WO2019001821A1

WO2019001821A1 - Microscope and method for microscopy of a specimen under a variable mechanical parameter

Info

Publication number: WO2019001821A1
Application number: PCT/EP2018/062184
Authority: WO
Inventors: Alexander Gaiduk; Detlef Hein; Pavlos ILIOPOULOS
Original assignee: Carl Zeiss Microscopy Gmbh
Priority date: 2017-06-29
Filing date: 2018-05-11
Publication date: 2019-01-03
Also published as: CN110799879A; DE102017114562A1

Abstract

The invention relates to a microscope and to a method for microscopy of a specimen, wherein in the microscope a mechanical parameter (θ) can be varied. In one step of the method, a first value (θtarget) of the mechanical parameter (θ) to be adjusted is selected. An electrically controllable actuating element (04) is activated in order to adjust the first value (θtarget) of the mechanical parameter (θ) to be adjusted. A first microscopic image of the specimen is recorded under the adjusted first value (θtarget) of the mechanical parameter (θ). In a further step, a second value (θtarget) of the mechanical parameter (θ) to be adjusted is selected. The electrically controllable actuating element (04) is activated in order to adjust the second value (θtarget) of the mechanical parameter (θ) to be adjusted. A second microscopic image of the specimen is recorded under the adjusted second value (θtarget) of the mechanical parameter (θ).

Description

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

The present invention relates to a microscope and a

Method for microscopying a sample, wherein the

Microscope a mechanical parameter is changeable. The mechanical parameter is preferably formed by an angle of inclination, which in at least one spatial direction between a

Lens of the microscope and a stage of the microscope is changeable. This tilt angle describes an inclination between the lens and the stage, which is also referred to as tilt.

JP 2001059599 A2 and JP 2010102344 A2 show

Swivel arm stand for a digital microscope. The

Schwenkarmstativ includes a pivotable about a horizontal axis of rotation pivot arm.

From DE 10 2013 005 999 AI and DE 10 2013 222 295 AI a Schwenkarmstativ for a digital microscope is known in which a Schwenkarmbewegung is blocked by a arranged around a rotation axis High Torque magnetic brake. The blocking may be for the duration of a key press

Push buttons by releasing the high torque magnetic brake

To get picked up. The button is arranged on the swivel arm so that it can be comfortably pressed with at least one finger of the same hand.

The US 6,642,686 Bl shows a controllable rotary arm, with which supports in particular medical operations can be. The rotary arm can, for example, carry a microscope as an instrument.

The object of the present invention, starting from the prior art is the application possibilities for a

Microscope, in which a mechanical parameter, such as a tilt angle between a lens and a stage is changeable to expand. The above object is achieved by a method according to the appended claim 1 and by a microscope according to the attached independent claim 15.

The method according to the invention is used for microscopying a sample with a microscope, in particular with a digital microscope

Microscope. In the digital microscope is preferably an electronic image conversion, wherein the recorded image processed in the form of digital data and brought to display on an electronic image display device. The microscope preferably comprises at least one objective and preferably an image sensor for converting an image imaged directly or indirectly on the image sensor from the objective. At the microscope, a mechanical parameter is changeable.

The mechanical parameter is preferably formed by a position, by an angle or by a distance, which on the sample, on a stage, on the image sensor, on the lens, on a lens component, on a filter and / or on a Illumination source of the

Microscope is related. Thus, the mechanical parameter, for example, by a height and / or a lateral position of the stage, by a distance from the image sensor or be formed by a zoom setting. The mechanical parameter is alternatively preferably formed by a position, by an angle or by a distance of a setting element, which serves to set a physical size on the microscope. For example, it may be a

rotatable adjusting element, with which, for example, a wavelength, a brightness on the microscope or a

Polarization or phase in a lighting or

Detection channel of the microscope can be adjusted.

The mechanical parameter is preferred by a

Inclination formed, which is changeable in at least one spatial direction between the lens of the microscope and the stage of the microscope. The microscope is the

Tilt angle between the lens and the sample

modifying object table of the microscope changeable. The

Object table serves to arrange the sample. The inclination angle is defined between an optical axis of the objective and a perpendicular on the stage. The inclination angle describes a tilt between the lens and the

Object table, which is also referred to as tilt. A

Inclination axis is formed by a rotation axis of the inclination, which is aligned perpendicular to the optical axis. The angle of inclination is in at least one spatial direction

but can also be defined, for example, in

Spherical coordinates can be defined by two values.

The method includes a step of selecting a first value of the mechanical parameter to be set. The first value to set the mechanical

Parameters defines the extent of the mechanical parameters after adjustment. In a further step, an electrically controllable electromechanical Actuator driven to set the first set value of the mechanical parameter. The

Actuator serves to set the mechanical parameter to the first value to be set. In a further step, a first microscopic image of the sample is taken by means of the microscope while the first value of the mechanical parameter is set.

In a further step, a second value of the mechanical parameter to be set is selected. The second value is different from the first value. The second value of the mechanical parameter to be set defines the extent of the inclination angle after adjustment. In a further step, the electrically controllable

actuated electromechanical actuator to set the second set value of the mechanical parameter. In a further step, a second microscopic image of the sample is made using the microscope

recorded while the second value of the mechanical

Parameters is set.

A particular advantage of the method according to the invention is that the microscopic images are each below a defined value of the mechanical parameter,

for example, below a defined value of

Tilt angle can be recorded so that on this basis a further microscopic examination of the sample is made possible. The inventive method allows, for example, an improved further investigation of low-contrast samples. As the operator through the

The method according to the invention is assisted in setting the defined values of the mechanical parameter is the examination of samples under different values of the mechanical parameter far less complex and less error prone.

Preferred embodiments of the method according to the invention further comprise a step, each after the

Actuate the electrically controllable actuator is carried out. In this step, the sample is focused; in particular a refocusing of the sample, since the change of the mechanical parameter,

For example, by changing the inclination angle or by changing the height of the stage, the sample no longer necessarily be focused through the lens. Alternatively or additionally, this is done again

Centering of the sample in the picture to be recorded, for which the

Object table is shifted in height and / or laterally.

In a first preferred embodiment, this is

Actuator by one with an electric motor

drivable actuator formed by which the mechanical

Parameter is adjustable and adjustable. By energizing the electric motor, the mechanical parameter can be adjusted. In a second preferred embodiment of the

mechanical parameters manually changeable. The operator of the microscope adjusts the mechanical parameter with his human strength, for which he prefers first a

Must operate unlocking device. The actuating element is formed by an electrically controllable locking device, with which the mechanical parameters can be determined. The locking device is preferably by an electrically controllable mechanical brake or by a high-torque Magnetic brake formed. In this second preferred

Embodiment, the steps of driving the electrically controllable actuator each comprise a sub-step in which a time course of a manually effected change in the value of the mechanical parameter is measured. For this purpose, an angle sensor or a distance sensor is preferably used. In a further sub-step, a point in time is predicted in which the previously determined chronological course of the manual change of the value of the mechanical parameter will lead to the respective value of the mechanical parameter to be set. Accordingly, the electrically operable locking device is driven at the predetermined time, so that it is ensured that the value to be set in each case of the mechanical

Parameters, such as the value to be set in each case of the inclination angle is actually achieved, and

For example, a dependence of the accuracy of the size of the angle or the distance between the lens and the sample is excluded. By driving the electrically actuated locking device is the mechanical

Parameter fixed, so that it is no longer manually changeable.

In embodiments in which the mechanical parameter is formed by the angle of inclination, the step of

Forecasting the time in which the previously

certain temporal course of the manual change in the value of the angle of inclination to the respective value to be set 0 _Z i _e i will lead the inclination angle, preferably several

Partial steps. In a sub-step, an initial value 0 _{start of} the inclination angle is measured, for which purpose preferably an angle sensor is used. In a further step, a

Angular velocity ω and angular acceleration the previously determined temporal course of the manual

Change in the value of the angle of inclination determined. In a further sub-step, based on the after t

solves the equation Θ _ζ ι _θ ι = 0 _Sta r _t + ω · t + Ή · · t ² using t predicts the time in which the previously determined time course of the manual change of the value of the inclination angle to the value to be set 0 _Z i _e i will lead the inclination angle. Predicting the time in which the previously determined time course of the manual change of the value of the mechanical parameter to the respective

is to be set value of the mechanical parameter is preferably continuously repeated until the manual

Changing the value of the mechanical parameter has led to each set value of the mechanical parameter. The continuous repetition achieves a high accuracy of the prognosis. In embodiments in which the mechanical parameter is formed by the inclination angle, the forecasting of the

Timing, in which the previously determined time course of the manual change of the value of the inclination angle will lead to the value to be set 0 _Z i _e i of the inclination angle, preferably repeated continuously until the manual change of the value of the inclination angle to the respective value to be set 0 _Z i _e i led the inclination angle.

In embodiments in which the mechanical parameter is formed by the angle of inclination, the

Forecast the time in which the previously

certain temporal course of the manual change of the value of the angle of inclination will lead to the respective value to be set 0 _Z i _e i of the inclination angle, preferably further a Mass distribution on the microscope and / or a manual for

Changing the value of the angle of inclination applied force taken into account. In this way, the time at which the previously determined time course of the manual change in the value of the angle of inclination to the respectively to be set

Value 0 _Z i _e i will lead to the inclination angle, more precisely

be predicted.

In embodiments in which the mechanical parameter is formed by the angle of inclination, the objective is preferably attached to a pivoting arm of the microscope. The swivel arm may be a component of a

Swing arm tripod act. The swivel arm can also as

Tiltarm be called. The swivel arm is pivotable, whereby the angle of inclination between the lens the

Object table is changeable.

Alternatively or additionally, the object table is preferably rotatable. The object table is preferably rotatable in an axis which is arranged parallel to the extension plane of the object table or particularly preferably in the

Extension plane of the object table is located. By rotating the object table in this axis, the angle of inclination between the lens of the stage is changeable. The

Object table can also be rotatable about an axis, the

is aligned perpendicular to the plane of extension of the object table. A rotation about this axis leads to a change in the angle of inclination, when the sample also extends on the support in the height.

In particularly preferred embodiments, this includes

Method a further step in which a response is generated to the operator when the value of the mechanical Parameter was changed by a predefined measure and / or if the value of the mechanical

Parameters was set. As a result, the operation of the microscope is facilitated because the operator can quickly detect whether the value of the mechanical parameter to be set has already been achieved. In addition, the operator can each receive a feedback when the value of the mechanical

Parameters changed by a predefined measure;

for example, if the value of the angle of inclination is

For example, 5 ° has been changed. The feedback is

preferably perceivable visually, acoustically and / or haptically.

The feedback to the operator is particularly haptic perceptible and is preferably generated with the electrically operable locking device. For this, in the embodiments in which the mechanical parameter is formed by the inclination angle, the change in the value of the inclination angle, i. H. especially the

Pivoting movement of the swivel arm or the rotation of the

rotatable object table, preferably interrupted in their temporal change briefly, what the operator can feel, so that the operator becomes aware that the value of the inclination angle has been changed by a predefined amount. The interruptions of the temporal change of the value of the inclination angle are generated in particular by the fact that the electrically operable locking device in the form of a brake interrupts the pivoting movement of the pivoting arm or the rotation of the rotatable object table for a very short period of time.

The feedback to the operator is alternatively or additionally preferred acoustically perceptible and is preferred with a buzzer or a comparable acoustic signal generator generated. Through the acoustic signals, the operator learns that the value of the mechanical parameter is a

predefined dimension has been changed or that the respectively set value of the mechanical parameter has been set.

In preferred embodiments of the invention

In the method, the steps of selecting the value of the mechanical parameter to be respectively set are manually performed by the operator. The operator specifies two or more mechanical parameter values to be set, which he preferably inputs via a user interface.

In further preferred embodiments of the

The method according to the invention is to be set in each case

Value of the mechanical parameter selected from several stored values. The stored values are preferably selected during the performance of the inventive method for a previously microscoped sample and thereafter

saved. Thus, the same values of the mechanical parameter are selected for both samples to be microscoped and corresponding microscopic images

recorded, which, for example, an extended evaluation can be made by a comparison. The

stored values preferably represent pre-programming or a presetting to which during the

Performing the procedure is accessed.

In further preferred embodiments of the

According to the method of the invention, the respective value of the mechanical parameter to be set is selected by incrementing the previously selected value of the mechanical parameter by one increment. In embodiments, at the mechanical parameter is formed by the angle of inclination, the sample is characterized in terms of

Inclination angle microscopically. The first value of the mechanical parameter represents a starting value.

More preferably, not only two values of the mechanical parameter but a plurality of the values of the mechanical parameter are selected and set.

Accordingly, the method preferably comprises a step in which another one of the other values of the

mechanical value of the mechanical parameter to be set is selected. In a further step, the electrically controllable drive takes place

Actuating element to set the further value to be set of the mechanical parameter. In another

Step is taking a further microscopic image of the sample below the set further value of the mechanical parameter. The above steps will be

preferably repeated several times for further values of the mechanical parameter.

The inventive method is preferred for

Microscopy of at least two samples formed. In this case, to record the microscopic images of the first sample and to record the microscopic images of the second

Each sample the same first values of the mechanical

Parameters, the same second values of the mechanical

Parameters and possibly also the same further values of the mechanical parameter selected and set. Thus, for the at least two samples, microscopic images will be under the same values of the mechanical parameter

recorded. In further preferred embodiments of the

According to the invention, different magnifications of the image are obtained for taking the first microscopic image of the sample and for taking up the second microscopic image and, if appropriate, for taking the further microscopic images

Lens selected and / or different lenses used. Thus, an adaptation to the spatial extent of the sample can be made or special areas of the sample, for example, with a higher magnification

be microscoped.

A preferred embodiment of the invention

Method comprises a further step in which a 2, 5-dimensional or three-dimensional representation of the sample is generated from the plurality of microscopic images.

Preferably, a plurality of the values of the mechanical parameter preferably formed by the tilt angle are selected and adjusted to produce a corresponding plurality of

take microscopic images and use them to generate the 2, 5-dimensional or 3-dimensional representation of the sample. Since the sample is microscopically examined under different values of the angle of inclination or a sample distance, microscopic images can also

Depth information can be obtained.

When taking the microscopic images of the sample at the various angles of inclination, one or more areas of the sample may be obscured. Therefore, further microscopic images of the sample are preferably taken at further set tilt angles to

to include those areas of the sample that are obscured at different angles of inclination. The microscope according to the invention is used for microscopy of a sample. It preferably comprises an objective for the optical

Imaging the sample. The microscope further preferably comprises a stage for placing the sample. A mechanical parameter on the microscope is changeable. As a mechanical parameter, an inclination angle between the

Objectively and the stage changeable. The microscope also comprises an electrically controllable actuating element for setting a value of the mechanical parameter and a control unit which is used to execute the

inventive method is configured. The

Control unit is preferred for execution preferred

Embodiments of the method according to the invention

configured. The microscope preferably also has features which are specified in connection with the method according to the invention.

Further details and developments of the invention will become apparent from the following description

Embodiments of the invention, with reference to the

Drawing. 1 shows a first preferred embodiment of a microscope according to the invention; a second preferred embodiment of the microscope according to the invention;

FIG. 3 shows the microscope shown in FIG. 1 in two states while carrying out a method according to the invention; FIG. and Fig. 4: The microscope shown in Fig. 2 in two states during the implementation of the method according to the invention.

Fig. 1 shows a first preferred embodiment of a microscope according to the invention. The microscope comprises a base Ol, on which a stage 02 is attached. At the base 01, a pivot arm 03 is further rotatably mounted, which with an electromotive actuator 04 relative to the

Stand 01 can be pivoted, so that it also with respect to the stage 02 and thereon a sample 06 (shown in Fig. 2) tilts. On the pivot arm 03, a lens 07 and an optical module 08 are attached. The

Pivoting the pivot arm 03 can be unlocked via an unlock key 09.

As an alternative to the electromotive actuator 04, an electrically actuated brake 11 (shown in Fig. 2) on

Swivel arm 03 can be used, which pivoting the

Swivel arm 03 relative to the base 01 and the stage 02 can lock by braking. The torque to

Pivoting the swivel arm 03 must therefore be applied by the operator. The electrically actuated brake 11 (shown in FIG. 2) of the swivel arm 03 is controlled such that a further manual swiveling of the swivel arm 03 is no longer possible as soon as a value of the swivel angle to be achieved has been reached. FIG. 2 shows a second preferred embodiment of the microscope according to the invention which, like the embodiment shown in FIG. 1, comprises the stage 02, the objective 07 and the optical module 08. In contrast to that in FIG. 1 shown embodiment, the lens 07 and the optical module 08 are not attached to a pivotable pivot arm, but arranged fixed. Instead, that is

Object table 02 rotatable. The rotational movement of the object table 02 can be braked with the electrically operable brake 11, so that the object table 02 can be fixed with regard to its rotatability. The embodiment shown in FIG. 2 comprises a further objective 12 and a further optical module 13, which are arranged below the stage 02.

Fig. 3 shows the microscope shown in Fig. 1 in two

States during the implementation of a method according to the invention. The left part of Fig. 3 shows the microscope in an initial situation in which the pivot arm 03 is not inclined, so that an inclination angle Θ is equal to zero.

According to the method, a value to be set Θ _ζ ι _θ ι des

Tilt angle Θ selected.

The right part of Fig. 3 shows the microscope in a

Situation, while the value to be set Θ _ζ ι _θ ι of

Inclination angle Θ is set according to the method, which is made possible by pressing the unlock key 09. The

Arm 03 was by driving the electromotive actuator 04 already up to a value 0 _aktue ii of

Tilt angle Θ swiveled. Driving the

electromotive actuator 04 is continued until the value to be set Θ _ζ ι _θ ι of the inclination angle Θ is reached.

Fig. 4 shows the microscope shown in Fig. 2 in two

States during the implementation of the invention

Process. The left part of Fig. 4 shows the microscope in an initial situation in which the stage 02 is not inclined, so that an inclination angle Θ is equal to zero. According to the method, a value to be set Θ _ζ ι _θ ι des

Tilt angle Θ selected.

The right part of Fig. 3 shows the microscope in a

Situation, while the value to be set Θ _ζ ι _θ ι of

Inclination angle Θ is set according to the procedure. The

Object table 02 is rotated manually, wherein the object table 02 has already been rotated to a value 0 _a ktueii of the inclination angle Θ. According to the method, the rotational movement of the object table 02 is measured and a time is predicted at which the value to be set Θ _ζ ι _θ ι of the inclination angle Θ is achieved. At this time, the electric

actuated brake 11 is controlled so that another

Rotary movement of the object table 02 is no longer possible.

LIST OF REFERENCE NUMBERS

01 stand

02 stage

03 swivel arm

04 electromotive actuator 05

06 sample

07 lens

08 optical module

09 unlock key

10

11 electrically actuated brake

12 additional lens

13 additional optical module

Claims

claims

1. A method for microscopy of a sample (06) with a

Microscope, wherein at the microscope a mechanical

Parameter (Θ) is variable, and wherein the method comprises at least the following steps:

Selecting a first value to be set (Θ _ζ ι _θ ι) of the mechanical parameter (Θ);

Driving an electrically controllable

Actuator (04; 11) to the first

to set the value (0 _Z i _e i) of the mechanical parameter (Θ) to be set;

Taking a first microscopic image of the sample (06) below the adjusted first value (0 _Z i _e i) of the mechanical parameter (Θ);

Selecting a second value to be set (0 _Z i _e i) of the mechanical parameter (Θ);

Driving the electrically controllable

Actuating element (04; 11) to the second

to set the value (0 _Z i _e i) of the mechanical parameter (Θ) to be set; and

Taking a second microscopic image of the sample (06) below the adjusted second value (0 _Z i _e i) of the mechanical parameter (Θ).

2. The method according to claim 1, characterized in that it comprises a further step, each after the

Driving the electrically controllable actuating element

(04; 11) and focusing the sample

(06).

3. The method according to claim 1 or 2, characterized in that the actuating element by a with a

Electric motor driven actuator (04) is formed by which the mechanical parameter (Θ) is adjustable.

4. The method according to any one of claims 1 to 3, characterized

characterized in that the mechanical parameter (Θ) is manually changeable and that the actuating element is formed by a locking device (11) with which the mechanical parameter (Θ) can be determined, and wherein the steps of driving the electrically controllable

Actuating element (11) each comprise the following sub-steps:

Measuring a time course of a manual

mechanical parameter (Θ);

Predicting a point in time at which the previously determined time course of the manual change of the value (0 _ak tueii) of the mechanical parameter (Θ) will lead to the respective set value (Θ _ζ ι _θ ι) of the mechanical parameter (Θ); and

electrically activating the electrically operable locking device (11) at the predetermined time.

5. The method according to claim 4, characterized in that the mechanical parameter is formed by an inclination angle (Θ), and in that the step of predicting the time in which the previously determined temporal

Course of manual change of the value (0 _ak tueii) of the inclination angle (Θ) to the value to be set (0 _z iei) of the inclination angle (Θ), the following

Sub-steps includes:

Measuring an output value 0 _S tart of the inclination angle; Determining an angular velocity ω and an angular acceleration from the previously determined

time course of the manual change of the value (0 _a ktueii) of the tilt angle (Θ); and

Predicting the point in time at which the previously determined time course of the manual change in the value (0 _ak tueii) of the inclination angle (Θ) will lead to the respectively to be set value Θ _ζ ι _θ ι of the angle of inclination, starting from the time t

solved equation 0 _Z i _e i = 0 _S tart + ω · t + Ή · · t ² .

6. The method according to claim 4 or 5, characterized in that the step of predicting a time in which the previously determined time course of the manual change of the value (0 _ak tueii) of the mechanical parameter (Θ) to the respective value to be set (Θ _ζ ι _θ ι) of

mechanical parameter (Θ), running

is repeated until the manual change of the value of the mechanical parameter (Θ) to the respective

value to be set (0 _Z i _e i) of the mechanical parameter

(Θ).

7. The method according to any one of claims 1 to 6, characterized

in that the mechanical parameter is defined by an angle of inclination (Θ) between a lens (07) of the

Microscope and a stage (02) of the microscope is formed; wherein the objective (07) is fastened to a pivotable swivel arm (03), or wherein the object table (02) is arranged in an axis parallel to the plane of extent of the Object table (02) is rotatable, whereby the inclination angle (Θ) between the lens (07) the object table (02) is variable.

8. The method according to any one of claims 1 to 7, characterized

characterized in that it comprises the following further step:

Generating feedback to the operator when the value of the mechanical parameter (Θ) changes by

predefined dimension has been changed and / or if the respectively set value (Θ _ζ ι _θ ι) of the mechanical parameter (Θ) has been set.

9. The method according to claim 8, characterized in that the feedback is haptic perceptible and with the

Electromagnetically actuated locking device (11) is generated.

10. The method according to any one of claims 1 to 9, characterized

characterized in that selecting the respective

to be set value (Θ _ζ ι _θ ι) of the mechanical parameter (Θ) is effected in that the previously selected value of the mechanical parameter (Θ) is incremented by one increment.

11. The method according to any one of claims 1 to 10, characterized

characterized in that it comprises the following further steps which are repeated several times:

Selecting another from the other values _(Z 0 iei) of the mechanical parameter (Θ) discriminating value to be set _{_(Θ} ζ ι θ ι) of the mechanical

Parameters (Θ); Driving the electrically controllable

Actuating element (04; 11) to the other

to set the value to be set (Θ _ζ ι _θ ι) of the mechanical parameter (Θ); and

Taking another microscopic image of the

Sample (06) below the set further value (0 _Z i _e i) of the mechanical parameter (Θ).

12. The method according to any one of claims 1 to 11, characterized

characterized in that for receiving the first

microscopic image of the sample (06) and for taking the second microscopic image of the sample (06)

different magnifications of a lens and / or different lenses are selected.

13. The method according to any one of claims 1 to 12, characterized

characterized in that it is designed for the microscopy of at least two samples (06), wherein for taking the microscopic images of the first sample (06) and the

Picking up the microscopic images of the second sample (06) the same first values (0 _Z i _e i) of the mechanical parameter (Θ) and the same second values (0 _Z i _e i) of the mechanical parameter (Θ) are selected and set ,

14. The method according to any one of claims 1 to 13, characterized

characterized in that it comprises the following further step: - generating a 2, 5-dimensional or 3-dimensional

Reproducing the sample (06) from the several

microscopic images. Microscope for microscopy of a sample (06), in which a mechanical parameter (Θ) is variable, comprising: an electrically controllable actuator (04; 11) for setting a value of the mechanical parameter (Θ); and

a control unit, which is used to execute a

Method according to one of claims 1 to 14 is formed.