Control device and method for controlling a microscope, microscope and computer program product

Abstract

The present disclosure relates to a control device (100) and method for controlling a microscope (101), such as an oblique plane microscope (102) or a SCAPE microscope (103), such a microscope (101) and a computer program product. In the art, a sample larger than the field of view (110a) of an optical system (110) can be scanned at high or low speed with a sample volume (105). As a result, the sample may be repositioned or even damaged, or the measurement time may increase. The improvement of the present invention over the prior art solutions lies in that the control device (100) is configured to adjust the sample volume (105) of the microscope (101) relative to the sample volume speed (104) of the microscope (101); and to adjust the light sheet (107) of the microscope (101) relative to the light sheet speed (106) of the microscope (101) depending on the sample volume speed (104). A corresponding microscope includes a light sheet generator; an optical system; a light sheet actuator; a sample actuator; a detection unit and the control device (100) of the present invention.

Description

Technical Field

[0001] The present invention relates to a control device and method for controlling a microscope such as an oblique plane microscope or a SCAPE microscope, to a microscope such as an oblique plane microscope or a SCAPE microscope, and to a computer program product. Background Art

[0002] In prior art solutions for oblique plane microscopes, the sample moves relative to a static light sheet. In those prior art solutions, a translation stage is typically applied. If the sample moves along the optical axis of the objective lens, a stack of images can be recorded in an oblique plane microscope.

[0003] In a SCAPE microscope, the light sheet moves through the sample within a scanning range by means of a scanner located in the illumination optical path and the observation optical path. Using SCAPE, it is more troublesome to record a large sample area wider than the field of view of the objective lens. Summary of the Invention

[0004] The present invention addresses the above problems for the control device described at the beginning in that the control device is configured to adjust the sample volume of the microscope relative to the sample volume speed of the microscope, and to adjust the light sheet of the microscope relative to the light sheet speed of the microscope depending on the sample volume speed.

[0005] The present invention solves the above problems of the microscope mentioned at the beginning in that the microscope includes a light sheet generator for generating a light sheet in the sample volume of the microscope, an optical system for transmitting light into the sample volume, a light sheet actuator for adjusting the light sheet speed, a sample actuator for adjusting the sample volume speed of the sample volume, a detection unit for detecting the light distribution in the sample volume, the light distribution being generated by the light sheet, and a control device for controlling at least one of the light sheet speed and the sample volume speed, wherein the control device is a control device according to the present invention.

[0006] The present invention further improves the prior art method for controlling a microscope, wherein the method includes adjusting the sample volume speed of the sample volume, and adjusting the light sheet speed of the light sheet depending on the sample volume speed.

[0007] If different parts of the sample are recorded by moving the sample, the required sample acceleration can be performed quickly to obtain high throughput or slowly and carefully. Both methods are disadvantageous because the high acceleration for fast scanning results in a large force acting on the sample, which may deform or reposition the sample, and the slow movement of the sample requires the frame rate of image recording to adapt to the acceleration of the sample. This may further increase the time required to complete the measurement.

[0008] Accordingly, one aspect of the present invention is to provide a control device, a method, and a microscope that facilitate recording a sample area larger than the field of view of the applied objective lens.

[0009] The computer program product includes program code for performing the method of the present invention when the computer program runs on a processor.

[0010] The control device, method, and microscope of the present invention can be further improved by the additional technical features described below. If the technical effects obtained by the omitted technical features are irrelevant to the present invention, these additional technical features can be combined arbitrarily with each other or omitted.

[0011] Preferably, it is the light sheet speed suitable for the sample volume speed. This has the advantage that the light sheet speed can be adjusted quickly and easily according to the (varying) sample volume speed. Adjusting the light sheet speed, for example, by the light sheet actuator of the microscope of the present invention, is easier and faster than adjusting the sample volume speed as the corresponding device for adjusting the light sheet speed. For example, compared with the acceleration of the sample holder that holds the sample, the light sheet actuator has lower inertia (the resistance of an object to a change in speed) and lower sensitivity respectively.

[0012] In the present disclosure, speed is understood as a physical vector defined by magnitude and direction. In some embodiments, where, for example, moving the sample is performed only along one axis, the speed can be regarded as a signed quantity.

[0013] Furthermore, in the present disclosure, an oblique plane microscope should be understood as a microscope, preferably a light sheet microscope including a light sheet, where the light sheet is oriented in an inclined manner in the sample volume, where the light sheet can be moved, preferably in the lateral direction, and where the sample, more generally, the sample volume, can be moved relative to the optical assembly, particularly relative to the objective lens. The control device, method, computer program product, and microscope can particularly refer to or be applied to such a light sheet microscope.

[0014] To allow scanning of the sample, an embodiment of the control device of the present invention is configured to adjust the light sheet speed relative to the microscope to be different from the sample volume speed relative to the microscope in at least one of direction and absolute value. This ensures that the light sheet and the sample volume do not move in the same direction with the same absolute value. If this is the case, static planar illumination of the sample via the light sheet will occur, which does not allow recording an image stack or scanning the sample.

[0015] The control device can be configured to adjust the scanning speed of the light sheet to be constant by adjusting one of a) the light sheet speed depending on the sample volume speed and b) the sample volume speed, where the scanning speed is the light sheet speed relative to the sample volume.

[0016] The scanning speed can be the superposition of the light sheet speed and the sample volume speed, where the scanning speed is referenced to the sample volume. Thus, if the light sheet does not move relative to the microscope and the sample volume moves at a first speed +v 1 then the scanning speed of the light sheet relative to the sample volume, v sc is equal to v sc =-v 1 .

[0017] Preferably, the control device can be configured to adjust the light sheet speed to provide a constant scanning speed, where the sample volume speed can also be adjusted, which in turn results in an adjustment of the light sheet speed.

[0018] To perform the adjustment step, an embodiment of the microscope according to the invention may further include a speed detector unit for determining at least one of the sample volume speed, the light sheet speed, and the superposition of the sample volume speed and the light sheet speed.

[0019] Thus, one of the speeds can be measured online or determined, for example, by means of a look-up table, where the progression of the speed (light sheet and / or sample volume) can be stored as a function of the time elapsed since the start of the measurement.

[0020] Thus, the control device can be configured to measure or determine at least one of the sample volume speed, the light sheet speed, and the scanning speed.

[0021] In one embodiment of the control device according to the invention, the control device can thus be configured to reduce the light sheet speed if the sample volume speed increases and / or to increase the light sheet speed if the sample volume speed decreases.

[0022] This increase or decrease is to be understood as an increase or decrease in the absolute value of the respective speed.

[0023] For example, if a target scanning speed of 10 (arbitrary unit) is to be achieved along the x-axis in the sample volume, i.e., v sc,t =+10, then a sample volume speed of v sv =-1 results in a first scanning speed v sc,1 =+1, where the control device adjusts the first light sheet speed v ls,1 to v ls,1 =+9, such that the adjusted scanning speed v sc,adj =+10 (arbitrary unit) results from the superposition of the two speeds.

[0024] Similarly, a sample volume speed of -5 will cause the control device to adjust the first light sheet speed to +5 to maintain a constant scanning speed.

[0025] In a further advantageous embodiment of the control device according to the invention, the control device may be configured to adjust the acceleration of the light sheet based on the acceleration of the sample volume.

[0026] Thus, if the sample volume accelerates, for example, in order not to deform or reposition the sample provided therein, the light sheet moves at an initially higher light sheet speed (adjusted by the control device such that the scanning speed remains constant at a predetermined target value), wherein, with further acceleration of the sample volume, the light sheet decelerates. As the speed of the sample volume increases (i.e., the absolute value of the speed increases), the light sheet speed decreases at a rate corresponding to the increase in the speed of the sample volume (which corresponds to deceleration, i.e., the rate of change of speed over time).

[0027] Thus, even if the speed of the sample is not constant, but for example during acceleration or deceleration, the scanning speed, i.e., the speed at which the light sheet scans through the sample volume, can be kept constant by the control device.

[0028] Thus, the control device, method, microscope, and computer program product according to the invention have the advantage that larger volumes, i.e., volumes or samples larger than the field of view of the objective lens, can be inspected completely and quickly, wherein in particular a three-dimensional image stack of the entire sample can be recorded. Each image of the image stack is recorded at an equidistant step between each image, without exposing the sample to excessive forces or accelerations, and without the need to adopt the frame rate of the detector, such as a camera, which would otherwise be necessary to keep the distance between the images equidistant if the sample volume speed were to change.

[0029] The control device and the microscope may include an adjustment range within which the light sheet can move. This adjustment range should be understood as the range within which the light sheet can be repositioned (oblique plane microscope) or the actual scanning range of the light sheet (SCAPE), in particular moved, in order to keep the scanning speed constant. The adjustment range may be defined by the light sheet actuator member for moving the light sheet. The ends of the adjustment range may be defined by the maximum and minimum deflection angles of the light sheet actuator member or by the orientation or position of the light sheet actuator member. The adjustment range may also be limited and thus defined by the available aperture of the optical system, such as the objective lens, via which the light of the light sheet is provided into the sample volume. In addition, the adjustment range may be defined by providing a hard aperture in the optical system. The light sheet actuator member may be a tiltable element that provides deflection or a linearly movable element that provides lateral displacement of the light sheet.

[0030] The control device may be configured to adjust the sample volume speed based on the position of the light sheet within the adjustment range.

[0031] Preferably, if the light sheet is located in a predetermined region at the boundary of the adjustment range, the control device may be configured to adjust the sample volume speed such that the first direction is equal to the second direction.

[0032] Thus, it is possible to detect whether the light sheet is close to the end, limit or boundary of the adjustment range, whereupon detection, the control device or the microscope can initiate overcompensation as a countermeasure by adjusting the sample volume velocity such that the light sheet and the sample volume move in the same direction. This overcompensation allows returning the light sheet beyond a predetermined region at the limit of the adjustment range, preferably to the center of the adjustment range. The predetermined region can represent a safety buffer ensuring that it is still possible to adjust the light sheet velocity. The angular range in which the light sheet enters the predetermined region can correspond to 20% of the entire adjustment range. It can also correspond to 10% or 5% of the adjustment range.

[0033] In the present disclosure, the position of the light sheet should be understood as corresponding to the position of the scanning range in the case of a SCAPE microscope. To perform the scanning and adjustment of the light sheet, the same or another optical element can be applied. Exemplarily, the same tilt mirror used for scanning the sample volume with the light sheet can be used to adjust the position of the light sheet.

[0034] In a further embodiment, the control device can be configured to adjust the sample volume velocity such that a first direction, which is the direction of the sample volume velocity, is oriented in a direction opposite to a second direction, which is the direction of the light sheet velocity. In other words, using the microscope as a reference frame, the direction of movement of the light sheet and the direction of movement of the sample volume point in opposite directions, i.e., they are anti-parallel.

[0035] In a further advantageous embodiment, the control device can be configured to adjust the sample volume velocity such that a first direction, which is the direction of the sample volume velocity, is equal to a second direction, which is the direction of the light sheet velocity. In other words, relative to the microscope, the sample volume and the light sheet move in the same direction.

[0036] Continuing with the above example, the target scanning velocity is 10 (arbitrary unit), and this occurs if the sample volume velocity reaches -15, i.e., v sv,1 =-15. In this case, if the light sheet does not move relative to the microscope, a first scanning velocity of v sc,1 = +15 will be obtained, which is higher than the target scanning velocity. In this case, the control device adjusts the light sheet velocity to v ls,1 = -5, thereby maintaining the scanning velocity at +10. In this case, the sample volume velocity and the light sheet velocity have the same direction.

[0037] The advantage of this embodiment is that once the light sheet is moved in a first adjustment direction to keep the scanning velocity constant, the direction of movement of the light sheet can be reversed to remain within the adjustment range. If the sample volume amount is greater than the amount of the required scanning velocity, the direction of the light sheet can be reversed and moved back to its initial position. This allows keeping the light sheet within its adjustment range.

[0038] In a further advantageous embodiment, the control device may be configured to adjust the sample volume velocity such that if the light sheet approaches the limit of the adjustable range in which the light sheet is movable, the first direction is equal to the second direction. In this embodiment, the light sheet is controlled in such a way as to remain within its adjustable range.

[0039] Accordingly, two control steps are applied here, namely adjusting the light sheet velocity depending on the sample volume velocity and controlling whether the light sheet velocity can still be adjusted in more than one direction. This method may be referred to as overcompensation and allows the light sheet to be repositioned to its initial position, preferably, this initial position corresponding to the center of the adjustable range. The second control step may trigger feedback to the first control step, for example, starting to change the sample volume velocity.

[0040] The control device may be configured to change the orientation of the first direction relative to the second direction from a parallel orientation to an anti-parallel orientation and / or from an anti-parallel orientation to a parallel orientation. During a first time period, the light sheet and the sample volume may move in the same direction, while during a second time period, preferably a subsequent time period, the light sheet and the sample volume may move in opposite directions (using the microscope as a reference frame in both cases). During one measurement, many of these time periods may alternate with each other.

[0041] The control device of the present invention may be configured to move the light sheet in a direction perpendicular to the light sheet, i.e., along its normal vector. The light sheet may also be located only within the sample volume and move in a direction at an angle to its normal vector.

[0042] The control device of the present invention may in particular be configured to adjust one of the light sheet velocity and the sample volume velocity to position the light sheet at the center of the adjustable range. If the magnitude of the sample volume velocity is greater than the magnitude of the target scan velocity (where the two velocities point in opposite directions), i.e., by overcompensating the sample volume velocity, such positioning at the center of the adjustable range can be achieved.

[0043] The microscope of the present invention may include a further optical system for transmitting light from the sample volume to the detection unit, wherein the light sheet actuator may be configured to translate the light sheet in a direction parallel to the optical axis of the further optical system.

[0044] The computer program product may be any conventional magnetic or optical storage medium or any read-only memory (ROM) or random access memory (RAM) based on a non-transitory storage medium. The program code may be executed on a personal computer, any field-programmable gate array, microcontroller, etc., in short, any integrated circuit configured to store the program code.

[0045] The sample volume velocity can vary linearly or non-linearly with time, or remain constant. In any case, depending on the sample volume velocity, the light sheet velocity can be adjusted by a control device so as to preferably obtain a constant scanning velocity. In different examples, it may be desirable to have a corrected scanning velocity in certain regions of the sample volume. For example, if a region of interest is defined to require a predetermined resolution, i.e., the distance between adjacent images acquired in the sample, where outside the said region of interest, a lower resolution may be sufficient. This will allow for an overview scan outside the region of interest and a more detailed scan, i.e., a scan with a higher resolution, within the region of interest. In such a case, the target scanning velocity can be set higher outside the region of interest and lower inside the region of interest.

[0046] In these two different parts of the sample (inside and outside the region of interest), the scanning velocity can be set to be constant.

[0047] Another example is the so-called multi-well plate (microplate), where different volumes (in the wells) are arranged in a pattern. The wells are separated by intermediate regions between them. In this case, there is no single sample, but rather different parts, i.e., regions of interest. The present disclosure may be beneficial for obtaining an overview of all well contents in the shortest possible time. The movement of the light sheet from one well to an adjacent well can be performed rather quickly, i.e., over the intermediate region, where the movement of the sample volume within the well, i.e., within the sample, is performed at a lower scanning velocity. This results in a permanent acceleration and deceleration of the multi-well plate, where the deceleration of the sample volume (when the light sheet enters the region of interest) and the acceleration of the sample volume (when the light sheet moves from the region of interest to the intermediate region between two adjacent wells) can be compensated for by the present disclosure.

[0048] Furthermore, within each well, in addition to the adjustment when entering or leaving the well, the light sheet velocity can be adjusted according to the sample volume velocity. The present invention can also be applied in a microscope using remote focusing, where the focus within the sample volume is changed by additional telescopes in the illumination optical path and / or the observation optical path. In this case, the remote focusing module can determine the light sheet velocity, which can be adapted to the possible velocity of the sample volume along the optical axis of the objective.

[0049] The microscope of the present invention, preferably a light sheet microscope, more preferably an oblique plane microscope or a SCAPE microscope, can optionally perform detection of the observation light emitted or generated in the sample volume by the same optical component (objective) that performs the illumination. Thus, one and the same optical component (objective) can be used as an optical illumination component and an optical observation component.

[0050] Furthermore, the optical observation assembly may generate a virtual image of a planar portion of the observation volume illuminated by the light sheet. The virtual image may be inclined, i.e., inclined with respect to the optical axis of the optical observation assembly. The microscope according to the invention may include a further optical assembly which is configured to be oriented at an angle with respect to the optical observation assembly such that the optical axis of the further optical assembly is substantially perpendicular to the virtual image. This allows a further image of the virtual image to be generated in a focal plane oriented perpendicular to the optical axis of the further optical assembly. A detector may be arranged within this focal plane. Such a further optical assembly (German: Aufrichteeinheit) thus creates an inclined virtual image. Alternatively, the inclination of the virtual image may be compensated for by a computer program product comprising code which, when run on a computer, calculates a corrected image.

[0051] The SCAPE microscope is a particularly advantageous embodiment of an inclined plane microscope, in which the light sheet is scanned through the sample volume.

[0052] The inclined plane microscope may be provided with a single objective (i.e., similar to the SCAPE microscope), in which an inclined image of the inclined light sheet is imaged onto the detector in a planar manner by increasing the depth of focus rather than by an objective at an angle to the illumination or detection objective.

[0053] The movement of the sample volume and the corresponding adjustment of the light sheet movement may be repeated periodically. Thus, after a certain time (cycle duration or period time), the light sheet may return to its initial position. Since it has to remain within its adjustment range, it may perform a periodic movement around the center of the adjustment range. During this time, the light sheet may reach its initial position once, twice or more times.

[0054] In a preferred embodiment, the present disclosure allows a constant scanning speed and a periodic movement of the light sheet as well as a periodic movement of the sample volume. This allows any large sample to be scanned.

[0055] As an alternative, images may be captured only during a specific time span, in particular until the light sheet reaches its maximum position within its adjustment range, after which image capture stops and the light sheet is moved back to its initial position. Then, the system may continue to move the sample volume.

[0056] The simplest case is to adjust the speed of the light sheet and the speed of the sample volume such that, after the measurement is completed, the light sheet has not reached the boundary of its adjustment range. Thus, after the measurement is ended, the light sheet may return to its initial position.

[0057] Although some aspects have been described in the context of an apparatus, it is clear that these aspects also represent a description of the corresponding method, where a block or device corresponds to a method step or a feature of a method step. Similarly, aspects described in the context of method steps also represent a description of corresponding blocks or items or features of a corresponding apparatus. Some or all of the method steps may be performed by (or using) a hardware apparatus, such as a processor, a microprocessor, a programmable computer, or an electronic circuit. In some embodiments, some one or more of the most important method steps may be performed by such an apparatus.

[0058] Depending on certain implementation requirements, embodiments of the present invention may be implemented in hardware or software. Such an implementation may be carried out using a non-transitory storage medium, such as a digital storage medium, for example, a floppy disk, a DVD, a Blu-ray disc, a CD, a ROM, a PROM, an EPROM, an EEPROM, or a flash memory, on which electronically readable control signals are stored, which cooperate (or are capable of cooperating) with a programmable computer system to perform the corresponding method. Thus, the digital storage medium may be computer-readable.

[0059] Some embodiments according to the present invention include a data carrier having electronically readable control signals, which is capable of cooperating with a programmable computer system so as to perform one of the methods described herein.

[0060] In general, embodiments of the present invention may be implemented as a computer program product having program code that is operable to perform one of the methods when the computer program product is run on a computer. For example, the program code may be stored on a machine-readable carrier.

[0061] Other embodiments include a computer program stored on a machine-readable carrier for performing one of the methods described herein.

[0062] In other words, thus, embodiments of the present invention are computer programs having program code for performing one of the methods described herein when the computer program is run on a computer.

[0063] Thus, a further embodiment of the present invention is a storage medium (or data carrier, or computer-readable medium) that includes a computer program stored thereon for performing one of the methods described herein when executed by a processor. The data carrier, digital storage medium, or recorded medium is generally tangible and / or non-transitory. A further embodiment of the present invention is an apparatus as described herein, including a processor and a storage medium.

[0064] Thus, a further embodiment of the present invention is a data stream or a signal sequence representing a computer program for performing one of the methods described herein. The data stream or signal sequence may be configured, for example, to be transmitted via a data communication connection, such as via the Internet.

[0065] Further embodiments include a processing device, such as a computer or a programmable logic device, configured to or adapted to perform one of the methods described herein.

[0066] Further embodiments include a computer having a computer program installed thereon for performing one of the methods described herein.

[0067] Further embodiments may include a device or system configured to transmit (e.g., electronically or optically) a computer program for performing one of the methods described herein to a receiver. For example, the receiver may be a computer, a mobile device, a memory device, etc. For example, the device or system may include a file server for transmitting the computer program to the receiver.

[0068] In some embodiments, a programmable logic device (e.g., a field programmable gate array) may be used to perform some or all of the functions of the methods described herein. In some embodiments, a field programmable gate array may cooperate with a microprocessor to perform one of the methods described herein. Generally, these methods are preferably performed by any hardware device. BRIEF DESCRIPTION OF THE DRAWINGS

[0069] Hereinafter, examples of the present disclosure will be shown with reference to the drawings. Only exemplary embodiments will be described, which are not intended to limit the possible scope of protection defined by the claims. In the exemplary embodiments, if the technical effects of the omitted technical features are irrelevant to the subject matter of the present disclosure, the technical features may be omitted.

[0070] The drawings show:

[0071] Figure 1 is a schematic diagram of the microscope and the control device of the present invention;

[0072] Figure 2 is a schematic diagram of the working principle of a non-constant acceleration control device;

[0073] Figure 3 is a schematic diagram of the working principle of a constant acceleration control device;

[0074] Figure 4 is a schematic example of possible movement patterns and corresponding adjustments performed by the control device;

[0075] Figure 5 is a further schematic example of another possible movement pattern and corresponding adjustments performed by the control device;

[0076] Figure 6 is a further schematic example of another possible movement pattern and corresponding adjustments performed by the control device; DETAILED DESCRIPTION

[0077] In the following description of the figures, the elements introduced in the figures are denoted by the preceding numbers corresponding to the figures in which the features are introduced. Figure 2 Technical features first occurring in Figure 4 are thus denoted by reference numerals 2xx. Identical features may be referred to in subsequent figures, where the features are then denoted by reference numerals 3xx, 4xx, 5xx, etc. Similarly, technical features first described in Figure 1 and denoted by reference numeral 4yy may also be addressed in Figure 1 where they are referred to as 1yy. In the list of reference numerals, a technical feature is given only by one reference numeral. Exemplarily, the velocity of the sample volume is given by reference numeral 217, where if the velocity of the sample volume is addressed in Figure 1 、 3 、4, 5 or 6, this reference numeral also includes reference numerals 117, 317, 417, 517 and 617. The same applies to the other features given in the list of reference numerals.

[0078] Figure 1 is a schematic illustration of microscope 101 of the present invention. The illustrated microscope 101 may be embodied as an inclined plane microscope 102 or a SCAPE microscope 103.

[0079] Figure 1 Schematically shown is a light sheet generator 107a that generates a light sheet 107 in a sample volume 105 of microscope 101.

[0080] Microscope 11 further includes an optical system 110 for transmitting light 107b from the light sheet generator 107a through the optical system 110 into the sample volume 105.

[0081] Figure 1 Further shown is a light sheet actuator 107c, which is shown exemplarily as a transmissive device but may also operate in reflection. The light sheet actuator 107c allows adjustment of the light sheet velocity 106.

[0082] The light sheet velocity 106 is characterized by a direction 106a and an absolute value 106b, i.e., they represent a connector unit 111.

[0083] Figure 1 Further shown is a detection unit 112 that detects the light distribution 113 in the sample volume 105, where the light distribution 113 is generated by the light sheet 107 and transmitted (in the illustrated embodiment) to the detection unit 112 via the optical system 110. Thus, the observation light 114 is transmitted to the detection unit 112 via the same optical system 110.

[0084] The microscope 101 further includes a control device 100, which is connected to the light sheet actuator 107c and the sample actuator 115. The sample actuator 115 is configured to adjust the sample volume velocity 104 of the sample volume 105.

[0085] In Figure 1 the embodiment shown, the light sheet actuator 107c and the sample actuator 115 are included in the microscope 101, where in different embodiments, these features may be included in the control device 100.

[0086] Figure 1 Three positions 116 of the sample volume 105 relative to the microscope 101 are shown. This time series of positions 116 describes the movement of the sample volume 105 with the sample volume velocity 104 in the direction of the sample volume velocity 104a having the absolute value of the sample volume velocity 104b.

[0087] Due to the superposition 109 of the light sheet velocity 106 and the sample volume velocity 104, the light sheet 107 moves relative to the sample volume 105 at the scanning velocity 108.

[0088] Since the scanning velocity 108 is considered with the sample volume 105 as the reference frame, and the directions 104a and 106a of the sample volume velocity and the light sheet velocity are respectively opposite to each other, the absolute values 104b and 106b of the sample volume velocity and the light sheet velocity are respectively added (added) to produce the absolute value of the scanning velocity 108b. The direction of the scanning velocity 108a is equal to the direction of the light sheet velocity 106a.

[0089] The control device 100 may include a velocity evaluation module 100a, which may be coupled to a velocity detector 100b, only schematically shown for clarity. In different embodiments, the velocity evaluation module 100a and the velocity detector 100b may be part of the microscope 101 and may directly provide velocity data (not shown) to a calculation module 100c. The calculation module 100c may be directly connected to the sample actuator 115 and / or the light sheet actuator 107c, thereby allowing the control device 100 to adjust the sample volume velocity 104 and / or the light sheet velocity 106. The control device 100 may further include a storage module 100d, in which calculation rules, movement pattern data, or further parameters of the control device or the microscope may be stored.

[0090] For example, the velocity of the light sheet 107 or the sample volume 105 may not be measured, but determined by reading a look-up table that may be stored in the storage module 100d.

[0091] Figure 1The field of view 110a of the optical system 110 is also shown. The recording of the image of the sample volume 105 is thus limited to this field of view 110a, so that it is necessary to move the sample volume 105 to examine the entire sample volume 105.

[0092] At Figure 1 the position 116 on the right side, the situation where the sample volume velocity becomes the second sample volume velocity 104f is shown. According to this changed second sample volume velocity 104f, the control device 100 corrects the light sheet velocity to the second light sheet velocity 106f. The superposition 109 of the two again results in a scanning velocity 108, which remains constant at all three positions 116 shown.

[0093] At Figure 2 and Figure 3 the velocities v related to the time t are shown. The figure shows the velocities of the sample volumes 217, 317 and the velocities of the light sheets 218, 318. The velocities 217, 317 and 218, 318 are respectively understood as the signed values of the corresponding absolute values 104b and 106b of the sample volume velocity and the light sheet velocity (see Figure 1 ).

[0094] In the figure, the scanning velocities 219, 319 are defined, the values of which correspond to the absolute value of the scanning velocity 108b, where the direction of the scanning velocity 108a is considered by mathematical symbols (this is a matter of the positive direction, i.e., the signs of the velocities shown may be opposite). The moving direction of the sample volume (not shown) is defined as positive. Therefore, the reverse movement of the light sheet results in negative velocities for the light sheets 218, 38. Since the scanning velocity 108 is considered to be opposite to the sample volume velocity 104, the scanning velocities 219, 319 are also negative.

[0095] The scanning velocities 219, 319 are predetermined values 219a, 319a. At Figure 2 the velocity of the sample volume 217 increases non-linearly from zero to the final value 217a. The control device 100 (not shown) adjusts the velocity of the light sheet 218 depending on the velocity of the sample volume 217, so as to obtain a constant scanning velocity 219.

[0096] At Figure 3 the velocity of the sample volume 317 increases with a constant acceleration 320, where, as Figure 2 at

[0097] the velocity of the light sheet 318 is adjusted accordingly to achieve a constant scanning velocity 319, which corresponds to the predetermined value 319a. Figures 4 to 6 A further possible movement pattern is shown at

[0098] All Figures 4 to 6 show the velocity v over time t.

[0099] In Figure 4 it, a periodic movement pattern 420 is shown for the speed of the sample volume 417 and the speed of the light sheet 418. The scanning speed 419 is constant (which is achieved by a control device 100 (not shown) that adjusts the speed of the light sheet 418 according to the speed of the sample volume 417) and is counted as -1 in any unit.

[0100] This pattern can be applied in the case of inspecting a multi-well plate 421. Such a multi-well plate 421 includes wells 422 and an intermediate region 423.

[0101] Generally, those multi-well plates 421 can be larger than the field of view 110a of the optical system 110, where measurements are not of interest in the intermediate region 423. Within the intermediate region 423, the sample volume, i.e., the entire multi-well plate 421, can move faster. In this figure, a first scanning range 424 and a second scanning range 425 are shown. In the first scanning range 424, the field of view 110a moves (i.e., faster) over the intermediate region 423, where in the second scanning range 425, the field of view is located in the wells 422. In the second scanning range 425, the speed of the sample volume 417 is lower, thus introducing less interference to the sample.

[0102] This figure also shows a shaded region 426, which corresponds to the distance 427 traveled by the light sheet 107.

[0103] Between the first scanning range 424 and the second scanning range 425, the speed of the light sheet 418 changes its sign. At these intersection points 428, the speed of the sample volume 417 corresponds in absolute value to a predetermined value 419a of the scanning speed 419. That is, at the intersection points 428, the light sheet 107 does not translate and the speed of the sample volume 417 causes the static light sheet 107 to scan through the sample volume 105 at the scanning speed 419 (when taking the microscope 101 as a reference).

[0104] In the first scan 424, the light sheet 107 moves in the positive direction, where in the second scanning range 425, the light sheet 107 moves in the negative direction (negative speed of the light sheet 418). Therefore, the movement of the light sheet 107 in the second scanning range 425 compensates for the movement of the light sheet 107 in the first scanning range 424.

[0105] Overall, the light sheet 107 oscillates around a center or initial position (not shown), which is located at the center of the adjustment range.

[0106] The adjustment range 129 is as Figure 1 shown.

[0107] In Figure 5 and Figure 6 a non-periodic movement pattern is shown.

[0108] In Figure 5 a situation is shown in which the porous plate 521 includes holes 522, where the sample 529 is located at the center of the hole 522. The sample 529 defines a region of interest 530 where high-resolution measurements are required. Therefore, Figure 5 the first scan range 524 of Figure 5 covers the intermediate region 523 and the part of the hole 522 outside the region of interest 530 to quickly move past the lower or uninteresting region (intermediate region 523).

[0109] In the second scan range 525, the speed of the sample volume 517 is reduced to improve the measurement in the region of interest 530. The speed of the light sheet 518 is adjusted to keep the scan speed 519 constant.

[0110] During the movement mode, the light sheet 107 reaches the return point 531 again at the initial position 532 of the light sheet 107.

[0111] Such a return point 531 is achieved again, for example, after a period of time 533. The light sheet 107 thus only moves around its initial position 532 and remains within its adjustment range 129. In Figure 5 it, the period of time 533 is plotted twice, where in each period of time 533, the light sheet 107 starts moving from its initial position 532, returns to its initial position 532, and finally returns to the initial position 532 again. In any case, after the period of time 533, the light sheet returns to the initial position 532. It is also possible to reach the initial position once, twice, or even several times within the period of time 533.

[0112] Figure 6 is shown similar to Figure 5 that shown in Figure 5 The difference from the movement mode shown in Figure 5 is that the third scan range 634 is included between the divided first scan ranges 624.

[0113] In this third scan range 634, the light sheet 107 remains stationary with the microscope 101 as the reference frame, where the speed of the sample volume 617 in this third scan range 634 results in a constant scan speed 619. In the third scan range 634, the light sheet 107 is at its initial position 632. This is also the case at the return 631 after the period of time 633.

[0114] The exemplary chart shown can be arbitrarily modified, for example, if during the scan in the porous plate near the end of the plate, where in the transitional movement described by the curve to position the field of view of the optical component to a position translated perpendicular to the scan direction, for example, the speed of the sample volume can be reduced so that the sample does not shift due to its inertia.

[0115] Similarly, during the transition of the field of view of the optical component over the middle region of the porous plate, the scanning speed can also be increased. This is not relevant in the middle region as no relevant content is provided there, but this increase in speed helps to scan the porous plate more effectively and faster.

[0116] Reference numeral

[0117] 100 Control device

[0118] 100a Speed evaluation module

[0119] 100b Speed detector

[0120] 100c Calculation module

[0121] 100d Storage module

[0122] 101 Microscope

[0123] 101a Light sheet generator

[0124] 101b Light

[0125] 101c Light sheet actuator

[0126] 102 Oblique plane microscope

[0127] 103 SCAPE microscope

[0128] 104 Sample volume velocity

[0129] 104a Direction of sample volume velocity

[0130] 104b Absolute value of sample volume velocity

[0131] 104f Second sample volume velocity

[0132] 105 Sample volume

[0133] 106 Light sheet velocity

[0134] 106f Second light sheet velocity

[0135] 106a Direction of light sheet velocity

[0136] 106b Absolute value of light sheet velocity

[0137] 107 Light sheet

[0138] 108 Scanning speed

[0139] 108a Direction of scanning speed

[0140] 108b Absolute value of scanning speed

[0141] 109 Superposition

[0142] 110 Optical system

[0143] 110a Field of view

[0144] 111 Vector unit

[0145] 112 Detector unit

[0146] 113 Light distribution

[0147] 114 Observation light

[0148] 115 Sample actuator

[0149] 116 Position

[0150] 129 Adjustment range 129

[0151] 217 Velocity of sample volume

[0152] 217a Final value

[0153] 218 Velocity of light sheet

[0154] 219 Scanning speed

[0155] 219a Predetermined value

[0156] 220 Acceleration

[0157] 420 Periodic movement mode

[0158] 421 Microplate

[0159] 422 Well

[0160] 423 Intermediate region

[0161] 424 First scanning range

[0162] 425 Second scanning range

[0163] 426 Shadow region

[0164] 427 Distance

[0165] 428 Intersection point

[0166] 529 Sample

[0167] 530 Region of interest

[0168] 531 Return point

[0169] 532 Initial position

[0170] 533 Time period

[0171] 634 Third scanning range

[0172] v Velocity

[0173] t Time

Claims

1. A control device (100) for controlling a microscope (101), wherein the microscope (101) is a SCAPE microscope (103) having an identical optical component serving as both an optical illumination component and an optical observation component, and including a light sheet generator for generating a light sheet (107) in a sample volume (105) of the microscope (101), a light sheet actuator (107c) for adjusting the light sheet speed (106), and a sample actuator (115) for adjusting a sample volume speed (104) of the sample volume (105); wherein the control device (100) is configured to - adjust the sample volume (105) of the microscope (101) relative to the sample volume speed (104) of the microscope (101); and - depending on the sample volume speed (104), adjust the light sheet (107) of the microscope (101) relative to the light sheet speed (106) of the microscope (101); wherein the control device (100) is further configured to perform at least one of the following: - adjust the sample volume speed (104) such that a first direction, which is the direction (104a) of the sample volume speed (104), is oriented in a direction opposite to a second direction, which is the direction (106a) of the light sheet speed (106); or - adjust the sample volume speed (104) such that a first direction, which is the direction (104a) of the sample volume speed (104), is equal to a second direction, which is the direction (106a) of the light sheet speed (106), wherein, the control device (100) is configured to adjust a scanning speed (108) of the light sheet (107) to be constant by adjusting one of the light sheet speed (106) and the sample volume speed (104) depending on the sample volume speed (104), wherein the scanning speed (108) is the speed of the light sheet (107) relative to the sample volume (105); wherein the control device (100) is configured to at least one of the following: decrease the light sheet speed (106) if the sample volume speed (104) increases, and increase the light sheet speed (106) if the sample volume speed (104) decreases.

2. The control device (100) according to claim 1, wherein, the control device (100) is configured to adjust the light sheet speed (106) relative to the microscope (101) to be different from the sample volume speed (104) relative to the microscope (101) in at least one of a direction (104a, 106a) and an absolute value (104b, 106b).

3. The control device (100) according to claim 1 or 2, wherein, the control device (100) is configured to adjust the sample volume speed (104) according to the position of the light sheet (107) within an adjustment range (129).

4. The control device (100) according to claim 3, wherein, the control device (100) is configured to adjust the sample volume velocity (104) such that if the light sheet (107) is located in a predetermined region at the boundary of the adjustment range (129), the first direction is equal to the second direction.

5. The control device (100) according to claim 1 or 2, wherein, the control device (100) is configured to change the orientation of the first direction relative to the second direction from a parallel orientation to an anti-parallel orientation and / or from an anti-parallel orientation to a parallel orientation.

6. The control device (100) according to claim 1 or 2, wherein, the control device (100) is configured to adjust the acceleration of the light sheet (107) depending on the acceleration of the sample volume (105).

7. The control device (100) according to claim 1 or 2, wherein, the control device (100) is configured to measure or determine at least one of the sample volume velocity (104), the light sheet velocity (106), and the scanning velocity (108).

8. The control device (100) according to claim 1 or 2, wherein, the control device (100) is configured to move the light sheet (107) in a direction perpendicular to the light sheet (107).

9. The control device (100) according to claim 3, wherein, the control device (100) is configured to adjust one of the light sheet velocity (106) and the sample volume velocity (104) to position the light sheet (107) at the center of the adjustment range (129).

10. A microscope (101), wherein, the microscope (101) is a SCAPE microscope (103) and has an identical optical component serving as both an optical illumination component and an optical observation component; wherein the microscope (101) includes - a light sheet generator for generating a light sheet in the sample volume (105) of the microscope (101); - an optical system (110) for transmitting light (107b) into the sample volume (105); - a light sheet actuator (107c) for adjusting the light sheet velocity (106); - a sample actuator (115) for adjusting the sample volume velocity (104) of the sample volume (105); - a detection unit (112) for detecting the light distribution (113) in the sample volume (105), the light distribution being generated by the light sheet (107); and - a control device (100) for controlling at least one of the light sheet velocity (106) and the sample volume velocity (104), wherein the control device (100) is the control device (100) according to any one of claims 1 to 9.

11. The microscope (101) according to claim 10, further comprising a velocity detector unit (100b) for determining at least one of the sample volume velocity (104), the light sheet velocity (106), and the superposition of the sample volume velocity (104) and the light sheet velocity (106).

12. The microscope (101) according to claim 10 or 11, comprising another optical system for transmitting light from the sample volume (105) to the detection unit (112), wherein the light sheet actuator is configured to translate the light sheet (107) in a direction parallel to the optical axis of the another optical system.

13. A method for controlling a microscope (101), wherein the microscope (101) is a SCAPE microscope (103) and has an identical optical component serving as both an optical illumination component and an optical observation component; wherein the method comprises - adjusting the sample volume velocity (104) of the sample volume (105); and - adjusting the light sheet velocity (106) of the light sheet (107) depending on the sample volume velocity (104); wherein the method further comprises at least one of the following: - adjusting the sample volume velocity (104) such that a first direction, which is the direction (104a) of the sample volume velocity (104), is oriented in a direction opposite to a second direction, which is the direction (106a) of the light sheet velocity (106); or - adjusting the sample volume velocity (104) such that a first direction, which is the direction (104a) of the sample volume velocity (104), is equal to a second direction, which is the direction (106a) of the light sheet velocity (106), wherein, the method further comprises: adjusting the scanning velocity (108) of the light sheet (107) to be constant by adjusting one of the light sheet velocity (106) and the sample volume velocity (104) depending on the sample volume velocity (104), wherein the scanning velocity (108) is the velocity of the light sheet (107) relative to the sample volume (105); wherein the method further comprises at least one of the following: decreasing the light sheet velocity (106) if the sample volume velocity (104) increases, and increasing the light sheet velocity (106) if the sample volume velocity (104) decreases.

14. A computer program product having program code for performing the method for controlling a microscope (101) according to claim 13 when the computer program runs on a processor. ​

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