WO2008011849A1

WO2008011849A1 - Method for operating a measuring arrangement having a scanning probe microscope device and a measuring arrangement

Info

Publication number: WO2008011849A1
Application number: PCT/DE2006/001648
Authority: WO
Inventors: Kate Poole; Detlef Knebel; Torsten JÄHNKE
Original assignee: Jpk Instruments Ag
Priority date: 2006-07-25
Filing date: 2006-09-19
Publication date: 2008-01-31

Abstract

The invention relates to a method for operating a measuring arrangement having a scanning probe microscope device, in which method a measuring probe (1), loaded in a measuring probe section with at least one probe substance (10, 20), of the scanning probe microscope device and a retainer for a probe (4; 30), with which the at least one probe substance (10, 20) interacts in a scanning probe microscope measurement, are moved in relation to one another using a displacement device (50, 52) within a displacement area which is defined by a plurality of relative positions, which can be assumed using the displacement device (50, 52), of measuring probe (1) and retainer, and the measuring probe (1) is cleaned in a manner in which it remains in the displacement area in the scanning probe microscope measurement after use.

Description

A method of operating a scanning probe microscope device

Measuring arrangement and measuring arrangement

The invention relates to a method for operating a measuring arrangement having a scanning probe microscope device and to a measuring arrangement with a scanning probe microscope device.

Background of the invention

Scanning Probe Microscopy (SPM) is a measurement and analysis technique in which a probe is scanned over a sample of a test medium to be examined and in which a topography of the sample is determined by a distance-dependent interaction between the probe and the sample However, it is also possible to obtain material constants or other sample information The most prominent representatives of this technology are the Atomic Force Microscope (AFM) and the Scanning Tunneling Microscope (STM) the Scanning Near Field Microscope (SNOM) and Scanning Photon Force Microscope (SPIiM).

In distance spectroscopy, the probe is displaced relative to the surface of the sample, for example, in a direction vertical to the sample surface, and the interaction between probe and sample is measured to measure the distance-dependent interaction between probe and sample. Alternatively, the sample can also be moved. It may also be provided a relative movement between the probe and sample in which both the probe and the sample are moved. In scanning probe microscopy, for example, this distance spectroscopy is used to measure the interaction between probe and sample to measure forces between molecules by binding one molecule to the probe and another molecule to the sample.

It can then measure the interaction between the two bound molecules. But it can also be measured intramolecular forces, for example, by lowering the probe to the sample and this is waiting for a bond. After that the probe can be removed again from the sample, in which case forces acting on the probe are recorded. In addition, further measurements are possible in which an interaction is measured that correlates with an associated distance to two or more locations.

As a probe in atomic force microscopy usually a component is used, which is also referred to as a cantilever. Without limitation of generality, reference is made in the following explanations to a cantilever. The statements apply correspondingly to other forms of probes in scanning probe microscopy.

It is known to use both untreated and pretreated cantilevers for distance spectroscopy. In the case of an untreated cantilever, binding of the sample to the measurement is nonspecific. For example, this involves drawing molecules from their surrounding medium by binding to the cantilever in order to measure the interaction of the molecules with the surrounding medium. In this case, however, it is also possible to characterize more accurately the molecules in which the blood is drawn. For example, DNA molecules show a specific spectroscopy curve due to an internal conformational transformation.

In particular, specific bindings can be investigated with a pretreated cantilever. Such an investigation may be advantageous if the formation of undesired bonds, which under certain circumstances can hardly be separated from one another, should be prevented during the measurement. Thus, it is common practice to bind one or more molecules to the probe designed as a cantilever which then forms a receptor-ligand system with the bound molecule (s). It is also known to bind whole cells to a measuring probe formed as a cantilever and to bring this system into interaction with a sample, for example a biomaterial, or with other cells. Pretreatments of measuring probes, in particular of cantilevers, are known in various embodiments. for example in the form of hydrophobizing the probe.

Known possibilities for the pretreatment of the cantilever generally lead to a coating of the measuring probe, at least in some areas. Thus, a cell attached to the cantilever coats a portion of the surface of the cantilever. It may be provided here, the first cantilever in the context of the pretreatment with a coating provided, in particular an adhesion-promoting coating on which then a substance to be measured is applied. Generally, in the following, the material applied in the course of the pretreatment to the measuring probe, in particular the cantilever, is referred to as a probe substance, be it a single material or a combination of several materials comprising, for example, an adhesion-promoting base and a substance arranged thereon and to be examined. A (base) coating applied as part of the pretreatment and covered by the probe substance is also referred to as probe coating.

If a pretreated cantilever is used in distance spectroscopy, several handling problems arise in practice. By each mechanical contact with the sample, the probe substance applied to the probe is usually loaded, so that after one or more individual experiments an aging of the probe substance occurs. The term "aging" is used in the present application quite generally for a change of a desired state of the probe substance which was induced at the beginning of the distance-spectroscopic measurements for the purpose of the measurement, but aging can not take place merely due to the execution of measurements. but even without such a measurement, for example, characterized in that there are given in a designed as a cell probe substance before or during the measurement no sufficient for the cell physiological conditions.

The aging of the probe substance can lead to fewer and fewer specific bonds taking place when carrying out the distance spectroscopic measurement, as a result of which the number of individual experiments must be substantially increased. In the case of a less specific individual measurement, care must be taken that the probe still has the probe substance in the required manner in each experiment. For example, it would be possible for a probe substance designed as a hydrophobic coating to be slowly converted into a hydrophilic coating, which would severely distort the measurement results. In the execution of the probe substance as a cell coating, often only one cell fits on the cantilever, it may happen that the cell dies and already changed very much before the measurement experiment, so that the actual measurement can not be performed. Again, there may be false results if this process goes unnoticed. In an untreated cantilever, it may happen that an adsorbate is formed by the contact with the sample, so that even non-specific bonds are no longer possible. If a measurement disturbing or even preventing aging of the measuring probe, pretreated or untreated, has occurred, the measuring probe is usually replaced. This change of the probe may well be associated with problems. The measuring probe is usually integrated in a Meßsondenaufhahme, which is held in the scanning probe microscope. The various implementations include, for example, a holder with a spring or a holder by means of vacuum. This holder must now be solved when changing the probe to install a new probe can. Usually, the replacement is done manually. After changing the probe, a new calibration must be performed. In the case of a force measurement to be performed with the cantilever, for example, the spring constant of the new cantilever must be determined. However, the methods available for this are only in an insufficient manner, so deviations of up to 20% or more are possible. A common method is the method of thermal noise. Here, a force-distance curve is recorded on a hard surface and then performed a measurement of the movement of the cantilever. As a rule, however, the sample to be examined does not constitute a hard base, so that in addition to the change of the measuring probe in this method, the sample also has to be exchanged in the meantime. In addition to the rather complicated handling, a major disadvantage is that the spring constant is not known exactly. Depending on the probe used, other deviations are thus obtained from experiment to experiment, which make interpretation of the measurement data considerably more difficult, in particular if, for example, a statistical evaluation is to be carried out.

It was alternatively proposed not to throw away the cantilever after removal from the scanning probe microscope, but to clean and re-coat with a probe substance. This should be achieved, for example, by cleaning the cantilever in a plasma. As a result, the cantilever is indeed cleaned, but then completely unprepared. This means that a base coating provided as part of the application of the probe substance must also be repeated. Again, this may affect the spring constant, so that must be recalibrated for safety in each case. In addition, the time required for this method is considerable. Summary of the invention

Based on the prior art, it is an object of the invention to provide a method for operating a scanning probe microscope having a measuring arrangement and a measuring arrangement with a scanning probe microscope in which the multiple use of a probe while maintaining the highest possible accuracy of the measurements is possible, even if a Aging of the probe occurs with a probe substance arranged thereon.

According to one aspect of the invention, there is provided a method of operating a measuring device comprising a scanning probe microscope device, wherein in the method with a displacement device a probe of the scanning probe microscope device loaded with at least one probe substance in a probe section and a receptacle for a sample with which the at least one probe Probe substance in a rastersondenmi- microscopic measurement interacts within one of several, with the help of the displacement ingestible relative positions of probe and recording spanned displacement range are moved relative to each other and the probe is cleaned after use in the scanning probe microscopic measurement in the displacement area remaining.

According to a further aspect of the invention there is provided a measuring arrangement comprising a scanning probe microscope device having a displacement device, the displacement device being configured to comprise a probe loaded with at least one probe substance in a probe section and a probe for a sample with which the at least one probe substance is at a scanning probe microscopy Measurement interacts to move relative to each other within one of a plurality of displaceable relative positions of the probe and receptacle for the sample spanned by the displacer, and wherein a probe cleaning device is configured to be the probe after scanning probe microscopy remaining in the transfer area to clean.

The invention includes the idea that the measuring probe used for a scanning probe microscopic measurement, which in turn is loaded with at least one probe substance, remains to be cleaned in a measuring environment after the scanning probe microscopic measurement. borrowed in a displacement range, which is spanned by a plurality of displacement positions, which are accessible to the probe and the Probenaufhahme relative to each other by means of the displacement device. In this way, a kind of "in j-Yw" cleaning is made possible, the intervention in the arrangement used for the scanning probe microscopic examination being kept as small as possible, while the prior art provides a complete replacement of the measuring probe or the displacement of the measuring probe In the case of the removal of the probe in the displacement area during cleaning, disadvantageous effects with regard to the subsequent reuse of the measuring probe, in particular with regard to the accuracy and comparability of the measurement, can be minimized or even completely ruled out Relocation options available with the aid of the displacement device preferably allow a relative displacement of the measuring probe and the sample holder in three-dimensional space, but also a displacement only in one plane or only along one dimension n be provided.

A preferred embodiment of the invention provides that when cleaning the probe after the rastersondenmicroscopic measurement on the probe remaining part of the at least one probe substance is cleaned. This makes it possible to perform the cleaning to obtain at least a part of the probe substance.

In an expedient embodiment of the invention can be provided that at least partially removed from the probe during cleaning of the remaining after the scanning probe microscopic measurement on the probe part of the at least one probe substance, whereby, for example, spent parts of the probe substance, which is not made reusable by means of cleaning can be eliminated.

An advantageous embodiment of the invention provides that when cleaning the remaining on the probe after the scanning probe microscopic measurement part of the at least one probe substance remains substantially completely on the probe.

Preferably, a development of the invention provides that during cleaning, measurement residues are removed. In an advantageous embodiment of the invention can be provided that after cleaning and optionally already during cleaning, the probe is loaded with new Sondesubstanz.

A further development of the invention can provide that at least the measuring probe section during cleaning is arranged in a measuring chamber of the scanning probe microscope device used for the scanning probe microscopic measurement, optionally in that at least the measuring probe section remains in the measuring chamber.

A preferred development of the invention provides that at least the measuring probe section is arranged during cleaning of the measuring probe in a measuring fluid used for the scanning probe microscopic measurement, optionally by at least the measuring probe section remaining in the measuring fluid after the scanning probe microscopic measurement. As an alternative to this embodiment, provision may be made for at least the measuring probe section to leave the measuring fluid for cleaning, as a result of which, due to surface tensions, the remaining probe substance can be at least partially released. If the probe substance can not be released by pulling it out of the measurement fluid, this process design can be supplemented by immersing the probe in another fluid in which a chemical process is used for cleaning. This can then take place without consideration for the sample to be examined. Thus, for example, a digestion of the probe substance can be carried out, if this is a cell. This can then be done, even if the sample itself is also a cell.

In an expedient embodiment of the invention, provision may be made for the measuring probe and the receptacle for the sample to be moved relative to one another in coarse graduations with fine coarse displacement elements encompassed by the displacing device and with fine graduation elements encompassed by the displacing device. For example, stepping motors can be used as coarse displacement elements, whereas, for example, piezoelectrically operated actuators are available as fine displacement elements.

An advantageous embodiment of the invention provides that during loading of the measuring probe with the at least one probe substance, the at least one probe substance is applied as substance compound, which has an adhesion-promoting base and a substructure formed thereon. includes punching. It can be provided that the substance is substantially completely removed during cleaning, so that only the adhesion-promoting base remains on the probe, which can then be reloaded.

Preferably, a development of the invention provides that the probe is brought into contact with a waste surface during cleaning. The waste surface has, for example, a waste layer made of a material on which the probe substance likes to settle. If the probe is then brought into contact with the waste layer after the scanning probe microscopic measurement, in particular the probe substance remaining on the probe, a bond between the waste layer and the remaining probe substance can form, so that the probe substance remains at the waste layer when the probe is again detached from the probe Waste surface is removed. The bond is usually based on adhesion forces between probe substance and waste layer, whereby by establishing preferred environmental parameters, such as pressure or temperature, the formation and the strength of the bond can be supported. The probe substance then remains at least partially on the waste layer. The waste layer may be formed on an additional component, which may itself be moved relative to the measuring probe. In this way, the additional component with the waste layer can also be moved into a region between the measuring probe and the sample, in order then to be included in the cleaning of the measuring probe. In a simple embodiment, however, the waste surface can also simply be provided on a sample carrier, preferably the sample carrier on which the sample examined by means of the scanning probe microscopic measurement is also applied. It can be provided that a part of the probe substance settles on the waste layer and separates independently from the probe. For example, a cell will move to the waste area if there are more attractive living conditions there. The measuring probe is then available to the next experiment without any additional force. The risk of possible damage to a probe coating is very low here. Another embodiment may provide for making the waste layer more effective by changing physical or chemical parameters. The waste layer may also be formed aggressively and at least partially destroy and absorb the probe substance.

In an advantageous embodiment of the invention can be provided that the probe and the waste surface are moved relative to each other when the contact of the probe is formed with the waste surface. A further development of the invention can provide that, during cleaning, the measuring probe flows around with a fluid selected from the following group of fluids: cleaning fluid and measuring fluid of the scanning probe microscopic measurement. The probe substance or parts thereof are removed in this embodiment by means of a directed to the probe flow. Such a flow may be generated, for example, by means of a pipette having a small opening and being brought to the measuring probe, for example from the side. The ejected fluid at least partially ruptures the probe substance from the probe and carries it away from the probe. Not only in this type of cleaning, but also in the other optionally provided options for cleaning the probe, it may be provided to shift the probe for the course of cleaning in a position which is spaced from later to be examined sample sections, so that in particular, it is prevented that probe substance parts dissolved by the measuring probe during cleaning fall on the sample surface still to be examined.

A preferred embodiment of the invention provides that light beams are irradiated to the probe during cleaning. For example, laser light can be used, preferably in the form of short laser pulses with a sufficiently high selected energy. An expedient development may be to irradiate laser light from the side of the probe.

In an expedient embodiment of the invention can be provided that during cleaning a probe surface of the probe is sucked at least in some areas. For aspiration in a possible embodiment, a pipette can be used as in the patch clamp technique by the pipette is for example brought laterally or from below to the probe and docks to the probe substance via a negative pressure. The force exerted thereby is generally greater than the binding force between the probe substance and the measuring probe or between different parts of the probe substance, so that at least partial detachment of the probe substance from the measuring probe occurs. The suction can be limited to a surface area of the probe substance.

An advantageous embodiment of the invention provides that when cleaning for the probe an environmental parameter selected from the following group of environmental parameters is changed: temperature, pH and ambient fluid composition. through Changing one or more environmental parameters can ensure that the probe substance at least partially no longer adheres to the probe or at least easier solvable. It should be noted that when changing the or the environmental parameters as possible, a loosening of the sample or its damage can be avoided. This is achieved by probe-specific solution parameters. For example, in the case where the probe substance comprises a cell on an adhesion-promoting layer, there may be provided the addition of molecules which have a higher or at least comparable affinity for the molecules involved in cell binding in the probe substance than the molecules involved in the binding the cell. The cell will dissolve and the initial state of the probe substance can be restored by subsequent rinsing with a fluid. In the event that the probe substance comprises a cell on an adhesion-promoting layer whose physical properties are temperature-dependent, lowering the temperature may cause the adhesion-promoting layer to become more fluid and the unusable cell to be released into the solution. By subsequent temperature change, the adhesion-promoting layer becomes stronger again.

In the following, preferred embodiments of the measuring arrangement with the scanning probe microscope device will be explained in more detail.

Preferably, a further development of the invention provides that the probe cleaning device is configured to clean a part of the at least one probe substance remaining after the scanning probe microscopic measurement on the probe during cleaning.

In an advantageous embodiment of the invention can be provided that the Meßson- denreinigungseinrichtung is configured to remove residues during cleaning.

A preferred embodiment of the invention provides that the at least one probe substance is applied as a composite substance comprising an adhesion-promoting base and a substance formed thereon.

In an expedient embodiment of the invention, a waste surface can be provided. An advantageous embodiment of the invention provides that the waste surface is formed in a usable for the scanning probe microscopic measurement chamber. Preferably, a further development of the invention provides that the measuring probe cleaning device be configured. in order to move the probe and the waste surface relative to each other during cleaning, when a contact of the probe is formed with the waste surface.

In an advantageous embodiment of the invention can be provided that the Meßson- denreinigungseinrichtung has a flow device which is configured to flow around the probe during cleaning with a fluid selected from the following group of fluids: cleaning fluid and Meßfluid the scanning probe microscopic measurement.

A further development of the invention can provide that the probe cleaning device has a light source which is configured to irradiate light beams onto the measuring probe during cleaning.

A preferred embodiment of the invention provides that the Meßsondenreinigungseinrichtung has a suction device which is configured to suck during cleaning a Meßsonden- surface of the probe, optionally limited to a surface region of the probe substance, at least in some areas.

In an expedient embodiment of the invention, it can be provided that the probe cleaning device has an adjustment device that is configured to change an environmental parameter selected from the following group of environmental parameters when cleaning for the probe: temperature, pH and ambient fluid composition. For this example, temperature controls can be used as Peltier elements.

An advantageous embodiment of the invention provides that the displacement device has coarse displacement elements and fine displacement elements.

DESCRIPTION OF PREFERRED EMBODIMENTS OF THE INVENTION

The invention is explained below with reference to embodiments with reference to figures of a drawing. Hereby show:

Fig. La and b is a schematic representation of an arrangement with a measuring probe on which a probe substance is immobilized; Fig. 2a, b and c is a schematic representation of an arrangement with a measuring probe, wherein the probe is cleaned by means of contact with a waste surface;

3 a, b and c show a schematic representation of an arrangement with a measuring probe, wherein the measuring probe is cleaned by means of a flow directed onto the measuring probe; and

4a and b is a schematic representation of an arrangement with a measuring probe, wherein the measuring probe is cleaned by means of a suction device.

1 a shows a schematic representation of an arrangement with a measuring probe 1 designed as a cantilever, as used in atomic force microscopes. The measuring probe 1 is loaded at a free end 2 on a bottom 3 with a probe substance, which is formed in the illustrated embodiment of a layer 10 and a cell 20. The layer 10 serves as a bonding agent for the cell 20. The layer 10 and the cell 20 are applied as part of a pretreatment on the probe. The layer 10 is applied to the free end 2 of the measuring probe 1, since there the sensitivity of the measuring probe 1 is greatest. Here, too, migration of the cell 20 to a location where the cell 20 can not interact with a sample 30 to be examined in the scanning probe microscopic measurement is prevented. The probe 1 is attached to a probe base 5 for easier handling.

The measuring probe 1 is coupled together with the probe base 5 via a probe holder 7 to a measuring probe 1 associated displacement device 50, which is shown schematically in Fig. Ia as a block and in an actual measuring arrangement comprises one or more displacement elements with which the probe 1 can be brought into different displacement positions within the measuring arrangement. These are not only positions occupied by the probe 1 in performing the scanning probe microscopic measurement, but also displacement positions in the area of the gauge to which the probe 1 can be brought by the displacement means 50 for various purposes. This includes the movement of the probe 1 into a position for cleaning the probe 1 after the scanning probe microscopic measurement. The displacement elements usually allow a displacement in three-dimensional space, but also a displacement only in one plane or only along one dimension can be provided. Manually or electrically operable displacement elements, for example using stepper motors (coarse displacement), can be used. relocation As such, dements are known to the person skilled in the art in various designs and therefore do not require any further detailed description. It can be provided to use a plurality of nested displacement elements in a structure, for example, a displacement element for feinstufϊgen method of the probe 1, for a rastersondenmikroskopi- see measurement, for example in a distance spectroscopy, and another, driven via stepper coarse displacement element to the probe 1 in to bring a cleaning position.

In another embodiment, the layer 10 covered by the probe substance can be distributed over the entire underside 3 of the measuring probe and also via an underside 6 of the probe base 5.

In FIG. 1 b, for better understanding, an arrangement is shown in which both the measuring probe 1 and the sample 30 can be displaced. The displacement device 50 associated with the measuring probe 1 is configured such that a point 25 on the cell 20 can only perform a vertical movement with a specific deflection 51. Another displacement means 52 associated with the sample 30 is configured such that a point 35 on the sample 30 can perform a lateral movement in the plane 53, which is shown here only in section. From the perspective of the point 35, the point 25 of the cell 20 can now reach all places of the cuboid 54. This space is the above-mentioned, created by means of the displacement means displacement area. Bases of both displacement devices are attached to the frame. This is indicated schematically in FIG. 1 b by means of the reference numeral 70.

Figures 2a, b and c show a schematic representation of a measuring arrangement with a measuring probe for explaining a method for cleaning the measuring probe, in which the measuring probe is brought into contact with a waste surface. For the same features, the same reference numerals as in Fig. 1 are used in Figs. 2a, b and c. The displacement devices are omitted for simplicity of illustration.

According to FIG. 2 a, the measuring probe 1 with the probe substance immobilized thereon as part of a pretreatment with the layer 10 and the cell 20 is arranged in a container 40, which is filled with a measuring fluid 41. First, a scanning probe microscopic measurement is carried out in which the cell 20 interacts with a sample 30. The container 40 is as Part of the scanning probe microscope device executed. The measuring fluid 41 serves to form a measuring environment for the cell 20 in which the cell 20 survives for at least the duration of an experiment. For this purpose, for example, a buffer solution is used. The filling level of the measuring fluid 41 in the container 40 is expediently selected such that the measuring probe 1 including the probe base 5 is completely submerged. The probe holder 7, however, only partially immersed in the measuring fluid 41. Immersion of the displacement device 50 (see Fig. Ib) would be possible in principle, but is less suitable from a technical point of view, since the displacement device would be more complicated in their construction because of necessary seals.

According to FIG. 2 b, the cell 20, after having been aged, in particular because of the scanning probe microscopic measurement and thus rendered unusable, is brought into contact with a waste layer 31 in order to clean the measuring probe 1. Such a cleaning can also be provided if, after a certain number of scanning probe microscopic measurements, age-related cleaning is necessary for statistical reasons.

The necessary relative movement between the measuring probe 1 and the sample 30 and the waste surface 31 can be carried out by moving the measuring probe with the aid of the displacement device 50 assigned to it. Alternatively or in addition thereto, a movement of the container 40 by means of the displacement device 52 can be performed. When the measuring probe 1 is transferred from the region above the sample 30 into the region above the waste layer 31, the cell 20 is continuously held in the measuring fluid 41. The contact between the cell 20 and the waste layer 31, which is then formed according to FIG. 2b, can be monitored by various methods. On the one hand, a force recording is an option. It may also be provided to use a fluorescence signal for monitoring. Depending on the cell type or probe substance, different methods can be used.

According to FIG. 2 c, the cell 20 remains on the waste surface 31 when the measuring probe 1 is again moved away from the waste surface 31. Again, it is of course possible to perform this with any relative movement of waste surface 31 and probe 1 to each other. The layer 10 remaining on the measuring probe 1 is now available for further scanning probe microscopic measurements. 3 a, b and c show a schematic representation of an arrangement with a measuring probe, wherein the measuring probe is cleaned by means of a directed to the probe flow. For the same features, the same reference numerals as in the preceding Figs. 1 and 2 are used in Figs. 3 a, b and c.

According to Fig. 3a, a pipette 80 in the vicinity of the measuring probe 1 is arranged. With the pipette 80, a flow 81 is generated, which is directed to the cell 20. According to FIG. 3 b, the cell 20 is then detached from the layer 10 with the flow 81.

According to the illustrations in FIGS. 3 a and 3 b, the region of the probe substance with the cell 20 is arranged directly inside the flow 81. This can lead to damage of the layer 10 under certain circumstances. According to Fig. 3 c, another embodiment is therefore provided that the flow 81 extends below the cell 20, so that the outgoing of the flow forces can be metered onto the cell 20 at its replacement.

The pipette 80 is disposed below the cell 20 in the illustrated embodiments. Alternatively, the pipette 80 may be positioned in other positions relative to the probe substance, for example, directly from the side or from above.

4a and 4b show a schematic representation of an arrangement with a measuring probe, wherein the measuring probe is cleaned by means of a suction device. In Figs. 4a and 4b, the same reference numerals as in the preceding Figs. 1 to 3 are used for the same features.

A pump 60 and a flexible line 61 connected thereto are provided, which is coupled to a suction device 62, so that a negative pressure generated by the pump 60 is applied to the cell 20 via the suction device 62. According to FIG. 4 b, a part of the cell 21 is sucked in with the aid of the suction device 62 and the entire cell 20 can be detached from the layer 10. The layer 10 remains on the measuring probe 1 and can be used to bind a new cell for further scanning probe microscopic examinations. In order to be able to monitor the process of detachment with the aid of the suction device 62, a patch clamp measurement is provided with a measuring device 63. This can then be tested, for example, after the docking shown in Fig. 4a to the cell 20, whether the cell 20 is still vital. This gives one possibility, one of which Standspectroscopy independent measurement, whether the probe substance, namely the cell 20, actually needs to be replaced.

A further embodiment can provide that the suction device 62 is equipped with an independent displacement device 64 (see Fig. 4b), which makes it possible to move the suction device 62 independently. Thus, an exchange or external cleaning of the suction device 62 would be possible without interfering with the scanning probe microscopic measurement. The arrangement is located, as in FIGS. 2 and 3, in a container with a measuring fluid, which, for the sake of clarity, has not been included in the schematic drawing. Technically, the pump 60, the flexible conduit 61, the measuring device 63 and the independent displacement device 64 can preferably also be arranged outside the container 40.

The features of the invention disclosed in the foregoing description, in the claims and in the drawing may be of importance both individually and in any combination for the realization of the invention in its various embodiments.

Claims

claims

1. A method for operating a scanning probe microscope having a measuring device, wherein in the method with a Verlagerangseinrichtung (50, 52) one ne in a Meßsondenabschnitt with at least one probe substance (10, 20) loaded

Measuring probe (1) of the scanning probe microscope device and a receptacle for a sample (4; 30), with which the at least one probe substance (10, 20) interacts in a scanning probe microscopic measurement, within one of several, by means of the Verlagerangseinrichtung (50 , 52) are moved relatively movable relative to each other and the measuring probe (1) is cleaned after use in the scanning probe microscopic measurement in the Verlagerangsbereich remaining removable position of the measuring probe (1) and recording spanned Verlagerangsbereiches.

2. Method according to claim 1, characterized in that, during cleaning, a part of the at least one probe substance (10, 20) remaining on the measuring probe (1) after the scanning probe microscopic measurement is cleaned.

3. The method according to claim 2, characterized eke ee zei chn et, that when cleaning the scanning probe microscopic measurement on the probe (1) remaining part of the at least one probe substance (10, 20) at least partially solved by the measuring probe (1) becomes.

4. Method according to claim 2, characterized in that, when cleaning the part of the at least one probe substance (10, 20) remaining on the measuring probe (1) after the scanning probe microscopic measurement, substantially completely on the measuring probe

(1) remains.

5. The method according to any one of the preceding claims, characterized e e e e e n ez e i chn e t that during cleaning Meßräge be removed.

6. The method according to any one of the preceding claims, characterized geke nnz egg chn et, that after cleaning and optionally already during the cleaning, the probe (1) is loaded with new probe substance.

7. The method according to any one of the preceding claims, characterized in that at least the Meßsondenabschnitt during cleaning in a used for the rastersondenmi- microscopic measurement chamber (40) the Rastersondenmikroskopeinrich- device is arranged, optionally by at least the Meßsondenabschnitt remains in the measuring chamber (40) ,

8. The method according to any one of the preceding claims, characterized in that at least the measuring probe section during cleaning in a used for rastersondenmi- microscopic measurement fluid (41) is arranged, optionally by at least the probe section after the scanning probe microscopic measurement in the measurement fluid (41) remains ,

9. The method according to any one of the preceding claims, characterized in that the measuring probe (1) and the receptacle for the sample (4; 30) with coarse displacement elements covered by the displacement device (50, 52) in coarse increments and with the fine displacement elements included in the displacement device Fine gradations are moved relative to each other.

10. The method according to any one of the preceding claims, characterized in that during loading of the measuring probe (1) with the at least one probe substance (10, 20), the at least one probe substance (10, 20) is applied as a substance composite, the adhesion-promoting base (10 ) and a substance (20) formed thereon.

11. The method according to any one of the preceding claims, characterized in that during cleaning, the measuring probe (1) is brought into contact with a waste surface (31).

12. The method according to claim 11, characterized in that the measuring probe (1) and the waste surface (31) are moved relative to each other when the contact of the probe

(1) is formed with the waste surface (31).

13. The method according to any one of the preceding claims, characterized e e e e e e n e z e i chn et that during cleaning the probe (1) is flowed around with a fluid selected from the following group of fluids: cleaning fluid and Meßfiuid the scanning probe microscopic measurement.

14. The method according to any one of the preceding claims, characterized g ek e nnz ei chn et, that when cleaning light beams are irradiated to the probe (1).

15. The method according to any one of the preceding claims, characterized e e e e e n e e e chn et et that during cleaning a probe surface of the measuring probe (1) is sucked off, at least in some areas.

16. Method according to one of the preceding claims, characterized in that when cleaning for the measuring probe (1) an environmental parameter selected from the following group of environmental parameters is changed: temperature, pH value and

Ambient fluid composition.

17. A measuring arrangement comprising a scanning probe microscope device having a displacement device (50, 52), wherein the displacement device (50, 52) is configured to have a measuring probe (1) and at least one probe substance (10, 20) loaded in a measuring probe section a receptacle for a sample (4; 30), with which the at least one probe substance (10, 20) interacts in a scanning probe microscopic measurement, within one of a plurality of relative positions of the measuring probe (1, 2) which can be captured with the aid of the displacement device (50, 52) and a receptacle cleaning means (31; 80; 62) configured to leave the probe (1) in the displacement area after the scanning probe microscopic measurement to clean.

18. Measuring arrangement according to claim 17, characterized in that the probe cleaning device (31; 80; 62) is configured in order to obtain, during cleaning, a part of the at least one probe substance (10) remaining on the measuring probe (1) after the scanning probe microscopic measurement. 20).

19. Measuring arrangement according to claim 17 or 18, characterized in that the measuring probe cleaning device (31; 80; 62) is configured to remove residues during cleaning.

20. Measuring arrangement according to one of claims 17 to 19, characterized in that the at least one probe substance (10, 20) is applied as a compound substance comprising an adhesion-promoting base (10) and a substance (20) formed thereon.

21. Measuring arrangement according to one of claims 17 to 20, characterized by a waste surface (31).

22. Measuring arrangement according to claim 21, characterized in that the waste surface (31) is formed in a usable for the scanning probe microscopic measurement measuring chamber (40).

23. A measuring arrangement according to claim 21 or 22, characterized in that the Meßsondenreinigungseinrichtung is configured to move the probe (1) and the waste surface (31) relative to each other during cleaning, when a contact of the probe (1) with the waste surface (31 ) is formed.

24. A measuring arrangement according to any one of claims 17 to 23, characterized in that the probe cleaning means comprises a flow device (80) which is configured to flow around during cleaning the measuring probe (1) with a fluid selected from the following group of fluids: Reinigungsfmid and measuring fluid of the scanning probe microscopic measurement.

25. A measuring arrangement according to any one of claims 17 to 24, characterized in that the Meßsondenreinigungseinrichtung comprises a light source which is configured to irradiate during cleaning light beams on the measuring probe (1).

26. Measuring arrangement according to one of claims 17 to 25, characterized in that the Meßsondenremigungseinrichtung a suction device (60, 61, 62, 63) which is configured to suck during cleaning a probe surface of the probe (1) at least in some areas.

27. Measuring arrangement according to one of claims 17 to 26, characterized in that the Meßsondenreinigungseinrichtung has an adjusting device which is configured to change the cleaning parameter for the probe (1) an environmental parameter selected from the following group of environmental parameters: temperature, pH, Value and

Ambient fluid composition.

28. Measuring arrangement according to one of claims 17 to 27, characterized in that the displacement device has coarse displacement elements and fine displacement elements.