Atomic force microscope and corresponding method for obtaining a force spectrum measurement of a sample

Abstract

This invention relates to an atomic force microscope (1) for obtaining force spectral measurements of a sample, comprising a head (100) movably connected to a mounting, wherein the head (100) includes a receiving unit (106) configured to receive a cantilever (202) including a tip (203) for contacting a soft material sample. The invention also relates to a system including an atomic force microscope, components including a system having an atomic force microscope, a method for obtaining force spectral measurements of a sample, and a method for obtaining a reference position of a beam using an atomic force microscope.

Description

Technical Field

[0001] This invention relates to an atomic force microscope (AFM) for obtaining force spectrum measurements of a sample. The invention also relates to a corresponding method for obtaining force spectrum measurements of a sample. Summary of the Invention

[0002] A first aspect of the invention relates to an atomic force microscope for obtaining force spectrum measurements of a sample. The atomic force microscope includes a head movably connected to a mounting, wherein the head includes: The receiving unit is configured to receive a cantilever including a tip for contacting a soft material sample: A light source and focusing optics for generating light are configured to focus the light, and when the cantilever is positioned at the receiving unit, the first light beam is guided onto the cantilever along a first longitudinal axis. A reflector is configured to receive a second beam of light reflected by the cantilever when the cantilever is positioned at the receiving unit. The detector is configured to detect the position of a third beam reflected by a mirror and generate a deflection signal indicating the deflection of the cantilever along a first longitudinal axis based on the detected position of the third beam.

[0003] The atomic force microscope also includes an actuator configured to move the head along a first longitudinal axis, wherein the head is connected to a mounting via the actuator.

[0004] According to an embodiment, an atomic force microscope is configured to obtain force spectrum measurements of samples, particularly soft materials, especially tissue specimens.

[0005] According to an embodiment, AFM is configured to obtain the stiffness value of a sample, particularly a soft material, especially a tissue specimen.

[0006] According to an embodiment, AFM is configured to obtain the nanomechanical phenotype of a sample, particularly a soft material, especially a tissue specimen. According to an embodiment, AFM is configured to determine the Young's modulus of a sample, particularly a soft material, especially a tissue specimen.

[0007] According to an embodiment, AFM is configured to determine the adhesiveness of samples, particularly soft materials, especially tissue specimens.

[0008] According to an embodiment, AFM is configured to determine the dissipation of a sample, particularly soft material, especially a tissue specimen.

[0009] According to an embodiment, the AFM includes an AFM head that includes a clamping mechanism for holding an optically transparent cantilever holder. Specifically, the cantilever holder may be an optically transparent cantilever chip holder with an attached cantilever chip, the cantilever chip including at least one cantilever.

[0010] In one embodiment, the AFM includes a tray for arranging a sample holder below the AFM head. The sample holder is configured to hold soft material samples, particularly tissue samples. The tray can be a sample stage. The sample holder can be a petri dish. The sample holder can be a Piper plate. The sample holder can be a microplate. The sample holder can be a 6-well plate. The sample holder can be a sample support.

[0011] In one embodiment, the AFM head includes a recess for the AFM head. In another embodiment, the tray includes a slot for accommodating a cantilever retainer. In yet another embodiment, the tray includes a slot and is configured such that the cantilever retainer can be automatically inserted from the slot of the tray into the recess of the AFM head. In yet another embodiment, the sample holder is a 6-well plate, one well of which can be used to store a rinsing / cleaning solution into which the cantilever can be lowered for automatic rinsing / cleaning.

[0012] In an embodiment, the AFM includes a first linear actuator for linear movement of the AFM head, and particularly the entire AFM head, relative to the tray along the vertical z-direction. In the context of this application, the vertical z-direction is also referred to as the first direction. In an embodiment, the first linear actuator connects the AFM head to a mounting bracket.

[0013] In an embodiment, the AFM includes a second linear actuator for linear movement of the AFM head, and particularly the entire AFM head, relative to the tray along the horizontal z-direction. In the context of this application, the horizontal z-direction is also referred to as the second direction.

[0014] In an embodiment, the AFM includes a third linear actuator for linear movement of the tray relative to the AFM head along the horizontal y-direction, particularly perpendicular to the x-direction. In the context of this application, the horizontal y-direction is also referred to as the third direction.

[0015] According to an embodiment, the first linear driver, the second linear driver, and the third linear driver each include a coarse driver. The coarse driver may include or be composed of a voice coil motor. Alternatively, the coarse driver may include or be composed of a piezoelectric stick-slip motor.

[0016] According to an embodiment, the first linear driver, the second linear driver, and the third linear driver each include at least one precision driver. The at least one precision driver may include or be composed of a piezoelectric motor. The at least one precision driver may include or be composed of a piezoelectric LEGS motor. The at least one precision driver may include or be composed of a friction driver.

[0017] In this embodiment, the coarse driver is configured for coarse positioning of the AFM head and the sample holder.

[0018] In one embodiment, the precision actuator is configured for precise positioning of the AFM head and the sample holder.

[0019] According to an embodiment, the fine actuator of the first linear actuator is configured to bring the cantilever closer to and / or level the soft material sample (automatically and without vibration) during force spectrum (stiffness) measurement of the soft material sample (in conjunction with the fourth linear actuator).

[0020] In one embodiment, the coarse actuator is configured to be de-energized during fine positioning to avoid vibration.

[0021] In one embodiment, the coarse driver and the fine driver can be magnetically coupled at the desired location on the AFM head to avoid vibration.

[0022] In an embodiment, the coarse drive and the fine drive can be magnetically coupled at the desired location on the tray, particularly in the case of a third linear drive, to avoid vibration.

[0023] In one embodiment, the coarse actuator vibrates to a certain extent, but the fine actuator vibrates essentially without vibration. According to one embodiment, the precision of both the coarse and fine actuators is 100 nm. According to one embodiment, the driving range of the coarse actuator is 85 mm in the z-direction. According to one embodiment, the driving range of the coarse actuator is 230 mm in the x-direction. In one embodiment, the driving range of the coarse actuator is 230 mm in the y-direction. In one embodiment, the driving range of the fine actuator is 40 mm in the z-direction. In one embodiment, the driving range of the fine actuator is 80 mm in the x-direction. In one embodiment, the driving range of the fine actuator is 80 mm in the y-direction.

[0024] In one embodiment, the first linear driver includes two fine drivers located on either side of the coarse driver of the first linear driver along the x-axis.

[0025] In one embodiment, the AFM includes a fourth linear actuator for linear movement of the AFM head, particularly the entire AFM head, relative to the tray along the z-axis. In another embodiment, the fourth linear actuator includes a piezoelectric motor, particularly a piezoelectric stack actuator. In yet another embodiment, the travel range of the fourth linear actuator along the z-axis is 38 μm. In yet another embodiment, the resolution of the travel range of the fourth linear actuator along the z-axis is less than or equal to 1 nm. In yet another embodiment, the fourth linear actuator is configured to move the cantilever during force spectrum measurements, particularly automated force spectrum measurements.

[0026] In an embodiment, the AFM includes at least one, particularly two, constant-force magnetic springs that apply an upward force along the z-direction across the entire AFM head, thereby counteracting the weight of the head. In this embodiment, the constant-force magnetic springs are arranged and configured such that when the first linear actuator, particularly the coarse and / or fine actuators of the first linear actuator, is de-energized to move the cantilever away from the sample, the entire AFM head moves upward along the z-direction, particularly as a safety measure, because it prevents the cantilever from colliding with the sample if the first linear actuator is de-energized.

[0027] In an embodiment, the AFM includes a first imaging system. The first imaging system can be arranged and configured to image the sample from above along the z-direction. According to an embodiment, a second imaging system is configured to obtain a depth map of the sample, particularly a 3D reconstructed or topographic deep-focus image, particularly a true-color image with all pixels in focus, particularly having a z-resolution (resolution in the z-direction) of about 4 μm (deep-focus imaging) and an xy-resolution of about 1 μm.

[0028] In one embodiment, the AFM includes a second imaging system for imaging the sample in the sample holder, particularly when the sample holder is arranged on a tray. In another embodiment, the second imaging system is arranged and configured to capture an overview image of the sample in the sample holder, particularly when the sample holder is arranged on a tray. In yet another embodiment, a first imaging system is arranged and configured to read barcodes on the sample holder and / or on the cantilever holder from above along the z-direction.

[0029] In an embodiment, the AFM includes a third imaging system, which is an optical microscope. The third imaging system may be a bottom-light microscope. In an embodiment, the third imaging system is arranged and configured to image the cantilever from below. In an embodiment, the third imaging system is arranged and configured to image the cantilever from below to determine the dimensions of the cantilever, particularly its length, width, and / or height, to obtain the cantilever's spring constant. In an embodiment, the third imaging system is arranged and configured to determine the position of the cantilever's tip. In an embodiment, the third imaging system is arranged and configured to perform automatic beam alignment in the x, y, and z directions.

[0030] In one embodiment, the third imaging system is configured to acquire a depth map / deep-focus image of the cantilever to automatically determine the exact position of the tip in the x, y, and z directions. In another embodiment, the third imaging system is configured to acquire a depth map / deep-focus image of the cantilever to automatically determine the exact position of the tip in the x, y, and z directions, thereby driving the optics to that position and then optimally aligning the beam in the x and y directions, and particularly using a z-drive to focus the beam onto the cantilever.

[0031] In one embodiment, the third imaging system has a fixed position relative to the first imaging system, allowing for the precise determination of the tip-to-sample offset. In another embodiment, the third imaging system has a fixed position relative to the second imaging system.

[0032] In one embodiment, a measurement unit, including an AFM head, first, second, and third linear actuators, and first, second, and third imaging systems, is fixed to an active vibration compensation stage. In another embodiment, the measurement unit is enclosed in an acoustically isolated housing.

[0033] According to an embodiment, the AFM head includes a body comprising an upper portion and a lower portion, the upper portion having a rectangular shape in a cross-section perpendicular to the z-direction, and the lower portion having a circular shape in a cross-section perpendicular to the z-direction.

[0034] According to an embodiment, the AFM head includes a base plate connected to a lower portion. This base plate includes a circular recess for inserting a cantilever holder that holds a cantilever chip including at least one cantilever, specifically wherein the cantilever extends along the y-direction. In an embodiment, the cantilever holder is a transparent cantilever chip holder. In an embodiment, the base plate includes a central through-hole disposed within the recess, specifically allowing light beams to enter the transparent cantilever holder from above and, if the cantilever holder includes or is a cantilever chip holder, to be reflected by the cantilever, particularly by the cantilever on the chip. In an embodiment, the recess has a tapered edge for centering the cantilever holder when inserted into the recess.

[0035] In one embodiment, three ceramic hemispheres are positioned in the groove at an angle of 120° around the circumference of the through-hole. In another embodiment, the cantilever retainer contacts the hemispheres when inserted into the groove.

[0036] According to an embodiment, the AFM head includes a light source for generating a light beam, which includes a laser source or superluminescent LED, SLED (e.g., IR, 830nm), fiber optic cable, and focusing and collimating optics (particularly having a focal length of 50mm and / or a light layer length of approximately 150μm). In an embodiment, the focusing and collimating optics include a lens. In an embodiment, the light source and the focusing and collimating optics are arranged and configured to generate a light beam and guide it to the free end of the cantilever held by the cantilever holder when the cantilever holder is positioned at the AFM, particularly when the cantilever is inserted into the recess. The free end of the cantilever may be formed by a mechanical fastening of the cantilever to the cantilever holder disclosed herein, wherein the free end of the cantilever is the end section of the cantilever remote from its mechanical fastening to the cantilever holder.

[0037] In one embodiment, the focusing and collimating optics are mounted in a holder connected to a cross-table comprising a first bracket and a second bracket.

[0038] In one embodiment, the holder and displacement stage include a central through-hole to receive focusing and collimating optics.

[0039] In an embodiment, the first carriage of the displacement stage can be moved in the y-direction by a first light source linear actuator, particularly with a resolution of about 10 μm. The first light source linear actuator includes a piezoelectric actuator, particularly a piezoelectric LEGS or a piezoelectric triboelectric actuator.

[0040] In an embodiment, the second bracket of the displacement stage can be moved in the x-direction by a second light source linear actuator, particularly with a resolution of about 10 μm. The second light source linear actuator includes a piezoelectric actuator, particularly a piezoelectric LEGS or a piezoelectric stick-slip motor.

[0041] In one embodiment, the retainer includes a first plate and a second plate stacked on top of each other in the z-direction, wherein the retainer can be tilted by three set screws connecting the first plate and the second plate.

[0042] In an embodiment, the beam can be aligned with the cantilever, particularly with the free end of the cantilever, through the linear movement of the displacement stage and the tilting of the holder.

[0043] According to an embodiment, the AFM head includes an additional z-driver (third bracket) configured to move the light source and focusing / collimating optics in the z-direction to automatically and optimally focus the beam onto the cantilever.

[0044] According to an embodiment, the AFM head includes a third bracket and a third light source linear actuator, particularly a piezoelectric motor, especially a piezoelectric leg motor, the third light source linear actuator being configured to move the third bracket along the z-direction.

[0045] In an embodiment, the beam can be aligned with the cantilever, particularly with the free end of the cantilever, by means of linear movement of the displacement stage, and in particular by means of a first light source linear driver, a second light source linear driver and / or a third light source linear driver.

[0046] According to an embodiment, the AFM includes an optical target, specifically a grid line on a plate below the AFM head and next to the sample holder. In an embodiment, the light beam of the light source is configured to scan the grid line to automatically determine and calibrate the precise position of the AFM head in the X and Y directions. In an embodiment, the AFM is configured to automatically determine and calibrate the precise position of the AFM head in the X and Y directions without the cantilever holder being installed. In an embodiment, for determination and calibration, the light beam passes through the head, strikes the grid, and the reflected light returns to a collimator, which has a beam splitter in the optical fiber to detect the presence of grid lines or spaces. In an embodiment, the AFM is configured to automatically determine and calibrate the precise position of the AFM head in the X and Y directions with the cantilever holder installed.

[0047] In one embodiment, the AFM includes a reflector configured to reflect a light beam reflected from a cantilever, particularly a cantilever with a cantilever, onto a 4-quadrant photodiode, particularly having a pivot at a certain location such that light reflected in air and liquid reaches the photodiode at the same angle. In one embodiment, the reflector is glued to a mirror frame. In another embodiment, the mirror frame includes a recess for receiving the glue.

[0048] According to an embodiment, a four-quadrant photodiode is configured to detect the deflection of a reflected beam during a force spectrum measurement. In the context of this application, a four-quadrant photodiode is also referred to as a photodiode. It can also be... According to one embodiment, the photodiode is mounted on a carriage. The carriage can be moved linearly in the x-direction by a PD linear actuator, which includes a piezoelectric actuator, particularly a piezoelectric LEGS motor or a friction actuator. The carriage can be connected to the PD linear actuator via a connecting rod extending in the z-direction. In one embodiment, the photodiode can be tilted on the carriage to adjust its position using three set screws.

[0049] In one embodiment, the reflector, particularly the mirror frame, is tiltable. In another embodiment, the reflector, particularly the mirror frame, is rotatable.

[0050] In an embodiment, the AFM head includes a push rod that can be moved along the z-direction by a push rod linear actuator, wherein the push rod linear actuator includes a piezoelectric actuator, particularly a piezoelectric leg motor or a piezoelectric stick-slip motor.

[0051] In one embodiment, the push rod includes a pin configured to engage a slot in the connecting rod, specifically such that the connecting rod pivots in the yz plane when the push rod moves in the z direction.

[0052] In one embodiment, the mirror frame is rotatable about an axis extending in the x-direction, which is connected to a connecting rod such that when the push rod moves in the z-direction, the connecting rod pivots the mirror frame (particularly the mirror associated with the mirror frame) about this axis.

[0053] In one embodiment, the reflector is motorized to automatically compensate for the bending drift of the cantilever, thereby keeping the reflected beam within the optimal operating range of the photodiode, particularly in the z-direction.

[0054] According to an embodiment, the leg springs are arranged to have springs around an axis, and the legs are respectively attached to the post and the spring fastener to keep the mirror frame in a fixed position, in particular to avoid vibration.

[0055] In one embodiment, the cantilever retainer clamping mechanism includes two L-shaped clamping elements for clamping the cantilever retainer in a groove in the base plate. In another embodiment, each clamping element is connected to a corresponding bolt extending in the x-direction. In yet another embodiment, the bolt is arranged in a ball bearing fixed to the base plate of the body. In still another embodiment, the clamping element includes a set screw for adjusting its position.

[0056] According to an embodiment, each bolt is connected, and in particular glued (e.g., by Loctite glue), to a corresponding L-shaped actuating lever in the lower part of the body.

[0057] In an embodiment, a corresponding leg spring is attached to each bolt, one of the legs is attached to a post on a corresponding actuating lever, the leg spring holding the actuating lever in an upper position, forcing the clamping element in a clamped position, in which the clamping element applies an upward force (in the z direction) on the cantilever retainer, which secures the cantilever retainer in a groove (also referred to as a recess in the context of this application).

[0058] In one embodiment, when the push rod moves to the lower position along the z-direction, the push rod engages the actuating lever and pushes the actuating lever into the lower position, which forces the clamping element into the open position, in which the cantilever retainer can be inserted into or removed from the recess.

[0059] In one embodiment, the piezoelectric actuator connected to the push rod has a dual function: pivoting the reflector and opening / closing the cantilever retainer clamping mechanism.

[0060] In an embodiment, the AFM includes a magnet and a corresponding 3D magnetic position sensor, particularly a Hall sensor, to determine the position of the corresponding magnet arranged on the following: - The axis of the mirror frame is used to determine the rotational orientation of the mirror. - The bolts connecting the clamping elements are used to determine the rotational orientation of the bolts, specifically indicating whether the clamping elements are in the open or clamped position, whether the cantilever retainer is inserted into the groove, and whether the cantilever retainer is inserted in the correct position. -The upper part of the connecting rod that connects the photodiode carriage to the PD linear driver is used to determine the y-position of the photodiode. - The first bracket of the displacement stage, to determine its x-position, and / or - The second bracket of the displacement stage is used to determine its y-position.

[0061] In one embodiment, the temperature sensor is located on the base plate at the AFM head. In another embodiment, the temperature sensor is located near a recess in the AFM head. In yet another embodiment, the temperature sensor is located at the cantilever retainer. In a third embodiment, the temperature sensor is located at or near the cantilever retainer when the cantilever retainer is inserted into the recess.

[0062] In an embodiment, the temperature sensor is configured to determine the temperature, particularly the temperature of the cantilever and / or the temperature around the cantilever, and especially to determine the spring constant of the cantilever.

[0063] In one embodiment, a temperature sensor is arranged and configured to determine the temperature of the sample. In another embodiment, the temperature sensor is arranged and configured to determine the temperature around the sample, particularly the temperature of the buffer surrounding the sample.

[0064] In one embodiment, the AFM is configured to determine temperature measurements over time. In another embodiment, the AFM is configured to repeatedly determine temperature measurements. In yet another embodiment, the AFM is configured to continuously determine temperature measurements.

[0065] In one embodiment, the AFM includes a temperature control device configured to adjust the temperature if the measured temperature deviates from a predetermined temperature interval.

[0066] Advantageously, the viability of the sample is determined by measuring the temperature of the sample and / or the buffer surrounding the sample.

[0067] According to an embodiment, the AFM includes an AFM controller. The AFM controller may be made of a System-on-Module (SoM), which consists of an FPGA (Field Programmable Gate Array) for computation, a real-time Linux running on an ARM processor for computation, and a device controller, particularly for computation on a timescale of about 5 ns, particularly for computation on a timescale of about 50 μs, and particularly for a Linux operating system for device automation and / or user input.

[0068] In this embodiment, the controller's inputs come from and / or the controller's outputs are sent to: Photodiodes (Force Spectrum Photodiodes), especially via preamplifiers and analog-to-digital converters, Head reference photodiode, First imaging system, Second imaging system, Third imaging system, Magnetic positioning sensor Ambient temperature sensor, Pressure sensor, Humidity sensor, Voltage sensor, Sample temperature sensor, Vibration compensation table, The coarse driver of the first linear driver, The coarse driver of the second linear driver, The coarse driver of the third linear driver Position controller of the first linear drive Position controller of the second linear drive Position controller of the third linear drive The fine actuator of the first linear actuator (especially piezoelectric actuators, particularly piezoelectric LEG motors or friction actuators), The fine actuator of the second linear actuator (especially the piezoelectric actuator, particularly the piezoelectric LEG motor or friction actuator), The fine actuator of the third linear actuator (especially piezoelectric actuators, particularly piezoelectric LEG motors or friction actuators), Position controller of the first linear drive Position controller of the second linear drive Position controller of the third linear drive Piezoelectric actuators for PD linear actuators, particularly piezoelectric LEG and / or friction actuators for PD linear actuators. A piezoelectric actuator that moves a push rod to rotate a reflector, particularly a piezoelectric LEGS and / or friction actuator that moves a push rod to rotate a reflector. Piezoelectric actuators that move push rods to actuate clamping mechanisms, particularly piezoelectric LEGS and / or friction actuators that move push rods to actuate clamping mechanisms. A piezoelectric actuator that moves a push rod to rotate a reflector and actuate a clamping mechanism, particularly a piezoelectric LEGS and / or friction actuator that moves a push rod to rotate a reflector and actuate a clamping mechanism. Piezoelectric actuators, particularly piezoelectric LEGS actuators and / or piezoelectric stick-slip motors, actuate a displacement stage to align an illumination beam from a light source onto the cantilever. Light source controller SLED controller The controller is configured to control the fourth linear actuator, specifically the force spectrum actuator controller (piezoelectric stack motor 140). The light curtain of the friction actuator A light curtain is arranged and configured to monitor the status of the device doors (especially system doors) used for inserting cantilever holders and sample holders.

[0069] In one embodiment, some of the controller's circuit boards are positioned around the upper part of the AFM head body in a chip-like manner (3D, multiple boards perpendicular to each other), resulting in a compact architecture. In another embodiment, multiple circuit boards of the controller are arranged in a 3D manner around the upper part of the AFM head body, perpendicular to each other, resulting in a compact architecture.

[0070] According to an embodiment, the controller implements at least one of the following methods: Automatic loading and clamping of the cantilever retainer Automatic beam alignment on the cantilever. Automatic beam focusing on the cantilever. A third linear actuator agitates the tray, where the first and / or second imaging systems (top-down camera) monitor the sample's position within the sample holder on the tray, and the controller automatically determines whether the sample is firmly attached to the sample holder and not floating. This is used as "intake control" to determine if the sample is intact and well-shaped. Automatic determination of the force map of the sample. Automatic collision detection, especially for finding areas that the probe cannot measure due to geometric constraints. Automatic tilt detection, especially for regions where the sample is too steep to perform an effective force spectrum analysis. The automatic determination of the cantilever's spring constant, particularly through the measurement of the cantilever's dimensions using a third imaging system and temperature sensors, The system's deflection sensitivity is automatically determined, where deflection [nN] = deflection signal on the photodiode [V] * deflection sensitivity [nm / V] * spring constant [N / m]. Cantilever drift compensation, Automated force spectrum measurement, particularly including at least one of the following steps: Automatic selection of measurement points on samples, especially soft material samples, and selection of, for example, 20×20 measurement points. Automatic leveling of the cantilever tip on soft material samples. Automatic slope length optimization for force spectrum measurement The advanced safety mechanism, in particular, involves an AFM head that automatically moves upward in the z-direction away from the soft material sample upon detection of vibration or an obstruction; specifically, the vibration and / or obstruction is identified by a first imaging system, a second imaging system, a vibration compensation stage, and / or a seismic detector. Automatic force curve quality control When the quality of the automatic force curve is poor, especially when it is lower than the predetermined quality measurement, a new measurement point is automatically selected on the soft material sample.

[0071] In one embodiment, a particular method is implemented in hardware. In another embodiment, a particular method is implemented in software.

[0072] In one embodiment, the first imaging system is repeatedly and removably attached to the mounting and is in a fixed position relative to the head.

[0073] In one embodiment, a second imaging system is repeatedly and removably connected to the mount, positioned in a fixed location relative to the head. In another embodiment, a third imaging system is repeatedly and removably connected to the AFM, positioned in a fixed location relative to the head.

[0074] In one embodiment, the third imaging system is repeatedly and removably connected to the AFM, and is in a fixed position relative to the first imaging system.

[0075] In one embodiment, the third imaging system is repeatedly and removably connected to the AFM, and is in a fixed position relative to the second imaging system.

[0076] In one embodiment, a first imaging system is repeatedly and removably connected to the mount, positioned in a fixed location relative to the head, and a second imaging system is repeatedly and removably connected to the mount, also positioned in a fixed location relative to the head. The first imaging system, the second imaging system, and the head can be fixedly coupled to each other, particularly to each other in a fixed position.

[0077] In one embodiment, a first imaging system is repeatedly and removably connected to the mount and is in a fixed position relative to the head, a second imaging system is repeatedly and removably connected to the mount and is in a fixed position relative to the head, and a third imaging system is repeatedly and removably connected to the AFM and is in a fixed position relative to the first imaging system.

[0078] Advantageously, the position of each of the first imaging system, the second imaging system, the third imaging system, and / or the head relative to each other can be determined and fixed.

[0079] Advantageously, the reference positions of the first imaging system, the second imaging system, the third imaging system, and / or the head relative to each other can be determined.

[0080] Advantageously, external disturbances can affect each of the first imaging system, the second imaging system, the third imaging system, and / or the head equally, since they are in fixed positions relative to each other.

[0081] Tag scanners for scanning samples and / or probe holders can be arranged and configured as follows: Provide traceability of the sample holder, and / or Provides probe traceability.

[0082] Advantageously, this avoids the double use of consumables. It can improve the quality of measurements.

[0083] In this embodiment, the probe is calibrated on the production floor, and its characteristics are stored in a barcode that can be read by a label scanner. Advantageously, the calibration (spring constant) can be removed during the measurement process to save time.

[0084] In this embodiment, the cantilever width and / or length can be determined with high precision on-site to improve the accuracy of spring constant measurement. The width and / or length can be stored in a barcode that can be read by a label scanner.

[0085] In this embodiment, the tip height is measured on-site to ensure the tip is long enough for measurement. The tip height can be stored in a barcode that can be read by a label scanner.

[0086] A second aspect of the invention relates to a system comprising an atomic force microscope and a vibration isolation stage according to a first aspect of the invention, wherein the vibration isolation stage includes a vibration sensor.

[0087] According to an embodiment, the system includes an interface, particularly a human-machine interface, which includes a drawer unit configured to receive a shuttle unit for carrying a sample holder and / or a cantilever holder. The drawer unit is movable between a loading position for placing the shuttle unit on the drawer unit remotely from the sample stage and an unloading position at the sample stage. The drawer unit includes a mechanism for unloading the shuttle unit from the drawer unit, particularly transferring it to a tray, when the shuttle unit is placed on the drawer unit and the drawer unit is moved to the unloading position.

[0088] In one embodiment, the drawer unit includes a plurality of protrusions for placing the shuttle unit on the protrusions of the drawer unit.

[0089] In another embodiment, the drawer unit includes a slider, particularly a telescopic slider, for moving the drawer unit, particularly along a third longitudinal direction, between a loading position and an unloading position.

[0090] According to an embodiment, the height-adjustable structure is slidably supported on each slider by at least one pin extending from the slider into the recess of the height-adjustable structure, and wherein the height-adjustable structure includes the protrusion, and wherein the recess is shaped such that when the drawer unit is moved to the unloading position and the height-adjustable structure engages with the stop, the vertical position of the protrusion and the shuttle unit placed on the protrusion is lowered by moving the height-adjustable structure relative to at least one pin of the slider.

[0091] By reversing the movement of the aforementioned mechanism, the protrusion can be raised to pick up the shuttle unit when it is positioned on the tray, particularly when it is positioned on the support element of the tray.

[0092] Specifically, the recess may have two substantially horizontally extending sections, i.e., extending along a third longitudinal axis, wherein the two horizontal sections are arranged in different vertical positions along a first longitudinal axis, and wherein the two horizontal sections are connected by a ramp portion of the recess. In this way, the pin of the slider can be moved from one end of the recess to the other by vertical displacement of the height-adjustable structure relative to the slider.

[0093] In another embodiment, the sample stage includes a plurality of support elements, particularly three support elements, for supporting the shuttle unit, wherein, in the unloading position, a protrusion for placing the shuttle unit is arranged above the support elements such that the shuttle unit can be unloaded from the drawer unit to the sample stage by means of a height-adjustable structure relative to the slider.

[0094] According to an embodiment, the system includes a label scanner, particularly a label scanner for scanning barcodes, wherein the barcode encodes information about a sample, a sample holder, a probe holder, and / or a cantilever.

[0095] In another embodiment, the sample holder is a petri dish, a Piper dish, a microplate (especially a 6-well plate), or a sample support.

[0096] In another embodiment, the system includes a hydraulic wheel, wherein the hydraulic wheel is arranged and configured such that the system can move via the hydraulic wheel, and wherein the system includes a fixed support foot, wherein the fixed support foot is arranged and configured such that the system can stand stably and / or can be aligned via the support foot.

[0097] A third aspect of the invention relates to an assembly comprising a system and a package according to a second aspect of the invention, wherein the package is configured to receive the system, wherein the package includes a ramp configured to support release of the system from the package, and in particular, wherein the package includes a lid configured wedge-shaped and arranged such that the system can be released from the package via the wedge.

[0098] A fourth aspect of the invention relates to a method for obtaining force spectral measurements of a sample using an atomic force microscope according to any one of claims 1 to 24, or a system according to any one of claims 25 to 34, or components according to claim 34, the sample being particularly from soft materials, and more particularly biological tissue samples, the method comprising the following steps: The sample is contacted at the measurement point by the tip of the cantilever. During the loading step, particularly automatically, the head is moved towards the sample parallel to the first longitudinal axis by an actuator, causing the tip of the cantilever to be pressed into the sample. During the unloading step, particularly automatically, the head is moved away from the sample by an actuator parallel to the first longitudinal axis. During the loading and / or unloading steps, multiple deflection values ​​of the cantilever are determined from the deflection signals of the detector. From the determined deflection value, in particular automatically, the force spectrum measurement of the sample at the measurement point is determined.

[0099] In an embodiment, the method includes determining a plurality of force spectrum measurements, the plurality of force spectrum measurements indicating forces cantilevered on the sample at corresponding positions along a first longitudinal axis, wherein the force spectrum measurements of the sample are determined based on at least a subset of force values, particularly automatically, and in particular, wherein force-position relationships and / or force-position curves are obtained.

[0100] In an embodiment, the position of the sample relative to the head, and particularly relative to the cantilever, is changed by moving the head along a first longitudinal direction, particularly by a first linear actuator and / or a second longitudinal direction, particularly by a second linear actuator, and / or moving the sample, particularly the sample holder holding the sample, particularly by a third linear actuator along a third longitudinal direction.

[0101] In another embodiment, the detector is moved along a second longitudinal axis by a detector linear actuator, and / or the mirror frame, particularly the associated reflector, is pivoted by a mirror actuator, such that the reflector and the detector are aligned, and a third beam reflected by the reflector satisfies a predetermined detection area of ​​the detector, particularly the linear area of ​​the detector, and in particular, the detector and / or the mirror frame are automatically moved to keep the third beam in the detection area of ​​the detector, particularly the linear area of ​​the detector.

[0102] According to an embodiment, the shape, size, length, width, and / or height of the cantilever are determined by a third imaging system, particularly automatically, and / or the position of the tip of the cantilever is determined by the third imaging system, particularly automatically.

[0103] In another embodiment, prior to contacting the sample via the tip of the cantilever, the method includes a step of 3D profile analysis of the sample, using information obtained by a first imaging system and / or using information obtained by a second imaging system, particularly information related to depth-of-focus images and / or depth maps, and calculations, particularly wherein the calculations include a Laplacian regression step.

[0104] In another embodiment, before contacting the sample via the tip of the cantilever, the method includes an automated point selection step that provides a prediction of the measurement quality at the selected measurement point based on sample-related information, information obtained through 3D contour analysis of the sample, information obtained through depth-of-focus imaging, information obtained through the depth map, information about the shape of the cantilever, information about the size of the cantilever, and / or the position of the tip of the cantilever.

[0105] In another embodiment, before contacting the sample via the tip of the cantilever, a reference sample with a predetermined stiffness is contacted via the tip of the cantilever, and while the tip contacts the reference sample without pressing into it, the head is moved by an actuator through a travel distance, wherein a deflection signal of the detector is obtained at the travel distance, and wherein the deflection sensitivity of the detector is determined, particularly automatically, from the travel distance and the deflection signal, and / or wherein the deflection sensitivity of the detector is determined in a non-contact manner, particularly based on information obtained through a resonance curve, particularly based on the area below the resonance curve, wherein the resonance curve is obtained in air or in a liquid, and / or wherein the deflection sensitivity of the detector is determined in a depth-stepping standard manner, particularly wherein before contacting the sample via the tip of the cantilever, the surface of a standard sample including a recess with a predetermined depth is contacted via the tip of the cantilever, and the head is moved by an actuator through a travel distance, wherein a deflection signal of the detector is obtained at the travel distance, and wherein the deflection sensitivity of the detector is determined, particularly automatically, from the travel distance and the deflection signal.

[0106] In another embodiment, a deflection sensitivity is determined, wherein the device detects when the deflection sensitivity deviates from a predetermined interval and initiates recalibration by realigning the beam and performing additional deflection sensitivity measurements, particularly automatically.

[0107] In another embodiment, the cantilever is moved near the sample or to the measurement point, particularly wherein the cantilever is moved near the sample or to the measurement point in a non-approach manner, based on sample-related information and / or based on information obtained through 3D profile analysis of the sample.

[0108] In another embodiment, adaptive optimization is performed to move the cantilever near the sample or to the measurement point, wherein information considering the deviation between the detected previous measurement point and the proposed previous measurement point is used, particularly by applying the deviation to the proposed measurement point.

[0109] In one embodiment, a vibration signal is obtained from a vibration sensor, and wherein, when the signal indicates the presence of vibration, particularly when it indicates the presence of vibration with an amplitude or frequency higher than a predetermined threshold, the head is moved away from the sample along a first longitudinal axis, particularly automatically.

[0110] According to the embodiment, the spring constant of the cantilever is determined, in particular automatically, based on data provided by a third imaging system, particularly the length, width and / or thickness of the cantilever, the temperature value obtained by a temperature sensor, the ambient humidity and / or the ambient pressure obtained by an environmental sensor, the resonant frequency of the cantilever and / or the quality factor of the cantilever.

[0111] In another embodiment, the spring constant of the cantilever is repeatedly detected during the measurement and / or the value related to deflection sensitivity calibration is repeatedly detected during the measurement, wherein the cantilever is replaced and / or recalibrated when the spring constant of the cantilever exceeds a predetermined range, and / or wherein the cantilever is replaced and / or recalibrated when the value related to deflection sensitivity calibration exceeds a predetermined range, wherein the recalibration includes realigning the first bundle relative to the cantilever, particularly the free end of the cantilever, and additional measurement of the value related to deflection sensitivity, and / or wherein the mass value of the cantilever is repeatedly detected during the measurement, wherein the cantilever is replaced when the mass value of the cantilever exceeds a predetermined range.

[0112] In another embodiment, when the head determines a predetermined force value to be performed by the cantilever to be picked up, in particular by the cantilever holder to be picked up, the cantilever is picked up, in particular the cantilever holder, in particular from the sample stage, in particular from a second slot included in the sample stage for receiving the cantilever holder including the cantilever.

[0113] In one embodiment, a correction factor related to the buffer surrounding the sample is determined, wherein a deflection signal is detected as the cantilever moves toward the sample along a first longitudinal axis before the tip of the cantilever contacts the sample, wherein the position of the cantilever along the first longitudinal axis is determined when the deflection signal is lost.

[0114] According to an embodiment, an atomic force microscope, particularly an AFM controller, especially a field-programmable gate array, monitors at least one monitoring signal, wherein the monitoring signal is one of the following: The sum of the cantilever and deflection signals, Vibration signal, The accelerometer signal, The signal from the seismic detector Signals obtained from a microphone, in atomic force microscopy: When the monitoring signal leaves the predetermined range, any movement in the atomic force microscope is interrupted to protect the probe. When the monitoring signal leaves the predetermined range, especially in the presence of external vibration or external acoustic noise, the force spectrum measurement is interrupted. When the monitoring signal leaves the predetermined range, especially in the event of external vibration or external acoustic noise, the optical measurements performed by the first optical system and / or the second optical system and / or the third optical system are interrupted, and / or When vibration and noise levels are low, the monitoring signal is used as a “gate” to trigger measurements only.

[0115] In another embodiment, the deflection sensitivity in a liquid, particularly a buffer, is determined based on the deflection sensitivity determined in air and positional information regarding the tip of the cantilever, particularly regarding the position of the tip of the cantilever relative to a first longitudinal direction, a second longitudinal direction, and / or a third longitudinal direction. The fifth aspect of the invention relates to a method for obtaining a reference position of a light beam using an atomic force microscope according to any one of claims 1 to 24 or a system according to any one of claims 25 to 34, comprising the following steps: a. Guide the first beam along the first longitudinal axis to the first optical target. b. The intensity of the first reference beam reflected by the first optical target is detected by the controller. c. The controller detects the position of the head perpendicular to the first longitudinal axis. d. To capture an optical image of the first optical target using the first imaging system and / or the second imaging system. e. Determine the positions of the following items: The head relative to the first imaging system, The head relative to the second imaging system, The first imaging system, compared to the second imaging system, has a head... The part relative to the first beam, The first imaging system relative to the first beam, and / or The second imaging system is relative to the first beam. Attached Figure Description

[0116] Exemplary embodiments are described below in conjunction with the accompanying drawings. The drawings are appended to the claims and accompanied by text explaining the various features of the illustrated embodiments and aspects of the invention. Each individual feature shown in and / or mentioned in the text of the drawings may be incorporated (or individually) into the claims relating to the first, second, third, fourth, and / or fifth aspects of the invention.

[0117] Figure 1 An embodiment of an atomic force microscope according to the present invention is shown, wherein a cantilever holder is arranged on the head of the atomic force microscope; Figure 2 A cross-sectional view of the head of an atomic force microscope according to an embodiment of the present invention is shown; Figure 3 An embodiment of an atomic force microscope according to the invention is shown, which has a tray for receiving a sample holder arranged below the head of the atomic force microscope; Figure 4a b. Schematically depicts the optical path of light emitted from the light source and reflected from the cantilever and mirror to the detector. Figure 4a ), in which the transparent body is arranged in Figure 4b In the cantilever retainer shown; Figure 5 A front view of the lower part of the head of an atomic force microscope according to an embodiment of the present invention is shown, with a cantilever retainer connected to the head; Figure 6 It shows Figure 5 Side view; Figure 7 A bottom view of the head of an atomic force microscope is shown, which has a slot for inserting a cantilever retainer. Figure 8 A bottom view of the head of an atomic force microscope is shown, with the cantilever retainer inserted in a slot.

[0118] Figure 9 An embodiment of a mechanism for operating a clamping member for attaching a cantilever retainer to a head is shown; Figure 10a Figures 1 and 2 show an embodiment of an atomic force microscope according to the present invention, wherein focusing and collimating optics are arranged on the upper part of the head; Figure 11 An embodiment of an atomic force microscope according to the present invention is shown, which has a third imaging system for cantilever imaging; Figure 12 An embodiment of an atomic force microscope according to the present invention is shown, which has focusing and collimating optics movable by a piezoelectric stick-slip motor; Figure 13An embodiment of the system according to the invention is shown, which has an atomic force microscope and an interface for controlling the drawer unit; Figure 14a Figures b and c illustrate embodiments of a system according to the invention, comprising two extraction units for loading and unloading a sample holder and / or a cantilever holder; and Figure 15 An embodiment of an atomic force microscope 1 according to the present invention is shown, the atomic force microscope including a linear actuator having an optical scale for optically detecting the driving distance of a linear actuator. Detailed Implementation

[0119] Figure 1 An embodiment of an atomic force microscope 1 according to the present invention is shown. The atomic force microscope 1 includes a head 100 movably connected to a mount (not shown). In particular, the head 100 can be moved relative to the mount via an actuator 140 of the atomic force microscope 1. For this purpose, the head 100 can be connected to the mount via the actuator 140. By means of the actuator 140, the head 100 can be moved along a first longitudinal axis L1, which corresponds to the embodiment of the atomic force microscope 1 according to the present invention. Figure 1 The z-axis of the coordinate system shown.

[0120] The atomic force microscope 1 includes a light source 105 located at the top of the upper portion 104a of the body 104 of the head 100. The light source 105 is configured to generate light for optically detecting the displacement of a cantilever 202, which is attached to a cantilever chip 201 of a cantilever chip holder 310 arranged at the receiving unit 106 of the atomic force microscope 1. Alternatively, the cantilever 202 can be attached to a cantilever holder. The body 104 includes a mirror configured to receive light emitted from the light source and reflected from the cantilever, and a detector configured to detect the position of the beam reflected from the mirror to the detector, such as... Figure 2 This will be further explained in the text.

[0121] The light source 105 may include focusing and collimating optics, such as lenses, and is connected to the body 104 via a displacement stage 110 including a first bracket 110a, a second bracket 110b, and a third bracket 110c.

[0122] The first bracket 110a of the displacement stage 110 is driven by the first light source linear driver 111a, according to... Figure 1 The coordinate system shown is movable in the y-direction, particularly with a resolution of about 10 μm. The first light source linear actuator includes a piezoelectric actuator, particularly a piezoelectric LEGS or a piezoelectric friction actuator or a piezoelectric stick-slip motor.

[0123] The second bracket 110b of the displacement stage 110 is connected to the second light source linear driver 111b, according to... Figure 1 The coordinate system shown is movable in the x-direction, particularly with a resolution of about 10 μm. The second light source linear actuator includes a piezoelectric actuator, particularly a piezoelectric LEGS or a piezoelectric friction actuator or a piezoelectric stick-slip motor or a piezoelectric stick-slip motor.

[0124] The third bracket 110c of the displacement stage is driven by a second light source linear actuator, according to... Figure 1 The coordinate system shown is movable in the z-direction, particularly with a resolution of about 10 μm. The second light source linear actuator includes a piezoelectric actuator 111c, particularly a piezoelectric LEGS or a piezoelectric friction actuator or a piezoelectric stick-slip motor or a piezoelectric stick-slip motor.

[0125] By moving the light source 105 relative to the head 100, and in particular relative to the cantilever 202, with the aid of the displacement stage 110 and its brackets 110a, 110b, 110c, the spot position of the first beam B1 emitted from the light source 105 along the first longitudinal axis L1 toward the cantilever 202 can be aligned with and focused on the free end of the cantilever 202. This allows for a strong signal indicating the deflection of the cantilever 202 during measurement, and thus increases the measurement accuracy of the atomic force microscope 1.

[0126] Figure 2 A cross-sectional view is shown through the head 100 of an atomic force microscope 1 according to an embodiment of the invention. The cross-sectional view highlights components arranged inside the head 100, such as focusing optics 105a for light and for focusing the light onto a cantilever 202. Light can be reflected from the cantilever 202 toward a mirror 112 mounted on a mirror frame 114, which is configured to reflect the light toward a detector 113, at which the reflected light indicating the deflection of the cantilever 202 is detected. In this embodiment, the detector 113 is a four-quadrant photodiode arranged on a detector holder 116. The head 100 includes a detector linear driver 116a, particularly a piezoelectric motor, configured to move the detector holder 116 with the detector 113 along an x-direction perpendicular to the x-axis. Figure 2 The z and y directions are indicated in the image. For this purpose, the detector linear actuator 116a is connected to the detector holder 116 via a connecting rod extending in the z direction. Translational movement of the connecting rod 117 caused by the detector linear actuator 116 can thus be transmitted to the detector holder 116. The detector holder 116 can be tilted by three set screws 116b to adjust the orientation of the detector holder 116 relative to the incident light beam reflected from the reflector 112. A magnet-based position sensor 126a is arranged on the photodetector holder 116 such that the position of the photodetector holder 116 with the photodetector 113 can be determined.

[0127] The mirror frame 114 is rotatable about an axis 114a extending in the x-direction, which is perpendicular to the axis 114a. Figure 2 The coordinate system shown includes the y and z directions. A mirror actuator 114b (particularly a piezoelectric motor) forms a clamping actuator 1222, which is configured to move the device 1224 along a first longitudinal axis L1. The clamping element 122 for clamping the cantilever chip holder 310 to the head 100 of the atomic force microscope 1 via the moving device 1224 can be transferred between a receiving position and a clamping position. In the receiving position, the cantilever chip holder 310 can be arranged in the receiving unit 106; in the clamping position, the cantilever chip holder 310 is fixed to the receiving unit 106 by the clamping element 122.

[0128] A shaft 114a of the mirror frame 114 is mechanically connected to a connecting lever 121, which is mechanically connected to a push rod 119. The connecting lever 121 includes a groove 121a, and the push rod includes a pin 120 disposed in the groove. Thus, when the mirror actuator 114b moves the device 1224 along the z-direction, the pin 120 moves in the groove 121 due to the pivoting movement of the mirror 112 on the mirror frame 114. Therefore, the mirror actuator 114b is configured to achieve engagement of the clamping element 122 and pivoting of the mirror 112.

[0129] Figure 3 An embodiment of an atomic force microscope 1 according to the present invention is shown, which has a tray 300 for receiving a sample holder 430 arranged below the head 100 of the atomic force microscope 1.

[0130] The head 100 is schematically depicted as being arranged on a first linear actuator 101 for linear movement of the head 100 relative to the tray 300 along the vertical z-direction. The first linear actuator 101 includes a coarse actuator 101a for coarse movement of the head 100 along the z-direction and a fine actuator 101b for fine movement of the head 100 along the z-direction. The coarse actuator 101a may include or be composed of a voice coil motor. The fine actuator 101b may include or be composed of a piezoelectric motor.

[0131] The atomic force microscope according to this embodiment includes a constant-force magnetic spring 130 that applies an upward force on the head 100 along the z-direction (i.e., away from the tray 300), which counteracts the weight of the head 100. The magnetic spring 130 is configured to cause the head 100 to move upward along the z-direction when the first linear actuator 101 (particularly the coarse actuator 101a and / or fine actuator 101b of the first linear actuator 101) is de-energized to move the cantilever 202 away from the sample. This represents a safety measure, as it prevents the cantilever 202 from colliding with the sample when the first linear actuator 101 is de-energized.

[0132] A first linear actuator 101 is disposed between the head 100 and the second linear actuator 102 for linear movement of the head 100 relative to the tray 300 along the x-direction, which is perpendicular to the y and z directions. The second linear actuator 102 includes a coarse actuator 102a for coarse movement of the head 100 along the x-direction and a fine actuator 102b for fine movement of the head 100 along the x-direction. The coarse actuator 102a may include or be composed of a voice coil motor. The fine actuator 102b may include or be composed of a piezoelectric motor.

[0133] In this way, the head 100 can move relative to the tray 300 in two orthogonal directions by means of the first linear driver 101 and the second linear driver 102.

[0134] Furthermore, the atomic force microscope 1 according to this embodiment includes a third linear actuator 103 for linear movement of the tray 300 relative to the head 100 in the y-direction. The third linear actuator 103 includes a coarse actuator 103a for coarse movement of the tray 300 in the y-direction and a fine actuator 103b for fine movement of the tray 300 in the y-direction. The coarse actuator 103a may include or be composed of a voice coil motor. The fine actuator 103b may include or be composed of a piezoelectric motor.

[0135] Therefore, the head 100 and the tray 300 can be moved relative to each other in three orthogonal directions using the first linear actuator 101, the second linear actuator 102 and the third linear actuator 103, which allows the sample to be positioned precisely and flexibly relative to the cantilever 202 arranged on the head 100.

[0136] The atomic force microscope 1 according to this embodiment also includes a first imaging system 450 disposed on a first linear actuator 101. The first imaging system 450 is configured to image a sample disposed on a sample holder 430 from above along the z-direction. The first imaging system 450 is specifically configured to obtain a depth map of the sample, particularly a 3D reconstructed or deep-focus image of the topography, particularly a true-color image in which all pixels are in focus. For example, the z-resolution (i.e., the optical resolution in the z-direction) may be about 4 μm, and the xy-resolution (i.e., the optical resolution along the x and y directions) may be about 1 μm.

[0137] like Figure 3 As can be further seen, a second imaging system 460 is disposed on the head 100. The second imaging system 460 is configured to capture an overview image of the sample disposed in the sample holder 430. In addition, the first imaging system 450 may be disposed and configured to read the barcode on the sample holder and / or cantilever chip holder 310 from above (i.e., from the viewpoint of the head 100) along the z-direction.

[0138] Figure 3 Two shuttle units 650 arranged on tray 300 are also shown. The shuttle units 650 can be mounted on tray 300 and... Figure 14a The transfer between drawer units 620 is explained in the context of b and c. The shuttle unit 650 is configured to receive the sample holder 430 and / or cantilever chip holder 310 with the sample.

[0139] A third imaging system 440 is disposed on the tray 300. The third imaging system 440 is an optical microscope, such as a bottom-light microscope, configured to image the cantilever 202 from below (i.e., from the viewpoint of the tray 300). The third imaging system 440 can be configured to determine the dimensions of the cantilever 202, particularly its length, width, and / or height, in order to determine the spring constant of the cantilever 202. Additionally, the position of the tip of the cantilever 202 can be determined by the third imaging system 440. Thus, the third imaging system 440 can be used to automatically align the light beam to the tip of the cantilever 202 in the x, y, and z directions.

[0140] The third imaging system 440 can be configured to acquire depth maps and / or deep-focus images of the cantilever 202 to automatically determine the position of the tip in the x, y, and z directions.

[0141] The atomic force microscope 1 according to this embodiment also includes a controller 410. The controller 410 may be made of a system-on-module (SoM), which consists of an FPGA (Field Programmable Gate Array) for computation, a real-time Linux running on an ARM processor for computation, and a device controller, particularly for computation on a timescale of about 5 ns, particularly for computation on a timescale of about 50 μs, and particularly for a Linux operating system for device automation and / or user input.

[0142] The inputs to controller 410 may come from one or more of the following, and / or the outputs of controller 410 may go to one or more of the following: detector 113, particularly via a preamplifier and analog-to-digital converter; first imaging system 450, second imaging system 460 and third imaging system 440; magnetic positioning sensor 126a; ambient temperature sensor; ambient air pressure sensor; ambient air humidity sensor; voltage sensor; sample temperature sensor; vibration compensation stage on which atomic force microscope 1 is arranged; first linear actuator 101, second linear actuator 102 and third linear actuator 103, particularly their coarse actuators 101a, 102a, 103a and / or their fine actuators 101b, 102b, 103c; position controller for operating the first linear actuator, second linear actuator and / or third linear actuator 101, 102, 103; detector linear actuator 116a; mirror actuator 114b for pivoting mirror 112 and for engaging clamping element 122; light source controller for controlling light source 105; SLED controller.

[0143] like Figure 3 As can be further seen, the optical target 402 is arranged on the tray 300, below the head 100. The atomic force microscope 1 is configured to automatically determine and calibrate the position of the head 100 in the x and y directions when the cantilever chip holder 310 is not mounted on the head 100. For determination and calibration, a light beam emitted from the light source 105 passes through the light and focusing optics 105a of the head 100 and strikes the optical target 402. The light reflected from the optical target 402 returns towards the light and focusing optics 105a, which has a beam splitter connected to an optical fiber. The beam splitter is configured to calibrate the position of the head 100 relative to the tray 300 in the x and y directions based on the light reflected from the optical target 402.

[0144] Figure 4aThe optical path of light emitted from the light source 105 and detected by the detector 113 is schematically depicted. Light is emitted from the light source 105 as a first beam B1 to a cantilever 202 held by a cantilever holder 310. From the cantilever 202, particularly from its top surface facing the light source 105, the light is reflected as a second beam B2 to a reflector 112, which can be rotated by the mirror actuator 114b. See [reference needed]. Figure 2 From mirror 112, light is ultimately reflected as a third beam B3 to detector 113, which may be a four-quadrant photodiode. The optical path can be altered by moving light source 105 relative to cantilever 202 using displacement stage 110 with first, second, and third light source linear drivers 111a, 111b, and 111c along the x, y, and z directions, respectively. Furthermore, photodetector 113 can be moved along the x-direction (i.e., in the z-direction) by said detector linear driver 116a. Figure 4a (Outside the drawing plane shown) the movement. The deflection of the cantilever 202 indicates the distance between the tip 203 of the cantilever and the sample 3 arranged on the sample stage 300.

[0145] Figure 4a The diagram also schematically illustrates a first linear driver 101 having a coarse linear motor 101a and a fine linear motor 101b for moving a cantilever 202 held by a cantilever retainer 310 along a first longitudinal axis L1, and a second linear driver 102 having a coarse linear motor 102a and a fine linear motor 102b for moving the cantilever 202 held by the cantilever retainer 310 along a second longitudinal axis L2. Furthermore, the sample stage 300 can be moved along a third longitudinal axis L3 by a third linear driver 103 including a coarse linear motor 103a and a fine linear motor 103b.

[0146] Figure 4b An embodiment is shown in which a cantilever retainer 310 is configured to receive a transparent body 320. The transparent body 320 is formed of an optically transparent material, such as glass. Specifically, the material of the transparent body has a refractive index greater than that of air. In some embodiments, the material of the transparent body has a refractive index greater than that of the retainer material. For example, the transparent body has a refractive index of 1.6 to 2.2, more particularly about 1.9.

[0147] With the transparent body 320 provided, the light beam used to determine the deflection of the free end of the cantilever 202 passes through the transparent body 320 and reaches the cantilever 202, and the light beam reflected from the cantilever 202 travels through the transparent body 320 toward the mirror 112. At the interface of the transparent body 320, the light is refracted. The transparent body 320 improves the stability of the assembly formed by the transparent body 320 and the cantilever holder 310, allowing the cantilever holder 310 to be made longer in the z-direction, which simplifies the handling of the cantilever holder 310. Furthermore, when the material of the transparent body includes a higher refractive index than air, this allows the optics of the atomic force microscope 1 to be constructed more compactly, because the beam refraction at the transparent body 320 allows the beam to be refracted to the mirror 112, whereas without the transparent body 320, the light must be completely reflected from the cantilever 202 and guided from the cantilever 202 to the mirror, which requires the cantilever holder 310 to have a rather bulky shape, compared to Figure 4a and 4b The cantilever retainer 310 is understandable.

[0148] With the transparent body 320 provided, the light beam used to determine the deflection of the tip of the cantilever 202 passes through the transparent body 320 and reaches the cantilever 202, and the light beam reflected from the cantilever 202 travels through the transparent body 320 toward the mirror 112. At the interface of the transparent body 320, the light is refracted. The transparent body 320 improves the stability of the assembly formed by the transparent body 320 and the cantilever chip holder 310, allowing the cantilever chip holder 310 to be made longer in the z-direction, which simplifies the handling of the cantilever chip holder 310. Furthermore, when the material of the transparent body includes a higher refractive index than air, this allows the optics of the atomic force microscope 1 to be constructed more compactly, because the beam refraction at the transparent body 320 allows the beam to be refracted to the mirror 112, whereas without the transparent body 320, the light must be completely reflected from the cantilever 202 and guided from the cantilever 202 to the mirror, which requires the cantilever chip holder 310 to have a rather bulky shape, compared to Figure 4a and 4b The cantilever chip holder 310 is understandable.

[0149] Figure 5 A front view (corresponding to the xz plane) of a cantilever chip holder 310 according to an embodiment of the present invention is shown, which is arranged in the receiving unit 106 of the lower part 104b of the head 100 in the region extending through the longitudinal axis L of the head 100.

[0150] Figure 6 A side view (corresponding to the yz plane) of a cantilever chip holder 310 according to an embodiment of the present invention is shown, which is arranged at the receiving unit 106 at the lower portion 104b of the head 100. Specifically, Figure 6The clamping elements 122 are depicted for clamping the cantilever chip holder 310 to the head 100. Each clamping element 122 is connected to a direction perpendicular to the axis of rotation. Figure 6 The corresponding bolt 123 extends in the x-direction of the drawing plane, which will be in Figure 9 The text provides further explanation.

[0151] Figure 7 A bottom view of the head 100 of an atomic force microscope 1 according to an embodiment of the invention is shown. This view corresponds to a perspective view of the tray 300 below the head 100, facing the head 100, so that the receiving unit 106 for inserting the cantilever chip holder 310 is visible. In this embodiment, the receiving unit 106 forms a recess that frames a circular through-hole 107 arranged at the center of the receiving unit 106. Three alignment elements 108, particularly formed of ceramic hemispheres, are arranged in the recess around the periphery of the through-hole 107 for aligning, and particularly centering, the cantilever chip holder 310 when it is inserted into the recess. The edges around the recess may further taper gradually in the z-direction to simplify the insertion of the cantilever chip holder 310.

[0152] Figure 8 A cantilever chip holder 310 is shown in the receiving unit 106 of the head 100 of an insertion atomic force microscope 1 according to an embodiment of the present invention. The cantilever chip holder 310 is held in a clamped position by two clamping elements 122 arranged on opposite sides of a recess, the clamping elements engaging with the connecting surface 211 of the cantilever chip holder 310 to secure the cantilever chip holder 310 in the recess 106. Furthermore, the clamping elements 122 are connected to bolts 123 (e.g., rods), these pivoting members being pivotally mounted in bearings 124. One end of each bolt 123 is also connected to a lever 125. Each lever 125 includes a post 125a configured to attach a spring, such as a leaf spring, to bias the clamping element 122 to the clamped position. The lever 125 from Figure 8 The outer side of the head 100 shown extends to Figure 9 The inside of the head 100, as shown, allows the lever 125 to be operated by the push rod 119 disposed inside the head 100. Figure 9 The middle lever 125 is shown from inside the head 100. (See image) Figure 2 As shown, the head 100 also includes a push rod 119, which is configured to move along the longitudinal axis L, specifically via the mirror actuator 114b. When the push rod 119 moves from... Figure 2 When the position shown is moved downwards to the cantilever chip holder 310, the push rod 119 engages with the lever 125, as... Figure 9As shown, the lever 125 is pushed to the downward position against the force of the spring. Thus, the bolt 123 is pivoted, thereby forcing the clamping element 122 toward the open position (not shown), in which the cantilever chip holder 310 can be automatically inserted into or removed from the recess. Figure 9 Magnet 126 and position sensor 126a are also shown for determining the position of clamping element 122, which allows control and confirmation of whether clamping element 122 and thus cantilever chip holder 310 are arranged in the clamped or open position.

[0153] Figure 10a An embodiment of an atomic force microscope 1 according to the present invention is shown, wherein a light source 105 is arranged on the upper portion 104a of the head 100. For example, the light source 105 includes a laser source or a superluminescent LED. In particular, the light source 105 may include an optical fiber cable, wherein light generated by the laser source or superluminescent LED is guided from the laser source or superluminescent LED through the optical fiber cable toward the cantilever 202 into the head 100. The atomic force microscope 1 according to this embodiment may include focusing and collimating optics 105a, such as a lens, arranged between the light source 105 and the cantilever 202, for example... Figure 2 As shown. Cantilever 202 ( Figure 10a (Not shown) is fixedly attached to the lower part 104b of the head 100, adjacent to... Figure 10a The upper part 104a is shown in the diagram. To control the optical path of the beam between the light source 105 and the cantilever 202, the light source 105 is movably mounted on the upper part 104a of the head 100, allowing the beam emitted from the light source 105 to move relative to the cantilever 202. The light source 105 is mounted to the upper part 104a of the head 100 via a displacement stage 110 including a first bracket 110a, a second bracket 110b, and a third bracket 110c.

[0154] The first bracket 110a of the displacement stage 110 can be moved in the y-direction by a first light source linear actuator, particularly with a resolution of about 10 μm. The first light source linear actuator includes a piezoelectric actuator, particularly a piezoelectric LEGS or piezoelectric triboelectric actuator (111a). The second bracket 110b of the displacement stage 110 can be moved in the x-direction by a second light source linear actuator, particularly with a resolution of about 10 μm. The second light source linear actuator includes a piezoelectric actuator, particularly a piezoelectric LEGS or piezoelectric triboelectric actuator (111b). The third bracket 110c of the displacement stage 110 can be moved in the z-direction by a third light source linear actuator, particularly including a piezoelectric motor, particularly a piezoelectric leg motor, configured to move the third bracket 110c along the z-direction. In this way, the light source 105 can be independently displaced relative to the cantilever along the x, y, and z directions, which allows for simplified beam alignment and focusing on the cantilever 202.

[0155] To record the position of the light source 105 relative to the cantilever 202, each of the first, second, and third brackets 110a, 110b, and 110c includes a magnetic position sensor 126a. Specifically, the displacement of the light source 105 relative to the cantilever 202 can be automatically controlled using the controller 410 based on the position information of the light source 105 relative to the cantilever 202, so that the light beam can be automatically focused on and maintained on the cantilever 202 during measurement.

[0156] Optionally, such as Figure 10b As shown, the first bracket 110a can be formed as a first plate 109a, and the second bracket 110b can be formed as a second plate 109b, wherein the first plate 109a and the second plate 109b circumferentially surround the body 150 having the light source 105. The first plate 109a and the second plate 109b are stacked on top of each other along the z-direction and connected by three set screws 114b. By tightening and loosening the set screws 109c, the body 150 having the light source 105 can be tilted relative to the head 100 and the cantilever 202, which allows adjustment of the focus of the light beam emitted by the light source 105 on the cantilever 202 arranged on the head 100.

[0157] Figure 11 An embodiment of the atomic force microscope 1 according to the present invention is shown, which includes a third imaging system 440 arranged and configured to image the cantilever 202. Specifically, the third imaging system 440 can be arranged on a tray 300, for example as... Figure 3 As shown, the third imaging system 440 is configured to image the cantilever 202 arranged on the head 100 from below (i.e., from the view of the tray 300).

[0158] According to this embodiment, the third imaging system 440 includes a light source 444 arranged and configured to illuminate the cantilever 202, particularly the tip of the cantilever 202. Light emitted from the light source 444 can be reflected by a mirror included in the third imaging system 440 toward a target of the third imaging system 440, wherein the target faces the cantilever 202. In particular, the third imaging system 440 is an optical microscope 441.

[0159] Figure 12An embodiment of an atomic force microscope 1 according to the present invention is shown, which has a focusing and collimating optics 105a for focusing and collimating light emitted from a light source 105 onto a cantilever 202. The focusing and collimating optics 105a includes a collimator 151 for collimating light from the light source 105, and a focusing lens 152 for focusing the light collimated by the collimator 151 onto the cantilever 202. Light propagates through a body 150 between the collimator 151 and the focusing lens 152, in which the lens of the focusing and collimating optics 105a is arranged. The focusing and collimating optics 105a also includes an optical fiber connector 153 for connecting an optical fiber to the focusing and collimating optics 105a, such that light generated by the light source 105 can travel along the optical fiber... Figure 12 The light source 105 is guided in the z-direction, towards the cantilever 202, into the focusing and collimating optics 105a. For example, the light source 105 may include a laser source or a superluminescent LED. According to this embodiment, the focusing and collimating optics 105a includes a third light source linear driver 111c for moving the body 150, collimator 151, focusing lens 152, and fiber optic connector 153, up and down in the z-direction relative to the upper portion 104a of the head 100. Thus, by moving the focusing and collimating optics 105a relative to the cantilever 202 in the z-direction, the focused position of the light emitted by the light source 105 on the cantilever 202 can be changed. Figure 13 An embodiment of a system 600 according to a second aspect of the present invention is shown. System 600 includes an atomic force microscope 1 including a head 100 according to a first aspect of the present invention. System 600 according to this embodiment includes an interface 610, particularly a human-machine interface, comprising two drawer units 620. The functions of the drawer units 620 include: System 600 also includes a label scanner 612 for scanning labels disposed on or potentially disposed on the sample to be tested, sample holder 430, cantilever chip holder 310, and / or cantilever 202. Specifically, the label includes or is a barcode. Therefore, The system is configured to record the sample used in the measurement, sample holder 430, cantilever chip holder 310 and / or cantilever 202.

[0160] Figure 14a An embodiment of an interface 610 having two drawer units 620 is shown for a system 600 according to a second aspect of the present invention.

[0161] Interface 610 can form a human-machine interface for transferring a sample holder 430, with or without a sample to be tested, to or from the sample stage 300 via interface 610. Similarly, a cantilever holder or cantilever chip holder 310 can be transferred to or from the sample stage 300 via interface 610. The cantilever chip holder 310 and / or sample holder 430 can be placed and carried by a shuttle unit 650, which can be placed on a protrusion 634 of each drawer unit 620. Interface 610 can be part of the housing 700 of system 600, which frames the atomic force microscope 1, has the sample stage 300, and a head 100 movably connected to a mounting. The two drawer units 620 can be in a loading position away from the sample stage 300 along a third longitudinal axis L3 (e.g., Figure 14a (as shown) and the unloading position at sample stage 300 (as shown below) Figure 14c (Further shown) The drawer unit 620 moves between the loading and unloading positions. For this purpose, the interface 610 includes a motor for driving the drawer unit 620 between the loading and unloading positions.

[0162] According to this embodiment, each drawer unit 620 includes a slider 630, particularly a telescopic slider, for moving the corresponding drawer unit 620 between a loading position and an unloading position.

[0163] Each drawer unit 620 includes a mechanism for unloading the shuttle unit 650 from the drawer unit 620 onto the sample stage 300. For this purpose, as... Figure 14a As shown, and more specifically, as Figure 14b and Figure 14c As shown, the height-adjustable structure 626 is slidably supported on each slider 630 by at least one pin 628, which extends from the slider 630 through a recess 632 of the height-adjustable structure 626. (From comparison...) Figure 14b and Figure 14c It is understood that moving the drawer unit 650 from the loading position to the unloading position causes the height-adjustable structure 626 to engage with the stop 627 of the drawer unit 620, which in turn causes the height-adjustable structure 626 to move relative to the slider 630, wherein the pin 628 of the slot 630 moves from one end of the associated recess 632 to the other end. Since the shape of the recess 632 includes two horizontal portions, each extending substantially along a third longitudinal direction L3 at two different vertical positions along the first longitudinal axis L1, these two different vertical positions are connected by a ramp portion of the recess 632, when the height-adjustable structure 626 engages with the stop 627, the height-adjustable structure 626 and thus the shuttle unit 650, which is placed on the protrusion 634 of the shuttle unit 650, are lowered.

[0164] like Figure 14b and Figure 15As can be further seen in section b, the sample stage 300 is arranged below the drawer unit 620. The sample stage 300 includes multiple, particularly three, support elements 303 for supporting the shuttle unit 650 when it is lowered by the aforementioned mechanism. Figure 14c The image shows the shuttle unit 650 placed on the support element 303 of the sample stage 300, wherein... Figure 14b In comparison, the height-adjustable structure 626 is lower than the slider 630.

[0165] Drawer unit 620 may include a resilient device, such as a spring, once the height-adjustable structure 626 is activated by a motor. Figure 14a Move to the position shown Figure 14b In the position shown, the spring is biased. The height-adjustable structure 626 can resist the spring force of the biasing spring by the resistance applied by the motor used to drive the drawer unit 620, thereby maintaining the position. Figure 14b The shuttle unit 650 can be picked up again from the support element 303 of the sample stage 300 via the protrusion 634 of the height-adjustable structure 626 when the elastic device is relaxed, which allows the height-adjustable structure 626 to move upward along the first longitudinal axis L1. Figure 14a The location shown.

[0166] The aforementioned mechanism can be used to mechanically separate the interface 610 and its drawer unit 620 from the atomic force microscope 1, particularly from the sample stage 300 and the head 100. Specifically, the atomic force microscope 1, with the sample stage 300 and the head 100, is arranged on a vibration isolation table and does not mechanically contact the interface 610 and the drawer unit 620, particularly the housing 700 of the system 600. Thus, user operations at the interface 610 do not affect the measurements of the atomic force microscope 1, while allowing the sample holder 430 and / or the cantilever chip holder 310 to be transferred to and from the sample stage 300.

[0167] Figure 15 An embodiment of an atomic force microscope 1 according to the present invention is shown. This embodiment illustrates a linear actuator, particularly a fine linear actuator, such as a piezoelectric stick-slip actuator having an optical scale for optically detecting the driving distance of the linear actuator. The linear actuator according to this embodiment can specifically form at least one of the brackets 110a, 110b, 110c of the displacement stage 110 for moving the light source 105 relative to the cantilever 202, for example as... Figure 1 As stated above.

Claims

1. An atomic force microscope (1) for obtaining force spectrum measurements of a sample, comprising a head (100) movably connected to a mounting, wherein, The head (100) includes: The receiving unit (106) is configured to receive a cantilever (202) including a tip (203) for contacting a soft material sample. A light source (105) and a focusing optics (105a) for generating light are configured to focus the light, and when the cantilever (202) is positioned at the receiving unit (106), guide the first light beam (B1) along the first longitudinal axis (L1) onto the cantilever (202). A reflector (112) is configured to receive a second beam (B2) reflected by the cantilever (202) when the cantilever (202) is positioned at the receiving unit (106). The detector (113) is configured to detect the position of the third beam (B3) reflected by the mirror (112), and generate a deflection signal based on the detected position of the third beam (B3) indicating the deflection of the cantilever (202) along the first longitudinal axis (L1). The atomic force microscope (1) includes an actuator (140) configured to move the head (100) along the first longitudinal axis (L1), and in particular, the head (100) is connected to the mount via the actuator (140).

2. The atomic force microscope (1) according to claim 1, wherein, The atomic force microscope (1) includes a first imaging system (450), wherein the first imaging system (450) is movably connected to the mount in a fixed position relative to the head (100), and / or wherein the atomic force microscope (1) includes a second imaging system (460), wherein the second imaging system (460) is movably connected to the mount in a fixed position relative to the head (100), and / or wherein the atomic force microscope (1) includes a third imaging system (440), wherein the third imaging system (440) includes a fixed orientation relative to the head (100), relative to the first imaging system (450), and / or relative to the second imaging system (460).

3. The atomic force microscope (1) according to any one of claims 1 or 2, wherein, The atomic force microscope (1) includes a reference assembly, wherein the reference assembly includes a first optical target (402) and a controller (410), wherein the first optical target (402) includes a high-reflectance section and a low-reflectance section, wherein the controller (410) is configured to detect the intensity of a first reference beam reflected by the first optical target (402), wherein the controller (410) is also configured to detect the position of the head (100) perpendicular to the first longitudinal axis (L1), and wherein a first imaging system (450) is arranged and configured to capture an optical image of the first optical target (402), and / or wherein a second imaging system (460) is arranged and configured to capture an optical image of the first optical target (402), wherein the reference assembly is configured to determine: The position of the head (100) relative to the first imaging system (450), The position of the head (100) relative to the second imaging system (460), The position of the first imaging system (450) relative to the second imaging system (460), The position of the head (100) relative to the first beam (B1), The position of the first imaging system (450) relative to the first beam (B1), and / or The position of the second imaging system (460) relative to the first beam (B1).

4. The atomic force microscope (1) according to any one of claims 1 to 3, wherein, The atomic force microscope (1) includes a cantilever (202) with a tip (203) for contacting a soft material sample. The cantilever (202) is arranged at the receiving unit (106). Specifically, the cantilever (202) is arranged at the receiving unit (106) via a cantilever holder (310). The light source (105) is configured and arranged such that the first light beam (B1) is guided onto the cantilever (202) along the first longitudinal axis (L1).

5. The atomic force microscope (1) according to any one of claims 1 to 4, wherein, The atomic force microscope (1) includes at least one linear actuator (101, 102, 103), which is configured to position the cantilever (202) relative to the sample stage (300) along a first longitudinal axis (L1) and / or along a second longitudinal axis (L2) perpendicular to the first longitudinal axis (L1) and / or along a third longitudinal axis (L3) perpendicular to the first longitudinal axis (L1) and the second longitudinal axis (L2), wherein the at least one linear actuator (101, 102, 103) includes: a coarse linear motor (101a, 102a, 103a), particularly a voice coil motor, for phase The sample stage (300) is used to coarsely position the cantilever (202); and fine linear motors (101b, 102b, 103b), particularly piezoelectric motors, particularly piezoelectric stack motors, are used to finely position the cantilever (202) relative to the sample, wherein a first linear actuator (101) is configured to move the cantilever (202) along the first longitudinal axis (L1), and / or a second linear actuator (102) is configured to move the cantilever (202) along the second longitudinal axis (L2), and / or a third linear actuator (103) is configured to move the sample stage (300) along the third longitudinal axis (L3).

6. The atomic force microscope (1) according to any one of claims 1 to 5, wherein, The atomic force microscope (1) includes at least one linear actuator (101, 102) configured to position the head (100) relative to the mount. The at least one linear actuator (101, 102) includes a coarse linear motor (101a, 102a), particularly a voice coil motor, for coarse positioning of the head (100), and a fine linear motor (101b, 102b), particularly a piezoelectric motor, particularly a piezoelectric stack motor, for fine positioning of the head (100). The head (100) is connected to the mount via the at least one linear actuator (101, 102) and the actuator (140). More particularly, the actuator (140) is directly connected to the mount, and the at least one linear actuator (101, 102) connects the actuator (140) and the head (100).

7. The atomic force microscope (1) according to any one of claims 1 to 6, wherein, The atomic force microscope (1) includes at least one spring (130) connecting the head (100) to the mounting, wherein the at least one spring (130) is configured to apply a force on the head (100) along the first longitudinal axis (L1), and in particular, wherein the force remains constant when the spring (130) is compressed or extended along the first longitudinal axis (L1).

8. The atomic force microscope (1) according to any one of claims 1 to 7, wherein, The head (100) includes an alignment mechanism for aligning the first beam (B1), the alignment mechanism including a support (109a) for holding the light source (105) and the focusing optics (105a); a first bracket (110a) and a first light source linear actuator (111a), the first light source linear actuator particularly including a piezoelectric motor, particularly including a piezoelectric leg motor, the first light source linear actuator being configured to move the first bracket (110a) along a second longitudinal axis (L2) perpendicular to the first longitudinal axis (L1); The second bracket (110b) and the second light source linear actuator (111b) are configured to move the second bracket (110b) along the third longitudinal axis (L3) which is perpendicular to the first longitudinal axis (L1) and the second longitudinal axis (L2).

9. The atomic force microscope (1) according to any one of claims 1 to 8, wherein, The head (100), particularly the support (109a), further includes: a first plate (109b) configured to hold the light source (105) and the focusing optics (105a); a second plate (109c); and at least one fixing screw (109d) connecting the first plate (109b) and the second plate (109c), wherein the first plate (109b) is tiltable relative to the second plate (109c) by adjusting the fixing screw (109d) to align with the first beam (B1), or wherein the head (100) further includes a third bracket (110c) and a third light source linear actuator (111c), the third light source linear actuator particularly including a piezoelectric motor, especially a piezoelectric leg motor, the third light source linear actuator being configured to move the third bracket (110c) along the first longitudinal axis (L1).

10. The atomic force microscope (1) according to any one of claims 1 to 9, wherein, The head (100) includes a mirror frame (114), wherein a reflector (112) is connected to the mirror frame (114), and wherein the mirror frame (114) is pivotally connected to a mirror pivot axis (114a), wherein the head (100) further includes a mirror actuator (114b) configured to pivot the mirror frame (114) about the mirror pivot axis (114a), wherein, in particular, the mirror actuator (114b) is a linear actuator, particularly a piezoelectric motor, particularly a piezoelectric leg motor, and wherein the head (100) includes a push rod (119) and a connecting lever (121), the push rod including a pin (1... 20), the connecting lever includes a groove (121a), wherein the mirror actuator (114b) is configured to move the push rod (119) along the first longitudinal axis (L1), wherein the connecting lever (121) is connected to the mirror pivot axis (114a), and the pin (120) of the push rod (119) is arranged in the groove (121a) of the connecting lever (121) such that when the push rod (119) moves along the first longitudinal axis (L1) via the mirror actuator (114b), the mirror frame (114) pivots about the mirror pivot axis (114a) by means of the connecting lever (121).

11. The atomic force microscope (1) according to any one of claims 1 to 10, wherein, The head (100) includes a detector holder (116), wherein the detector (113) is connected to the detector holder (116), and wherein the head (100) includes a detector linear driver (116a), particularly a piezoelectric motor, particularly a piezoelectric leg motor, the detector linear driver being configured to move the detector holder (116) along a second longitudinal axis (L2) perpendicular to the first longitudinal axis (L1) to adjust the position of the detector (113).

12. The atomic force microscope (1) according to any one of claims 1 to 11, wherein, The head (100) includes at least one position sensor (126a), particularly a Hall sensor, said at least one position sensor being configured to determine: The rotational orientation of the mirror frame (114) on the mirror pivot axis (114a) The position of the detector holder (116) along the second longitudinal axis (L2), The position of the first bracket (110a) of the alignment mechanism along the second longitudinal axis (L2), The position of the second bracket (110b) of the alignment mechanism along the third longitudinal axis (L3), and / or The third bracket (110c) of the alignment mechanism is positioned along the first longitudinal axis (L1).

13. The atomic force microscope (1) according to any one of claims 1 to 12, wherein, The head (100) includes a temperature sensor, specifically located at or near the cantilever (202), enabling the temperature of the cantilever (202) to be determined based on a temperature value measured by the temperature sensor, and / or enabling the temperature of the sample to be determined based on a temperature value measured by the temperature sensor, specifically, wherein the temperature sensor is arranged and configured to repeatedly determine the temperature value, specifically, wherein the temperature sensor is arranged and configured to continuously determine the temperature value over time.

14. The atomic force microscope (1) according to any one of claims 1 to 13, wherein, The atomic force microscope (1), particularly the head (100), includes an environmental sensor configured to detect ambient humidity and / or ambient pressure at or near the cantilever (202).

15. The atomic force microscope (1) according to any one of claims 1 to 14, wherein, The atomic force microscope (1) further includes a sample stage (300) for receiving the sample, particularly a biological tissue sample. Specifically, the sample stage (300) is configured to receive a sample holder (430) configured to hold the sample. Specifically, the sample holder (430) is one of a petri dish, a Piper dish, a microplate, particularly a 6-well plate, or a sample support. More specifically, the sample stage (300) includes a first slot for receiving the sample holder (430). Even more specifically, the sample stage (300) includes a second slot for receiving a cantilever holder (310) including a cantilever (202). The atomic force microscope (1) is configured such that the cantilever holder (310) can be automatically mounted onto the head (100).

16. The atomic force microscope (1) according to any one of claims 1 to 15, wherein, The third imaging system (440) is arranged and configured to detect the cantilever (202), particularly the tip (203) of the cantilever (202). In particular, the third imaging system (440) includes a light source configured for coaxial illumination, and / or in particular, the atomic force microscope (1) includes a light source (444) arranged and configured for illuminating the cantilever (202), particularly the tip (203) of the cantilever (202). In particular, the third imaging system (440) is an optical microscope (441).

17. The atomic force microscope (1) according to any one of claims 1 to 16, wherein, The atomic force microscope (1) includes a vibration sensor, particularly wherein the vibration sensor is a vibrometer, accelerometer or seismograph, and / or wherein the atomic force microscope (1) includes an AFM controller, particularly wherein the AFM controller includes a field-programmable gate array, and / or wherein the AFM controller includes a device controller for automation of the atomic force microscope (1).

18. The atomic force microscope (1) according to any one of claims 1 to 17, wherein, The atomic force microscope (1) includes a clamping element (122), wherein the clamping element (122) is configured to be in a preloaded position, a clamped position, or a receiving position, wherein in the receiving position, the clamping element (122) is configured to receive a cantilever retainer (310), wherein in the clamped position, the clamping element (122) is configured to clamp the cantilever retainer (310) to the atomic force microscope (1), wherein the atomic force microscope (1) includes at least one spring configured to bias the clamping element (122) to the clamped position.

19. The atomic force microscope (1) according to any one of claims 1 to 18, wherein, The atomic force microscope (1) includes a magnetic sensor configured to sense the position of the clamping element (122), particularly sensing that the clamping element (122) is in the preloaded position, the clamped position, or the receiving position.

20. The atomic force microscope (1) according to any one of claims 1 to 19, wherein, The atomic force microscope (1) includes a clamping actuator (1222), particularly, wherein the clamping actuator (1222) is a linear actuator, especially a piezoelectric motor, particularly a piezoelectric leg motor, wherein the clamping actuator (1222) is configured to move a device (1224) along the first longitudinal axis (L1), wherein the device (1224) is configured to move the clamping element (122) from the preloaded position to the receiving position and / or from the clamping position to the receiving position when the device (1224) moves along the first longitudinal axis (L1), particularly, wherein the clamping actuator (1222) is the mirror actuator (114b).

21. The atomic force microscope (1) according to any one of claims 1 to 20, wherein, The atomic force microscope (1) includes a control device configured to control the actuator (140) such that the head (100) moves along the first longitudinal axis (L1), particularly such that the tip (203) of the cantilever (202) contacts the sample, wherein, in particular, the control device is configured to receive the deflection signal from the detector (113).

22. The atomic force microscope (1) according to claims 2 to 21, wherein, The atomic force microscope (1) includes a first imaging system (450) and a second imaging system (460), wherein the first imaging system (450) is configured to provide a deep-focus image and / or a depth map, and in particular, wherein the first imaging system (450) is configured to record a plurality of patches stacked along the first longitudinal direction (L1), particularly patches extending perpendicular to the first longitudinal direction (L1), to provide a deep-focus image and / or a depth map.

23. The atomic force microscope (1) according to any one of claims 1 to 22, wherein, The light source (105) is an optical fiber.

24. The atomic force microscope (1) according to any one of claims 1 to 23, wherein, The atomic force microscope (1) includes a power source, specifically a battery.

25. A system (600) comprising an atomic force microscope (1) according to any one of claims 1 to 24 and a vibration isolation stage, wherein, The vibration isolation table includes a vibration sensor.

26. The system (600) according to claim 25, wherein, The system (600) includes an interface (610), particularly a human-machine interface, which includes a drawer unit (620) configured to receive a shuttle unit (650) for carrying a sample holder (430) and / or a cantilever holder (310), wherein the drawer unit (620) is movable between a loading position for placing the shuttle unit (650) on the drawer unit (620) and an unloading position at the sample stage (300) remote from the sample stage (300), and wherein, when the shuttle unit (650) is placed on the drawer unit (650) and the drawer unit (650) is moved to the unloading position, the drawer unit (620) includes a mechanism for unloading the shuttle unit (650) from the drawer unit (650) onto the tray (300).

27. The system (600) according to claim 26, wherein, The drawer unit (620) includes a plurality of protrusions (634) for placing the shuttle unit (650) on the protrusions (634) of the drawer unit (620).

28. The system (600) according to claim 26 or 27, wherein, The drawer unit (620) includes a slider (630), particularly a telescopic slider, for moving the drawer unit (620) between the loading position and the unloading position, particularly along the third longitudinal direction (L3).

29. The system (600) according to claim 28, wherein, The height-adjustable structure (626) is slidably supported on each slider (630) by at least one pin (628) extending from the slider (630) into a recess (632) of the height-adjustable structure (626), and wherein the height-adjustable structure (626) includes the protrusion (634), and wherein the recess (632) is shaped such that when the drawer unit (650) moves to the unloading position and the height-adjustable structure (626) engages with the stop (627), the vertical position of the protrusion (634) and the shuttle unit (650) placed on the protrusion (634) is lowered by moving the height-adjustable structure (626) relative to at least one pin (628) of the slider (630).

30. The system (600) according to claim 29, wherein, The sample stage (300) includes a plurality of support elements (303), particularly three support elements (303) for supporting the shuttle unit (650), wherein, in the unloading position, the protrusion (634) for placing the shuttle unit (650) is arranged above the support elements (303) such that the shuttle unit (650) can be unloaded from the drawer unit (620) onto the sample stage (300) by lowering the height-adjustable structure (626) relative to the slider (630).

31. The system (600) according to any one of claims 25 to 30, wherein, The system (600) includes a label scanner, particularly a label scanner for scanning barcodes, wherein the barcodes encode information about the sample, the sample holder, the probe holder, and / or the cantilever.

32. The system (600) according to any one of claims 25 to 31, wherein, The sample holder (430) is a petri dish, a Piper dish, a microplate, particularly a 6-well plate, or a sample support.

33. The system (600) according to any one of claims 25 to 32, wherein, The system (600) includes hydraulic wheels, wherein the hydraulic wheels are arranged and configured such that the system (600) can move by means of the hydraulic wheels, and wherein the system (600) includes fixed support feet, wherein the fixed support feet are arranged and configured such that the system can stand stably and / or be aligned by means of the support feet.

34. A component comprising the system (600) according to any one of claims 25 to 33 and packaging, wherein, The package is configured to receive the system (600), wherein the package includes a ramp configured to support the release of the system from the package, and in particular, wherein the package includes a lid configured as a wedge and arranged such that the system can be released from the package via the wedge.

35. A method for obtaining force spectrum measurements of a sample using an atomic force microscope (1) according to any one of claims 1 to 24, or a system (600) according to any one of claims 25 to 34, or a component according to claim 34, said sample being particularly from soft materials, and more particularly biological tissue samples, said method comprising the steps of: The sample is contacted at the measurement point by the tip (203) of the cantilever (202). During the loading step, particularly automatically, the head (100) is moved toward the sample by the actuator (140) parallel to the first longitudinal axis (L1), such that the tip (203) of the cantilever (202) is pressed into the sample. During the unloading step, particularly automatically, the head (100) is moved away from the sample by the actuator (140) parallel to the first longitudinal axis (L1). During the loading and / or unloading steps, multiple deflection values ​​of the cantilever (202) are determined from the deflection signal of the detector (113). From the determined deflection value, in particular automatically, the force spectrum measurement of the sample at the measurement point is determined.

36. The method according to claim 35, wherein, The method includes determining a plurality of force spectrum measurements indicating the force of the cantilever (202) on the sample at corresponding positions along the first longitudinal axis (L1), wherein the force spectrum measurements of the sample are determined based on at least a subset of force values, particularly automatically, and specifically, wherein force-position relationships and / or force-position curves are obtained.

37. The method according to any one of claims 35 or 36, wherein, The position of the sample relative to the head (100), and particularly relative to the cantilever (202), is changed by moving the head (100) along the first longitudinal direction (L1), particularly by the first linear actuator (101) and / or the actuator (140), and / or along the second longitudinal direction (L2), particularly by the second linear actuator (102), and / or by moving the sample, particularly the sample holder (430) holding the sample, particularly by the third linear actuator (103) along the third longitudinal direction (L3).

38. The method according to any one of claims 35 to 37, wherein, The detector (113) is moved along the second longitudinal axis (L2) by the detector linear actuator (116a), and / or the mirror frame (114), in particular the associated reflector (112), is pivoted by the mirror actuator (114b), such that the reflector (112) and the detector (113) are aligned, such that the third beam (B3) reflected by the reflector (112) satisfies a predetermined detection area of ​​the detector (113), in particular the linear area of ​​the detector (113), and in particular the detector (113) and / or the mirror frame (114) are automatically moved to hold the third beam (B3) in the detection area of ​​the detector (113), in particular the linear area of ​​the detector (113).

39. The method according to any one of claims 35 to 38, wherein, The shape, size, length, width, and / or height of the cantilever (202) are determined by the third imaging system (450), particularly automatically, and / or wherein, The position of the tip (203) of the cantilever (202) is determined by the third imaging system (450), and in particular, is determined automatically.

40. The method according to any one of claims 35 to 39, wherein, Before contacting the sample via the tip (203) of the cantilever (202), the method includes a step of 3D profile analysis of the sample, using information obtained by the first imaging system (450) and / or using information obtained by the second imaging system (460), particularly information related to the depth-of-focus image and / or depth map, and calculations, particularly wherein the calculations include a Laplacian regression step.

41. The method according to any one of claims 35 to 40, wherein, Before contacting the sample via the tip (203) of the cantilever (202), the method includes an automated point selection step that provides a prediction of the measurement quality at the selected measurement point based on sample-related information, information obtained through 3D contour analysis of the sample, information obtained through depth-of-focus imaging, information obtained through the depth map, information about the shape of the cantilever (202), information about the size of the cantilever (202), and / or the position of the tip (203) of the cantilever (202).

42. The method according to any one of claims 35 to 41, wherein, Before contacting the sample via the tip (203) of the cantilever (202), the tip (203) of the cantilever (202) contacts a reference sample with a predetermined stiffness, and while the tip (203) contacts the reference sample without pressing into it, the head (100) is moved by the actuator (140) to travel a distance, wherein the deflection signal of the detector (113) is obtained at the distance, and wherein the deflection sensitivity of the detector (113) is determined, in particular automatically, from the distance and the deflection signal, and / or wherein the deflection sensitivity of the detector (113) is determined in a non-contact manner, particularly based on the resonance curve, especially the resonance curve Information obtained from the area below the line, wherein the resonance curve is obtained in air or in a liquid, and / or wherein the deflection sensitivity of the detector (113) is determined in a depth-stepping standard manner, particularly wherein the tip (203) of the cantilever (202) contacts the surface of a standard sample comprising a recess of a predetermined depth before contacting the sample via the tip (203) of the cantilever (202), and the head (100) is moved by the actuator (140) through a moving distance, and wherein the deflection signal of the detector (113) is obtained at the moving distance, and wherein the deflection sensitivity of the detector (113) is determined from the moving distance and the deflection signal, particularly automatically.

43. The method according to any one of claims 35 to 42, wherein, The deflection sensitivity is determined, wherein the device detects when the deflection sensitivity deviates from a predetermined interval and initiates recalibration by realigning the beam and performing additional deflection sensitivity measurements, particularly automatically.

44. The method according to any one of claims 35 to 43, wherein, The cantilever (202) is moved near the sample or to the measurement point, particularly wherein the cantilever (202) is moved near the sample or to the measurement point in a non-approach manner, based on sample-related information and / or based on information obtained through 3D profile analysis of the sample.

45. The method according to any one of claims 35 to 44, wherein, Adaptive optimization is performed to move the cantilever (202) near the sample or to the measurement point, wherein information considering the deviation between the detected previous measurement point and the proposed previous measurement point is used, particularly by applying the deviation to the proposed measurement point.

46. ​​The method according to any one of claims 35 to 45, wherein, A vibration signal is obtained from the vibration sensor, and wherein, when the signal indicates the presence of vibration, particularly when it indicates the presence of vibration with an amplitude or frequency higher than a predetermined threshold, the head (100) is moved away from the sample along the first longitudinal axis (L1), particularly automatically.

47. The method according to any one of claims 35 to 46, wherein, The spring constant of the cantilever (202) is determined, in particular automatically, based on data provided by the third imaging system (440), particularly the length, width and / or thickness of the cantilever (202), the temperature value obtained by the temperature sensor, the ambient humidity and / or the ambient pressure obtained by the ambient sensor, the resonant frequency of the cantilever (202) and / or the quality factor of the cantilever (202).

48. The method according to any one of claims 35 to 47, wherein, The spring constant of the cantilever (202) is repeatedly detected during the measurement and / or the value related to the deflection sensitivity calibration is repeatedly detected during the measurement, wherein the cantilever (202) is replaced and / or recalibrated when the spring constant of the cantilever (202) exceeds a predetermined range, and / or wherein the cantilever (202) is replaced and / or recalibrated when the value related to the deflection sensitivity calibration exceeds a predetermined range, wherein the recalibration includes realigning the first bundle relative to the cantilever, particularly the tip (203) of the cantilever, and additional measurement of the value related to the deflection sensitivity, and / or wherein the mass value of the cantilever is repeatedly detected during the measurement, wherein the cantilever (202) is replaced when the mass value of the cantilever (202) deviates from a predetermined range.

49. The method according to any one of claims 35 to 48, wherein, When the head (100) determines a predetermined force value to be performed by the cantilever (202) to be picked up, in particular by the cantilever holder (310) to be picked up, the cantilever (202) is picked up, in particular the cantilever holder (310), in particular from the sample stage (300), in particular from the second slot (302) included in the sample stage (300) for receiving the cantilever holder (310) including the cantilever (202).

50. The method according to any one of claims 35 to 49, wherein, A correction factor related to the buffer surrounding the sample is determined, wherein the deflection signal is detected as the cantilever (202) moves toward the sample along the first longitudinal axis (L1) before the tip (203) of the cantilever (202) contacts the sample, wherein the position of the cantilever (202) along the first longitudinal axis (L1) is determined when the deflection signal is lost.

51. The method according to any one of claims 35 to 50, wherein the atomic force microscope, particularly the AFM controller, particularly the field-programmable gate array, monitors at least one monitoring signal, wherein the monitoring signal is one of the following: The sum of the cantilever and deflection signals, Vibration signal, The accelerometer signal, The signal from the seismic detector The signal obtained by the microphone, in, The atomic force microscope: When the monitoring signal leaves the predetermined range, any movement in the atomic force microscope is interrupted to protect the probe. When the monitoring signal leaves the predetermined range, especially in the presence of external vibration or external acoustic noise, the force spectrum measurement is interrupted. When the monitoring signal leaves the predetermined range, especially in the event of external vibration or external acoustic noise, the optical measurements performed by the first optical system and / or the second optical system and / or the third optical system are interrupted, and / or When vibration and noise levels are low, the monitoring signal is used as a "gate" to trigger measurements only.

52. The method according to any one of claims 35 to 51, wherein, The deflection sensitivity in the liquid, particularly the buffer, is determined based on the deflection sensitivity determined in air and positional information regarding the tip (203) of the cantilever, particularly regarding the position of the tip (203) of the cantilever relative to the first longitudinal direction, the second longitudinal direction, and / or the third longitudinal direction.

53. A method for obtaining a reference position of a beam using an atomic force microscope (1) according to any one of claims 1 to 24 or a system (600) according to any one of claims 25 to 34, comprising the following steps: a. Guide the first beam (B1) along the first longitudinal axis (L1) to the first optical target (402). b. The intensity of the first reference beam reflected by the first optical target (402) is detected by the controller (410). c. The controller (410) detects the position of the head (100) perpendicular to the first longitudinal axis (L1). d. To capture an optical image of the first optical target (402) using the first imaging system (450) and / or the second imaging system (460). e. Determine the positions of the following items: The head (100) is relative to the first imaging system (450). The head (100) is relative to the second imaging system (460). The first imaging system (450) is relative to the second imaging system (460). The head (100) is positioned relative to the first beam (B1). The first imaging system (450) relative to the first beam (B1), and / or The second imaging system (460) is relative to the first beam (B1).

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