzuhalten and wait. This is accompanied by the further advantage of saving space. Furthermore can all Treatment methods cumulative, i. simultaneously or in succession carried out become. For example, could the ultrasonic treatment and the rotor / stator homogenization simultaneously carried out become. Through the sequential treatment sequence of all three methods could for example improved digestion of very difficult sample materials become.

The inventive device allows Advantageously, the digestion (lysis) of biological materials, such as cells or tissues. By destroying the membrane of the cells become intracellular released biological components (for example, nucleic acids and Proteins) following the cell digestion further processed, or can be purified or analyzed.

The biological sample containing the components to be lysed, such as the cells, is preferably processed in a liquid medium, in the manner of a suspension. The bead-mill procedure However, ren also allows a digestion of a sample with or without surrounding medium. The sample may be present as a "fresh" or frozen or freeze-dried material

In the most general versions the device according to the invention Hold the Take the sample container. The receptacle is preferably designed so that the container the recording can be taken out. In a preferred embodiment the receptacle is designed such that it has a container when he is inserted into the device, surrounds like a jaw. It has it turned out that the so held in the receptacle container very well fixed and, as far as a drive is provided for the movement of the recording, to a good transfer the movement on the container suitable is. Further leaves a container very easy to insert in a jaw-like recording. Under a jaw-like receptacle is in particular a U-shaped or comparable trained recording understood. A jaw is a name to be named Example. The receptacle can be designed, for example, that she a vertical 50ml Falcon tube can hold safely and by means of an adapter usually used for the "Bead Mill Disruption" method container can record. For example, the held by the recording Adapter be designed so that he when installed, one or more of the containers used in the "Bead Mill Disruption" method horizontal lying down. For example, in the "Bead Mill Disruption" method 2.0 ml Eppendorf reaction tubes used become. For example, 6 mm balls of glass, Stainless steel or tungsten carbide used. In the digestion of bacteria and Mushrooms are sometimes used even very small glass particles. For larger quantities Material (<< 1 g) come too Edelstahlschraubbehälter with a 2 cm ball for use.

In a preferred embodiment the drive is configured for the oscillating movement of the receptacle. It has been shown that the Treatment of the aufzuschließenden Sample is done particularly well when the container is oscillating by means of the drive is moved. This is especially true of a movement in the Suspension particles in the implementation of the so-called "Bead Mill Disruption" method.

In a preferred embodiment the movement of the recording caused by the drive takes place in a horizontal direction. This also serves a particularly good Treatment of the sample, especially when using the "Bead Mill Disruption" method.

The Homogenization of the sample can be achieved by holding it in the holder Mix element done. The mixing element is in particular after the so-called

Rotor / stator design formed and has a (stationary held) mixing rod (stator) and a rotating relative to the stator rotation element, for example is located at the end of the blender bar. The rotating element has preferably cutting up. For example, the Mixelement after the design of the "Tissueruptor" distributed by Qiagen GmbH, Hilden, Germany.

In The particularly preferred design is therefore by rotation of a arranged at the end of the stator rotor causes the desired lysis. The device according to the invention but also allows other, possibly complementary to this rotation procedures, to achieve a homogenization. For example, the complete Mixelement rotated and / or moved vertically and / or moved horizontally. Instead of a horizontal and / or vertical movement of the Mixelements For example, the container be moved horizontally and / or vertically by the recording so the same relative movement between mixing element and container too produce. The movement of the recording can therefore be in addition to the use at the "Bead Mill Disruption "method also when carrying out a Homogenization method can be used.

In a preferred embodiment the device has the option to choose between different types of homogenization. So can in a first program only a movement of recording be provided while in another, selectable Program an exclusive Movement of the rotor / stator element may be provided and, for example in a third program a simultaneous movement of the recording and the Mixelements be provided. In this way, the Possible uses of Device according to the invention elevated, since the user now not only between the type of treatment (Sonication, Homogenization, Bead Mill Disruption), but also within the method of homogenization different approaches choose can.

In a particularly preferred embodiment, the drive provided for the movement of the receptacle may also translate and / or rotationally move the holder of the mixel element, or transmit a rotational movement in such a way that a rotating element arranged at the end of a mixer bar held in the holder rotates in relation to the mixing bar can be driven. For this purpose, the drive may have a plurality of motors and / or a plurality of transmissions. This preferred off The design makes use of the synergy effects that result from the fact that the device is designed to carry out the three mentioned treatment methods, particularly well by allowing the saving of components.

In a particularly preferred embodiment is provided between the drive and the mixer a flexible shaft. Such a flexible shaft allows particularly good, a rotational movement of the drive to the holder of Mixelements to transfer. Furthermore can allow a flexible drive shaft feed movements, the performed with a mixing element Need to become, around the mixing element from the top into a vertically held in the receptacle tube immerse.

In a preferred embodiment the device has a sound horn whose sound output opening is such it is arranged that they on the container wall in the (if provided) driven or another Recordings held container is present when a container is inserted into the receptacle. It has been shown that on these Especially good is a sound wave in the sample to be split up can be generated.

In a preferred embodiment the sound generating device has an elastic element, with the one / the sound outlet opening against one / the container wall depressed can be. Especially when using a sonic horn, its sound output opening is arranged so that they on the container wall of the container held in the or another receptacle, it is advantageous to use this sound horn by means of an elastic Elements, such as a spring, against the container wall to press. In this way, the sound generating device has a special high efficiency.

The inventive device preferably has a container for receiving the biological sample. This container can in particular a tubular sample container such as for example, a 50 ml Falcon tube be. The container is preferably made of plastic.

In a preferred embodiment are in the container loose particles, the comminution of the sample and thus the digestion of the material, facilitate. These particles ("grinding balls") serve with the device a treatment of the suspension after the "Bead Mill Disruption 'procedure perform.

• – Aufschluß von Zellen und Gewebe für DNA und RNA Präparationen;
• – Aufschluß von forensischen Materialien (bspw Knochen) für DNA und RNA Präparationen;
• – Aufschluß von Zellen oder zum Aufschluß von anderen biologischen Materialien für Protein- und NA-Extraktionen.

The device according to the invention is particularly preferably used for the digestion of biological samples, and in particular preferably for:

• - disruption of cells and tissue for DNA and RNA preparations;
• - digestion of forensic materials (eg bones) for DNA and RNA preparations;
• - Digestion of cells or digestion of other biological materials for protein and NA extractions.

The inventive device has the further advantage that it a flexible size of the sample in terms the amount and volume of the sample. Leave with the device according to the invention processing both frozen and fresh samples.

following The invention is based on one, only one embodiment performing, drawing closer explained. Show in it

1 a schematic, perspective view of the device according to the invention for the digestion (lysis) of biological materials, in particular cell materials, in a structure, as it is particularly preferably used for homogenization,

2 a schematic, perspective view of the device according to 1 in a construction, as it is particularly preferred for the "Bead Mill Disruption" method is used and

3 a schematic, perspective view of an adapter that can accommodate several containers with aufzuschließendem biological material.

At the in 1 The apparatus shown is the sample to be treated in a container 1 held. The container 1 is in a jaw-like recording 2 arranged. The container 1 is conveniently a common vessel for receiving the biological sample, such as here a 50-ml Falcon tube. The device further comprises a holder 3 for a mixing element 4 on that in the recording 2 kept container 1 is introduced. There is also a drive 5 provided a motor 6 having. The drive is designed such that it receives 2 oscillating in a horizontal movement according to the double arrow A can move. A flexible shaft 7 connects the drive 5 with the bracket 3 and allows it over the bracket 3 a rotation of one at the end of the mixelement 4 provided rotor with cutting effect and also the holder - as indicated by the double arrow B - to translate. Finally, the device has a sound generating device 8th which can generate a sound wave in the sample suspension. The sound generating device has a sound horn 9 on, whose sound output opening is arranged such that they on the container wall in the receptacle 2 kept container 1 is applied. Not shown springs press the sound output opening of the sound horn 9 to the container wall.

The device allows the recording 2 to oscillate and in this way loose particles, such as. Mahlkugeln, which are in the container 1 can move and thereby achieve the digestion of the sample or support. Alternatively or additionally, via the drive 5 and the flexible shaft the mixing element 4 be moved rotationally or translatorily and thus also achieve a breakup of the samples. Finally, the sample may additionally or alternatively be treated by sound waves passing through the sound generating means into those in the container 1 sample is introduced (Sonication).

2 shows, while maintaining the same reference numerals for the same components, a second structure of the device according to the invention. Here, the sample to be treated is not stored in a vertical container 1 So, for example, a 50ml Falcon tube held but in a reaction vessel 20 equipped container, for example, a 2.0 ml Eppendorf reaction vessel (see. 3 ). An adapter 21 can several of these reaction vessels 20 in a lower part 22 with correspondingly shaped holes 23 take up. The lower part 22 is through a lid 24 is closed. In the reaction vessel 20 is the sample material 25 and the ball used for the Bead Mill Disruption method 26 represented ( 3 ). The adapter 21 is with horizontally lying reaction vessels 20 from the recording 2 held. The movement of the recording 2 in the direction of the double arrow A allows the "Bead Mill Disruption" method. Comparable adapters and reaction vessels are used, for example, in the product known from the product name "Tissuelyser" from Qiagen GmbH, Hilden, Germany.

1

container

2

admission

3

bracket

4

mixing element

5

drive

6

engine

7

flexible wave

8th

Sound generator

9

sound Horn

20

reaction vessel

21

adapter

22

lower part

23

drilling

24

cover

25

sample material

26

Bullet

arrow A

horizontal Movement of the recording

arrow B

translational Movement of the bracket

Claims (12)

Apparatus for digesting biological samples, comprising a sound generating device ( 8th ), which can generate a sound wave in the sample to be treated, characterized by - a recording ( 2 ) for a container ( 1 . 20 ), which can take the sample, - a mixer with a holder ( 3 ) for a mixing element ( 4 ) placed in the container ( 1 ) can be introduced when the container ( 1 ) in the recording ( 2 ) or is arranged in another receptacle and - a drive ( 5 ) for moving the recording ( 2 ) and / or the mixelement ( 4 ). Device according to claim 1, characterized in that the receptacle ( 2 ) the container ( 1 ) surrounds like a jaw. Device according to claim 1 or 2, characterized in that the drive ( 5 ) for the oscillating movement of the recording ( 2 ) is configured. Device according to one of claims 1 to 3, characterized in that the drive ( 5 ) for horizontal movement of the recording ( 2 ) is configured. Device according to one of claims 1 to 4, characterized in that in the holder ( 3 ) held mixing element ( 4 ) can be moved translationally and / or rotationally. Apparatus according to claim 5, characterized in that the drive ( 5 ) the holder ( 3 ) of the mixelement ( 4 ) can move translationally and / or rotationally. Device according to claim 6, characterized by a flexible shaft ( 7 ) between the drive ( 5 ) and the blender. Device according to one of claims 1 to 7, characterized by a sound horn ( 9 ), whose sound outlet opening is arranged such that it on the container wall of the in or another receptacle ( 2 ) container ( 1 ) is present. Device according to one of claims 1 to 8, characterized in that the sound generating device ( 8th ) has an elastic element, with the one / the sound outlet opening can be pressed against a / the container wall. Device according to one of claims 1 to 9, characterized by a container ( 1 . 20 ) for receiving the sample. Apparatus according to claim 10, characterized in that in the container ( 1 . 20 ) loose particles to assist digestion of the biological material. Device according to one of claims 1 to 11, characterized by a in the receptacle ( 2 ) usable adapter ( 21 ), the container ( 20 ).