WO2011047853A1

WO2011047853A1 - Biochip, associated examination appliance and corresponding operating method

Info

Publication number: WO2011047853A1
Application number: PCT/EP2010/006424
Authority: WO
Inventors: Andreas Lankenau
Original assignee: Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V.
Priority date: 2009-10-21
Filing date: 2010-10-20
Publication date: 2011-04-28
Also published as: DE102009050175A1

Abstract

The invention relates to a biochip (1) for examining a sample, wherein the biochip (1) has a plurality of reservoirs (7-13) for various reagents for examining the sample by means of various detection reactions, in which the sample is brought into contact with the various reagents. Furthermore, the invention comprises an associated examination appliance and a corresponding operating method.

Description

DESCRIPTION Biochip, associated examination device and operating procedure

The invention relates to a biochip for examining a sample and to an associated examination device and a corresponding operating method.

In medical diagnostics, so-called POC devices (POC: Point of Care) are known, which allow, for example, in medical practices quick tests for disease parameters. For example, such POC devices are used to determine the blood sugar level. As part of such POC diagnostics, a disposable article (eg, a test strip) is typically used which contains a reagent which is contacted with the sample (eg, blood, urine) and reacts with the sample, for example, a color change indicates the critical value of the disease parameter.

However, the known test strips allow only a single detection reaction between the sample and the reagent. Many analysis values, however, can not be detected immediately with such simple assays as they can only be determined by a chain of reaction steps. Examples include the lysis of pathogenic bacteria and their subsequent determination by DNA hybridization tests

(DNA: deoxyribonucleic acid) and the determination of surface antigens of living cells by immunological methods. At present, there are no POC assays for the determination of such analysis values, so that these analyzes are carried out by specialized personnel in appropriately equipped laboratories. need to be guided, which is associated with a high cost and time.

The invention is therefore based on the object, the

Field of application of the known POC devices to expand, so that analysis values can be determined, which can be determined only by a concatenation of reaction steps.

This object is achieved by a biochip according to the invention and an associated examination device as well as by a corresponding operating method according to the subsidiary claims.

The invention includes the general technical teaching of providing in a per se known biochip for examining a sample not only a single reagent for a single detection reaction, but several reagents for different detection reactions, wherein the different reagents are arranged in the biochip in separate reservoirs. For linking several reaction steps, the various reagents can then be emptied successively from the reservoirs and brought into contact with the sample so that the individual reagents react in succession with the sample.

The biochip according to the invention is particularly advantageous for medical diagnostics.

In a preferred embodiment of the biochip according to the invention, the reservoirs for the various reagents are formed as blisters, as are known, for example, from tablet packages, but with the difference that the blisters contain the different reagents for the different detection reactions instead of the tablets. The reagents can be emitted from the respective blister to carry out a detection reaction. be pressed to get in contact with the sample.

The individual blisters for the different reagents preferably protrude from the surface of the biochip, so that the reagents can be pressed out of the individual blisters by unrolling an ejection roller on the surface of the biochip. In the preferred embodiment, the individual blisters each have a compliant cover to facilitate squeezing out of the reagents. The resilient cover comprises in particular a film or membrane. In addition, the cover of the individual blisters is preferably transparent in order to be able to recognize the filling state of the biochip with a visual sample.

In contrast, in another embodiment of the invention, the reservoirs for the various reagents are chambers which are embedded in the surface of the biochip, the chambers preferably being provided at the top with a

collapsible cover are closed, so that the chambers can be emptied by the cover is pressed, wherein the in-chamber reagent is displaced. In addition, the cover of the individual chambers in this case preferably flush with the surface of the biochip. In this embodiment of the biochip according to the invention, there are various possibilities for emptying the individual chambers, which are briefly described below. One possibility for emptying the chambers is that an ejection roller is unrolled on the biochip, wherein the ejection roller in its mantle and rolling cam-like projections which push the covers of the chambers when rolling and penetrate into the chambers, wherein in the respective Chamber is displaced reagent.

Another way of emptying the chambers is that separate plunger break the cover of the chambers and penetrate into the chambers, wherein the located in the respective chambers reagent is displaced. The individual plungers can be operated independently of each other to empty the desired chambers.

In addition, to empty the chambers, it is possible that the entire biochip is made of a mechanically soft material and compressed to empty the chambers, for example with a smooth ejection roller. Upon compression of the soft biochip, the chambers therein are then also compressed, so that the reagent located in the chambers can escape and be brought into contact with the sample. In the preferred embodiment, the biochip has a reaction chamber in which the various detection reactions take place between the sample and the different reagents. For this purpose, the various reservoirs open directly or indirectly into the reaction chamber in order to bring the reagents together with the sample.

Moreover, in the preferred embodiment, the biochip according to the invention has a main line which opens into the reaction chamber, the reservoirs being used for various reagents along the main line one behind the other into the main line and from there into the reaction chamber. It is possible that the various reservoirs flow into the main conduit via channels for the reagents. Preferably, each channel connects a reservoir to the main line. Furthermore, the biochip according to the invention preferably comprises an inlet chamber into which the sample can be introduced.

In particular, the main line starts from the inlet chamber.

Preferably, the inlet chamber is provided for receiving a carrier liquid into which the sample can be introduced, the main line starting from the inlet chamber.

The inlet chamber of the biochip according to the invention preferably has a septum through which the sample can be injected into the inlet chamber by means of an injection needle and / or which after withdrawal of the injection needle automatically closes the opening in the septum produced by the injection needle.

Furthermore, the biochip according to the invention preferably also comprises a waste chamber for receiving the sample and / or the reagents after the detection reactions have been carried out, the waste chamber being arranged downstream of the reaction chambers. With regard to the structural design of the reaction chambers and the inlet chamber, in turn, there are the various possibilities mentioned above. These chambers can therefore be designed as blisters or be embedded as a chamber in the surface of the biochip.

Usually, due to the small height of known inlet chambers, it is difficult to inject a sample into the inlet chamber. Due to the low height, there is even the danger of damaging the biochip by means of the injection needle. Therefore, the inlet chamber of the biochip according to the invention may comprise a first portion and a second portion located downstream of the first portion, which are interconnected by means of a connection channel, the height of the first portion being greater than the height of the second portion, thereby allowing the sample can easily be injected into the first section and conveyed from there into the second section, preferably until the second section has been filled sufficiently (in particular completely).

The height of the second section may e.g. about 0.5mm, 1.0mm or at least 1.5mm. It is possible that the height of the first section is at least 1.5 times larger, at least 3.0 times larger or at least 4.0 times larger than the height of the second section.

It should also be mentioned that the inlet chamber, the reservoirs for the reagents and the reaction chamber are preferably arranged one behind the other along the biochip so that an emptying device, in particular at least one ejection roller when moving over the biochip, in particular during unrolling on the biochip one after the other Content of the admission chamber and the various reservoirs for the reagents in the reaction chamber expresses. The individual chambers are preferably arranged one behind the other along the main line. In this way, it is possible that the sample is brought into contact with the different reagents one after the other, so that the corresponding detection reactions take place sequentially.

It should also be mentioned that the reservoirs for the different reagents can have different shapes and / or different filling volumes, if this is necessary in the context of the respective detection reactions. However, within the scope of the invention it is also possible that the different reservoirs for the different reagents have the same filling volume and / or the same shape.

The biochip according to the invention makes it possible to automate examination protocols, which may comprise, for example, the following steps of an immunofluorescence analysis:

- fixation with formaldehyde

- Wash with phosphate buffered saline (PBS: Phosphate buffered saline)

- blocking non-specific interactions with bovine serum albumin (BSA: Bovine serum albumin)

- Incubation with a specific antibody

- Wash with PBS

- Incubation with a labeled secondary antibody - Wash with PBS

- Fluorescence detection. It is also possible that the reservoirs for the reagents in the order of their emptying contain the following reagents: formaldehyde,

- phosphate buffered saline,

bovine serum albumin,

a specific antibody,

- phosphate buffered saline,

a secondary antibody, and / or

- Phosphate buffered saline solution.

The formaldehyde, the PBS, the BSA and the antibodies thereby form the various reagents that are brought into contact with the sample in succession.

Furthermore, it is advantageous if the biochip has an identification mark, which can preferably be detected by a mark reader provided on an examination device, in order to be able to identify the biochip or the type of the biochip. The identification mark can be, for example, a bar code, a semiacode or an RFID transponder (RFID: radio frequency identification), but the invention is not limited to these types of identification markings. It is possible that the identification tag is automatically detected by the tag reader.

It should also be mentioned that the biochip according to the invention is preferably a flat disposable article which can only be used once.

It should also be mentioned that, with the exception of an inlet for introducing the sample, the biochip is preferably closed fluidically. is sen and no other inlets or outlets, which are provided in conventional biochips, for example, for pumping. The biochip may include an emptying device for expressing a sample from the inlet chamber and / or a reagent from at least one reservoir. The emptying device is preferably provided to be moved over the biochip.

The biochip works in a particularly advantageous manner if it can be ensured that the substance to be carried in the biochip liquid (for example, the sample and / or at least one Re ^¬ agent) are driven exclusively downstream or forward, and in particular in the direction of the reaction chamber. It should be avoided that the liquid forced out of the inlet chamber by means of the emptying device is partly returned to the inlet chamber. Similarly, it should be avoided that the liquid forced out of a reservoir by means of the emptying device is in part again returned to the reservoir.

Undesirable reflux may occur e.g. arise in two ways. If the emptying device has acted on a reservoir to push the reagent from this reservoir, the elasticity of the reservoir generates a restoring force and thus a negative pressure. As a result, the reagent which has been forced out of the reservoir is drawn towards this reservoir, ie pulled backwards or upwards, instead of forward or downstream in the direction of the reaction chamber, as desired. Furthermore, within a reservoir, reagent from the part lying in front of the emptying device (in particular under an ejection roller) into the already through

8th the emptying device emptied the back part of the reservoir to flow back. The reservoir would be emptied only incompletely, whereby the desired reaction and thus the reaction result would be adversely affected.

In a preferred embodiment of the invention, a substantially complete emptying of the inlet chamber and / or at least one reservoir is ensured. Furthermore, it can be at least temporarily reduced or prevented that a reagent of a reservoir after emptying or loading of this reservoir by means of the emptying device flows back in the direction of this reservoir. In addition, at least temporarily, it can be reduced or prevented that a sample introduced into the inlet chamber can be opened by emptying or pressurizing the inlet chamber

Emptying device flows back towards the inlet chamber. It can thus be ensured with particular advantage that the liquid to be conveyed in the biochip (the sample and / or at least one reagent, preferably all reagents) are conveyed exclusively in the direction of the reaction chamber and do not or only slightly flow back.

It is possible for the inlet chamber and / or at least one reservoir after depletion to remain depressed by means of an emptying device, in order to reduce or avoid an undesired reflux of the sample and / or at least one reagent. In particular, a surface of the biochip, which is preferably designed as a film or membrane and delimits or forms the inlet chamber and / or at least one reservoir, can remain depressed.

Preferably, a first mooring structure is provided internally in the inlet chamber and / or at least one reservoir, while at an opposite surface of the Biochips a corresponding second mooring structure is provided, wherein the first mooring structure and the second mooring structure are formed and brought into connection with each other, that the inlet chamber and / or the at least one reservoir after being depressed by an emptying device remains depressed, to an undesirable reflux to reduce or avoid the sample and / or at least one reagent. The first mooring structure is preferably arranged on a surface of the biochip which is designed as a foil or membrane and which supports the

Inlet chamber and / or at least one reservoir limits or forms.

It is possible that the surface preferably formed as a membrane or foil is made of an elastic material. The surface may define at least one of the following: the inlet chamber, the reaction chamber, the waste chamber, at least one reservoir, at least one conduit for the sample, and / or at least one reagent. It is possible that the surface comprises the crushable cover or the resilient cover. The surface is preferably made of plastic.

The mooring structures may be structured such that, after a single contact with each other, e.g. is generated by applying the surface by the emptying device, stick to each other.

The mooring structures can be formed, for example, in the form of a force-fit connection, form-locking connection or material connection, eg similar to a hook-and-loop fastener (hooks and fibers), in the form of interlocking hooks, in the form of pins, which can preferably be pressed into corresponding recesses with an exact fit. in the form of an adhesive Connection, in the form of a snap or snap connection or other suitable measures.

The biochip may, in particular, comprise at least one container filled with a fluid (eg a gas or a liquid), which is preferably designed as a pressure container, wherein the fluid flows into a preferably blind-ending, in particular outlet-free (ie without outlet) return reduction / prevention channel (hereinafter referred to for brevity as a backflow prevention channel) which can act on a conduit for the sample and / or for at least one reagent. It is possible that the return-preventing passage crosses the main line and / or is arranged in sections above and / or below the main line, thereby advantageously the effectiveness.

It is possible that, by loading the container through the emptying device, the return-preventing channel is deformed by means of the fluid such that the line for the sample and / or for the at least one reagent is closed, in particular squeezed, in order to prevent unwanted reflux of the sample and / or at least one reagent to reduce or avoid. Preferably, the return prevention channel substantially perpendicularly crosses the conduit for the sample and / or for the at least one reagent.

Furthermore, it is possible for the containers to be elongated and / or for at least two containers, which are arranged relative to the main channel on different sides of the biochip, to overlap. It is possible for the biochip to comprise a thickened film or membrane (for example a nubbed foil), which is preferably made of plastic. The thickened film may be a separate, for example, loosely placed on the biochip film or attached to the biochip or fastened.

In particular, the thickenings are formed and arranged to be e.g. by means of the emptying device, preferably together with the surface mentioned above, at least in sections, to be pressed accurately into the inlet chamber and / or at least one reservoir. Preferably, the inlet chamber and / or at least one reservoir is at least temporarily filled substantially completely by the thickenings.

Preferably, the chambers of the biochip are concave, i. formed inwardly curved while the thickenings convex, i. are formed curved outward.

The thickenings may be substantially rigid or deformable (elastic or plastic). The thickening may protrude on one or both sides of the film.

Advantages of the thickened film are e.g. the much easier manufacturability and better tightness due to higher precision and accurate fit of thickening and reservoirs.

The biochip may comprise at least one reflux reducing / preventing agent which, after being pressurized by means of the Emptying device preferably remains depressed together with the inlet chamber and / or at least one reservoir in order to reduce or avoid unwanted backflow of the sample and / or at least one reagent. The reflux reducing / preventing means is preferably plastically deformable and in particular a grid of metal or plastic. Preferably, the metal mesh may be made of stainless steel screen material, for example. The metal or plastic mesh may have meshes of about Ιμπι to about 1 mm and / or (wire) diameter of about lpm to about 100 m have.

Furthermore, it is possible for the biochip to comprise at least one viscous (preferably highly viscous) reflux reducing / preventing agent, which is arranged, for example, in the inlet chamber and / or at least one reservoir, by admission to a line for the sample by means of an emptying device and / or is depressible for at least one reagent, and which is provided to remain after the application by means of the emptying device substantially in the conduit for the sample and / or for at least one reagent or to close the conduit, whereby an undesirable reflux the sample and / or at least one reagent is reduced or avoided. The viscous reflux reducing / preventing agent preferably comprises a chemically neutral and / or viscous, especially high viscosity material (eg, silicone grease commonly used for vacuum applications). It is possible that the conduit for the sample and / or for at least one reagent comprises the main conduit. It is also possible that the conduit for the sample and / or for at least one reagent comprises at least one channel connecting a reservoir to the main conduit. In addition, the biochip may include at least one reflux reducing / preventing means configured as a valve.

Preferably, the valve is a self-medium actuated valve which can be actuated by means of the sample and / or at least one reagent. In particular, the valve is designed as a check valve, preferably as a flap-like or flap-like valve, which may comprise one or two flaps. Preferably, the valve is self-medium actuated in an open position to let pass the sample and / or at least one reagent, and / or in a closed position to reduce or avoid unwanted backflow of the sample and / or at least one reagent.

The aforementioned reflux reducing / preventing means may be arranged in or above the inlet chamber, in or above at least one reservoir, in or on a conduit for the sample and / or for at least one reagent.

It should also be mentioned that the means or methods described above for reducing or avoiding unwanted reflux of a sample and / or at least one reagent according to the invention also apply to the above-mentioned container or the above-mentioned return reduction / prevention channel or the fluid therein are applicable.

The invention is not limited to the above-described biochip according to the invention as a single product. Rather, the invention also includes a corresponding examination device for such a biochip. The examination device according to the invention therefore has a correspondingly adapted receptacle for the biochip, so that the biochip can be inserted or used for examination purposes in the examination device.

In addition, the examination device according to the invention preferably comprises an emptying device in order to empty the individual reservoirs with the different reagents.

By way of example, this emptying device may be the at least one rotatable ejection roller already mentioned above, which pushes the sample out of the inlet chamber and / or the reagents from the reservoirs of the biochip when the at least one ejection roller rolls over the biochip. In a preferred embodiment of the examination device according to the invention a plurality of ejection rollers are provided, one of which can be selected. The different squeezing rollers have different arrangements of squeezing cams, so that when unrolling on the biochip only certain reservoirs are emptied into which the squeezing cams encounter, while other reservoirs are not emptied. The arrangement of the Ausdrücknocken on the lateral surface of the individual Ausdrückwalzen so defined each a specific protocol that determines the selection and the timing of the emptying of the various reservoirs.

On the other hand, if the reservoirs of the biochip are arranged as blisters protruding from the surface of the biochip, the ejection rollers are for selectively expressing the blisters in their lateral surface instead of the Ausdrücknocken recesses into which the blisters can enter when rolling over, so that the blister in question is not expressed. The arrangement of the recesses in the lateral surface of the ejection rollers thus defines in each case a protocol for the associated detection reaction.

Furthermore, within the scope of the invention, the possibility exists that only a single ejection roller is provided, which, however, in its lateral surface has an adjustable arrangement of

Expressing cam or recesses has to be able to selectively empty the reservoirs. For a desired protocol, the squeezing cams or recesses in the mantle surface of the squeezing roller are then adjusted so that successively the desired reservoirs in the biochip are emptied.

This makes it possible for the same biochip to be used repeatedly in succession, with each of the successive uses of the biochip emptying a certain selection of the reagents from the associated reservoirs according to a specific protocol. The biochip according to the invention then enables successive different protocols for different detection reactions. In addition, the examination device according to the invention preferably also comprises a feed device which can roll the ejection roller over the biochip, wherein the rotational speed of the ejection roller is preferably adjustable in order to be able to control the supply of the various reagents to the sample in time.

In the embodiment of the biochip with blisters according to the invention described above, the ejection roller preferably has a smooth rolling surface. In the above In contrast, the described alternative embodiment of the biochip with recessed chambers for the various reagents, the Ausdrückwalze on the other hand in its rolling surface preferably at least one cam, which penetrates when overrunning a chamber in the chamber and pushes the reagent located in the chamber from the chamber.

In addition, the examination apparatus according to the invention preferably comprises a marker reader (e.g., bar code reader) for detecting the associated identification mark of the biochip. This identification of the respective biochip in the examination device offers the advantage that the examination device can individually control the time sequence of the subsequent examination as a function of the respective sample.

Furthermore, within the scope of the invention, a sensor for detecting the detection reactions in the biochip can be integrated into the examination device, which can be, for example, an optical sensor.

Furthermore, the examination device according to the invention can also have a display unit (for example a display) in order to visually display the examination result.

In addition, the examination device according to the invention preferably comprises a program memory which contains various control programs for the control unit. The control unit can then determine a specific function depending on the identification mark detected by the marker reader

Select control program. The emptying device of the examination device according to the invention is then controlled according to the selected control program, wherein the propulsion of the ejection roller can be variably adjusted. It is possible that an ejection roller comprises a raised ring, which is formed and arranged to partially close the main line when rolling over the biochip so that an undesired backflow of the sample

and / or at least one reagent is reduced or avoided.

In particular, a flow of the sample and / or at least one reagent located before, i. downstream of the ejection roller, back behind, i. be reduced or avoided upstream of the Ausdrückwalze.

Preferably, the ring has a width substantially equal to the width of the main conduit and / or a height substantially equal to the height of the main conduit.

It is possible that the emptying device comprises a circulating element.

In particular, the revolving element can enclose and / or connect at least two rotatable rollers.

The peripheral element in particular forms a running surface for acting on the biochip, as a result of which the circumferential element can act on the biochip in a planar manner.

The circulating element is preferably provided in order on the one hand to achieve an emptying of the inlet chamber and / or at least one reservoir and, on the other hand, to ensure that an undesired reflux of the sample and / or at least one reagent after emptying is reduced or avoided. In particular, the circulating element can be provided in order to access a previously through the emptying device Empty inlet chamber and / or act at least one previously emptied reservoir that an undesirable backflow of the sample and / or at least one reagent is reduced or avoided, preferably by further depressing the previously emptied inlet chamber and / or the previously emptied reservoir.

The at least two rollers are preferably arranged parallel to one another.

The circulating element is preferably designed in such a way that it simultaneously extends over the inlet chamber and / or a plurality of reservoirs spaced apart from one another in the flow direction.

The circulating element may be strip-shaped, chain-shaped, grid-shaped or otherwise suitable.

It can already be seen from the foregoing that the invention also encompasses an operating method for the biochip according to the invention and the associated examination apparatus, the individual steps of the operating method according to the invention being derived from the above description. Finally, the invention also encompasses the novel use of such a biochip for examining a sample.

The above features and embodiments of the invention can be combined with each other as desired.

Other advantageous developments of the invention are characterized in the subclaims or are explained in more detail below together with the description of the preferred embodiments of the invention with reference to FIGS. Show it: 1 shows a schematic representation of a biochip according to the invention with a plurality of reservoirs with reagents in the form of blisters,

FIGS. 2A-2C show various cross-sectional views of the biochip according to FIG. 1 with an ejection roller for emptying the blisters in different states,

Figures 3A-3C are various cross-sectional views of an alternative embodiment of a biochip according to the invention with recessed chambers for receiving the various reagents, a schematic representation of the inventive biochip with an associated examination device, an inventive operating method in the form of a flow chart, a flowchart showing an immunofluorescence analysis means of the biochip according to the invention illustrates various cross-sectional views of a portion of an alternative embodiment of a biochip according to the invention with a backflow prevention measure,

FIG. 8 shows a representation of another alternative

Embodiment of an inventive Biochips with a backflow prevention measure,

FIGS. 9A-9D are various illustrations of a portion of an alternative embodiment of a biochip according to the invention with a backflow prevention measure;

10A-10C show various cross-sectional views of a portion of an alternative embodiment of a biochip according to the invention with a thickened film,

IIA IIB different cross-sectional views of a portion of an alternative embodiment of a biochip according to the invention with a backflow prevention measure,

12A shows various cross-sectional views of a portion of another alternative embodiment of a biochip according to the invention and an emptying device according to the invention,

FIG. 13 a representation of a section of an alternative exemplary embodiment of a biochip according to the invention and an emptying device according to the invention,

FIGS. 14A-14C show various illustrations of a section of an alternative embodiment of a biochip according to the invention with a backflow prevention measure, and FIGS Figures 15A-15C are various views of a portion of an alternative embodiment of a biochip according to the invention.

1 shows a schematic representation of a biochip 1 according to the invention for the preferably diagnostic examination of a particularly medical sample (for example blood sample, urine sample) with a plurality of time-sequential detection reactions.

For receiving the sample, the biochip has an inlet chamber 2, which can be filled with a carrier liquid and is closed off by a septum 3. The sample may then be injected through an injection syringe 4 through the septum 3 into the inlet chamber 2, the hypodermic syringe 4 not shown to scale here for simplicity.

From the inlet chamber 2 is a main line 5, which opens into a reaction chamber 6, wherein the various detection reactions take place in the reaction chamber 6.

For this purpose, the biochip 1 has a plurality of reservoirs 7-13, which are filled with the various reagents for the different detection reactions and preferably via channels 16.1-16.7, which connect the reservoirs 7-13 to the main line 5, open into the main line 5.

Finally, the biochip 1 still has a waste chamber 14, in which the sample and the various reagents are collected after the detection reactions have been carried out, since the biochip 1 is a disposable product which can only be used once. After injecting the sample into the inlet chamber 2, the sample is first transferred from the inlet chamber 2 through the main conduit 5 into the reaction chamber 6.

For this purpose, one of a plurality of ejection rollers 15.1-15.3 can be selected, wherein each of the ejection rollers 15.1-15.3 defines a specific protocol which determines in which selection and in what time sequence the reservoirs 7-13 are emptied. For this purpose, the ejection rollers 15.1-15.3 in their lateral surface on different arrangements of recesses, which prevent the overrun of a blister that the respective blister is expressed.

However, there is also the alternative possibility that the inlet chamber 2, the reaction chamber 6 and the reservoirs 7-13 are arranged sunk in the biochip 1. In this case, the various ejector rollers 15.1-15.3 have different arrangements of squeezing cams in their lateral surface in order to empty a specific selection of the reservoirs 7-12 according to the desired protocol.

It should be mentioned here that the reservoirs 7-13 for the various reagents and the inlet chamber 2 are designed as blisters, as can be seen in particular from FIGS. 2A-2C. When the blisters are rolled over by the selected ejection roller 15.1, 15.2 or 15.3, the blisters are pressed together, so that the fluid (reagent or sample) in the blisters is displaced from the blister into a channel 16.1-16.7 and finally into the main line 5 , However, a part of the reservoirs 7-13 is not emptied when rolling over by the selected ejector roller 15.1, 15.2 or 15.3, since the ejector roller 15.1, 15.2 or 15.3 has corresponding recesses in its lateral surface. The arrangement of the recesses in the lateral surface of the Ausdrück- roller 15.1, 15.2 or 15.3 thus determines which of the reservoirs 7-13 are emptied.

When unrolling the ejection roller 15 in the arrow direction on the biochip 1, the blister of the inlet chamber 2 is first expressed, so that the sample from the inlet chamber 2 is transferred through the main line 5 into the reaction chamber 6.

Subsequently, the blisters of the reser- voire 7-13 are then successively expressed, so that the reagents contained therein are successively transferred into the reaction chamber 6 and there perform the desired detection reactions.

FIGS. 3A-3C show cross-sectional views of an alternative embodiment of a biochip according to the invention

1, this alternative embodiment being partially identical to the embodiment described above and illustrated in FIGS. 1 and 2A-2C, so that reference is made to the above description to avoid repetition, the same reference numerals being used for corresponding details.

A special feature of this exemplary embodiment is that the inlet chamber 2, the reaction chamber 6 and the servo valves 7-13 for the various reagents are not formed as blisters, but as chambers which are embedded in the biochip 1 and with the surface of the Completing the biochips 1 flush, the individual chambers are closed by a collapsible cover.

Another special feature of this embodiment is that the ejection roller 15 has no smooth surface. Rather, 15 cams 17 are attached to the lateral surface of the ejection roller, which when rolling on the biochip. 1 penetrate into the inlet chamber 2, the chamber-like reservoirs 7-13 and the reaction chamber 6 and to displace the fluids contained therein (sample or reagents). FIG. 4 shows a schematic representation of a POC examination system according to the invention which consists of the above-described biochip 1 according to the invention and an associated examination device 18. The examination device 18 has a receptacle into which the biochip 1 can be inserted in order to examine the sample injected into the biochip 1.

To identify the biochip 1, the examination unit 18 has a bar code reader 19, which detects a bar code 20 attached to the biochip 1 and can thereby identify the biochip 1.

The bar code reader 19 forwards the read bar code 20 to a control unit 21, which from a program memory 22, a specific control program according to the

Barcode 20 of the biochip 1 selects. In addition, the control unit 21 selects one of the ejection rollers 15.1-15.3 corresponding to the barcode 20 of the biochip 1, whereby a specific protocol for a particular detection reaction is determined. The control unit 21 then controls according to the read control program a propulsion 23 for the selected ejection roller 15.1, 15.2 and 15.3. According to the selected control program then the individual reagents from the various reservoirs 7-13 in a specific Time sequence and speed transferred to the reaction chamber 6.

In addition, the examination device 18 has an optical sensor 24 in order to be able to detect a color change of the sample in the reaction chamber 6, for example. The reaction chamber 6 is transparent to this, so that the sensor 24 can detect any color change. Finally, the examination device also has an optical display unit 25 in order to be able to visually display the examination result.

FIG. 5 shows the operating method of the examination device 18 according to the invention in the form of a flow chart.

In a first step S1, the sample is first injected by means of the injection syringe 4 through the septum 3 into the inlet chamber 2 of the biochip 1.

In a further step S2, the biochip 1 is then inserted into the examination device 18.

In a step S3, the bar code reader 19 then reads the bar code 20 on the biochip 1 in order to identify the biochip 1.

In a next step S4, the control unit 21 then selects a control program from the program memory 22 as a function of the barcode 20 of the biochip 1 in order to tailor the examination method individually to the respective biochip 1. Furthermore, in a step S5, the control unit 21 selects one of the ejection rollers 15.1-15.3, wherein the selection takes place as a function of the barcode 20 of the biochip 1. In addition, the control unit 21 controls the propulsion 23 for the selected ejection roller 15.1, 15.2 or 15.3 according to the selected control program, so that the reagents from the various reservoirs 7-13 are transferred into the reaction chamber 6 according to a predetermined time.

Subsequently, the sensor 24 then detects the detection reaction in a step S7 and the control unit 21 outputs the examination result via the display unit 25 in a step S8.

Subsequently, the biochip 1 can then be removed from the examination device 18 in a step S9 and disposed of in a step S9.

FIG. 6 shows an immunofluorescence analysis in the form of a flow chart, wherein the immunofluorescence analysis can be carried out with the examination device 18 and the biochip 1.

In a first step S1, the sample is first fixed with formaldehyde. For this purpose, the formaldehyde is transferred from the reservoir 7 into the reaction chamber 6, in which the sample is already located.

In a second step S2, the sample is then washed with a phosphate-buffered saline solution (PBS), wherein the phosphate-buffered saline solution is transferred from the reservoir 8 into the reaction chamber 6. In a step S3, non-specific interactions are then blocked, in which bovine serum albumin (BSA) is transferred from the reservoir 9 into the reaction chamber 6 of the biochip 1.

A subsequent step S4 then provides for incubation with a specific antibody, which is transferred from the reservoir 10 into the reaction chamber 6.

In a further step S5, the sample is then washed again with the phosphate-buffered saline solution (PBS), which is transferred from the reservoir 11 into the reaction chamber 6. A further step S6 then provides for incubation with a labeled secondary antibody, which is transferred from the reservoir 12 into the reaction chamber 6.

A further step S7 then provides for washing the sample again with a phosphate-buffered saline solution, which is transferred from the reservoir 13 into the reaction chamber 6.

Finally, a fluorescence detection of the sample in the reaction chamber 6 then takes place in a step S8.

Figures 7A-7B show various cross-sectional views of a portion of an alternative embodiment of a biochip 1 according to the invention with a backflow prevention measure, this embodiment is partially consistent with the embodiments described above, so reference is made to avoid repetition of the above description, wherein for corresponding Details the same reference numerals are used. FIG. 7A shows a reservoir formed by a blister 7 before emptying by means of the emptying device 15, while FIG. 7B shows the blister 7 after emptying by means of the emptying device 15. Furthermore, in FIGS. 7A and 7B, the channel 16.1 connecting the blister 7 to the main line 5 can be seen, and the surface 100 defining or forming the blister 7 at least in sections is formed as a film or membrane.

A peculiarity of this exemplary embodiment is that a mooring structure 41 is provided on the blister 7, in particular on the surface 100, and a corresponding mooring structure 42 is provided on an opposite surface of the biochip 1, which is designed and arranged in such a way that the Blister 7, in particular the blister 7 delimiting surface 100 after being acted upon by the emptying device 15 to empty the blister 7, remains depressed to reduce or prevent the reagent of the blister 7 after the impingement of the blister 7 back into Direction of the

Blister 7 is flowing. The inlet chamber 2 may be provided substantially identically with two opposing mooring structures and function in substantially the same manner.

FIG. 8 shows a representation of another alternative embodiment of a biochip 1 according to the invention with a backflow prevention measure, this alternative embodiment being partially identical to the above-described embodiments, so that reference is made to the above description to avoid repetition the same reference numerals are used. The biochip 1 shown in FIG. 8 comprises five containers 51-55 filled with a fluid (gas or liquid), which may be raised or formed in the surface of the biochip 1, wherein the fluid flows into a blind-ending, outlet-free return-preventing channel 51.1-55.1 is depressible.

The return-preventing channel 51.1 intersects the main line 5 between the inlet chamber 2 and the first reservoir 7 in the downstream direction (see arrow direction), and after the container 51 has been acted upon by the emptying device 250, the return-preventing channel 51.1 is deformed such that the main line 5 is at least temporarily (until towards the end of the loading of the container 51), in particular squeezed, in order to reduce or avoid an undesired flow of the sample introduced from the inlet chamber 2 into the main line 5 back into the direction of the inlet chamber 2. The return flow prevention channels 52.1-55.1 each intersect a channel 16.1-16-5 connecting a reservoir 7-11 to the main line 5, the respective return flow prevention channel 52.1-55.1 being deformed after loading of a respective container 52-55 by the emptying device 250 such that the respective channel 16.1-16.5 connecting a reservoir 7-11 to the main conduit 5 is at least temporarily closed, in particular squeezed, in order to reduce or avoid an undesirable reflux of the reagent in the direction of the respective reservoir 7-11.

As further shown in FIG. 8, the respective backflow prevention channels 51.1-55.1 substantially perpendicularly intersect the main pipe 5 and the respective channels 16.1-16.5 connecting the reservoirs 7-11 to the main pipe 5. alternative However, the intersection can be executed at any angle. Furthermore, it is possible to arrange the return-prevention channels 51.1-55.1 preferably in sections over and / or under the main line 5, preferably in parallel, so as to achieve a larger overlapping distance and thus an improved squeezing effect.

FIGS. 9A-9D show various representations of a portion of an alternative embodiment of a biochip 1 according to the invention with a backflow prevention measure, this embodiment being partially identical to the embodiments described above, so that reference is made to the above description to avoid repetition the same reference characters are used for corresponding details.

FIGS. 9A-9D show the reservoir 7, the channel 16.1 connecting the reservoir 7 to the main line 5, and the emptying device 15. The emptying device 15 is rolled in the direction of the arrow via the reservoir 7 and the channel 16.1 connecting the reservoir 7 to the main line 5 , wherein resulting states the different figures 9A-9D can be removed. A special feature of this embodiment is that the biochip 1 comprises a backflow prevention means 61, which is formed of a highly viscous, chemically neutral material and disposed in the reservoir 7, by applying by means of the emptying device 15 into which the reservoir 7 with the main line. 5 connecting channel

16.1 can be pressed, and which is provided to remain after being acted upon by the emptying device 15 substantially in the channel connecting the reservoir 7 to the main line 5 16.1, whereby an undesirable reflux of the reagent in the direction of the reservoir 7 is reduced or avoided. Similarly, the inlet chamber 5 may be provided with a backflow prevention means 61 which can be pressed into the main pipe 5 by the discharge device 15 and which is provided to remain substantially in the main pipe 5 after being subjected to the discharge device 15, whereby an undesired backflow of the sample in the direction of the inlet chamber 5 is reduced or avoided.

FIGS. 10A-10C show various cross-sectional views of a portion of an alternative embodiment of a biochip 1 according to the invention with a foil 70 having a thickening 71, this embodiment partially coinciding with the exemplary embodiments described above, so that reference is made to the above description to avoid repetition , wherein the same reference numerals are used for corresponding details.

FIGS. 10A-10C show a reservoir formed by a chamber 7, a reservoir formed as a chamber 8, the channels 16.1-16.2 connecting the chambers 7 and 8 to the main conduit 5, the emptying device 15 and the surface 100 formed as a film or membrane which covers the inlet chamber 5 and the chambered reservoirs of the biochip 1 (of which only the chambers 7 and 8 are seen in Figures 10A-10C). The emptying device 15 is moved in the direction of the arrow, with it being possible to take the resulting states from the different FIGS. 10A-10C.

A special feature of this embodiment is that the biochip 1 is attached to the biochip 1 or placeable, thickening 71 comprising film or membrane 70 comprises. The thickenings 71 are formed and arranged to be pressed into the inlet chamber 5 and the chambers 7-8. The thickenings 71 protrude on both sides of the film 70.

As can be seen in FIGS. 10A-10C, at least a portion of the thickenings 71 (the lower part of the thickened portions 71 in FIGS. 10A-10C) can be pressed into the chambers 7-8 with an exact fit. The thickenings 71 comprise a convex part, while the chambers 7-8 comprise a corresponding concave part. The thickenings 71 may be formed from a substantially rigid or deformable (elastic or plastic) material.

11A-11B show various cross-sectional views of a portion of an alternative embodiment of a biochip 1 according to the invention with a backflow prevention measure, this embodiment being partially in accordance with the embodiments described above, so reference is made to the above description to avoid repetition, wherein for corresponding Details the same reference numerals are used.

FIG. 11A shows a reservoir formed by a blister 7 before emptying by means of the emptying device 15, while FIG. IIB shows the blister 7 after emptying by means of the emptying device 15. Further, in Figures IIA and IIB of the blister 7 to the main line 5 connecting channel 16.1 to see and the blister 7 bounding or forming surface 100, which is formed as a film or membrane. A peculiarity of this embodiment is that the biochip 1 comprises a lattice-shaped, plastically deformable backflow prevention means 31, which over the

Blisters (of which in Figures 11A-11B only the blister 7 can be seen) is arranged and after being acted upon by the emptying device 15 together with the blister 7 (or the blister 7 delimiting surface 100) remains depressed to an undesirable reflux of To reduce or avoid reagent in the direction of the respective blister.

Similarly, the backflow prevention means 31 is disposed above the inlet chamber 2 and, after being acted upon by the emptying device 15 together with the inlet chamber 2 (or the inlet chamber 2 delimiting surface 100) depressed to an undesirable back flow of the sample in To reduce or avoid the direction of the inlet chamber 2.

12A-12B show cross-sectional views of a portion of an alternative embodiment of a biochip 1 according to the invention and an emptying device 150 according to the invention, this embodiment being partially in accordance with the embodiments described above, so referred to in the foregoing description, for avoiding repetition, wherein corresponding details the same reference numerals are used.

FIGS. 12A-12B show three reservoirs formed by blisters 7-9 and the channels 16.1-16.3 connecting the blisters 7-9 to the main channel 5. Figure 12A shows the three

Blister 7-9 before emptying by means of the emptying device 150, while the figure 12B shows the blister 7-8 after emptying by means of the emptying device 150. Further 12A-12B show the blister of the biochip 1 bounding or forming surface 100, which as a foil or

Membrane is formed. A special feature of this exemplary embodiment is the emptying device 150 for emptying the inlet chamber 2 and the blisters of the biochip 1. The emptying device 150 comprises six rotatable rollers 151 arranged parallel to one another, which are provided by a revolving element 152 (eg a belt, a chain, etc.). connected to each other. Circumferential element 152 forms a tread surface for biochip 1, thereby allowing peripheral element 152 to act on a plurality of reservoirs of biochip 1 (in FIGS. 12A and 12B, chambers 7-9) arranged one behind the other in the flow direction To reduce or avoid an undesirable backflow of the sample in the direction of the inlet chamber 2 and a reagent in the direction of a respective reservoir. FIG. 13 shows an illustration of another alternative exemplary embodiment of a biochip 1 according to the invention and an emptying device 250 according to the invention, this embodiment being partially identical to the exemplary embodiments described above, so that reference is made to the above description to avoid repetition, with the same being given for corresponding details Reference signs are used.

In particular, the exemplary embodiment shown in FIG. 13 corresponds to the embodiment shown in FIG. 8, but without the containers 51-55 and without the backflow prevention channels 51.1-55.1. A special feature of this embodiment is the emptying device 250 for emptying the inlet chamber 2 and the reservoirs 7-11 of the biochip 1. The emptying device 250 comprises a raised ring 81, which is designed and arranged to close the main line 5 in sections, when the emptying device 250 is rolled over the biochip 1, whereby an undesirable backflow of the sample in the direction of the inlet chamber 2 and an undesired backflow of a reagent in the direction of a respective Reservoirs 7-11 is reduced or avoided.

Figures 14A-14C show various representations of a portion of an alternative embodiment of a biochip 1 according to the invention with a backflow prevention measure, this embodiment is partially in accordance with the embodiments described above, so reference is made to avoid repetition of the above description, wherein for corresponding Details the same reference numerals are used.

FIGS. 14A-14C show the reservoir 7, the channel 16.1 connecting the reservoir 7 to the main line 5, and the emptying device 15. The emptying device 15 is shown in FIGS. 14A-14C from left to right via the reservoir 7 and the reservoir 7 with the main line 5 connecting channel 16.1 rolled, wherein resulting states the different figures 14A-14C can be removed. A special feature of this exemplary embodiment is that the backflow prevention means is designed as a self-medium-actuated (in particular sample and / or reagent-operated) non-return valve 350. The check valve 350 is designed as a flap or wing flap return valve. det and arranged in the channel 16.1. The check valve may comprise two, as shown in Figures 14A-14C, or only a flap or wing flap. As a comparison of Figures 14A-14C shows, the check valve 350 passes self-medium actuated in an open position, when the reservoir 7 is acted upon by the emptying device 15 for emptying to let pass the reagent located in the reservoir 7, while Eigenmediumbetät igt after emptying of the Reservoirs 7 enters a closed position to reduce or avoid unwanted backflow of the reagent in the direction of the reservoir 7. The check valve 350 may be arranged in an identical manner in further lines for the sample and / or for the reagent (eg the main line 5 and further channels connecting the reservoirs to the main line 5).

Figure 15A shows a plan view and Figures 15B-15C a portion of an alternative embodiment of a biochip 1 according to the invention, this embodiment being partially in accordance with the embodiments described above, so reference is made to avoid repetition of the above description, wherein for corresponding details the same reference numerals are used.

Figures 15A-15C show a septum 3, a main channel 5 and an injection needle or injection syringe 4. A special feature of this embodiment is that the inlet chamber 2 comprises a first section 2.1 and a downstream second section 2.2, which by means of a connecting channel 2.3 with connected to the first section 2.1. The height hl of the first section 2.1 is larger as the height h.2 of the second section 2.2, whereby the sample can be easily injected into the first section 2.1 by means of the injection needle 4. As a comparison of Figures 15A-15C shows, a sample is injected by means of the injection needle 4 through the septum 3 in the first section 2.1 until the second section 2.2 is sufficiently filled. If the second section 2.2 is sufficiently filled, the injection needle 4 can be pulled out of the septum 3, which then automatically closes.

The invention is not limited to the above-described be ^¬ preferred embodiments. Rather, a variety of variants and modifications is possible, which also make use of the inventive idea and therefore fall within the scope. In particular, the de Gegenstän ^¬ the subclaims independently of the features of the preceding and the referenced claims realizable bar.

biochip

inlet chamber

Portions of the inlet chamber

connecting channel

septum

syringe

main

Reaction kämmer

reservoirs

waste chamber

drain

squeeze rollers

channel

cam

Monitoring equipment

Bar code reader

barcode

control unit

program memory

jacking

sensor

display unit

Reflux Reducing / Preventive Mooring Structure

Mooring structure

fluid container

Backflow Reduction / Prevention Channel Backflow Reducing / Preventing Agent Foil / Membrane

thickening

ring

Discharge device

rotatable rollers

revolving element

Entleerungsvorriehtung

Reflux reducing / preventing means

Claims

CLAIMS 1. Biochip (1) for assaying a sample characterized by several reservoirs (7-13) for different reagents for assaying the sample with different detection reactions by contacting the sample with the different reagents.

2. biochip (1) according to claim 1, characterized in that a) that the reservoirs (7-13) for the reagents are blisters, wherein the reagents can be pushed out of the blisters to make the contact with the sample and with the sample to react, and / or b) that the blisters from the surface of the biochip (1) protrude, and / or

c) that the blisters have a compliant cover to facilitate squeezing out of the reagents, and / or

d) that the cover of the blister is transparent to the

To be able to recognize the filling state of the biochip (1) with a visual sample.

3. biochip (1) according to claim 1, characterized in that a) that the reservoirs (7-13) for the reagents are chambers, which are embedded in the surface of the biochip (1), and / or

b) that the chambers are closed at the top with a depressible cover, and / or

c) that the cover of the chambers with the surface of the

Biochips (1) flush.

4. biochip (1) according to claim 3, characterized in that: a) that the biochip (1) consists of a resilient material, so that the reagents from the chambers of the biochip (1) can be pressed out by the entire biochip ( 1) is compressed, or

b) that the biochip (1) consists of a rigid material, so that the reagents can be pushed out of the chambers of the biochip (1) by a cam (17) is pressed into one of the chambers, thereby displacing the reagent therein ,

5. biochip (1) according to any one of the preceding claims, characterized by

a) a reaction chamber (6) for carrying out the detection reactions in the reaction chamber (6), wherein the reservoirs (7-13) for the reagents open into the reaction chamber (6) in order to introduce the reagents into the reaction chamber (6), and or

b) a main line (5), which opens into the reaction chamber (6), wherein the reservoirs (7-13) for the reagents along the main line (5) in succession in the main line (5) and from there into the reaction chamber (6) lead, and / or

c) an inlet chamber (2) into which the sample can be introduced, the main line (5) starting from the inlet chamber (2), and / or

d) a septum (3) through which the sample can be injected into the inlet chamber (2) by means of an injection needle, and / or

e) a waste chamber (14) for receiving the sample and / or the reagents after the detection reaction in the reaction chamber (6), wherein the waste chamber (14) downstream of the reaction chamber (6) is arranged.

6. biochip (1) according to claim 5, characterized in that the inlet chamber (2) comprises a first portion (2.1) and a downstream behind the first portion (2.1) arranged second portion (2.2), which by means of a connection channel ( 2.3) are interconnected, wherein the

Height (hl) of the first section (2.1) is greater than the height (h2) of the second section (2.2), whereby an injection of the sample into the first section (2.1) is facilitated.

7. biochip (1) according to claim 5 or 6, characterized in that the inlet chamber (2), the reservoirs (7-13) for the reagents and the reaction chamber (6) along the biochip (1) are arranged one behind the other, so that an emptying device when moving over the biochip (1) successively expresses the contents of the inlet chamber (2) and the various reservoirs (7-13) for the reagents in the reaction chamber (6).

8. biochip (1) according to any one of the preceding claims, characterized

a) that the biochip (1) an identification mark

(20) for identifying the biochip (1) or the type of biochip (1), in particular a bar code, a semacode or an RFID transponder,

and or

b) that the biochip (1) is flat, and / or

c) that biochip (1) is a disposable article that can be used only once, and / or

d) that biochip (1) except for an inlet, in particular in the form of the septum (3), is fluidically closed and has no other inlets or outlets.

9. biochip (1) according to any one of claims 5 to 8, characterized in that the inlet chamber (2) and / or at least one reservoir (7-13) remains depressed after emptying by means of an emptying device, in order to reduce or avoid an undesired backflow of the sample and / or at least one reagent.

10. biochip (1) according to any one of claims 5 to 9,

characterized in that in the inlet chamber (2)

and / or at least one reservoir (7-13) is provided internally a mooring structure (41) and on a ^gegenüber¬ lying surface of the biochip (1) a corresponding mooring structure (42) is provided, which are formed and connectable with each other, that the a ^¬ let chamber (2) and / or the at least one reservoir (7-13) remains pressed down after emptying by means of an emptying device in order to reduce an undesired backflow of the sample and / or at least one reagent or avoid.

11. biochip (1) according to any one of the preceding claims, characterized in that the biochip (1) comprises at least one container (51-55) filled with a fluid, wherein the fluid in at least one reflux reducing / preventing channel (51.1- 55.1) can be pressed, which can act on a line for the sample and / or for at least one reagent, by means of loading the container (51-55), the reflux reducing / preventing channel (51.1-55.1) is deformable so that the Closed line for the sample and / or for at least one reagent to reduce or avoid unwanted backflow of the sample and / or at least one reagent.

12. biochip (1) according to any one of the preceding claims, characterized in that the biochip (1) comprises a film (70) having thickened portions (71), wherein the thickenings (71) are formed and arranged to enter into the inlet chamber (2) and / or at least one reservoir (7-13). to be pressed.

13. biochip (1) according to claim 12,

characterized in that

a) the thickenings (71) project on one or both sides of the foil (70), and / or

b) a portion of the thickenings (71) fit into the

Inlet chamber (2) and / or at least one reservoir (7- 13) is insertable.

14. biochip (1) according to any one of the preceding claims, characterized in that the biochip (1) comprises at least one reflux reducing / preventing means (31) which remains depressed after exposure by means of an emptying device in order to prevent undesired reflux of the pro- be and / or at least reduce or avoid a reagent.

15. biochip (1) according to claim 14, characterized in that the reflux reducing / preventing means (31) a) is plastically deformable, and / or

b) comprises a metal or plastic grid.

16. biochip (1) according to any one of the preceding claims, characterized in that the biochip (1) comprises at least one viscous reflux reducing / preventing means (61),

a) which is arranged in the inlet chamber (2) and / or at least one reservoir (7-13), b) can be pressed by applying an emptying device in a line for the sample and / or for at least one reagent, and

c) is provided to remain after the application by means of the emptying device substantially in the line for the sample and / or for at least one reagent, whereby an undesirable backflow of the sample and / or at least one reagent is reduced or avoided.

17. biochip (1) according to any one of claims 11 to 16,

characterized in that:

a) the line for the sample and / or for at least one reagent comprises the main line (5), and / or

b) the lead for the sample and / or for at least one

Reagent comprises at least one a reservoir (7-13) with the main line (5) connecting channel (16.1-16.7).

A biochip according to any one of the preceding claims, characterized in that the reflux reducing / preventing means (350) is at least one of the following:

a) a self-medium actuated valve,

b) a check valve,

c) a wing flap-like valve.

The biochip (1) according to claim 18, characterized in that the reflux reducing / preventing means (350) is sample and / or reagent operated

a) enters an open position to allow the sample and / or at least one reagent to pass, and / or b) enters a closed position to reduce or avoid unwanted backflow of the sample and / or at least one reagent.

20. biochip (1) according to any one of the preceding claims, characterized in that the reflux reducing / preventing means is arranged according to at least one of the following:

a) in or above the inlet chamber (2),

b) in or via at least one reservoir (7-13),

c) in or on a conduit for the sample and / or for at least one reagent.

21. Examination device (18) for examining a sample, characterized by a receptacle for the biochip (1) according to one of the preceding claims.

22. examination device (18) according to claim 21,

characterized

a) that the examination device (18) for emptying the individual reservoirs (7-13) in the biochip (1) has an emptying device (15.1-15.3, 23, 150, 250), and / or

b) that the emptying device (15.1-15.3, 23, 150,

250) has at least one rotatable ejection roller (15.1-15.3, 150, 250) for pushing out the reagents from the reservoirs (7-13) of the biochip (1) when the ejection roller (15.1-15.3, 150, 250) has the reservoirs (7 -13) rolls over, and / or

c) that the ejection roller (15.1-15.3) has recesses in its lateral surface in order to be able to avoid raised reservoirs when rolling over, and / or

d) that the examination device (18) comprises a feed device (23) for unrolling the ejection roller (15.1-15.3, 150, 250) via the biochip (1), and / or e) that the ejection roller (15.1-15.3) has a smooth rolling surface without surveys, or f) that the ejector roller (15.1-15.3) has in its rolling surface at least one cam (17) which penetrates into the chamber when Ü overrunning a chamber and presses the reagent located in the chamber out of the chamber.

23. examination device according to claim 22,

characterized,

a) that the emptying device several Ausdrückwalzen

(15.1-15.3) with different arrangements of Ausdrücknocken, or

b) that the ejection roller has an adjustable arrangement of Ausdrücknocken.

24. examination device (18) according to any one of claims 21 to 23, characterized by

a) a marker reader (19) for detecting the identification mark (20) of the biochip (1), and / or b) a sensor (24) for detecting the detection reactions in the biochip (1), in particular for immunofluorescence analysis, and / or

c) a display unit (25) for displaying an examination result, and / or

d) a control unit (21) which controls the operation of the examination device (18), and / or

e) a program memory (22) containing at least one control program for the control unit (21).

25. Examination device (18) according to claim 24, characterized in that the control unit (21) performs at least one of the following functions:

a) selecting one of the control programs stored in the program memory (22) in accordance with the control program cation reader (19) detected identification mark (20) of the biochip (1),

b) control of the emptying device (15.1-15.3, 23) according to the selected control program,

c) Variable adjustment of the propulsion of the ejection roller

(15.1-15.3).

26. examination device (18) according to any one of claims 21 to

25, characterized in that an ejection roller (250) comprises a raised ring (81) formed and arranged to close the main conduit (5) so as to reduce unwanted backflow of the sample and / or at least one reagent or avoided.

27. examination device (18) according to any one of claims 21 to

26, characterized in that an emptying device (150) comprises a peripheral element (152) which connects at least two rotatable rollers (151) and

a) which is provided to apply the biochip (1) area, and / or

b) which is provided to further act on a deflated inlet chamber (2) and / or at least one empty reservoir (7-13) so that an undesired backflow of the sample and / or at least one reagent is reduced or avoided.

28. A method of operating a biochip (1) according to any one of claims 1 to 20, comprising the steps of:

a) introducing the sample into the biochip (1),

b) Merging the sample with the various reagents in the reservoirs (7-13).

29. Operating method according to claim 28, characterized by the following steps: a) injecting the sample by means of an injection needle through the septum (3) into the inlet chamber (2),

b) passing the sample from the inlet chamber (2) into the

Reaction chamber (6),

c) emptying the reservoirs (7-13) with the reagents into the reaction chamber (6) so that the individual reagents react with the sample, whereby the reservoirs (7-13) are emptied successively.

30. Operating method according to one of claims 28 to 29, characterized:

a) that the same biochip (1) is used repeatedly in succession, and

b) that in each of the successive uses of the biochip (1) a certain selection of the reagents from the associated reservoirs is emptied according to a particular protocol.

31. Use of the biochip (1) according to any one of claims 1 to 20 for the examination of a sample.