System for microscopic examination of incubated samples

Abstract

To provide an improved system in microscopic examination.SOLUTION: A microscope 100 comprises: a microscope housing 102; and an integrated type sample chamber 106 which is provided in the microscope housing, and is formed of a separated housing section 104 in the microscope housing. The housing section comprises a microscope interface 108 which is configured to connect an incubation environment adjusting unit 110 to a sample chamber and / or a stage top chamber which is configured to receive the sample 120 and is arranged in the sample chamber. In a first incubation mode, the sample chamber is incubated by supply of first incubation atmosphere by the incubation environment adjusting unit, in a second incubation mode, the stage top chamber is incubated by supply of second incubation atmosphere by the incubation environment adjusting unit.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The concept of the present invention relates to a system for microscopic examination of a sample, comprising a microscope and an incubation environment control unit connected to said microscope, in particular a system for examination of a sample that has been incubated in an incubation atmosphere adapted to the sample. [Background technology]

[0002] Particularly in the field of microscopy of living samples, e.g., cells, it is of great interest to maintain the samples for as long as possible under favorable, stress-free environmental conditions. To achieve this goal, incubators are used in order to generate a microclimate adapted to the sample being examined. Existing incubators can be distinguished between, on the one hand, stage-top incubators and, on the other hand, cage incubators.

[0003] Cage incubators are mounted on standard microscopes, preferably upright microscopes, and include a large climate chamber that encloses the microscope's main components, such as the nosepiece, a microscope stage containing a sample carrier, and a concentrator. This requires a large incubation volume. Access to the work area for placing or manipulating samples is provided within the cage incubator and can be reached through dedicated openings in the cage incubator wall. These handling openings, in particular, increase the cage incubator's dimensions, causing it to protrude significantly beyond the microscope stand. Therefore, it is impossible to provide a microscope with a cage incubator in a space-saving manner. On the other hand, stage-top incubators enclose only the sample itself and are positioned above the microscope stage, resulting in a smaller incubation volume. Even if stage-top incubators have minimal space requirements, access to the sample is minimal because the sample is enclosed by a sealed box, which must be opened, destroying the incubation atmosphere within the box. Access to the sample to introduce additional instruments for sample manipulation is difficult, if not impossible, due to the narrow space limitations of the stage-top incubator module. While cage incubators have high energy and gas consumption, stage-top incubators provide a small, enclosed incubation chamber with connected supply conduits to provide the desired incubation atmosphere. On the other hand, stage-top incubator modules can quickly equilibrate any disturbances in the incubation atmosphere (e.g., after opening the module) or achieve the desired setting, because the air volume exchanged per hour in a cage incubator typically exceeds the volume of a stage-top incubator.

[0004] Stage-top incubators allow precise control of the incubation environment directly surrounding the sample, reducing the regulated volume to a minimum. This allows for rapid changes to the incubation environment. However, access to the sample itself is very limited, and traditional stage-top solutions add significant complexity and cost to the customer's system. Cage incubators, on the other hand, allow easier access to the sample, but are slower to change environmental conditions or reach a set point for a given incubation atmosphere.

[0005] Currently, customers need to install / purchase two separate systems, i.e., a stage-top incubator system and a cage incubator, to be able to flexibly test various samples under different incubation environments and to use the benefits of both systems, which adds cost, complexity, and is not beneficial to the availability of the overall system, as each system has its own control unit and software. Summary of the Invention [Problem to be solved by the invention]

[0006] In view of the above drawbacks and problems, there is a need for improved incubation solutions in microscopy. [Means for solving the problem]

[0007] An embodiment of the inventive concept provides a system for microscopic examination (also understood as imaging) of a sample, comprising a microscope and an incubation environment control unit connected to the microscope, according to claim 1. The microscope of the system comprises a microscope housing enclosing illumination optics, a microscope stage, and imaging optics, and further comprises an integrated sample chamber located within the microscope housing and formed by a separated housing section within the microscope housing, the housing section comprising a microscope interface configured to connect the incubation environment control unit to the sample chamber and / or a stage top chamber disposed within the sample chamber and configured to receive a sample. The system provides a first incubation mode and a second incubation mode, wherein in the first incubation mode the sample chamber is incubated by providing a first incubation atmosphere, and in the second incubation mode the stage top chamber is incubated by providing a second incubation atmosphere by the incubation environment control unit.

[0008] Thus, the present invention provides a microscope with a relatively large sample chamber formed by a separate housing section within the microscope housing, which can be incubated by a connected incubation environment conditioning unit without the large volume associated with the high energy and gas consumption of a cage incubator. Because the dedicated sample chamber is formed by a separate housing section, a larger incubation space can be designed compared to known stage-top incubators, while avoiding the bulky cage incubator configuration. Therefore, this incubation solution is also referred to herein as a "sample chamber incubator." At the same time, the system provides a stage-top chamber disposed within the sample chamber and configured to receive a sample. The term "disposed within the sample chamber" can also be used to mean "formed / constructed within the sample chamber," as described in more detail below. The microscope interface of the system is also configured to connect the incubation environment conditioning unit to the stage-top chamber, so that, in a second incubation mode, the stage-top chamber is incubated with a second incubation atmosphere provided by the conditioning unit.

[0009] Thus, the present invention provides customers with two incubation solutions implemented in the same system. In the first incubation mode, the incubated volume is reduced to the volume of a separate housing section within the microscope housing, specifically designed for the needs and intended application of each microscope, thereby minimizing the internal volume. The sample chamber provides sufficient space for all necessary sample manipulations required for the application, leaving sufficient space for additional equipment. Furthermore, the system allows users to utilize a stage-top incubator without switching to a different system. The present invention combines the stage-top incubator and the sample chamber incubator into a single system, sharing common components. The system preferably provides a sample chamber incubator as the default incubation solution. The incubation environment conditioning unit is the central shared component for providing the first and second incubation atmospheres in the first and second incubation modes, respectively.

[0010] In an advantageous embodiment, the incubation environment conditioning unit includes a pump control unit interface for connecting a first pump control unit and / or a second pump control unit, the first pump control unit configured to supply a first incubation atmosphere having a first group of parameters in a first incubation mode, and the second pump control unit configured to supply a second incubation atmosphere having a second group of parameters in a second incubation mode. In this embodiment, the pump control unit is configured to supply a corresponding incubation atmosphere to the sample chamber or the stage top chamber via the microscope interface in each incubation mode. Each incubation atmosphere is defined by a parameter group including, in particular, flow rate, temperature, and at least one of H2O content, CO2 content, N2 content, and / or O2 content. For imaging of live cells under a microscope, the incubation environment conditioning unit typically needs to be able to control the temperature, humidity, and CO2 content of the incubation atmosphere. To achieve this goal, connections must be present to supply H2O and CO2, with the remainder being air. On the other hand, it may be desirable to perform hypoxic experiments, in which low levels of oxygen are present in the incubation atmosphere. Such oxygen-deficient conditions are typically created by replacing oxygen in the incubation chamber with nitrogen (N2). The desired incubation atmosphere with the corresponding group of parameters is supplied by one of the pump control units.

[0011] The pump control unit interface can be configured to connect both the first and second pump control units. In this case, depending on the selected incubation mode, one of the pump control units needs to be handled by the system. On the other hand, the pump control unit interface can be configured to connect the first pump control unit and, after the first pump control unit is removed, to connect the second pump control unit instead, or vice versa. In this case, the system is preferably configured to automatically switch to the respective incubation mode depending on which pump control unit is connected to the pump control unit interface.

[0012] The housing section of the microscope housing advantageously includes a lid that provides direct access to the microscope stage for accessing the sample in the sample chamber. In other words, the housing section includes an opening or recess that can be closed by a "lid," which should be understood to mean a door, hood, window, or other equivalent means for opening and closing. In particular, the lid is a hinged lid. The housing section of the microscope housing is also advantageously configured so that the sample chamber is sealed when the lid is closed. Sealing the sample chamber minimizes loss of the incubation atmosphere due to leakage. By opening the lid, a sample can be placed in the sample chamber, particularly on the microscope stage, and a stage-top chamber can be placed in the sample chamber, allowing the sample to be replaced and / or manipulated. Instead of a lid that provides direct access to the microscope stage, other solutions can be considered, such as an automatic sample delivery means for transporting the sample onto the microscope stage.

[0013] In an advantageous embodiment, the microscope, particularly the housing section, includes a common atmosphere conditioning module for controlling a first group of parameters of the first incubation atmosphere in the first incubation mode and a second group of parameters of the second incubation atmosphere in the second incubation mode. Such a common atmosphere conditioning module, or more generally, any of the atmosphere conditioning modules described further below, can be used for further adjustment / fine-tuning of the incubation atmospheres. To achieve this goal, sensors can be provided in the atmosphere conditioning module, or sensor signals representing the values ​​of at least some of the parameters of each incubation atmosphere can be provided in the atmosphere conditioning module. If the actual value of such a parameter deviates from the setpoint for that parameter, the atmosphere conditioning module can adjust / readjust the respective parameter. For this purpose, it is advantageous for the common atmosphere conditioning module to communicate with a first pump control unit in the first incubation mode and with a second pump control unit in the second incubation mode. As already pointed out, the same applies to other types of atmosphere conditioning modules described further below. In this embodiment, if the temperature or humidity of the incubation atmosphere deviates from the corresponding set point, the atmosphere conditioning module can send a request to the corresponding pump control unit to adjust / readjust the temperature or humidity to the correct set point. Meanwhile, the atmosphere conditioning module can provide corresponding means for such adjustment / readjustment. For example, the atmosphere conditioning module can include a heater / cooler and / or a humidifier to adjust the temperature and / or humidity of the incubation atmosphere by itself without communicating with the corresponding pump control unit.

[0014] In another advantageous embodiment, the microscope, particularly the housing section, comprises an atmosphere-conditioning module interface for connecting a first atmosphere-conditioning module and / or a second atmosphere-conditioning module, the first atmosphere-conditioning module being configured to control a first group of parameters of a first incubation atmosphere in a first incubation mode, and the second atmosphere-conditioning module being configured to control a second group of parameters of a second incubation atmosphere in a second incubation mode. In contrast to a common atmosphere-conditioning module, this embodiment provides separate atmosphere-conditioning modules for adjusting / readjusting the first or second incubation atmosphere in the first or second incubation mode. All other features described above with respect to the common atmosphere-conditioning module apply equally to the first and second atmosphere-conditioning modules.

[0015] The atmospheric-conditioning module interface may be configured to simultaneously connect a first and a second atmospheric-conditioning module. In this case, depending on the selected incubation mode, one of the first and second atmospheric-conditioning modules must be handled by the system. Alternatively, the atmospheric-conditioning module interface may be configured to connect a first atmospheric-conditioning module and, after the first atmospheric-conditioning module is removed, to connect a second atmospheric-conditioning module in its place, or vice versa. In this case, the system is preferably configured to automatically switch to the corresponding incubation mode depending on which atmospheric-conditioning module is connected to the interface. In embodiments in which first and second pump control units and first and second atmospheric-conditioning modules are present, two distinct configurations exist for realizing a first incubation mode ("sample chamber incubation") and a second incubation mode ("stage-top incubation"). In the first incubation mode, the first pump control unit is connected to the pump control unit interface of the incubation environmental control unit and paired with the first atmosphere control module, which is connected to the atmosphere control module interface on the microscope. For example, this configuration can control the temperature, percentage CO2 level, and relative humidity in the sample chamber. The overall setup allows free access to the sample and the use of additional modules, such as a plate loader, pipetting solutions, etc.

[0016] In a second incubation mode, the first pump control unit is replaced with a second pump control unit, and the first atmospheric conditioning module is replaced with a second atmospheric conditioning module (either automatically or manually by the customer). Additionally, a stage-top chamber is positioned or formed within the sample chamber, and a sample is placed within the stage-top chamber for testing. In this configuration, for example, the temperature, percentage of CO2, percentage of O2, and relative humidity within the small-volume stage-top chamber can be controlled. An oxygen-depleted condition is achieved by replacing oxygen with nitrogen (N2) and humidity (HO).

[0017] Such a system, providing the above two configurations for realizing the first and second incubation modes, includes numerous advantages. The first and second pump control units are connected to the same conditioning unit interface in both configurations. The same applies to the first and second atmosphere-conditioning modules, which are connected to the same atmosphere-conditioning module interface. Furthermore, the second incubation mode can use the same power supply, temperature control, and heat distribution means, as well as the same mechanical interface, as the first incubation mode. Furthermore, both configurations are advantageously controlled by a common software interface, as discussed further below.

[0018] Conventionally, oxygen-depletion experiments in cage incubators are technically difficult to achieve because of the enormous amount of nitrogen required to flush oxygen out of the large incubation volume. This leads to high overall system complexity to ensure airtightness and requires additional safety measures on the customer's side (e.g., gas alarms in the lab, customer inspection before installation, additional ventilation measures in the lab). In particular, the inventive concept of combining a sample chamber incubator with a stage-top incubator for oxygen-depletion experiments helps to reduce overall system and operation costs, overall system complexity, and customer risks.

[0019] In the above example, it is advantageous for the first atmosphere adjustment module (or the common atmosphere adjustment module) to include sensors for measuring the temperature and / or HO content and / or CO content in the incubated sample chamber. Additionally or alternatively, corresponding sensors are disposed in the sample chamber and connected to the first atmosphere adjustment module for sending corresponding sensor signals to the first atmosphere adjustment module. Similarly, it is advantageous for the second atmosphere adjustment module to include sensors for measuring the temperature and / or O content and / or HO content and / or CO content in the incubated stage top chamber. Additionally or alternatively, such sensors are disposed in the incubated stage top chamber and connected to the second atmosphere adjustment module for providing corresponding sensor signals to the second atmosphere adjustment module. These embodiments enable the first and second atmosphere adjustment modules to better adjust / readjust the respective incubation atmospheres to setpoint parameters.

[0020] As already mentioned above, it may be advantageous if the atmosphere conditioning module, in particular the second atmosphere conditioning module, further comprises a humidifier for humidifying the second incubation atmosphere, independent of the incubation environment conditioning unit. By introducing HO into the stage top chamber, a high relative humidity is achieved.

[0021] In general, it is advantageous for the system to share as many components as possible between the first and second incubation modes, and in particular it is advantageous for the system to include a common water pump configured to supply HO to both the first and second pump control units, thereby providing a unified user interface.

[0022] In another advantageous embodiment, in the second incubation mode, the system is configured to use the same temperature control and / or method for heat dissipation as in the first incubation mode by controlling the temperature in the stage top chamber via the temperature in the sample chamber. Because the temperature in the relatively small stage top chamber is primarily determined by the temperature in the surrounding sample chamber, the temperature in the stage top chamber can be controlled by controlling the temperature in the sample chamber. To achieve this goal, the incubation environment conditioning unit can, for example, provide a dedicated airflow at a desired temperature into the sample chamber, while simultaneously providing a second incubation atmosphere in the stage top chamber.

[0023] In a particularly advantageous embodiment, the system comprises a universal software interface for communicating with the microscope controller on the one hand and with the incubation environmental conditioning unit on the other hand, providing an incubation environmental conditioning unit interface for communicating with the incubation environmental conditioning unit. The microscope controller is configured to control the microscope's functional components, such as the imaging optics, microscope stage, and / or illumination optics, for controlling microscopy / sample imaging and the experimental workflow. The universal software interface communicates with the microscope controller and the incubation environmental conditioning unit so that both the microscope and the incubation environmental conditioning unit can be controlled / handled by the universal software interface. It is particularly advantageous for the universal software interface to provide a corresponding graphical user interface (GUI) for the customer / user, which is displayed on a display screen. The GUI is preferably adapted to the various possible types of experiments of the system according to the inventive concept. In the above example, the user can set, for example, the desired temperature, humidity, and CO2 content of the incubation atmosphere (and the desired O2 content in the case of hypoxic experiments) by selecting the corresponding buttons in the GUI. Preferably, default values ​​for each parameter are shown in the GUI and can be changed by the user. Furthermore, parameters of the microscope's functional components, such as illumination wavelength / intensity, camera parameters, such as sensitivity, gain, etc., can be set through the same GUI.

[0024] As already mentioned above, the universal software interface is configured to automatically switch the system to the first incubation mode when a first pump control unit is connected to the pump control unit interface, and to automatically switch to the second incubation mode when a second pump control unit is connected to the pump control unit interface. The universal software interface may be further configured to detect whether a suitable atmosphere conditioning module is present in the microscope, for example, whether a first atmosphere conditioning module is connected to a corresponding module interface or whether a common atmosphere conditioning module is activated in the first incubation mode.

[0025] Advantageously, the system further includes a sample holder for holding a sample in a first incubation mode, the sample holder being adapted for attachment of a cover to form a stage-top chamber used in a second incubation mode. Such a configuration is particularly useful for avoiding cumbersome sample handling, such as removal from the sample chamber, introduction into the stage-top chamber, and return to the sample chamber, or exchange of one sample for another within the stage-top chamber. Rather, in this embodiment, the sample can be inspected / imaged first in the first incubation mode, where it is placed on the (universal) sample holder, and then in the second incubation mode, where a cover is attached onto the sample holder to form a stage-top chamber surrounding the sample. Advantageously, the sample holder includes a connection for introducing a gas / incubation atmosphere, allowing a second incubation atmosphere to be introduced into the stage-top chamber.

[0026] As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items and may be abbreviated as " / ".

[0027] It should be noted that each feature of the above examples and the examples described below can be combined, in whole or in part, with other examples not explicitly mentioned herein but which are part of this disclosure. [Brief explanation of the drawings]

[0028] [Figure 1] FIG. 1 is a schematic diagram illustrating a perspective view of an embodiment of a system according to the inventive concept. [Figure 2A] 1 is a diagram illustrating a schematic configuration of an embodiment of a system according to the concept of the present invention; [Figure 2B] 1A-1D are schematic diagrams illustrating different configurations of an embodiment of a system according to the inventive concept; [Figure 2C] 1A-1D are schematic diagrams illustrating different configurations of an embodiment of a system according to the inventive concept; [Figure 3] FIG. 1 is a schematic diagram illustrating an embodiment of a system overview according to the inventive concept. [Figure 4] FIG. 10 is a schematic diagram of an embodiment of a stage-top chamber for use in a system according to the concepts of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0029] FIG. 1 schematically illustrates a perspective view of an embodiment of a microscope 100 for microscopic examination of a sample 120 disposed on a microscope stage 116. A microscope housing 102 encloses illumination optics 118, a microscope stage 116, and imaging optics 124. An integrated sample chamber 106 is located within the microscope housing 102 and is formed by a separate housing section 104 within the microscope housing 102. The housing section 104 includes a hinged lid 109 that places the sample 120 on the microscope stage 116 within the sample chamber 106 and provides direct access to the microscope stage 116 when the lid is open for replacing and / or manipulating the sample 120. The embodiment illustrated in FIG. 1 is an inverted transmitted light microscope 100. In this case, the transmitted light illumination optics 118 are disposed within the housing section 104, while the imaging optics are located below the microscope stage 116 in a second housing section 122. The imaging optics 124 typically include a microscope objective and an image detector as its main components. The image detector usually comprises a camera that produces a microscope image that is typically displayed on a display screen 142 outside the microscope housing 102 .

[0030] The structure of the microscope housing section 104 allows for the formation of a dedicated sample chamber 106, which, after closing the lid 109, constitutes an incubatable, enclosed space, such that a preferably living sample 120, e.g., a cell, can be maintained under favorable and stress-free environmental conditions during microscopic examination / imaging. To achieve this goal, the housing section 104 comprises an interface 108 for connecting an external incubation environmental conditioning unit 110 to the sample chamber 106. The interface 108 is configured to provide a connection between the conditioning unit 110 and the sample chamber 106. As a result, when the conditioning unit 110 is connected to the interface 108, the environmental conditions within the sample chamber 106 can be controlled. This configuration realizes a first incubation mode ("sample chamber incubation").

[0031] In the illustrated embodiment, the interface 108, part of the housing section 104, includes two openings 112 on the rear surface of the housing section 104. Each opening 112 is configured to receive a conduit 114. An incubation atmosphere can be introduced into the sample chamber 106 through at least one of the conduits 114. Depending on leakage in the sample chamber 106, a portion of the incubation atmosphere can escape from the sample chamber 106. Alternatively, a portion of the incubation atmosphere can be withdrawn from the sample chamber 106 through another conduit 114. A suitable incubation atmosphere includes air containing a predetermined content of H2O (relative humidity) and a predetermined content of CO2 (carbon dioxide). It may also be desirable to perform hypoxic experiments in which the atmosphere is depleted of oxygen. Typically, the temperature of the incubation atmosphere can be set from ambient temperature of approximately 20°C to 50°C, particularly in the range of 25-37°C. The CO2 range is set to 0.5-20%, and the O2 range is set to 1-18%. Humidity should be adjusted to avoid potential condensation or at least ensure that it does not adversely affect either the microscope 100 or the conditioning unit 110 or the sample itself. It is preferable to control at least the temperature, humidity, and CO content. In hypoxic experiments, the O content is controlled by N introduction. Such experiments are preferably performed in a second incubation mode ("stage-top chamber incubation"), as further described below.

[0032] To control the above parameters, sensors are preferably located within the conduit 114 and / or within the sample chamber 106 and / or near the sample 120 on the microscope stage 116. In a preferred embodiment, at least some of the sensors are integrated into the conduit 114 for supplying an incubation atmosphere to the sample chamber 106.

[0033] 1, the lower side of the housing section 104 is defined by a work surface 107 that includes the upper side of the microscope stage, i.e., the microscope stage tabletop. This configuration provides user-friendly access to a work area for placing and manipulating a sample 120. On the opposite side, the housing section is defined by the inside of a lid 109 and the back side of the housing section 104 itself.

[0034] In order to extend the lifetime of the imaging optics 124, it is preferable to air-condition and / or temperature-control the second housing section 122 when using an immersion objective. This can be done either by the same conditioning unit 110 and its corresponding expansion interface 108, or more preferably by a separate air-conditioning unit.

[0035] In a preferred embodiment, the microscope 100 comprises a control unit 140, which can be or can include functional components for controlling the microscopy / imaging of the sample 120, such as a microscope controller 350 (see FIG. 3 ) that controls the illumination optics 118, the microscope stage 116, and the imaging optics 124 of the microscope 100. Typically, a graphical user interface (GUI) is displayed on a display screen 142 for user-friendly operation of the microscope 100. The control unit 140 is further configured to control the operation of the incubation environment conditioning unit 110 when the conditioning unit 110 is coupled to the interface 108. Cables and lines for communication between the microscope control unit 140 / microscope controller 350 and the conditioning unit controller (not shown) can be routed through one or more of the above-mentioned openings / conduits in the interface 108. In this case, the same GUI can display corresponding buttons for setting desired values ​​for the above-mentioned incubation atmosphere parameters. After it is confirmed that the lid 109 is closed and the sample to be examined is placed on the table of the microscope stage 116, the generation of a user-defined incubation atmosphere begins automatically, and once the desired atmosphere is created and the sample 120 is in the appropriate position, the sample is examined / imaged.

[0036] As shown in FIG. 1 , the default configuration is the first incubation mode, i.e., in this embodiment, the sample chamber incubation mode. As will be described in more detail below in conjunction with the drawings, the incubation environment conditioning unit 110 includes a pump control unit interface for connecting the first pump control unit 212 shown in FIG. 1 . At the rear of the housing section 104, there is an atmosphere conditioning module interface to which the first atmosphere conditioning module 216 is connected as shown in FIG. 1 . The above-mentioned sensors for detecting one or more incubation atmosphere parameters are preferably located within the sample chamber 106, particularly within or in the first atmosphere conditioning module 216. The first atmosphere conditioning module 216 is connected to the incubation environment conditioning unit 110 via a communication line. In this way, in response to the sensor signals corresponding to the parameter values, the first atmosphere conditioning module 216, together with the first pump control unit 212, can provide feedback control for setting a first group of parameters of the first incubation atmosphere to desired set points. This process is further described below in connection with the figures.

[0037] Different configurations of an embodiment of a system according to the inventive concept are shown diagrammatically in Figure 2. The figures are collectively described and like reference numerals indicate the same or at least functionally the same components.

[0038] FIG. 2A corresponds to the configuration shown in FIG. 1. A microscope 100 is shown on the right side of FIG. 2A. The microscope 100 includes, as its main components, illumination optics 118, a microscope stage 116, and imaging optics 124, with the microscope objective defining an optical axis 126. A sample 120 is placed on the microscope stage 116 within a sample chamber 106 formed by a separate housing section 104 within a microscope housing 102 of the microscope 100. The imaging optics, or at least a portion thereof, is located within a second separate housing section 122. An incubation environmental conditioning unit 110 is shown on the left side of FIG. 2A and is connected to the microscope 100 via an interface 108 within the housing section 104 of the microscope 100 and by a conduit 114 extending from the conditioning unit 110 into the sample chamber 106.

[0039] The incubation environmental conditioning unit 110 includes a first pump control unit 212 connected to a pump control unit interface 210 and to a common / universal software interface 250 and a common water pump 240. The common software interface 250 and common water pump 240 can also be used in a second incubation mode, as described below. The incubation environmental conditioning unit 110 is designed to provide a first incubation atmosphere with a predetermined flow rate, a predetermined temperature, and predetermined H2O and CO2 contents. The power supply of the temperature control and conditioning unit 110 can also be used in the second incubation mode. The first incubation atmosphere is introduced into the sample chamber 106 via the interface 108. A portion of the incubation atmosphere can also be exhausted through the interface 108 and conduit 114, as indicated by the arrows. A first atmosphere conditioning module 216 is located within the sample chamber 106 and is connected to an atmosphere conditioning module interface 220. As already mentioned above, the first atmosphere conditioning module 216 comprises sensors for detecting the temperature and the CO2 and HO contents in the incubation atmosphere. Depending on the actual values ​​of these parameters, the first atmosphere conditioning module 216 communicates with the first pump control unit 212 to set the set values ​​of these parameters, thereby enabling feedback control.

[0040] FIG. 2B illustrates an embodiment of system 130 in a second incubation mode. Below, only the differences from the embodiment illustrated in FIG. 2A will be described. As illustrated in FIG. 2B, a stage-top chamber 230 is placed or constructed within sample chamber 106. Stage-top chamber 230 forms a relatively small volume around sample 120 located within stage-top chamber 230. First pump control unit 212 is replaced by a second pump control unit 214 connected to the same interface 210 as illustrated in FIG. 2A. Second pump control unit 214 shares the same common software interface 250 and the same common water pump 240 as first pump control unit 212. In the embodiment illustrated in FIG. 2B, a second atmospheric conditioning module 218 replaces first atmospheric conditioning module 216 and is connected to the same interface 220 as illustrated in FIG. 2A. In this configuration, system 130 can operate in a second incubation mode.

[0041] In the second incubation mode, in this embodiment, the incubation environment conditioning unit 110 generates a second incubation atmosphere with a predetermined flow rate, a predetermined temperature, and predetermined HO, CO, and O contents. The second incubation atmosphere is introduced into the stage-top chamber 230 via the interface 108 and the second atmosphere conditioning module 218. Corresponding supply lines are provided, as shown in FIG. 2B . The second atmosphere conditioning module 218 includes sensors for detecting the values ​​of one or more parameters of the second incubation atmosphere—in this embodiment, sensors for detecting temperature, CO, HO, and O contents. A temperature sensor can also be placed in the stage-top chamber 230 to more precisely detect the temperature within the stage-top chamber 230. Again, the corresponding sensor signals enable feedback control to supply the second incubation atmosphere with the corresponding parameter setpoints into the stage-top chamber 230. Typically, hypoxic experiments are performed in the second incubation mode. In this case, only the small volume stage top chamber 230 needs to be flushed with N2, as opposed to the large volume sample chamber 106 in the first incubation mode.

[0042] As can be seen from Figures 2A and 2B, a single system is provided for operation in two different incubation modes without the need to modify or convert / rebuild the system to switch from one incubation mode to the other.

[0043] FIG. 2C illustrates another embodiment of a system 130 according to the inventive concept. Again, like reference numerals refer to identical or at least functionally similar components. Only the differences relative to FIGS. 2A and 2B will be described in more detail. As can be seen in FIG. 2C, a common atmosphere-conditioning module 219 is connected to an atmosphere-conditioning module interface 220. The common atmosphere-conditioning module 219 is configured to control a first group of parameters of a first incubation atmosphere in a first incubation mode and a second group of parameters of a second incubation atmosphere in a second incubation mode. It should be noted that the common atmosphere-conditioning module 219 can be used in both the embodiment according to FIG. 2A and the embodiment according to FIG. 2B. This eliminates the need to replace the first and second atmosphere-conditioning modules when switching from one incubation mode to the other.

[0044] The incubation environmental conditioning unit 110 includes a pump control unit interface 210 for connecting both the first and second pump control units 212, 214, as shown in FIG. 2C. The pump control units 212, 214 are each connected to a common software interface 250 and a common water pump 240. The configuration shown in FIG. 2C eliminates the need for any replacement of the pump control units and atmosphere conditioning modules. The system can automatically detect the presence of the stage-top chamber 230 within the sample chamber 106 and automatically switch to the second incubation mode. Alternatively, the user selects the first or second incubation mode via a GUI. For remaining features and advantages of the configuration shown in FIG. 2C, please refer to the previous discussion of FIGS. 2A and 2B.

[0045] An embodiment of a system overview according to the concepts of the present invention is shown schematically in Figure 3. In Figure 3, a legend is provided in the lower right corner, and the meaning of the symbols is explained below. Black oval symbols denote external interfaces, black triangles denote common interfaces, rectangles denote internal components, and dashed hatched rectangles denote common internal components. Figure 3 shows an embodiment of a complete integrated system 130.

[0046] The universal software interface 250 is configured to control the entire system 130. To achieve this goal, the universal software interface 250 provides an incubation environment control unit interface 360, which in turn is connected to a microscope controller 350. Thus, all functions and settings of the microscope 100 and the incubation environment control unit 110 can be controlled through the universal software interface 250. The universal software interface 250 typically provides a graphical user interface (GUI) that can be operated by a user.

[0047] The incubation environment conditioning unit 110 includes an interface, which is the pump control unit interface 210 shown in FIG. 2. The first and / or second pump control units 212, 214 can be connected to this interface. The conditioning unit 110 further includes a mechanical system interface 301, a main control PCB 302, a heating module 303a, and a power supply 303b as common internal components. These components are shared between the first and second incubation modes. Finally, the conditioning unit 110 includes a power interface 304, a USB interface 305, and a water pump control interface 306 as common interfaces.

[0048] The first pump control unit 212 includes, as internal components, an external interface for an HO supply, an external interface for a CO supply, a control PCB 307, and a gas humidifier 308. The second pump control unit 214 includes, as internal components, an external interface for an HO supply, an external interface for a CO supply, and an external interface for an N supply, and a communication connection (not shown) to the incubation environment conditioning unit 110. Further, as internal components, the second pump control unit 214 includes, as internal components, a control PCB 307, a gas humidifier 308, and a gas mixing module 309. The water pump 240 is connectable to the first and / or second pump control units 212, 214, and includes, as internal components, a water reservoir 310 and a water pump 311.

[0049] In the embodiment shown in Figure 3, two incubation modes can be realized as already described above with reference to Figures 1 and 2. As many components as possible are shared between the two incubation modes. The second pump control unit 214 is provided with means for humidifying the gas mixture in addition to or independently of the incubation environmental conditioning unit 110.

[0050] 1 and 2, the incubation environmental conditioning unit 110 communicates with the first and second atmosphere conditioning modules 216 and 218. The first atmosphere conditioning module 216 comprises a gas sensor 312 as an internal component and an external interface 314 for an ambient temperature sensor. Optionally, an external interface 315 for a sample temperature sensor can be provided. The interface 314 is connected to a temperature sensor in the sample chamber 106, while the interface 315 is connected to a temperature sensor in the vicinity of the sample 120 itself. The second atmosphere conditioning module 218 comprises a gas sensor 313 as an internal component and the same external interfaces 314 and 315 as the module 216. It should be noted that, as already mentioned above, in another embodiment, the atmosphere conditioning modules 216 and 218 can alternatively comprise a temperature sensor and / or be connected to a gas sensor located in the sample chamber or the stage top chamber via a corresponding external interface. Gas sensors 312 and 313 include sensors for detecting first and second groups of parameters of the first and second incubation atmospheres, respectively. In the absence of external interface 315 of second atmosphere conditioning module 218, the temperature in stage top chamber 230 is controlled solely by the temperature in the surrounding sample chamber 106.

[0051] The conditioned / reconditioned first incubation atmosphere is introduced into the sample chamber 106, which includes an ambient temperature sensor 316 connected to the interface 314 of the module 216, while the conditioned / reconditioned second incubation atmosphere is introduced into the stage top chamber 230, which may include a sample temperature sensor 317 connected to the interface 315 of the module 218. Alternatively, as described above, the ambient temperature sensor 316 of the sample chamber 106 is connected to the interface 314 of the module 218.

[0052] In another embodiment, the second atmosphere-conditioning module 218 can comprise an external interface for a water supply connected to a gas humidifier as an internal component. In this case, module 218 can additionally and independently regulate / readjust the relative humidity in the second incubation atmosphere. In another alternative embodiment already discussed above, modules 216 and 218 are combined into a common atmosphere-conditioning module 219 (see FIG. 2C).

[0053] FIG. 4 shows a schematic cross-section of an embodiment of a stage-top chamber 230 preferably used in a system consistent with the concepts of the present invention. Traditionally, in a sample chamber incubation mode, a sample is placed in a sample holder on the microscope stage 116. To operate in the second incubation mode, a separate sample holder in the form of a stage-top incubator must be placed on the microscope stage. The stage-top chamber 230 shown in FIG. 4 overcomes this drawback by providing a universal sample holder 410 onto which the sample 120 is placed in both the first and second incubation modes. The universal sample holder 410 is located in or on the microscope stage 116. A gas connector 430 is flanged on the side of the universal sample holder 410. To provide a sealed stage-top chamber 230, a cover 420 is attached. During the second incubation mode, a second incubation atmosphere is introduced into the stage-top chamber 230 via a conduit connected to the gas connector 430. [Explanation of symbols]

[0054] 100 microscopes 102 Microscope housing 104 Separated Enclosure Sections 106 Sample Chamber 107 Work Surface 108 Microscope Interface 109 Lid 110 Incubation Environment Control Unit 112 Aperture 114 Conduit 116 Microscope Stage 118 Illumination optical system 120 samples 122 Second Enclosure Section 124 Imaging Optical System 126 Optical axis 130 Systems 140 Control Unit 142 displays 210 Pump Control Unit Interface 212 first pump control unit 214 Second pump control unit 216 First Atmosphere Control Module 218 Second Atmosphere Control Module 219 Common Atmosphere Control Module 220 Atmosphere Control Module Interface 230 Stage Top Chamber 240 Water Pump 250 Universal Software Interface 301 Mechanical System Interface 302 Main control PCB 303a Heating Module 303b power supply 304 Power Interface 305 USB interface 306 Pump Control Interface 307 Control PCB 308 Gas Humidifier 309 Gas Mixing Module 310 Water storage section 311 Water Pump 312 Gas Sensor 313 Gas Sensor 314 Ambient Temperature Sensor Interface 315 Sample Temperature Sensor Interface 316 Ambient Temperature Sensor 317 Sample Temperature Sensor 350 Microscope Controller 360 Incubation Environment Control Unit Interface 410 Universal Sample Holder 420 Cover 430 Gas Connector

Claims

1. A system (130) for microscopic examination of a sample (120), comprising: The system (130) comprises a microscope (100) and an incubation environment control unit (110) connected to the microscope (100); The microscope (100) a microscope housing (102) enclosing an illumination optics (118), a microscope stage (116), and an imaging optics (124); an integrated sample chamber (106) located within the microscope housing (102) and formed by a separate housing section (104) within the microscope housing (102), the integrated sample chamber containing the illumination optics (118); Equipped with the housing section (104) comprises a microscope interface (108) configured to connect the incubation environmental conditioning unit (110) to the sample chamber (106) and / or a stage top chamber (230) disposed within the sample chamber (106) and configured to receive a sample (120); The system provides a first incubation mode and a second incubation mode, in which the sample chamber (106) is incubated by supplying a first incubation atmosphere by the incubation environment adjustment unit (110), and in which the stage top chamber (230) is incubated by supplying a second incubation atmosphere by the incubation environment adjustment unit (110). system.

2. the incubation environment conditioning unit (110) comprises a pump control unit interface (210) for connecting a first pump control unit (212) and / or a second pump control unit (214); the first pump control unit (212) is configured to supply the first incubation atmosphere having a first group of parameters in the first incubation mode, and the second pump control unit (214) is configured to supply the second incubation atmosphere having a second group of parameters in the second incubation mode. The system of claim 1 .

3. the pump control unit interface (210) is configured to connect the first pump control unit (212) and, after the first pump control unit (212) is removed, to connect the second pump control unit (214) in place of the first pump control unit (212); The system of claim 2.

4. the pump control unit interface (210) is configured to connect both the first pump control unit (212) and the second pump control unit (214); The system of claim 2. Claim 5: The housing section (104) comprises a common atmosphere regulation module (219) for controlling the first group of parameters of the first incubation atmosphere in the first incubation mode and the second group of parameters of the second incubation atmosphere in the second incubation mode. A system according to any one of claims 2 to 4.

6. the common atmosphere conditioning module (219) communicates with the first pump control unit (212) in the first incubation mode and with the second pump control unit (214) in the second incubation mode; The system of claim 5.

7. The housing section (104) comprises an atmosphere conditioning module interface (220) for connecting a first atmosphere conditioning module (216) and / or a second atmosphere conditioning module (218); The first atmosphere adjustment module (216) is configured to control the first group of parameters of the first incubation atmosphere in the first incubation mode, and the second atmosphere adjustment module (218) is configured to control the second group of parameters of the second incubation atmosphere in the second incubation mode. A system according to any one of claims 2 to 4.

8. the atmosphere-conditioning module interface (220) is configured to connect the first atmosphere-conditioning module (216) and, after the first atmosphere-conditioning module (216) is removed, to connect the second atmosphere-conditioning module (218) in place of the first atmosphere-conditioning module (216); The system of claim 7.

9. The first and second groups of parameters of the first and second incubation atmospheres are flow rate, temperature, and H ２ O content and / or CO ２ Content and / or N ２ Content and / or O ２ at least one of the following contents: A system according to any one of claims 1 to 8, with reference to claim 2.

10. The first atmosphere control module (216) controls the temperature and / or H in the sample chamber (106) to be incubated. ２ O content and / or CO ２ a sensor for measuring the content; A system according to any one of claims 1 to 9, with reference to claim 7.

11. The second atmosphere control module (218) controls the temperature and / or O 2 in the incubated stage top chamber (230). ２ Content and / or H ２ O content and / or CO ２ a sensor for measuring the content; A system according to any one of claims 1 to 10, with reference to claim 7.

12. the second atmosphere conditioning module (218) further comprises a humidifier for humidifying the second incubation atmosphere; A system according to any one of claims 1 to 11, with reference to claim 7.

13. The system includes a H for both the first and second pump control units (212, 214). ２ a common water pump (240) configured to supply O; A system according to any one of claims 2 to 12.

14. In the second incubation mode, the system is configured to use the same temperature control as in the first incubation mode by controlling the temperature in the stage top chamber (230) via the temperature in the sample chamber (106). A system according to any one of claims 1 to 13.

15. The system comprises a universal software interface (250) for communicating with a microscope controller (350); the microscope controller (350) is configured to control functional components (116, 118, 124) of the microscope (100) for controlling microscopic examination of the sample (120) and to provide an incubation environment conditioning unit interface (360) for communication with the incubation environment conditioning unit (110), such that both the microscope (100) and the incubation environment conditioning unit (110) can be controlled by the universal software interface (250); A system according to any one of claims 1 to 14.

16. the universal software interface (250) is configured such that when the first pump control unit (212) is connected to the pump control unit interface (210), the system automatically switches to the first incubation mode, and when the second pump control unit (214) is connected to the pump control unit interface (210), the system automatically switches to the second incubation mode. The system according to claim 15, which recites claim 3.

17. the system comprising a sample holder (410) for holding the sample in the first incubation mode; the sample holder (410) is adapted to receive a cover (420) to form the stage top chamber (230) used in the second incubation mode; 17. A system according to any one of claims 1 to 16.

18. the housing section (104) comprises a lid (109) providing direct access to the microscope stage (116) for placing the sample (120) and / or the stage top chamber (230) in the sample chamber (106); 18. A system according to any one of claims 1 to 17.

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