Microscope and method for microscopy of a sample under a variable mechanical parameter

Abstract

Method for microscopying a sample (06) with a microscope, wherein a mechanical parameter (Θ) on the microscope is variable, and wherein the method comprises at least the following steps: - Selecting a first value to be set (Θ Ziel ) of the mechanical parameter (Θ); - Actuating an electrically controlled actuator (04; 11) to set the first value to be set (Θ Ziel ) of the mechanical parameter (Θ); - Taking a first microscopic image of the sample (06) at the set first value (Θ) Ziel ) of the mechanical parameter (Θ); - Selecting a second value to be set (Θ Ziel ) of the mechanical parameter (Θ); - Actuating the electrically controlled actuator (04; 11) to set the second value to be set (Θ Ziel ) of the mechanical parameter (Θ); and - Taking a second microscopic image of the sample (06) at the set second value (Θ) Ziel ) of the mechanical parameter (Θ); wherein the mechanical parameter (Θ) is manually changeable; wherein the actuating element is formed by a locking device (11) with which the mechanical parameter (Θ) can be locked; and wherein the steps of actuating the electrically controllable actuating element (11) each comprise the following sub-steps: - Measuring the time course of a manually induced change in value (Θ) aktuell ) of the mechanical parameter (Θ); - Predicting a point in time at which the previously determined time course of the manual change of the value (Θ) aktuell ) of the mechanical parameter (Θ) to the value to be set (Θ) Ziel ) of the mechanical parameter (Θ); and - electrical activation of the electrically actuated locking device (11) at the previously determined time.

Description

[0001] The present invention relates to a microscope and a method for microscopically examining a sample, wherein a mechanical parameter of the microscope is variable. The mechanical parameter is preferably formed by an angle of inclination that is variable in at least one spatial direction between an objective lens of the microscope and a stage of the microscope. This angle of inclination describes an inclination between the objective lens and the stage, which is also referred to as tilt.

[0002] JP 2001-059 599 A and JP 2010-102 344 A show a swivel arm stand for a digital microscope. The swivel arm stand includes a swivel arm that can be pivoted about a horizontal axis.

[0003] From DE 10 2013 005 999 A1, a swivel arm stand for a digital microscope is known in which a swivel arm movement can be blocked by a high-torque magnetic brake arranged around a rotary axis. The blockage can be released for the duration of a button press by releasing the high-torque magnetic brake. The button is arranged on the swivel arm such that it can be conveniently pressed with at least one finger of the same hand.

[0004] JP 2013-88490 A shows a microscope in which the optics can be swiveled relative to a sample. This swiveling movement can be electrically driven.

[0005] US patent 2003 / 0223111 A1 shows an analytical instrument with a eucentric goniometer. The analytical instrument may be a microscope. The goniometer can be adjusted to allow examinations at different angles.

[0006] EP 3 006 980 A1 describes a eucentric digital microscope with a pivotally mounted swivel unit. The microscope includes a brake unit for locking the swivel unit. The swivel unit includes an actuating element for releasing the brake unit. The actuating element can be electrically coupled to a brake element of the brake unit.

[0007] From DE 10 2013 222 295 A1, a digital microscope with a swivel stand, a method for its calibration, and a method for automatic focus and image center tracking are known. The swivel stand includes an angle sensor for determining the current swivel angle of the swivel arm. The current swivel angle is processed in the control unit to perform automatic focus tracking and / or center tracking when the swivel arm is actuated. Calibration is performed using two swivel angles, whereby differing focus and image center positions are determined, and a swivel-angle-dependent function for the focus and image center position is derived from this.

[0008] US Patent 2005 / 0237604 A1 describes an in vivo examination device for examining live animals. The device includes a lens unit for optically imaging the animal under examination. The lens unit is pivotable by means of a rotary mechanism. This pivoting movement is powered by a motor.

[0009] US Patent 6,642,686 B1 discloses a swivel arm with a mounting device for an instrument. The swivel arm has at least one pivot point, allowing it to be moved. At least one sensor is located at the pivot point to detect the position and / or movement of the pivot point. A controllable, passive actuator at the pivot point can slow down and / or block movement of the pivot point. A control device is used to control the passive actuator based on position and / or motion data transmitted by the sensor, as well as data from the target object. The passive actuator can be guided such that an instrument connected to the swivel arm can perform only one or more predefined movements.

[0010] German patent DE 10 2012 215 307 A1 discloses a magnifying observation device, in particular a microscope. An imaging unit of the microscope is tiltable, and this tilt can be fixed with a locking knob.

[0011] Starting from the prior art, the object of the present invention is to expand the application possibilities for a microscope in which a mechanical parameter, such as an angle of inclination between an objective and a stage, can be changed.

[0012] The aforementioned problem is solved by a method according to the attached claim 1 and by a microscope according to the attached dependent claim 9.

[0013] The method according to the invention is used for microscopy of a sample with a microscope, in particular with a digital microscope. In the digital microscope, electronic image conversion preferably takes place, wherein the captured image is further processed in the form of digital data and displayed on an electronic image display device. The microscope preferably comprises at least one objective lens and preferably an image sensor for converting an image projected directly or indirectly from the objective lens onto the image sensor.

[0014] The microscope has a variable mechanical parameter. This mechanical parameter is preferably defined by a position, angle, or distance relative to the sample, a stage, the image sensor, the objective lens, an objective component, a filter, and / or an illumination source of the microscope. Thus, the mechanical parameter can be defined, for example, by the height and / or lateral position of the stage, a distance to the image sensor, or a zoom setting. Alternatively, the mechanical parameter is preferably defined by the position, angle, or distance of an adjustment element used to set a physical quantity on the microscope.It could, for example, be a rotatable adjustment element with which, for example, a wavelength, a brightness on the microscope, or a polarization or phase in an illumination or detection channel of the microscope can be adjusted.

[0015] The mechanical parameter is preferably defined by an angle of inclination that can be varied in at least one spatial direction between the microscope objective and the microscope stage. In this microscope, the angle of inclination between the objective and the stage, which supports the sample, is variable. The stage serves to position the sample. The angle of inclination is defined between an optical axis of the objective and a perpendicular line on the stage. The angle of inclination describes an inclination between the objective and the stage, which is also referred to as tilt. An axis of inclination is formed by a rotation axis of the inclination that is oriented perpendicular to the optical axis. The angle of inclination is defined in at least one spatial direction but can also be defined, for example, in spherical coordinates by two values.

[0016] The procedure comprises a step in which an initial value for the mechanical parameter to be set is selected. This initial value defines the desired measurement of the mechanical parameter after adjustment. In a further step, an electrically controlled electromechanical actuator is activated to set this initial value. The actuator ensures that the mechanical parameter is set to this initial value. Finally, a microscopic image of the sample is taken using a microscope while the initial value of the mechanical parameter is set.

[0017] In a further step, a second adjustable value for the mechanical parameter is selected. This second value differs from the first. The second adjustable value of the mechanical parameter defines the desired angle of inclination after adjustment. In a further step, the electrically controlled electromechanical actuator is activated to set this second adjustable value of the mechanical parameter. Finally, a second microscopic image of the sample is taken using a microscope while the second value of the mechanical parameter is set.

[0018] A particular advantage of the method according to the invention is that the microscopic images are each acquired at a defined value of the mechanical parameter, for example, at a defined value of the tilt angle, thus enabling further microscopic examination of the sample. The method according to the invention allows, for example, improved further examination of samples with low contrast. Since the operator is supported by the method according to the invention in setting the defined values ​​of the mechanical parameter, the examination of samples at different values ​​of the mechanical parameter is far less time-consuming and less prone to errors.

[0019] Preferred embodiments of the method according to the invention further comprise a step that is to be carried out after the electrically controlled actuating element has been actuated. In this step, the sample is focused; in particular, the sample is refocused, since the sample no longer necessarily needs to be in focus through the objective lens due to a change in the mechanical parameter, for example, by changing the angle of inclination or the height of the stage. Alternatively or additionally, the sample is re-centered in the image to be recorded, for which the stage is moved vertically and / or laterally.

[0020] In a first preferred embodiment, the actuating element is formed by an actuator driven by an electric motor, by which the mechanical parameter can be adjusted. The mechanical parameter can be changed by energizing the electric motor.

[0021] In a second preferred embodiment, the mechanical parameter can be changed manually. The operator of the microscope adjusts the mechanical parameter using their own strength, preferably after first actuating a release mechanism. The actuating element is formed by an electrically controlled locking device with which the mechanical parameter can be locked. The locking device is preferably formed by an electrically controlled mechanical brake or by a high-torque magnetic brake. In this second preferred embodiment, the steps of actuating the electrically controlled actuating element each include a sub-step in which the time course of a manually induced change in the value of the mechanical parameter is measured. An angle sensor or a distance sensor is preferably used for this purpose.In a further step, a time is predicted at which the previously determined time course of the manual change in the value of the mechanical parameter will result in the desired value of the mechanical parameter being set. Accordingly, the electrically actuated locking device is activated at the previously determined time, ensuring that the desired value of the mechanical parameter, such as the desired tilt angle, is actually achieved and eliminating any dependence of the accuracy on the size of the angle or the distance between the lens and the sample. Activating the electrically actuated locking device fixes the mechanical parameter, making it no longer manually adjustable.

[0022] In embodiments where the mechanical parameter is formed by the inclination angle, the step of predicting the time at which the previously determined temporal course of the manual change of the value of the inclination angle to the respective value to be set Θ comprises Ziel The angle of inclination will be determined, preferably in several sub-steps. In one sub-step, an initial value Θ is set. Start The tilt angle is measured, preferably using an angle sensor. In a further step, an angular velocity ω and an angular acceleration α are determined from the previously determined time course of the manual change in the value of the tilt angle. In a further sub-step, Θ is calculated based on the equation solved for t. Ziel = Θ Start + ω · t + ½ · α · t 2Using t, the time is predicted at which the previously determined time course of the manual change of the value of the inclination angle to the respective value to be set Θ will be reached. Ziel of the angle of inclination will lead.

[0023] Predicting the time at which the previously determined time course of the manual change in the value of the mechanical parameter will lead to the respective set value of the mechanical parameter is preferably repeated continuously until the manual change in the value of the mechanical parameter has reached the respective set value. This continuous repetition ensures high prediction accuracy. In embodiments where the mechanical parameter is defined by the inclination angle, the prediction of the time at which the previously determined time course of the manual change in the value of the inclination angle will reach the respective set value Θ is performed accordingly. Ziel of the inclination angle, preferably repeatedly, until the manual change of the inclination angle value to the value to be set Θ Zielof the angle of inclination.

[0024] In embodiments where the mechanical parameter is formed by the inclination angle, the time at which the previously determined temporal course of the manual change of the value of the inclination angle corresponds to the value to be set is predicted. Ziel The tilt angle will preferably be further determined by considering a mass distribution on the microscope and / or a force applied manually to change the tilt angle value. This allows the time at which the previously determined time course of the manual change in the tilt angle value corresponds to the value Θ to be set. Ziel the angle of inclination will be predicted more accurately.

[0025] In embodiments where the mechanical parameter is determined by the tilt angle, the objective lens is preferably attached to a swivel arm of the microscope. The swivel arm can be a component of a swivel arm stand. The swivel arm can also be referred to as a tilt arm. The swivel arm is pivotable, thus allowing the tilt angle between the objective lens and the stage to be changed.

[0026] Alternatively or additionally, the specimen stage is preferably rotatable. The specimen stage is preferably rotatable about an axis that is parallel to the plane of extension of the specimen stage, or, more preferably, lies within the plane of extension of the specimen stage. By rotating the specimen stage about this axis, the angle of inclination between the objective lens and the specimen stage can be changed. The specimen stage can also be rotatable about an axis that is oriented perpendicular to the plane of extension of the specimen stage. Rotation about this axis leads to a change in the angle of inclination if the specimen on the support also extends vertically.

[0027] In particularly preferred embodiments, the method comprises a further step in which feedback is generated for the operator when the value of the mechanical parameter has changed by a predefined amount and / or when the respective value of the mechanical parameter to be set has been reached. This facilitates the operation of the microscope, as the operator can quickly ascertain whether the respective value of the mechanical parameter to be set has already been achieved. Furthermore, the operator can receive feedback whenever the value of the mechanical parameter has changed by a predefined amount; for example, when the value of the tilt angle has changed by, say, 5°. The feedback is preferably perceptible visually, audibly, and / or haptically.

[0028] The feedback to the operator is preferably haptic and is preferably generated by the electrically actuated locking device. In embodiments where the mechanical parameter is the tilt angle, the change in the value of the tilt angle, i.e., in particular the pivoting movement of the swivel arm or the rotation of the rotatable stage, is preferably briefly interrupted in its temporal change, which the operator can feel, thus informing them that the value of the tilt angle has changed by a predefined amount. These interruptions in the temporal change of the tilt angle value are generated, in particular, by the electrically actuated locking device, in the form of a brake, interrupting the pivoting movement of the swivel arm or the rotation of the rotatable stage for a very short period of time.

[0029] The feedback to the operator is preferably audible, either alternatively or additionally, and is preferably generated by a buzzer or a comparable acoustic signaling device. The acoustic signals inform the operator that the value of the mechanical parameter has changed by a predefined amount or that the respective value of the mechanical parameter has been set.

[0030] In preferred embodiments of the method according to the invention, the steps of selecting the respective value of the mechanical parameter to be set are performed manually by the operator. The operator specifies two or more values ​​of the mechanical parameter to be set, preferably via a user interface.

[0031] In further preferred embodiments of the method according to the invention, the value of the mechanical parameter to be set is selected from several stored values. The stored values ​​are preferably selected and saved during the execution of the method according to the invention for a previously microscopically examined sample. Thus, the same values ​​of the mechanical parameter are selected for both samples to be microscopically examined, and corresponding microscopic images are taken, allowing, for example, an extended evaluation by comparison. The stored values ​​preferably represent pre-programming or a preset value that is accessed during the execution of the method.

[0032] In further preferred embodiments of the method according to the invention, the selection of the respective value of the mechanical parameter to be set is achieved by incrementing the previously selected value of the mechanical parameter by one increment. In embodiments in which the mechanical parameter is defined by the inclination angle, the sample is thereby microscopically scanned with respect to the inclination angle in a grid-like manner. The first value of the mechanical parameter represents a starting value.

[0033] Preferably, not just two values ​​of the mechanical parameter, but a plurality of values ​​of the mechanical parameter are selected and set. Accordingly, the method preferably comprises a step in which another value of the mechanical parameter, different from the other values, is selected. In a further step, the electrically controlled actuating element is activated to set the additional value of the mechanical parameter. In a further step, another microscopic image of the sample is acquired at the set value of the additional mechanical parameter. The aforementioned steps are preferably repeated multiple times for further values ​​of the mechanical parameter.

[0034] The method according to the invention is preferably designed for microscopy of at least two samples. For taking microscopic images of the first sample and for taking microscopic images of the second sample, the same first values ​​of the mechanical parameter, the same second values ​​of the mechanical parameter, and optionally also the same further values ​​of the mechanical parameter are selected and set. Thus, microscopic images of the at least two samples are taken under the same values ​​of the mechanical parameter.

[0035] In further preferred embodiments of the method according to the invention, different magnifications of the objective lens are selected and / or different objectives are used for capturing the first microscopic image of the sample, for capturing the second microscopic image, and optionally for capturing further microscopic images. This allows for adaptation to the spatial extent of the sample, or for specific areas of the sample to be examined at a higher magnification.

[0036] A preferred embodiment of the method according to the invention comprises a further step in which a 2.5-dimensional or three-dimensional representation of the sample is generated from the multiple microscopic images. Preferably, a plurality of values ​​of the mechanical parameter, preferably formed by the inclination angle, is selected and set in order to acquire a corresponding plurality of microscopic images and to use them for generating the 2.5-dimensional or three-dimensional representation of the sample. Since the sample is microscopically examined at different values ​​of the inclination angle or sample distance, depth information can also be obtained from the microscopic images.

[0037] When taking microscopic images of the sample at different angles of inclination, one or more areas of the sample may be obscured. Therefore, it is preferable to take further microscopic images of the sample at different angles of inclination to capture those areas of the sample that are obscured at other angles.

[0038] The microscope according to the invention is used for microscopic examination of a sample. It preferably comprises an objective lens for optically imaging the sample. The microscope further preferably comprises a stage for arranging the sample. A mechanical parameter of the microscope is adjustable. Preferably, the adjustable mechanical parameter is the angle of inclination between the objective lens and the stage. The microscope also comprises an electrically controlled actuating element for setting a value of the mechanical parameter and a control unit configured to carry out the method according to the invention. The control unit is preferably configured to carry out preferred embodiments of the method according to the invention. The microscope preferably also has features that are specified in connection with the method according to the invention.

[0039] Further details and developments of the invention will become apparent from the following description of preferred embodiments of the invention, with reference to the drawing. The drawing shows: Fig. 1: a first preferred embodiment of a microscope according to the invention; Fig. 2: a second preferred embodiment of the microscope according to the invention; Fig. 3: the in Fig. 1 Microscope shown in two states during the execution of a method according to the invention; and Fig. 4: the in Fig. 2 Microscope shown in two states during the execution of the method according to the invention.

[0040] Fig. Figure 1 shows a first preferred embodiment of a microscope according to the invention. The microscope comprises a base 01 on which a stage 02 is mounted. A swivel arm 03 is rotatably mounted on the base 01 and can be swivelled relative to the base 01 by means of an electrically driven actuator 04, so that it can also swivel relative to the stage 02 and a sample 06 located thereon (shown in Figure 1). Fig. 2) tilts. A lens 07 and an optical module 08 are attached to the swivel arm 03. The swivel arm 03 can be unlocked via an unlock button 09.

[0041] As an alternative to the electrically motorized actuator 04, an electrically operated brake 11 (shown in Fig. 2) can be used on the swivel arm 03, which can lock the swivel arm 03 from swiveling relative to the base 01 and the stage 02 by braking. The torque for swiveling the swivel arm 03 must therefore be applied by the operator. The electrically operated brake 11 (shown in Fig. 2) of the swivel arm 03 is controlled in such a way that further manual swiveling of the swivel arm 03 is no longer possible once a desired swivel angle value has been reached.

[0042] Fig. Figure 2 shows a second preferred embodiment of the microscope according to the invention, which is similar to the one in Fig. The embodiment shown in 1 comprises the object stage 02, the lens 07 and the optical module 08. In contrast to the embodiment shown in Fig. In the embodiment shown in Figure 1, the lens 07 and the optical module 08 are not attached to a pivoting swivel arm, but are fixedly arranged. Instead, the stage 02 is rotatable. The rotational movement of the stage 02 can be slowed down by the electrically actuated brake 11, so that the stage 02 can be fixed with respect to its rotation. The in Fig. 2 The embodiment shown comprises a further lens 12 and a further optical module 13, which are arranged under the object stage 02.

[0043] Fig. 3 shows this in Fig. 1 Microscope shown in two states during the execution of a method according to the invention. The left part of the Fig. Figure 3 shows the microscope in an initial position in which the swivel arm 03 is not tilted, so that the tilt angle Θ is zero. A value Θ is set according to the procedure. Ziel of the inclination angle Θ selected.

[0044] The right part of the Fig. Figure 3 shows the microscope in a situation where the value to be set Θ Ziel The tilt angle Θ is set according to the procedure, which is made possible by pressing the unlock button 09. The swivel arm 03 has already been moved up to a value Θ by driving the electrically motorized actuator 04. aktuell The tilt angle Θ is pivoted. The driving of the electrically motorized actuator 04 continues until the value to be set Θ is reached. Ziel of the inclination angle Θ is reached.

[0045] Fig. 4 shows this in Fig. 2 Microscope shown in two states during the execution of the method according to the invention. The left part of the Fig. Figure 4 shows the microscope in an initial position in which the stage 02 is not tilted, so that the tilt angle Θ is zero. A value Θ is set according to the procedure. Ziel of the inclination angle Θ selected.

[0046] The right part of the Fig. Figure 3 shows the microscope in a situation where the value to be set Θ Ziel The tilt angle Θ is set according to the procedure. The object stage 02 is rotated manually, whereby the object stage 02 is already set to a value Θ. aktuell The tilt angle Θ was rotated. The rotational movement of the object stage 02 is measured according to the procedure, and a time is predicted at which the value to be set Θ will be reached. Ziel the tilt angle Θ is reached. At this point, the electrically operated brake 11 is activated, so that further rotation of the object stage 02 is no longer possible. Reference symbol list 01 Base 02 Object table 03 Swivel arm 04 electrically motorized actuator 05 - 06 Sample 07 Lens 08 optical module 09 Unlock button 10 - 11 electrically operated brakes 12 additional lenses 13 additional optical modules

Claims

[1] Method for microscopying a sample (06) with a microscope, wherein a mechanical parameter (Θ) of the microscope is variable, and wherein the method comprises at least the following steps: - Selecting a first value to be set (Θ Ziel ) of the mechanical parameter (Θ); - Actuating an electrically controlled actuator (04; 11) to set the first value to be set (Θ Ziel ) of the mechanical parameter (Θ); - Taking a first microscopic image of the sample (06) at the set first value (Θ) Ziel ) of the mechanical parameter (Θ); - Selecting a second value to be set (Θ Ziel ) of the mechanical parameter (Θ); - Actuating the electrically controlled actuator (04; 11) to set the second value to be set (Θ Ziel ) of the mechanical parameter (Θ); and - Taking a second microscopic image of the sample (06) at the set second value (Θ) Ziel ) of the mechanical parameter (Θ); wherein the mechanical parameter (Θ) is manually changeable; wherein the actuating element is formed by a locking device (11) with which the mechanical parameter (Θ) can be locked; and wherein the steps of actuating the electrically controllable actuating element (11) each comprise the following sub-steps: - Measuring the time course of a manually induced change in value (Θ) aktuell ) of the mechanical parameter (Θ); - Predicting a point in time at which the previously determined time course of the manual change of the value (Θ) aktuell ) of the mechanical parameter (Θ) to the value to be set (Θ) Ziel ) of the mechanical parameter (Θ); and - electrical activation of the electrically actuated locking device (11) at the previously determined time. [2] Method according to claim 1, characterized by , that the actuating element is formed by an actuator (04) which can be driven by an electric motor and by which the mechanical parameter (Θ) can be adjusted. [3] Method according to claim 1 or 2, characterized by , that the mechanical parameter is formed by an angle of inclination (Θ) between an objective (07) of the microscope and a stage (02) of the microscope; wherein the objective (07) is attached to a pivotable swivel arm (03), or wherein the stage (02) is rotatable in an axis parallel to the extension plane of the stage (02), whereby the angle of inclination (Θ) between the objective (07) and the stage (02) can be changed. [4] Method according to any one of claims 1 to 3, characterized by that it includes the following further step: - Generating feedback to the operator when the value of the mechanical parameter (Θ) has been changed by a predefined amount and / or when the value to be set (Θ) Ziel ) of the mechanical parameter (Θ) was set. [5] Method according to claim 4, characterized by , that the feedback is haptically perceptible and is generated by the electromagnetically actuated locking device (11). [6] Method according to any one of claims 1 to 5, characterized by that it includes the following further steps, which are repeated several times: - Selecting another value that differs from the other values ​​(Θ Ziel ) of the mechanical parameter (Θ) differentiating value to be set (Θ) Ziel ) of the mechanical parameter (Θ); - Actuating the electrically controlled actuating element (04; 11) to set the next value to be set (Θ Ziel ) of the mechanical parameter (Θ); and - Taking another microscopic image of the sample (06) at the set further value (Θ) Ziel ) of the mechanical parameter (Θ). [7] Method according to any one of claims 1 to 6, characterized by , that it is designed for microscopy of at least two samples (06), wherein the same first values ​​(Θ) are used for taking the microscopic images of the first sample (06) and for taking the microscopic images of the second sample (06). Ziel ) of the mechanical parameter (Θ) and the same second values ​​(Θ Ziel ) of the mechanical parameter (Θ) can be selected and set. [8] Method according to any one of claims 1 to 7, characterized by that it includes the following further step: - Generating a 2.5-dimensional or three-dimensional representation of the sample (06) from the multiple microscopic images. [9] Microscope for microscopy of a sample (06) in which a mechanical parameter (Θ) is variable, comprising: - an electrically controlled actuating element (04; 11) for setting a value of the mechanical parameter (Θ); and - a control unit configured to execute a method according to any one of claims 1 to 8.

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