Aids for removing and removing immersion liquid from a sample chamber of a microscope and methods for their use

Abstract

The invention relates to aids (10, 20, 30) for removing immersion fluid (40) from a sample chamber of a microscope, as well as to methods for using the same and a microscope. Each of the proposed aids (10, 20, 30) comprises a sorbent material (14, 24, 34) with which immersion fluid residues (40) can be absorbed.

Description

[0001] The invention relates to aids for ablating and removing immersion liquid from a sample chamber of a microscope, as well as methods for using the same and a microscope.

[0002] Current practice requires manual wiping of immersion medium residues, for example, with a cloth. This requires human intervention in the microscope system, which is also time-consuming. However, this is not possible or difficult to achieve with fully automated 24 / 7 use of a microscope stand or with a completely enclosed microscope stand.

[0003] Previous automated solutions absorb the remaining immersion fluid at designated locations on the stage, for example, using sponges (see JP2006123844A) or wipes (see WO 2021 / 124282 A1). The residual fluid thus remains within the system and is not deliberately removed from it.

[0004] Another approach is to aspirate immersion fluid from the objective. However, a certain amount of immersion fluid always remains due to the wetting behavior during immersion and can accumulate over time in unwanted, critical areas of the system or be transported to unpredictable locations by the motorized movement of the objective nosepiece.

[0005] The object of the invention is therefore, among other things, to propose solutions for removing immersion liquid from the sample space, both for cleaning an objective and the sample, for example for further investigations.

[0006] The object of the invention is achieved with the aids, the microscope, and the methods according to the independent claims. Further preferred embodiments of the invention emerge from the remaining features mentioned in the subclaims.

[0007] The following introductory explanations apply equally to the aids, the microscope and the methods according to the invention.

[0008] The disclosed aids are intended to be used to remove immersion fluid from a sample chamber of a microscope, and preferably from the microscope itself. This should be done as completely as possible; in particular, at least 95 percent of the remaining immersion fluid in the sample chamber of the microscope should be removed.

[0009] The sample chamber refers to the immediate environment of the sample during an examination. Depending on the chosen tool and / or method, the immersion fluid is removed from a wetted objective lens and / or from a wetted sample or sample carrier. By removing the immersion fluid from the objective lens or sample, the sample can be used for further examinations without the need for manual cleaning or replacement.

[0010] The aids, methods, and microscope according to the invention encompass both inverted and upright microscopes. The method can be manual or motorized, in particular automated. A prerequisite for an automated method is a motorized sample stage that can be moved in the x- and y-directions. Depending on the design, the sample stage should also be movable manually or by motor in the z-direction, and / or a motorized focus drive of the objective should be present to move the objective in the z-direction. The control of the sample stage and, if applicable, the focus drive of the microscope is carried out, as is customary in the art, by appropriately controlling the microscope through the implementation of suitable methods.

[0011] A first aid according to the invention for removing and ablating immersion liquid from a sample chamber of a microscope is formed with a first element with a sorbing material, which is to be inserted into a sample stage of the microscope or placed on the latter and is to be aligned such that by moving an immersion objective with an immersion agent wetting towards the first element and / or by moving the sample stage in the direction of the immersion objective with the immersion agent wetting, the sorbing material of the first element comes into contact with the immersion agent wetting and removes it.

[0012] The first aid therefore comprises a first element with a sorbent material, i.e., a material capable of absorbing immersion medium upon contact with it. "Sorption" encompasses both the processes of absorption and adsorption.

[0013] If the sorbing material is sufficiently inherently stable or dimensionally stable, it can form the first aid alone or at most surrounded by a frame, i.e. in particular without a carrier material or a base for the sorbing material.

[0014] The above-described first element should be shaped and dimensioned such that it can be inserted into or placed on a sample stage of a microscope. It should be inserted into the designated receptacles or placed on the designated storage surfaces. In the simplest case, this is achieved by the first element corresponding in shape and size to a sample or sample carrier. However, deviations from this are expressly possible and within the scope of the invention.

[0015] The first aid device should also be aligned when inserting and / or placing it on the microscope so that contact can be achieved between the objective of the microscope and the first element. This should be achieved by moving the immersion objective, i.e. the objective wetted with an immersion medium, towards the first element using a motorized focus drive and / or by moving the sample stage in the x, y and / or z direction with the first aid device towards the immersion medium objective. When the first element of the first aid comes into contact with the immersion medium, the latter is absorbed by capillary forces and thus removed from the wetted surface of the immersion objective. It can now be removed from the sample chamber and from the microscope. This can be done manually or automatically, for example using a robotic arm.

[0016] The distance from the microscope is particularly important when the application is in an enclosed or closed microscopy system, because this is the only way to ensure that the absorbed immersion liquid does not evaporate and thus unintentionally influence the relative humidity in such an enclosed system.

[0017] The insertion or placement of the first aid and its removal can be done manually and / or automatically, for example using a robotic arm.

[0018] By using a first aid device that is formed over its entire surface with a sorbent material or where the first element or the sorbent material alone forms the first aid device (hereinafter also referred to as “direct application”), a particularly high absorption capacity of the first aid device is achieved and it can be used for a service mode of the microscope in which immersion liquid is to be drained from the microscope, for example for use during the so-called “priming” process. During priming, an increased volume of immersion liquid is transported through the hose towards the sample chamber (for example 500 µl per priming process) in order to flush air pockets out of the hose of the immersion module. The formation of air pockets is due to the permeability of the hose material and occurs particularly after extended periods of inactivity and / or elevated temperatures.This requires repeated use of the priming process, for example, as a service function. The aid can be used for at least one priming process. If only water was used as the immersion fluid, the aid can be used multiple times after successful drying.

[0019] If the first element lacks inherent stability, the first aid can be formed according to a first embodiment with a dimensionally stable carrier material, a sample carrier, or a sample vessel, and is inserted into the sample table or placed on it by means of the same. This is also referred to below as indirect application.

[0020] The sorbent material of the first element is arranged on a dimensionally stable support material, such as a plastic plate or on a sample carrier. The use of a sample carrier eliminates the need for a separate support material, as these are usually available and selected to suit the microscope used. Such a sample carrier can be one-piece or multi-piece, as is known from so-called insert frames for microscope stages. The first material can also be arranged in and / or on a sample vessel, or underneath it in the case of inverted microscopes.

[0021] By using a dimensionally stable base for the first element, variable sorbent materials can be used which alone are not sufficiently stiff to fulfil the previously described function of the first aid.

[0022] According to the invention, a second aid for removing immersion liquid from a sample chamber of a microscope is also provided, comprising a second element with a sorbing material which is to be introduced into an objective changing device and is to be aligned such that by moving the second element towards a sample carrier with an immersion agent wetting and / or by moving the sample stage in the direction of the second element with sorbing material, the latter comes into contact with the immersion agent wetting of the sample carrier and removes it.

[0023] The second aid is therefore not used instead of or with a sample to clean an objective, but rather it is inserted into an objective changing device instead of an objective in order to clean a sample wetted with an immersion agent or to remove the immersion agent. For this purpose, the second aid, which is formed with the second element, is inserted manually or automatically into the objective changing device. As already explained with reference to the first element, the second element can, if it has sufficient inherent stability and a suitable shape and size, be formed solely from or with the sorbing material, or it can, in addition to the sorbing material, also have a carrier, a frame or the like, by means of which it can be inserted into the objective changing device and fixed in place.

[0024] The immersion agent wetting is removed in a similar manner to the first aid: the second element is moved toward the sample carrier or sample, and / or the sample stage is moved toward the second aid until the sorbent material of the second aid and the immersion agent-wetted surface of the sample touch each other. Capillary forces then cause the immersion agent to be absorbed into the sorbent material, removing it from the wetted surface. The used aid can then be removed from the microscope or microscope system.

[0025] A third aid for removing immersion liquid from a sample chamber of a microscope is formed by a third element with a sorbing material that is arranged on a stationary component of the microscope and aligned so that • by moving an immersion objective with an immersion agent wetting to the third element and / or • by moving a sample table with a sample carrier with immersion agent wetting to the third element, the sorbing material of the third element comes into contact with the immersion agent wetting and removes it.

[0026] Such a third aid can be used alternatively or in addition to the first and / or second aid. For this purpose, it can be attached immovably to or in the microscope, preferably to its stand, by means of a holder such as a rod, in an area outside the observation position of the sample. This position must, however, be selected such that the sample stage and the wetted sample therein and / or the wetted immersion objective can reach it in such a way that contact can be achieved between the sorbing material of the third aid and the wetted surface. With regard to the objective, this can be a position in which the objective, optionally by adjustment using a focus drive, "slides" over the sorbing material of the third element when changing and is cleaned in the process. A similar situation is provided for a sample wetted with immersion medium.

[0027] As with the first and second aids, the third aid comprises a third element formed from a sorbent material and optionally with a frame, a mount, or the like. As in the first case, this depends, among other things, on the shape, size, and inherent stability of the sorbent material. The third aid, or at least the sorbent material of the third element, should be detachably attached to it so that it can be removed from the sample chamber. Reversible closure techniques such as form-fitting fasteners, hook-and-loop fasteners, or magnetic fastenings are possible for this purpose. Removal can be carried out manually by a user or automatically using a robotic arm.

[0028] The embodiments explained below can apply to each of the three aids according to the invention. If several of these aids are provided in a microscope or microscope system, they can apply to one, two, or all three of the aids.

[0029] For example, the sorbent material can be fixed in position on or in its element magnetically, adhesively, or by form-fitting. If a first aid device is formed with a sample carrier or a sample vessel, it can be useful for the aid to be adhesively attached to it. In this way, it can point downwards in the use position and be used to clean an objective in an inverted microscope. A magnetic attachment can work in the same way, for example by one or more magnets on the sorbent material and on or in its element. A magnetic or adhesive attachment also serves as protection against loss. In the case of an adhesive attachment, the sorbent material in the respective aid can be self-adhesive in order to achieve the adhesive attachment, whereby the use of additional adhesives can be avoided.

[0030] With a form-fitting attachment of the sorbent material in or on its respective element, the sorbent material is shaped in such a way that it fits into a corresponding receptacle, which can also be formed by the edge of a sample carrier, within its element. As a result of the shape, it is fixed in position and aligned. This facilitates the use of such a device and thus also the replacement of the sorbent material within a device.

[0031] The sorbent material of the aids can be a sponge, a cloth, in particular a cloth made of cotton, bamboo or bamboo viscose, a polyester fleece, a polypropylene fleece, hemp, wool, or clay. The selection of the appropriate material depends on the immersion medium to be used and the amount to be absorbed, and may also depend on the available space for the aid. For example, a small footprint of the sorbent material may require a greater thickness to absorb the desired amount of immersion medium. In such a case, a sponge would be a suitable sorbent material. For example, if a first aid is formed with a carrier over whose entire surface a sorbent material can be applied, a cloth, in particular a cotton cloth, may be a suitable choice.

[0032] Natural materials such as bamboo, bamboo viscose, hemp, or wool can absorb large amounts of moisture relative to their specific weight and are also readily available. Clay is a material that can absorb liquids and release them again when dried. Clay also has antibacterial properties and can be shaped or modified. Nonwovens such as polyester or polypropylene nonwovens are particularly suitable for absorbing oily liquids.

[0033] If water is used as the immersion medium, the respective sorbent materials can be dried and reused.

[0034] In one embodiment, the sorbent material can be formed in layers in a stack, the layers of which can be removed individually. This applies in particular to flat sorbent materials such as paper or wipes. The individual layers of such a sorbent material are arranged in a stack and can be connected to one another, for example with an adhesive edge in a stack of paper layers. After use, i.e. after absorbing immersion agent to clean a wetted surface, one or more of the moistened layers can be removed if necessary. Several layers can also be removed simultaneously if desired or necessary. In the aforementioned example with paper and an adhesive edge, one or more layers can be retracted like a pad of paper.

[0035] A further embodiment of one or more of the aids according to the invention provides that the shape and / or size of the element and in particular of the sorbing material is adapted to the shape and / or size of the immersion objective, a sample vessel and / or the sample carrier, in particular to standardized sample vessels and / or standardized sample vessels.

[0036] The first, the second and / or the third element and especially the respective sorbing material should therefore be adapted in terms of shape and / or size to the immersion objective, the sample vessel and / or the sample carrier, i.e. the object to be cleaned or freed of immersion agent.

[0037] A first element as a component of a first aid for cleaning an immersion objective should therefore be adapted in shape and size to the shape and size of the immersion objective or the surface to be wetted. This allows for almost full-surface contact between the immersion objective and the sorbent material of the first aid, allowing the immersion agent to be almost completely removed.

[0038] For a second device for absorbing the immersion medium from a sample carrier, the second element or the sorbent material therein would have to be adapted to the latter in terms of shape and / or size. For example, with a rectangular sample carrier, it is advisable to use a rectangular element or a rectangular sorbent material to absorb immersion medium in corner areas as well. For round sample vessels, a round element or a round-shaped sorbent material may be more suitable for reaching the edge areas as well.

[0039] Ideally, the shape and / or size of the aid or the sorbent material are adapted to standardized sample vessels and / or sample carriers in order to enable quick and easy use by a user when using standardized vessels or sample carriers, such as Petri dishes or well plates.

[0040] Especially if the first aid is used without its own support but with sufficient inherent stability, it is helpful if it has standardized dimensions and / or a standardized size, so that it can be placed on or inserted into a sample table without additional effort, for example, to be used as an immersion medium holder for a priming process. This also prevents the immersion medium from running down the sides of the aid.

[0041] According to a further embodiment, the absorbent material of an aid can be loaded with a solvent to increase its absorption capacity for immersion media. For this purpose, it is loaded or impregnated with a solvent such as isopropanol or ethanol during its manufacture or before being placed in its intended use position, so that upon contact with the immersion media to be absorbed, its surface tension is reduced, making it easier for the aid to absorb it.

[0042] If the sorbent material is loaded with the solvent during its manufacture, it should subsequently be sealed or packaged, for example, with a film, to keep volatile solvents contained in the sorbent material until use. Once the tool is ready to be used, it can be unpacked manually by a user or, in the case of an automated microscope or microscope system, by a dedicated robot.

[0043] Alternatively, in an arrangement in which a second or third device with sorbent material is arranged below a sample vessel, a solvent can be applied to the sorbent material using a suitably prepared sample vessel. For this purpose, a sample vessel can, for example, have fine holes through which the solvent can escape downwards. A lid can be provided on the sample vessel to prevent the solvent from evaporating. Preferably, the sample vessel is filled only after it has reached its position above the sorbent material and, if necessary, also covered, for example, using the aforementioned robot arm.

[0044] Advantageously, an aid can be further developed such that the associated element has an indicator for absorbed liquid, which is arranged at least partially in contact with the sorbent material. Such indicators are known, for example, as humidity indicators. They are usually formed from a paper or cardboard strip loaded with a humidity indicator, such as cobalt dibromide or cobalt dichloride. If the indicator becomes moist or wet, a color change occurs, so that the color of the indicator can be recognized that it has come into contact with moisture.This can be applied similarly to the sorbent material of one or more of the disclosed aids: by placing an indicator in contact with the sorbent material, at least in part, it comes into contact with the immersion medium absorbed there and indicates this by a color change. The use of colored paper that darkens in this area upon contact with moisture, as can be observed, for example, with a blotting paper moistened with water, can also implement such an indicator.

[0045] Such an indicator can advantageously be reversible, meaning it can return to its original state when dried and can then be reused.

[0046] Preferably, the indicator can be arranged or attached as a fully or partially circumferential strip on the edge of the sorbent material. It can also be arranged on a side of the aid or the sorbent material that is opposite or facing away from the contact surface with the immersion medium. In both cases, this ensures that the indicator only comes into contact with the immersion medium when the sorbent material has absorbed a sufficient amount of immersion medium.

[0047] By positioning the indicator at a sufficient distance from the initial wetting point of the sorbent material when absorbing the immersion medium, it can be ensured that the indicator only indicates contact with the immersion liquid once the sorbent material has absorbed a defined amount of immersion medium. This is naturally also influenced by the type of sorbent material, its size, and shape, so the indicator's placement must be adapted accordingly.

[0048] If an indicator in the form of a strip is placed on or in the direction of propagation of the absorbed immersion medium, the position or fill level of the sorbent material can be roughly estimated by the recolored area of ​​the strip.

[0049] If the indicator is loaded with an indicator for the absorbed moisture, the time of the color change can be made dependent on the fill level by the selection and quantity of the indicator, so that a color change only occurs when a predetermined moisture level is reached.

[0050] Detecting a liquid volume aspirated and / or a specified liquid volume is particularly relevant when this should or must be verified, for example, during a priming procedure, where it must be ensured that a sufficient amount of immersion medium has been flushed out to flush bubbles and the like from the immersion system, especially the supply tube from the pump to the tip. A safety factor can also be provided for this purpose, further increasing the target liquid volume to be achieved.

[0051] The detection itself can be carried out by the user, but it can also be done (especially in automated systems) by means of a camera that captures the sample area and whose images are analyzed with regard to the color change.

[0052] The invention also provides a microscope equipped with a device for automatically applying a first element containing sorbent material to a sample stage and removing it therefrom. Such a microscope can primarily be an automated device and, in particular, an enclosed microscope.

[0053] The device can, in particular, be a robotic arm that is controlled by a user or automatically in such a way that it picks up the first element from a position where it is stored or provided, moves it to the sample stage, and removes it from there after removing the immersion agent wetting. The used first element can be moved to a position where it is easily removed by a user, or to an output position that can be separated from the sample chamber so that the absorbed moisture does not spread uncontrollably inside the microscope during drying.

[0054] Such a device can also be advantageously designed to peel off individual layers of the sorbent material and remove them from the sample chamber if it is layered. If the sorbent material is sealed with a film or similar material to prevent solvent evaporation, the device should also be able to peel this off.

[0055] The aforementioned microscope may comprise or interact with a camera for capturing the sample area and an indicator for displaying the absorbed moisture.

[0056] Such a camera should capture the sample area and, above all, detect the indicator described above. The images should then be evaluated to determine whether and to what extent immersion medium has been absorbed and indicated by the indicator. This allows us to determine whether any immersion medium has been absorbed, and if so, whether a predefined or at least sufficient amount has been absorbed.

[0057] The camera can be the microscope's own camera, either an overview camera or a camera specifically designed for examining the sample. The camera can also be any external camera capable of capturing the area of ​​interest with sufficient accuracy, such as a webcam. The microscope can therefore incorporate the camera or interact with it, with images captured by a separate camera being transmitted to the microscope and evaluated by a computer on the microscope.

[0058] To improve the imaging conditions for the camera, an illumination device can be provided, either integrated with the microscope or in conjunction with the microscope. This is intended to better illuminate the area around the sample and / or minimize the effect of stray light, which could otherwise distort the image.

[0059] According to the invention, a method for removing immersion liquid from a sample chamber of a microscope using a first aid device is also provided, comprising the following steps: • Inserting or placing a first aid in or on a sample table of the microscope, • Moving the immersion objective and / or the sample stage in the z-direction until the sorbing material and the immersion objective come into contact and the immersion medium is absorbed, • Moving the sample table to an output position, and • Remove this first aid.

[0060] For this procedure, the first aid is used as a standalone aid and not as or with a sample. Its sole purpose is to absorb the immersion medium and thus clean the objective or perform the priming process described above. The procedure can be performed manually by a user or automated.

[0061] For this purpose, a first aid, preferably one formed exclusively with a first element containing sorbent material as described above, is inserted into or placed on a sample stage. The (used) immersion objective and / or the sample stage are then moved in the z-direction until the immersion objective and the first aid come into contact, so that the immersion medium is absorbed by the sorbent material. The immersion objective can be moved in the z-direction using the focus drive.

[0062] If the sorbent material is designed with a surface area larger than that of the surface to be cleaned, alignment of the immersion objective and the first auxiliary device in the x- and y-directions may be omitted. Otherwise, this can be performed as an additional optional step before moving the immersion objective and / or sample stage in the z-direction.

[0063] Once the immersion medium has been absorbed, the sample stage is moved to a position where the first aid device used can be removed manually or automatically, or where, in the case of a stacked version of the sorbent material, one or more layers can be removed. Meanwhile, the immersion objective can be moved to a suitable position or even replaced.

[0064] In another method for removing and ablating immersion liquid from a sample chamber of a microscope using at least one of the disclosed aids, the following steps are provided: • Inserting or placing a sample carrier or a sample container with a sample in or on a sample stage of the microscope, and • Examine the sample with an immersion objective, and optionally o.1) if the sample carrier or sample vessel is part of a first aid device with a sample carrier or sample vessel: o.2) Positioning the first aid by moving the sample table in the x-direction and y-direction in relation to the immersion objective so that the sorbing material and the immersion objective are aligned one above the other, and o.3) Moving the immersion objective and / or the sample stage in the z-direction until the sorbing material and the immersion objective come into contact and the immersion medium is absorbed, and / or p.1) Changing from an immersion objective to a second aid according to one of the preceding claims by means of an objective changing device, p.2) Positioning the sample carrier or sample vessel by moving the sample table in the x-direction and y-direction so that the sorbent material of the second aid and the immersion medium wetting of the sample carrier or sample vessel are aligned with each other, p.3) Moving the second tool and / or the sample stage in the z-direction until the sorbent material and the immersion agent wetting come into contact and are removed.

[0065] This method counts towards the remaining cleaning tasks, where the first element of the first aid does not only comprise the sorbent material, but can also comprise a carrier material, a sample carrier, or a sample vessel (steps o.1 to o.3), but alternatively or additionally also the cleaning of the sample with a second aid (steps p.1 to p.3).

[0066] First, a sample carrier or sample container is placed on a microscope stage and examined. It goes without saying from the context that this is also done using an immersion medium, since otherwise subsequent cleaning would be unnecessary.

[0067] Afterwards, the sequence of steps o.1 to o.3 can be followed to clean the objective and / or the sequence of steps p.1 to p.3 to clean the sample. This can be selected depending on the subsequent planned examinations.

[0068] As already indicated, cleaning of the immersion objective can be performed according to this procedure if the sample carrier or sample vessel is part of a first auxiliary device or its first element. Otherwise, the previously described procedure could be used instead. Thus, step o.1 is more of a requirement than a procedural step.

[0069] If this condition is met, the first aid is moved relative to the immersion objective by moving the sample stage in the x-direction and y-direction, so that the sorbing material of the first aid and the immersion objective are approximately aligned (step o.2). In step o.3, the sample stage and / or the immersion objective are moved in the z-direction until contact is established between the sorbing material and the immersion objective, thereby absorbing the immersion medium present on it. Steps o.2 and o.3 can be performed sequentially or simultaneously.

[0070] Alternatively or additionally, the sample can be cleaned, for example because it is to be reused and examined with a different objective. To do this, in step p.1, the immersion objective is exchanged using an objective changing device, and a second aid is inserted or pivoted into the beam path. For this purpose, it was previously inserted into the objective changing device (slider, turret). The sample carrier or sample vessel is then aligned with respect to the second aid by moving the sample stage in the x- and y-directions such that the sorbent material of the second aid and the immersion medium wetting of the sample carrier or sample vessel are approximately aligned one above the other (step p.2).Thereafter or simultaneously, the second aid and / or the sample table is moved in the z-direction until a contact is established between the sorbing material of the second aid and the immersion agent wetting, through which the immersion agent is absorbed (step p.3).

[0071] Yet another method is used to ablate and remove immersion fluid from a sample chamber of a microscope using a third aid, and is carried out with the following steps: • Moving an immersion objective with an immersion agent wetting to the third element and / or • Moving a sample table with a sample carrier with immersion agent wetting to the third element, so that the sorbent material of the third element comes into contact with the immersion agent wetting and removes it.

[0072] The third aid is, as explained above, formed with a third element with a sorbing material, which is arranged on a stationary component of the microscope and is aligned in such a way that an immersion objective and / or a sample stage can move towards the third aid and the sorbing material of the third aid can be brought into contact with the immersion agent wetting of the immersion objective or the sample carrier in the sample stage, so that the immersion agent is absorbed by the sorbing material.

[0073] The associated procedure involves moving the immersion objective and / or sample stage toward the third device, where the contact of the sorbent material with the immersion agent wetting agent occurs. This movement can be done manually or automatically.

[0074] With respect to an immersion objective, the third aid can be arranged at a position on the microscope where one objective is moved over the third aid when changing to another, creating a wiping motion. This movement can also include an intermediate stop, during which the immersion objective remains in a position for a defined time, ensuring contact between the front lens of the immersion objective and the sorbent material of the third aid in order to absorb a larger amount of the immersion medium.

[0075] The same applies to the sample stage. This can be moved by moving the sample stage in the x- and y-directions so that the sample carrier with immersion agent wetting and the sorbent material of the third aid are aligned, and then moved in the z-direction until contact is established, through which the immersion agent is absorbed. This can be combined with a brief stop at the position of the sample stage, as described previously.

[0076] In order to remove the moisture or the aid(s) used from the sample chamber, one or more of the following steps can be carried out after the immersion medium has been absorbed, depending on the design of the process. • the sample table is moved to a dispensing position in step o.4 and the first aid is removed, or • the position of the second aid is changed in a step p.4 by means of the lens changing device to an output position where the second aid is removed, or • if the sorbent material is formed in layers in a stack, one or more layers of the sorbent material are removed, or • if the immersion medium was water, the sorbent material of the aid dries.

[0077] For the process or process steps in which the sample carrier or sample container is part of the first tool (process steps o.1 to o.3), the sample table can be moved manually or automatically to an output position in step o.4, where the first tool with the sample can be removed manually or automatically and, if necessary, a new sample or a new first tool can be inserted. Automated removal can be performed, for example, using a robotic arm.

[0078] When changing from an immersion objective to a second auxiliary device (process steps p.1 to p.3), in step p.4, the second auxiliary device is to be moved to an output position using the objective changing device. It is thus moved from the position in which cleaning was performed and placed in a position where the second auxiliary device can be removed manually or automatically.

[0079] In both cases, an dispensing position is a location where a user can access the respective aid without any modifications or dismantling, i.e., without significant effort. In particular, the dispensing position can be located outside the actual sample room where the sample is observed.

[0080] If the sorbent material is stacked in layers, one or more layers should be removed, which are now wetted and saturated with the immersion medium. This can also be done manually or automatically, as already explained. Automated, this can again be done using a robot arm. For manual removal, it is recommended to move the respective tool to a position where the user can remove one or more layers of the sorbent material. This can be the dispensing position described above.

[0081] In cases where the immersion medium was water, and preferably no further investigations are immediately carried out that require the use of immersion medium and cleaning of the wetted surface(s), the respective aid (first, second, and / or third aid), especially in an unenclosed microscope or microscopy system, can dry over time and then be reused. In enclosed systems, the aid can be moved out of the sample chamber, for example, into the dispensing position, and dried there. Drying outside the microscope is also covered by this design.

[0082] The methods described above can additionally provide that during contact between the immersion objective and the sorbent material of the first aid, the sorbent material of the second aid with the sample carrier, and / or the sorbent material of the third aid with the sample carrier, the sample stage is moved in the x-direction and / or y-direction to generate a wiping motion. Such a wiping motion can absorb further immersion agent residues, especially if the wetted area is larger than the contact area between the sorbent material of the aid and the surface to be cleaned.

[0083] The aids, microscope, and method according to the invention allow for the controlled removal of immersion medium from the sample chamber, but above all, the surfaces wetted with it can be dried or cleaned, allowing, for example, further examination of the sample. For this purpose, the respective aid is inserted and brought into contact with the immersion medium coating, so that it is absorbed. The optional configurations allow adaptation to the existing conditions of the microscope or further optimization.

[0084] The various embodiments of the invention mentioned in this application can be advantageously combined with one another, unless otherwise stated in the individual case.

[0085] The invention is explained below in exemplary embodiments with reference to the accompanying drawings. They show: Fig. 1 a first example of a first aid in a schematic diagram of the application situation, Fig. 2 a second example of a first aid in a schematic diagram of the application situation, Fig. 3 a third example of a first aid in a schematic diagram of the application situation, Fig. 4 and Fig. 5 examples of a first aid in plan view and elevation, Fig. 6 an example of a second aid in a schematic diagram of the application situation, and Fig. 7 an example of a third aid in a schematic diagram of the application situation.

[0086] The Fig. 1, Fig. 2 and Fig. 3 (a and b respectively) show an exemplary embodiment of the first aid. First, Fig. 1a are explained in detail, for the Fig. 1b, Fig. 2a and Fig. 2b as well as 3a and 3b primarily highlight the differences.

[0087] In Fig. Figure 1a shows a highly simplified and sketched illustration of a first aid device 10 for removing immersion fluid 40 from the specimen chamber of a microscope. The device is shown in an inverted microscope, but this is not intended to be limiting. The immersion objective 42 is mounted on a lens turret 44.

[0088] The first aid device 10 is formed with a first element 12, which in this exemplary embodiment is a sample carrier 52, and a sorbent material 14 attached thereto. The sample carrier 52 is, in particular, a sample carrier 52 in a standard shape and size, and is inserted into the sample table 50. The sorbent material 14 is arranged such that a sample 56 can be inserted into the sample carrier 52 and viewed.

[0089] Nevertheless, the arrangement of the sorbing material 14 on the first element 12 or sample carrier 52 is such that, within the scope of the displaceability of the sample stage 50 and, if applicable, the objective 42, an alignment of the sorbing material 14 of the first auxiliary device 10 and the immersion medium residue 40 on the immersion objective 42 is possible one above the other. The mobility of the sample stage 50 is indicated by the arrows drawn to the right.

[0090] If the sample stage 50 and thus the sorbent material 14 of the first auxiliary device 10 and the immersion objective 52 with the immersion agent residue 40 are aligned one above the other, one or both of the aforementioned components can be moved toward each other in the z-direction, so that contact is established between the sorbent material 14 and the immersion agent residue 40, which is absorbed. Optionally, the sample stage can be moved in the x-direction and / or y-direction, resulting in a wiping motion that improves the cleaning effect on the immersion objective 42.

[0091] The alignment of sample table 50 with first auxiliary means 10 and immersion objective 42 with immersion medium residue 40 one above the other can be done manually, but also motor-driven and in particular automated.

[0092] In this case, the sorbent material 14 is to be adhesively attached, i.e., glued, to the first element 12. In this embodiment, it is a sponge, which can optionally be moistened with a solvent such as isopropanol to increase the absorption capacity of the sorbent material.

[0093] The advantage of such a first aid is that the sample 56 does not have to be removed from the sample chamber to clean the immersion objective 42.

[0094] According to an embodiment of the first aid according to the invention, an indicator 16 can be attached to the sorbing material 14, as shown in Fig. 1b (sectional view). The indicator 16 here is a paper strip loaded with cobalt dichloride and applied circumferentially around the sorbent material 14 of the first aid device 10. Using an overview camera (not shown), which captures the contact area between the first aid device 10 and the immersion objective 42, it is possible to monitor when the color change of the indicator 16 indicates the absorption of the immersion liquid 40. The indicator strip 16 can also be arranged only at the edge near the sample carrier, so that it is only moistened and indicates this when the sorbent material 14 itself is sufficiently moistened, and not immediately upon contact with the immersion liquid 40 on the immersion objective 42.

[0095] Fig. 2a and Fig. Figure 2b shows an embodiment in which a sample carrier 52 acts as the first element 12 of the first auxiliary device 10, but is not used with a sample 56. Rather, the sorbent material 14 of this first auxiliary device 10 is arranged centrally on the sample carrier 52. This can be advantageous if the range of motion of the sample stage 50 is limited. In this case, a sample carrier with a sample must be removed from the sample chamber and replaced with this first auxiliary device 10 in order to clean the immersion objective 42.

[0096] Fig. 2b shows the same structure as Fig. 2a, but supplemented by an indicator 16, which shows the absorption of the immersion liquid into the sorbing material 14. The same applies here as already stated with regard to Fig. 1b What has been said.

[0097] In the Fig. 3a and Fig. 3b shows a first aid 10 in which the sorbing material 14 has sufficient inherent stability to also be the first element 12 without any additional carrier material. The first aid 10 in Fig. 3a and Fig. 3b is therefore formed solely by the sorbent material 14 and is referred to by the inventor as a “priming slide” since it can be used in particular in the process of flushing the immersion device, as already explained above, in which contaminants and / or air bubbles are to be flushed out and which is referred to by the inventor as “priming”.

[0098] The sorbing material 14 is designed in a shape and size corresponding to those of a standardized sample carrier, so that it can be used without additional effort.

[0099] Exemplary embodiments of a first aid 10 according to Fig. 3a and Fig. 3b are the Fig. 4a, Fig. 4b and Fig. 4c, where a top view is shown on the left and a side view on the right. It can be seen that a first aid device 10, in which the first element 12 is formed entirely with the sorbing material 14, can have the form of a sample carrier 52 ( Fig. 4a), in which case it can be called a “priming slide.” According to Fig. 4b, it can have the shape and size of a sample vessel 54 such as a Petri dish, and would then also be called a “priming dish”. Fig. Figure 4c is intended to show the first aid 10 in the shape and size of a microtiter plate (also known as a well plate), which could be called a “priming plate”.

[0100] To use a first aid 10 formed in this way, a sample carrier or a sample vessel is removed from the sample chamber of the microscope, and the first aid 10 is inserted into or placed on the sample stage 50. The first aid is then moved in the x-direction and y-direction so that it is aligned as centrally as possible with respect to the immersion objective 42. This would not be absolutely necessary in the present embodiment due to the size of the first aid 10 and the sorbent material 14 provided thereby. By bringing the sample stage 15 and the immersion objective 4 closer together, contact is created between the sorbent material 14 and the immersion agent residue 40 on the immersion objective 42, so that the latter is absorbed. Here, too, a wiping motion by moving the sample stage 50 in the x-direction and / or y-direction can improve the cleaning result.

[0101] In Fig. 3b, an indicator 16 is also arranged on the side of the first aid 10 facing away from the immersion lens 42. This is only moistened with immersion agent 40 once a larger amount of it has already been absorbed by the sorbing material 14. This makes it possible to estimate whether contact for absorbing the immersion agent 40 has occurred and has lasted for a sufficient length of time. If the indicator 16 is designed to indicate the absorption of a predefined amount of liquid or to indicate the amount of liquid absorbed, this can be identified by its color. A user can check this visually themselves or capture it using an overview camera and have it displayed on a display device.

[0102] After cleaning the immersion objective 40 with a first aid 10 according to the embodiments of the figures described above, this first aid 10 can be moved to an output position of the microscope (not shown), where it can be easily removed. This output position can of course also act as an input position through which the first aid 10 can be picked up and moved to the sample chamber. The removed first aid 10 can, if the immersion medium 40 was water, be dried and reused at a later time. If there is no need for further use and the microscope is not being used for any other purpose, a first aid 10 can also remain in place and dry, particularly in an open, unenclosed system, if the immersion medium 40 was water.

[0103] Fig. 5a, Fig. 5b and Fig. Figure 5c shows embodiments of first aids 10, in which a carrier material 18 forms the first element 12 and on which the sorbent material 14 is attached or into which it can be inserted (not shown) so that it is held by a form fit. The designation and use can be analogous to those in Fig. 4a, Fig. 4b and Fig. 4c.

[0104] The embodiment in which the sorbent material 14 is arranged on a carrier material 18 as the first element 12 is particularly suitable when a sorbent material 14 without sufficient inherent stability is to be used, such as paper, a cloth, or the like. This embodiment is also suitable for a sorbent material 14 that is stacked in layers and is to be removed in single or multiple layers, as may be the case with cloths or paper.

[0105] Fig. Figure 6 shows a second aid 20 according to the present invention. This aid, comprising a sorbent material 24 and a second element 22 that receives and holds the sorbent material 24, is inserted into an objective changing device 44, such as a lens turret, and can be moved within the scope of the latter's mobility and the available travel of the focus drive.

[0106] The second aid 20 can be used alone to free a sample 56 or a sample carrier 52 from any immersion medium residue 40 or immersion medium wetting. For this purpose, the sample carrier 52 or sample 56 are aligned one above the other by moving the sample stage 50 and / or the second aid 20. Thereafter, or simultaneously, an approach in the z-direction can occur until there is contact between the immersion medium 40 and the sample 56 or the sample carrier 52, through which the immersion medium 40 can be absorbed. The objective changing device can then be moved so that the second aid 20 can be removed. Here, too, it can be dried and reused if water is used as the immersion medium 40.

[0107] In particular, a combined application of the first aid 10 and the second aid 20 can be carried out, as described in this Fig. 6 shows. The first tool 10 there corresponds to the one in Fig. 1a, therefore, the above statement applies. Preferably, after observing the sample 56, the first tool 10 should first be used to clean the immersion lens (not shown). This is then swung out, and the second tool 20 is swung in and used as explained above. As a result, both the sample and the immersion lens can be reused thereafter.

[0108] Fig. Figure 7 shows a third tool 30, which is arranged on a stationary component 36 of the microscope, for example, its stand, by means of a holder forming the third element 32. A sorbent material 34 is arranged at the free end of the third tool 32. The third tool can be used alone or in combination with one or both of the other tools 10 or 20. This can be decided at the discretion of the user or based on the available movement paths of the immersion objective 42 and / or sample stage 50. Fig. 7 shows a combined application with a first aid as in Fig. 1a.

[0109] After examining a sample 56, the sample stage 50 can now be moved so that it is positioned above the sorbent material 14 of the third auxiliary device 30 and then lowered onto it until contact is established, which absorbs the immersion medium 40. Alternatively or additionally, the immersion objective 42 can be guided past the third auxiliary device 30 during pivoting by means of the objective changing device 44 such that it comes into contact with the sorbent material 34 and is thus cleaned. This requires that the third auxiliary device is positioned at a suitable location along the movement path of the immersion objective 42.

[0110] For the second and third aids 20, 30, the same applies as for the first aid 10, as far as optional configurations such as indicators, stacked arrangement of sorbing material, etc. List of reference symbols 10 first aid 12 first element 14 sorbent material 16 Indicator 18 Carrier material 20 second aid 22 second element 24 sorbent material 30 third aid 32 third element 34 sorbing material 36 immovable components 40 immersion agents 42 Immersion lens 44 Lens changing device 50 sample table 52 sample carriers 54 sample vessels 56 Sample 60 100 microscope QUOTES CONTAINED IN THE DESCRIPTION

[0000] This list of documents submitted by the applicant was generated automatically and is included solely for the convenience of the reader. This list is not part of the German patent or utility model application. The DPMA assumes no liability for any errors or omissions. Cited patent literature

[0000] JP 2006123844A

[0003] WO 2021 / 124282 A1

[0003]

Claims

[1] First aid for removing immersion liquid from a sample chamber of a microscope with a first element with a sorbing material which is to be placed in a sample stage of the microscope or placed on it and aligned in such a way that by moving an immersion objective with an immersion agent wetting towards the first element and / or a movement of the sample stage in the direction of the immersion objective with the immersion agent wetting, the sorbing material of the first element comes into contact with the immersion agent wetting and removes it. [2] Aid according to claim 1, characterized by that the first element is formed with a dimensionally stable carrier material, a sample carrier or a sample vessel and is inserted into the sample table or placed on it by means of the same. [3] Aid according to claim 2, characterized by that the sample carrier is made up of several parts. [4] Second aid for removing immersion liquid from a sample chamber of a microscope with a second element with a sorbing material, which is to be introduced into an objective changing device and aligned in such a way that by moving the second element towards a sample carrier with an immersion agent wetting and / or a movement of the sample table in the direction of the second element with sorbing material, the latter comes into contact with the immersion agent wetting of the sample carrier and removes it. [5] Third aid for removing and removing immersion liquid from a sample chamber of a microscope with a third element with a sorbing material arranged on a stationary component of the microscope and aligned so that - by moving an immersion objective with an immersion agent wetting to the third element and / or - by moving a sample table with a sample carrier with immersion agent wetting to the third element, the sorbing material of the third element comes into contact with the immersion agent wetting and removes it. [6] Aid according to one of claims 2 to 5, characterized by that the sorbing material is fixed in its position on or in its element magnetically, adhesively or by form fit. [7] Aid according to one of the preceding claims, characterized by that the sorbent material is a sponge, a cloth, in particular a cloth made of cotton, bamboo or bamboo viscose, a polyester fleece, a polypropylene fleece, hemp, wool or clay. [8] Aid according to one of the preceding claims, characterized by that the sorbing material is formed in layers in a stack, the layers of which are to be removed individually. [9] Aid according to one of the preceding claims, characterized by that the shape and / or size of the element and in particular of the sorbing material is adapted to the shape and / or size of the immersion objective, a sample vessel and / or the sample carrier, in particular to standardized sample vessels and / or standardized sample carriers. [10] Aid according to one of the preceding claims, characterized by that the sorbent material is loaded with a solvent to increase the absorption capacity for immersion agents. [11] Aid according to one of the preceding claims, characterized by that the element has an indicator for absorbed liquid which is arranged at least partially in contact with the sorbent material. [12] Aid according to claim 11, characterized by that the indicator is arranged and / or designed to indicate the amount of liquid taken up and / or the reaching of a predetermined amount of liquid. [13] A microscope comprising a device for automatically applying a first element comprising sorbing material according to any one of the preceding claims to and removing from a sample stage. [14] Microscope according to claim 13, wherein it comprises or cooperates with a camera for detecting the sample area and an indicator according to claim 11 or 12. [15] Microscope according to claim 14, wherein it comprises or cooperates with an illumination device. [16] Method for removing and removing immersion liquid from a sample chamber of a microscope with a first aid according to one of the preceding claims, comprising the following steps: • Inserting or placing a first aid in or on a sample table of the microscope, • Moving the immersion objective and / or the sample stage in the z-direction until the sorbing material and the immersion objective come into contact and the immersion medium is absorbed, • Moving the sample table to an output position, and • Remove this first aid. [17] Method for removing and removing immersion liquid from a sample chamber of a microscope with at least one of the aids according to one of the preceding claims, comprising the following steps: • Inserting or placing a sample carrier or a sample container with a sample in or on a sample stage of the microscope, and • Examine the sample with an immersion objective, and optionally o.1) if the sample carrier or the sample vessel is part of a first aid according to at least claim 2 with a sample carrier or a sample vessel: o.2) Positioning the first aid by moving the sample table in the x-direction and y-direction in relation to the immersion objective so that the sorbing material and the immersion objective are aligned one above the other, and o.3) Moving the immersion objective and / or the sample stage in the z-direction until the sorbing material and the immersion objective come into contact and the immersion medium is absorbed, and / or p.1) Changing from an immersion objective to a second aid according to one of the preceding claims by means of an objective changing device, p.2) Positioning the sample carrier or sample vessel by moving the sample table in the x-direction and y-direction so that the sorbent material of the second aid and the immersion medium wetting of the sample carrier or sample vessel are aligned with each other, p.3) Moving the second tool and / or the sample stage in the z-direction until the sorbent material and the immersion agent wetting come into contact and are removed. [18] Method for removing and removing immersion liquid from a sample chamber of a microscope with a third aid according to one of claims 5 to 12, comprising the following steps: • Moving an immersion objective with an immersion agent wetting to the third element and / or • Moving a sample table with a sample carrier with immersion agent wetting to the third element, so that the sorbent material of the third element comes into contact with the immersion agent wetting and removes it. [19] Method according to claim 17 or 18, characterized by that after absorption of the immersion medium • the sample table is moved to a dispensing position in a step o.4 and the first aid is removed, or • the position of the second aid is changed in a step p.4 by means of the lens changing device to an output position where the second aid is removed, or • if the sorbent material is formed in layers in a stack, one or more layers of the sorbent material are removed, or • if the immersion medium was water, the sorbent material of the aid dries. [20] Method according to claim 16 or any one of claims 17 to 19, characterized bythat during the contact of the immersion objective and the sorbing material of the first aid, the sorbing material of the second aid with the sample carrier and / or the sorbing material of the third aid with the sample carrier, the sample table is moved in the x-direction and / or y-direction in order to generate a wiping movement.

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