A method of sample preparation using a charged particle beam device

Abstract

The present invention provides a method of preparing a sample using a device with at least one charged particle beam, said device comprising at least one column with a charged particle source producing charged particles and with elements for forming and directing charged particles into a charged particle beam, which is connected to a chamber comprising a stage for placing at least one substrate, a manipulator for transferring the sample, a device for mounting a sample holder, a sample holder for holding the sample placed in the device for mounting the sample holder, wherein the device optionally further comprises at least one device generating a photon beam connected to the chamber, a substrate with at least one sample placed on the stage, and an evaluation unit, the method comprising the following sequentially performed steps: - obtaining an image of the manipulator using the device with at least one charged particle beam, - moving the manipulator to a distance of at least 0 μm and at most 50 μm from the sample, - creating a connection between the manipulator and the sample using material deposition or redeposition, - separating the sample from the substrate, - removing the sample from the substrate using the manipulator, - transferring the sample to a distance of at least 0 μm and at most 50 μm from the sample holder, using the manipulator, - creating a connection between the sample and the sample holder using material deposition or redeposition, - separating the manipulator from the sample using a beam of charged particles or optionally using a photon beam, characterized in that the step of separating the manipulator from the sample using a beam of charged particles or optionally a photon beam comprises the following steps: - obtaining an image of the sample connected to the manipulator using the device with at least one charged particle beam, - comparing the image of the manipulator with the image of the sample connected to the manipulator, using an evaluation unit, - determining the edge of the manipulator adjacent to the sample, using the evaluation unit, based on the result of comparing the image of the manipulator with the image of the sample connected to the manipulator, - separating the manipulator from the sample using a beam of charged particles or optionally a photon beam guided along the edge of the manipulator adjacent to the sample determined in the preceding step.

Description

[0001] A method of sample preparation using a charged particle beam device

[0002] Field of Art

[0003] The invention relates to a method of automatized sample preparation for use in electron microscopy using a device with a focused charged particle beam or photon beam.

[0004] Background Art

[0005] To examine the internal structure of samples, for example, using a transmission electron microscope, it is necessary to have a very thin sample, the so-called lamella, which is a part of the substrate containing the area of interest. Such samples are usually created using a device with a focused ion beam (FIB), usually in combination with a scanning electron microscope (SEM) for observing the process of sample preparation. The device may also contain a device generating a photon beam. The process usually proceeds as follows: a substrate containing the area of interest is inserted into the device. The area of interest is found using the REM and then a sample is created from the substrate using the FIB or using the photon beam. The sample thus created is transferred using a manipulator to a sample holder (so-called grid), where it can subsequently be further processed, for example using a charged particle beam.

[0006] A manipulator is a movable device located in the electron microscope chamber, and its end is formed by a needle which is a separate part detachably connected to the rest of the manipulator or is an integral part of the manipulator. The sample is attached to the needle in various ways, e.g. by material deposition. In this method, various types of gases containing the desired material are supplied through one or more nozzles to the sample, which is still connected to the substrate, and are deposited under a beam of charged particles. A pure element or a combination of elements is then attached to the sample and the manipulator needle, forming a solid connection between the sample and the needle. Subsequently, the sample is separated from the substrate, e.g. by FIB, and transferred by the manipulator to a grid, to which it is again attached, e.g. by FIB and material deposition. The manipulator needle is then separated from the sample by FIB. This method is described, e.g. in US9040908B2.

[0007] The process of preparing a lamella is time-consuming and technically demanding and requires a constant presence of qualified staff who must monitor and control the process. There are efforts to develop a solution for preparing a lamella that would be performed automatically, reliably and at the same time efficiently by the device, without the need for permanent qualified staff.

[0008] In the prior art, automation currently only applies to some steps of preparing a lamella, such as the step of attaching the lamella to the manipulator needle before removing it from the sample and the subsequent separation (so-called cutting) of the lamella from the needle after transferring the sample to the grid. Automating this process not only saves time and money in terms of operating the device, but also in terms of material. The manipulator needle is an expensive device that is very susceptible to damage. In case of minor damage, the shape of the needle can be repaired using FIB, which is a time-consuming process, or it is necessary to use a new needle. Therefore, it is desirable that the attachment of the sample to the needle and its subsequent cutting take place without damaging the needle and the sample, ideally so that the shape of the needle and the sample does not change at all.

[0009] In the prior art, for example, in US9601313B2, there are described solutions for attaching the sample to the needle, wherein the needle is placed in close proximity to the sample, or directly touches it. Subsequently, a solid connection between the needle and the sample is created using material deposition. After transferring the sample to the grid and attaching it to the grid, it is necessary to separate the sample from the needle, for example, using an FIB beam. The disadvantage of this method is the fact that after cutting, part of the needle tip remains on the sample, or part of the sample on the needle tip, or deposited material remains on the needle or sample. In both cases, the shape of the needle is changed or deformed after each use, and therefore it is necessary to adjust the needle tip using FIB after each use, or use a new needle.

[0010] For this reason, solutions have been developed, described, for example, in patent US9620333B2, which aim to minimize the damage to the tip of the manipulator needle. These solutions consist in leaving a gap between the needle and the sample and depositing the material into this gap. When the sample is subsequently separated from the needle, the FIB beam is guided through the deposited material, which does not damage the needle tip. The disadvantage of this method is the fact that even in this case a thin layer of deposited material remains on the needle, which deforms the shape of the needle, and therefore it is necessary to remove the layer of deposited material after each use or after several uses.

[0011] Another option is the method described, for example, in US2023215683A1 , wherein an intermediate piece made of material that can be easily sputtered using a beam of charged particles is attached to the manipulator needle. The material sputtered from this intermediate piece creates a connection between the sample and the intermediate piece, and therefore the manipulator needle, and then, after being transferred to the holder, creates a connection between the sample and the sample holder in the same way. The needle is then separated from the sample in the area of the manipulator needle tip, so that the needle tip including the intermediate piece remains connected to the sample. The disadvantage of this method is that the shape of the needle tip changes.

[0012] To monitor the sample preparation process, an image created using various devices and techniques is used, the aim of which is to accurately display the position of the manipulator needle and the sample. The resulting image can be further processed using an appropriate software. To automate the lamella preparation process, it is necessary for the software to recognize in the image the position of the needle tip, the substrate position and the position of the sample in the substrate. For this software to function properly, as described, for example, in US10825651 B2, it is required that the sample and the needle have a shape that can be clearly recognized in the image. Based on information from such an image, the device controls the movement of the manipulator needle. It is undesirable for the shape of the needle tip to change, because in such a case the device could determine the position of the needle tip incorrectly, as a result of which a collision with the sample could occur and the needle or the sample could be destroyed.

[0013] In the prior art described, for example, in patent US10692688B2, an image created from the absorbed current is used. The disadvantage of this method is that the device must also include an absorption current detector, the signal of which is converted by the computer into an image of the needle tip and the sample. The connection between the sample and the needle is verified using an electric current that is fed into the manipulator needle. If the measured value of the electric current passing through the needle and the sample exceeds a specified limit, it is considered that the connection between the needle and the lamella is sufficient. The disadvantage of this method is that the sample must be electrically conductive, or must be provided with a sufficient layer of electrically conductive material. Another disadvantage is that in the step of separating the sample attached to the holder from the needle, the position of the needle and the sample cannot be displayed completely accurately. The software then determines the location for separation using a beam of charged particles based on a calculation derived from the sample size entered into the software by the user. This method is not accurate and, after cutting the needle, results in leaving part of the sample or layer of deposited material on the needle or leaving part of the needle on the sample.

[0014] It would therefore be desirable to provide a solution for attaching the lamella to the tip of the manipulator needle and its subsequent cutting off from the needle, which could be performed automatically and which would not lead to a change in the shape of the manipulator needle or to a change in the shape of the sample.

[0015] Disclosure of the Invention

[0016] The above aims are achieved by a method of preparing a sample using a device with at least one charged particle beam, said device comprising at least one column with a charged particle source producing charged particles and with elements for forming and directing charged particles into a charged particle beam, which is connected to a chamber comprising a stage for placing at least one substrate, a manipulator for transferring the sample, a device for mounting a sample holder, a sample holder for holding the sample placed in the device for mounting the sample holder, wherein the device further comprises a substrate with at least one sample placed on the stage, and an evaluation unit, the method comprising the following sequentially performed steps:

[0017] - obtaining an image of the manipulator using the device with at least one charged particle beam,

[0018] - moving the manipulator to a distance of at least 0 pm and at most 50 pm from the sample,

[0019] - creating a connection between the manipulator and the sample using material deposition or redeposition,

[0020] - separating the sample from the substrate,

[0021] - removing the sample from the substrate using the manipulator,

[0022] - transferring the sample to a distance of at least 0 pm and at most 50 pm from the sample holder, using the manipulator,

[0023] - creating a connection between the sample and the sample holder using material deposition or redeposition,

[0024] - separating the manipulator from the sample using a beam of charged particles.

[0025] The method according to the present invention solves the technical problem by the step of separating the manipulator from the sample using a beam of charged particles comprising the following steps:

[0026] - obtaining an image of the sample connected to the manipulator using the device with at least one charged particle beam,

[0027] - comparing the image of the manipulator with the image of the sample connected to the manipulator, using an evaluation unit,

[0028] - determining the edge of the manipulator adjacent to the sample, using the evaluation unit, based on the result of comparing the image of the manipulator with the image of the sample connected to the manipulator,

[0029] - separating the manipulator from the sample using a beam of charged particles guided along the edge of the manipulator adjacent to the sample determined in the preceding step (i.e. determined by means of the evaluation unit based on the result of comparing the image of the manipulator with the image of the sample connected to the manipulator).

[0030] The above aims are also achieved by a method of preparing a sample using a device with at least one charged particle beam, said device comprising at least one column with a charged particle source producing charged particles and with elements for forming and directing charged particles into a charged particle beam, which is connected to a chamber comprising a stage for placing at least one substrate, a manipulator for transferring the sample, a device for mounting a sample holder, a sample holder for holding the sample placed in the device for mounting the sample holder, wherein the device further comprises at least one device generating a photon beam connected to the chamber, a substrate with at least one sample placed on the stage, and an evaluation unit, the method comprising the following sequentially performed steps:

[0031] - obtaining an image of the manipulator using the device with at least one charged particle beam,

[0032] - moving the manipulator to a distance of at least 0 pm and at most 50 pm from the sample,

[0033] - creating a connection between the manipulator and the sample using material deposition or redeposition,

[0034] - separating the sample from the substrate,

[0035] - removing the sample from the substrate using the manipulator,

[0036] - transferring the sample to a distance of at least 0 pm and at most 50 pm from the sample holder, using the manipulator,

[0037] - creating a connection between the sample and the sample holder using material deposition or redeposition,

[0038] - separating the manipulator from the sample using a beam of charged particles or using a photon beam.

[0039] The second method according to the present invention solves the technical problem by the step of separating the manipulator from the sample using a beam of charged particles or a photon beam comprising the following steps:

[0040] - obtaining an image of the sample connected to the manipulator using the device with at least one charged particle beam,

[0041] - comparing the image of the manipulator with the image of the sample connected to the manipulator, using an evaluation unit,

[0042] - determining the edge of the manipulator adjacent to the sample, using the evaluation unit, based on the result of comparing the image of the manipulator with the image of the sample connected to the manipulator,

[0043] - separating the manipulator from the sample using a beam of charged particles or a photon beam guided along the edge of the manipulator adjacent to the sample determined in the preceding step (i.e. determined by means of the evaluation unit based on the result of comparing the image of the manipulator with the image of the sample connected to the manipulator).

[0044] The advantage of both methods described herein, both using the comparison of the image of the manipulator with the image of the sample connected to the manipulator, is a precise determination of the edge of the manipulator in the image, thanks to which it is possible to guide a beam of charged particles or a beam of photons along this edge and use the beam to separate the sample from the manipulator. This means that no layer of deposited material remains on the manipulator and no material of the manipulator is removed, i.e. the shape of the manipulator does not change, and hence the manipulator can be used repeatedly without the need to adjust the shape of the manipulator or to replace the manipulator. In one aspect, the present invention provides a computer programme configured for performing one or both methods described herein above.

[0045] In one aspect, the present invention provides an evaluation unit comprising a computer programme as described herein above loaded in the memory or processor of the evaluation unit.

[0046] Brief description of drawings

[0047] The invention is further explained by examplary embodiments, which are described with reference to the attached drawings. For better clarity, only those parts of the device that are important from the point of view of the principle of the present invention are shown in the drawings.

[0048] Fig. 1 is a schematic representation of the arrangement of the device with one beam of charged particles,

[0049] Fig. 2 is a schematic representation of the arrangement of the device with two beams of charged particles,

[0050] Fig. 3 is a schematic representation of the image of the manipulator,

[0051] Fig. 4 is a schematic representation of the top view of the manipulator in the vicinity of the sample, Fig. 5 is a schematic representation of the top view of the manipulator connected to the sample by means of material deposition or redeposition, when the sample is separated from the substrate, Fig. 6 is a schematic representation of the top view of the sample connected to the manipulator as well as to the sample holder,

[0052] Fig. 7 is a schematic representation of the top view of the separation of the manipulator from the sample.

[0053] Examples of carrying out the invention

[0054] The embodiments shown illustrate exemplary embodiments of the invention, which, however, shall not be construed as limiting the scope of protection.

[0055] The method of preparing a sample from a substrate is carried out using a device with at least one charged particle beam.

[0056] In a first exemplary embodiment of the device, the device is shown in Fig. 1. This device comprises one column 1 with a charged particle source 2 producing charged particles and with elements 3 for forming and directing the charged particles into a charged particle beam, the column being connected to a chamber 5.

[0057] In a second exemplary embodiment of the device, the device is shown in Fig. 2. This device comprises two columns 1, 11 with charged particle sources 2, 12, the columns being connected to a chamber 5 so that the optical axes 4, 13 of these columns , 11 form an angle with respect to each other of greater than 0° and less than 180°.

[0058] In a first exemplary embodiment of the charged particle sources 2, 12, the charged particles are electrons, in the second exemplary embodiment of the charged particle sources 2, 12, the charged particles are ions.

[0059] The first exemplary embodiment of the device may include any of the above-mentioned charged particle sources 2, 12. The second exemplary embodiment of the device may include the same or different charged particle sources 2, 12 or a combination thereof.

[0060] In an alternative exemplary embodiment of the device, the device includes three or more columns , 11 connected to the chamber 5, and various combinations of the charged particle sources 2, 12 located in the columns.

[0061] In the column , 11 with the charged particle source 2, 12, there are always elements 3, 14 for shaping and directing the charged particles into a charged particle beam, for example lenses, stigmators, electrodes on a potential, apertures and other commonly used particle optics components.

[0062] In a third exemplary embodiment of the device, the device comprises one or more columns , 11 with charged particle sources 2, 12 and at least one device generating a photon beam connected to the chamber 5.

[0063] The device further comprises a chamber 5 to which the columns 1_, 11 and the optional device generating a photon beam are connected. In the chamber s, there is a movable stage 8 for placing at least one substrate 7. The stage 8 is movable along at least two axes, which are perpendicular to each other in an exemplary embodiment of the stage 8, and further is rotatable around at least one axis and tiltable around at least one axis. A substrate 7 is placed on the stage 8, in the substrate 7 at least one sample 17 was created, for example, using a focused ion beam or using a photon beam. The sample 17 is a part of the substrate 7 containing the region of interest. The sample 17 may be a lamella or a block larger than a lamella.

[0064] The chamber 5 further comprises a movable manipulator 6. In an exemplary embodiment of the manipulator 6, the manipulator 6 is terminated by a needle 15, which is detachably connected to the manipulator 6. In an alternative embodiment of the manipulator 6, the end of the manipulator 6 is shaped into the shape of a needle 15. The manipulator 6 is movable along three axes, which are perpendicular to each other in the exemplary embodiment, and rotatable around one axis.

[0065] The chamber 5 further comprises a device 10 for mounting a sample holder and a sample holder 9.

[0066] The device further comprises at least one detector of signal particles. The signal particles are in particular secondary or back-reflected particles, or other particles emitted by the sample due to interaction with an incident charged particle beam.

[0067] The device further comprises an evaluation unit containing software for performing the method. The device uses a beam of charged particles or a beam of photons to create at least one sample 17 in the substrate 7, wherein the sample 17 remains connected to the substrate 7 by a small portion. It is also possible to place the substrate 7 with the samples 17 already prepared in this way in the device.

[0068] Subsequently, the device uses a beam of charged particles to create an image of the manipulator 6 by creating an image of the field of view without the manipulator 6 in the field of view and an image of the manipulator 6 in the field of view. The image of the manipulator 6 is created as the difference between the created images, for example using a software using any of the commonly used techniques.

[0069] Furthermore, in the image of the manipulator 6, the tip 16 of the needle 15 of the manipulator 6 is identified, for example using a software.

[0070] In the next step, the tip 16 of the needle 15 of the manipulator 6 is moved towards the sample 17 to a distance of at least 0 pm and at most 50 pm from the sample 17. To move the manipulator 6, it is necessary to know its position in the chamber space. For this reason, it is necessary to create two images of the manipulator 6 at different angles using a beam of charged particles. In the case where the device is a device according to the first exemplary embodiment of the device, a first image of the manipulator 6 is obtained at a first angle and subsequently the stage 8 with the substrate 7 is tilted so that the plane of the stage 8 forms an angle different from the first angle with the optical axis 4 of the column 1_. The manipulator 6 is tilted together with the stage 8, so that the end of the manipulator 6 with the needle 15 is in the same position and at the same angle of inclination relative to the plane of the stage 8 as at the moment of obtaining the first image of the manipulator 6 at the first angle. Subsequently, a second image of the manipulator 6 is created. When the device is a device according to the second exemplary embodiment of the device, in which the device comprises two columns 1_, 11 , the optical axes of which 4, 13 form an angle greater than 0° and less than 180°, the first image of the manipulator 6 is obtained using a beam of charged particles produced by the source 2 of the first column 1_ and the second image of the manipulator 6 is obtained using a beam of charged particles produced by the source 12 of the second column 11 (and the tilting step is not necessary). Since the images of the manipulator are created at different angles, it is possible to determine the position of the manipulator in space using the evaluation unit.

[0071] In the following step, the manipulator 6 is connected to the sample 17 by redeposition or by material deposition. Redeposition is carried out, for example, in such a way that in the step of preparing the sample 17 in the substrate 7, a layer of molecules is deposited on the surface of the sample 17, and after irradiation with a charged particle beam, wherein the charged particles can be e.g. ions or electrons, the molecules move and thus create a connection between the sample 17 and the manipulator 6. In an alternative case, instead of a deposited layer of molecules, the material of the substrate 7 is sputtered, which creates a connection between the sample 17 and the manipulator 6. Material deposition can be carried out e.g. using a gaseous precursor, which, under a charged particle beam, wherein the charged particles can be e.g. ions or electrons, decomposes into gaseous molecules, which are pumped out of the chamber 5, and solid molecules, which settle on the surface, e.g. on the surface of the sample 17 or between the sample 17 and the manipulator 6, thereby creating a connection between them. The gaseous precursor is supplied to the chamber 5 using a gas injection system (GIS), which is a system of one or more capillaries that supply the gaseous precursor to the surface of the sample 17.

[0072] The connection point of the manipulator 6 and the sample 17 (so-called welding point 18) is located in an exemplary embodiment on the tip 16 of the needle 15 of the manipulator 6 and on the edge of the sample 17. In an alternative exemplary embodiment, the connection point of the manipulator 6 and the sample 17 is located in the immediate vicinity of the tip 16 of the manipulator 6 and anywhere on the sample 17 where this is possible with respect to the shape of the sample 17 and the manipulator 6. The exemplary embodiments of the connection point of the manipulator 6 and the sample 17 can also be combined.

[0073] In the following step, the sample 17 is separated from the substrate 7. The separation can be performed, for example, using a charged particle beam or a photon beam directed at the part of the sample 17 by which the sample 17 is connected to the substrate 7, or mechanically, e.g. by breaking off at the location of the part of the sample 17 by which the sample 17 is connected to the substrate 7. Subsequently, the sample 17 is removed from the substrate 7 by the manipulator 6 and transferred to the sample holder 9 so that the edge of the sample 17 is at a distance of at least 0 pm and at most 50 pm from the edge of the sample holder 9.

[0074] In the following step, the sample 17 is connected to the edge of the sample holder 9 by means of material deposition or redeposition.

[0075] In the following step, the device separates the manipulator 6 from the sample 17 using a charged particle beam or a photon beam, such that the beam is guided to the edge of the manipulator 6 adjacent to the sample 17. In the first exemplary embodiment of determining the edge of the manipulator 6 adjacent to the sample 17, the device creates an image of the sample 17 connected to the manipulator 6 before the step of connecting the sample 17 to the edge of the sample holder 9. In the second exemplary embodiment of determining the edge of the manipulator 6 adjacent to the sample 17, the device creates an image of the sample 17 connected to the manipulator 6 and simultaneously connected to the sample holder 9. In both exemplary embodiments of determining the edge of the manipulator 6 adjacent to the sample 17, the edge of the manipulator 6 adjacent to the sample 17 is determined using a software based on the created images using image analysis based on searching for patterns (so-called template matching), wherein the image of the manipulator 6 without the attached sample 17 obtained by the method described above is used as a pattern (template), which is searched for in the image of the sample 17 connected to the manipulator 6 or in the image of the sample 17 connected to the manipulators and simultaneously connected to the sample holder 9.

[0076] Since the charged particle beam or the photon beam is guided exactly along the edge of the manipulator 6, the shape of the manipulator 6 does not change and it can therefore be used repeatedly.

[0077] In a first specific embodiment, the device comprises one scanning electron microscope (SEM) column , in which a particle source 2 produces electrons, and one focused ion beam (FIB) column 11 , in which a particle source 12 produces ions. The optical axes 4, 13 of the SEM and FIB columns 1, 11 form an angle of 45° with respect to each other. A substrate 7 with prepared samples 17 is placed on a stage 8 in the chamber 5 of the device.

[0078] In the first step, an image of a manipulator 6 is created by using the FIB to create an image of the field of view without the manipulator 6 in the field of view and then an image of the manipulator 6 in the field of view. Based on the difference between these images, the software creates an image of the manipulator 6. Next, the tip 16 of the needle 15 of the manipulator 6 is determined in the image of the manipulator 6 using a software.

[0079] In the following step, two images of the sample 17 are created, the first using the SEM, the second using the FIB. Due to the fact that the optical axes 4, 13 of the SEM and FIB columns 1, 11 form an angle of 45° with respect to each other, the sample 17 can be viewed from two angles. From the combination of these images, two edges of the sample 17 are detected by the software. At the intersection of these edges, the software determines the welding point 18 for creating a connection between the tip 16 of the needle 15 of the manipulator 6_and the sample 17.

[0080] Subsequently, the manipulator 6 is moved towards the welding point 18 using the determined tip 16 of the needle 15 and the images of the sample 17 so that the distance between the tip 16 of the needle 15 and the welding point 18 is 1 pm. The position of the manipulator within the space of the chamber 5 is determined by the evaluation unit based on the combination of images taken simultaneously by the SEM and the FIB.

[0081] Subsequently, while simultaneously supplying a gaseous precursor using GIS, the FIB is directed between the tip 16 of the needle 15 of the manipulator 6 and the welding point 18, and a connection is created between the needle 15 of the manipulator 6 and the sample 17 by the deposited material.

[0082] In the following step, the FIB is directed to the place where the sample 17 is connected to the substrate 7, and using this beam, the sample 17 is separated from the substrate 7. Subsequently, the sample 17 is removed from the substrate 7 using the manipulator 6.

[0083] In the following step, the manipulator 6 transfers the sample 17 to the sample holder 9 located in the device 10 for mounting the sample holder 9, at a distance of 1 pm. Subsequently, while simultaneously supplying a gaseous precursor using the GIS, the FIB is directed between the sample 17 and the sample holder 9, and a connection is created between the sample 17 and the sample holder 9 by the deposited material.

[0084] In the following step, an image of the sample 17 connected to the manipulator 6 and also connected to the sample holder 9 is created using the FIB. This image is compared with the image of the manipulator 6 without the attached sample 17, which is used as a template, on the basis of which the evaluation unit determines in the image obtained in this step the edge of the manipulator 6 adjacent to the sample 17.

[0085] Subsequently, the software sets the trajectory 19 along which the sample 17 will be separated from the needle 15 of the manipulator 6 using the FIB, so that this trajectory 19 is located on the designated edge of the needle 15 of the manipulator 6 adjacent to the sample 17.

[0086] In the following step, the sample 17 is separated from the needle 15 of the manipulator 6 by means of the FIB following the trajectory 19 determined in the previous step.

[0087] In the second specific example, the device contains one column 1_ with a source 2 of ions (FIB), connected to the chamber 5. In the chamber 5, there is a stage 8 movable in three mutually perpendicular axes and rotatable around one axis and tiltable around one axis. A substrate 7 with prepared samples 17 is placed on the stage 8.

[0088] In the first step, an image of the manipulator 6 is created by creating an image of the field of view without the manipulator 6 in the field of view using the FIB and then an image of the manipulator 6 in the field of view. Based on the difference between these images, the software creates an image of the manipulator 6. Furthermore, the tip 16 of the needle 15 of the manipulator 6 is determined in this image of the manipulator 6 using a software.

[0089] In the next step, two images of the sample 17 are created. The first image of the sample 17 is created using the FIB so that the optical axis 4 of the FIB forms an angle of 90° with the plane of the stage 8. Subsequently, the stage 8 is tilted about the tilt axis so that the plane of the stage 8 forms an angle of 45° with the optical axis 4 of the FIB, and a second image of the sample 17 is created using the FIB. By combining these two images, two edges of the sample 17 are detected using the software. At the intersection of these edges, the software determines the welding point 18 for creating a connection between the tip 16 of the needle 15 of the manipulator 6 and the sample 17.

[0090] Subsequently, the manipulator 6 is moved towards the welding point 18 using the determined tip 16 of the needle 15 and the images of the sample 17 so that the distance between the tip 16 of the needle 15 and the welding point 18 is 1 pm. The spatial position of the manipulator is determined based on images taken using the FIB, and these images are created at the moment when the optical axis 4 of the FIB forms an angle of 90° with the plane of the stage 8, and further at the moment when the stage 8 is tilted so that the plane of the stage 8 forms an angle of 45° with the optical axis 4 of the FIB, while the manipulator 6 is also tilted so that the end of the manipulator 6 with the needle 15 is in the same position and at the same angle of inclination with respect to the plane of the stage 8.

[0091] Subsequently, while simultaneously supplying gaseous precursor using GIS, the FIB is directed between the tip 16 of the needle 15 of the manipulator 6 and the welding point 18, and a connection is created by the deposited material between the needle 15 of the manipulator 6 and the sample 17.

[0092] In the next step, the FIB is directed to the place where the sample 17 is connected to the substrate 7, and using this beam, the sample 17 is separated from the substrate 7. Subsequently, the sample 17 is picked up from the substrate 7 by the manipulator 6.

[0093] In the next step, the manipulator 6 transfers the sample 17 to the sample holder 9 located in the device 10 for mounting the sample holder 9, at a distance of 1 pm.

[0094] Subsequently, while simultaneously supplying a gaseous precursor using the GIS, the FIB is directed between the sample 17 and the sample holder 9, and a connection is created between the sample 17 and the sample holder 9 by the deposited material.

[0095] In the next step, an image of the sample 17 connected to the manipulator 6 and also connected to the sample holder 9 is created by the FIB. This image is compared with the image of the manipulator 6 without the attached sample 17, which is used as a template, on the basis of which the evaluation unit determines in the image obtained in this step the edge of the manipulator 6 adjacent to the sample 17.

[0096] Subsequently, the software sets the trajectory 19, along which the sample 17 will be separated from the needle 15 of the manipulator 6 using the FIB, so that this trajectory 19 is located on the specified edge of the needle 15 of the manipulator 6 adjacent to the sample 17.

[0097] In the next step, the sample 17 is separated from the needle 15 of the manipulator 6 using the FIB following the trajectory 19 determined in the preceding step.

[0098] List of reference symbols:

[0099] 1 - column

[0100] 2 - charged particle source

[0101] 3 - elements for shaping and directing charged particles

[0102] 4 - optical axis of the column

[0103] 5 - chamber

[0104] 6 - manipulator

[0105] 7 - substrate

[0106] 8 - stage

[0107] 9 - sample holder

[0108] 10 - device for mounting the sample holder

[0109] 11 - column 12 - charged particle source

[0110] 13 - optical axis of the column

[0111] 14 - elements for shaping and directing charged particles

[0112] 15 - needle 16 - needle tip

[0113] 17 - sample

[0114] 18 - welding point

[0115] 19 - trajectory for separating the sample from the manipulator

Claims

CLAIMS1. A method of preparing a sample using a device with at least one charged particle beam, said device comprising at least one column with a charged particle source producing charged particles and with elements for forming and directing charged particles into a charged particle beam, which is connected to a chamber comprising a stage for placing at least one substrate, a manipulator for transferring the sample, a device for mounting a sample holder, a sample holder for holding the sample placed in the device for mounting the sample holder, wherein the device optionally further comprises at least one device generating a photon beam connected to the chamber, a substrate with at least one sample placed on the stage, and an evaluation unit, the method comprising the following sequentially performed steps:- obtaining an image of the manipulator using the device with at least one charged particle beam,- moving the manipulator to a distance of at least 0 pm and at most 50 pm from the sample,- creating a connection between the manipulator and the sample using material deposition or redeposition,- separating the sample from the substrate,- removing the sample from the substrate using the manipulator,- transferring the sample to a distance of at least 0 pm and at most 50 pm from the sample holder, using the manipulator,- creating a connection between the sample and the sample holder using material deposition or redeposition,- separating the manipulator from the sample using a beam of charged particles or optionally using a photon beam, characterized in that the step of separating the manipulator from the sample using a beam of charged particles or optionally a photon beam comprises the following steps:- obtaining an image of the sample connected to the manipulator using the device with at least one charged particle beam,- comparing the image of the manipulator with the image of the sample connected to the manipulator, using an evaluation unit,- determining the edge of the manipulator adjacent to the sample, using the evaluation unit, based on the result of comparing the image of the manipulator with the image of the sample connected to the manipulator,- separating the manipulator from the sample using a beam of charged particles or optionally a photon beam guided along the edge of the manipulator adjacent to the sample determined in the preceding step.

2. The method of preparing a sample using a device with at least one charged particle beam according to claim 1 , wherein the said device comprises at least one column with a charged particle source producing charged particles and with elements for forming and directing charged particles into a charged particle beam, which is connected to a chamber comprising a stage for placing at least one substrate, a manipulator for transferring the sample, a device for mounting a sample holder, a sample holder for holding the sample placed in the device for mounting the sample holder, wherein the device further comprises a substrate with at least one sample placed on the stage, and an evaluation unit, the method comprising the following sequentially performed steps:- obtaining an image of the manipulator using the device with at least one charged particle beam,- moving the manipulator to a distance of at least 0 pm and at most 50 pm from the sample,- creating a connection between the manipulator and the sample using material deposition or redeposition,- separating the sample from the substrate,- removing the sample from the substrate using the manipulator,- transferring the sample to a distance of at least 0 pm and at most 50 pm from the sample holder, using the manipulator,- creating a connection between the sample and the sample holder using material deposition or redeposition,- separating the manipulator from the sample using a beam of charged particles, characterized in that the step of separating the manipulator from the sample using a beam of charged particles comprises the following steps:- obtaining an image of the sample connected to the manipulator using the device with at least one charged particle beam,- comparing the image of the manipulator with the image of the sample connected to the manipulator, using an evaluation unit,- determining the edge of the manipulator adjacent to the sample, using the evaluation unit, based on the result of comparing the image of the manipulator with the image of the sample connected to the manipulator,- separating the manipulator from the sample using a beam of charged particles guided along the edge of the manipulator adjacent to the sample determined in the preceding step.

3. The method of preparing a sample using a device with at least one charged particle beam according to claim 1 , wherein said device comprises at least one column with a charged particle source producing charged particles and with elements for forming and directing charged particles into a charged particle beam, which is connected to a chamber comprising a stage for placing atleast one substrate, a manipulator for transferring the sample, a device for mounting a sample holder, a sample holder for holding the sample placed in the device for mounting the sample holder, wherein the device further comprises at least one device generating a photon beam connected to the chamber, a substrate with at least one sample placed on the stage, and an evaluation unit, the method comprising the following sequentially performed steps:- obtaining an image of the manipulator using the device with at least one charged particle beam,- moving the manipulator to a distance of at least 0 pm and at most 50 pm from the sample,- creating a connection between the manipulator and the sample using material deposition or redeposition,- separating the sample from the substrate,- removing the sample from the substrate using the manipulator,- transferring the sample to a distance of at least 0 pm and at most 50 pm from the sample holder, using the manipulator,- creating a connection between the sample and the sample holder using material deposition or redeposition,- separating the manipulator from the sample using a beam of charged particles or using a photon beam, characterized in that the step of separating the manipulator from the sample using a beam of charged particles or a photon beam comprises the following steps:- obtaining an image of the sample connected to the manipulator using the device with at least one charged particle beam,- comparing the image of the manipulator with the image of the sample connected to the manipulator, using an evaluation unit,- determining the edge of the manipulator adjacent to the sample, using the evaluation unit, based on the result of comparing the image of the manipulator with the image of the sample connected to the manipulator,- separating the manipulator from the sample using a beam of charged particles or a photon beam guided along the edge of the manipulator adjacent to the sample determined in the preceding step.

4. A computer programme configured for performing the method according to any one of claims 1 to 3.

5. An evaluation unit comprising a computer programme according to claim 4 loaded in the memory or processor of the evaluation unit.

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