EP1438560A1

EP1438560A1 - Laser microdissection system

Info

Publication number: EP1438560A1
Application number: EP02779452A
Authority: EP
Inventors: Karin SCHÜTZE
Original assignee: PALM Microlaser Technologies GmbH
Current assignee: PALM Microlaser Technologies GmbH
Priority date: 2001-10-24
Filing date: 2002-10-02
Publication date: 2004-07-21
Also published as: DE10152404B4; US20040252291A1; DE10152404C5; JP4112493B2; DE10152404A1; US6930764B2; CA2463388A1; JP2005506546A; WO2003036266A1

Abstract

The invention relates to a laser microdissection system provided, in particular, for processing a biological material (14) located on a support (3) with a laser beam. Said system comprises a laser (4), a microscope (1) with a camera for generating a microscope or video image of the biological material located on the support (3), and a display screen (8) for displaying the recorded video image. Appropriate input means (9, 10) are used in order to be able to select, on the video image displayed on the display screen (8), a desired biological object (15) to be processed with a laser beam and to assign it to a corresponding object group. In particular, it is possible to mark the biological objects (15) on the video image in different colors, whereby the biological objects marked with one and the same color form a common object group. The control unit (7) of the laser microdissection system, which can be provided in the form of a computer, automatically generates a list in which the individual selected objects and a comprehensive representation (29) of the object groups is contained. The objects contained in the list can be individually selected for a laser processing. A separate selection of individual object groups is likewise possible in order to be able to process the objects, which are combined to form the corresponding object groups, in succession with an appropriate laser function.

Description

LASER microdissection

The present invention relates to a laser microdissection system for processing a biological or non-biological mass, in particular a laser microdissection system for processing, separating and / or obtaining microscopic biological and / or non-biological objects of a biological or not biological mass.

Such a conventional laser microdissection system from the applicant is known for example from WO 97/29355 A or WO 01/73398 A. With the laser microdissection systems described in these documents, individual biological or non-biological objects which are arranged on a planar carrier can be selected with the aid of a computer and processed with a laser beam. In this case, a selected object can be separated from the surrounding mass, for example with the aid of a laser, with the aid of a computer, in order to freely prepare the respectively selected object from the surrounding mass. The freely prepared object can then be catapulted from the carrier to a collecting device by means of a laser-induced transport process with the aid of a laser shot, which is aimed at the freely prepared object. For example, a polymer film can be used as the carrier.

The method described above enables the separation, sorting and extraction of both biological objects and also of non-biological objects. In the context of the present patent application, the term “biological objects” is understood to mean primarily living or fixed biological cells or cell components which are part of a liquid or solid biological material, such as a cell tissue, a smear or a cell culture etc. With the help According to the method described above, the selected objects can be specifically loaded with a selected substance by means of contactless laser micro-projection and the successfully injected biological objects can then be sorted out.The biological objects can be applied next to one another on a solid planar carrier, the process of secreting being carried out within a short time The survivability or the morphology of the biological objects is guaranteed, ie the biological objects are created by the micro-projection process and by the separation and catapulting process ss not damaged or impaired.

In principle, the previously explained laser-induced transport process, i.e. the catapulting out of individual previously selected objects from the surrounding mass, even without prior free preparation of the respectively selected object, if the laser energy and / or the laser focus is selected or selected at the moment the separate laser shot is set in such a way that the resulting the resulting impulse force of this laser shot is sufficient for removing the corresponding object from the surrounding mass and for the transport process to the collecting device.

Since the above-described method can only be carried out manually with relatively great effort and with the desired precision, the laser microdissection systems of the above publications are computer-aided, ie the Cutting and / or catapulting a selected object is computer-aided, so that the laser light source, which generates the laser beam used for cutting and / or catapulting, is automatically controlled and the relative movement required for cutting and / or catapulting between the laser beam and the biological or power - Carrier containing biological objects is automatically brought about and controlled. In particular, computer-aided selection or marking of the desired objects located on the carrier is possible, so that these can subsequently be processed automatically with the laser microdissection system. For this purpose, the laser microdissection system comprises a screen or monitor, on which a video image of the material on the carrier recorded by a digital camera is displayed. The user can draw a desired cutting curve on the screen or the video image, for example with the aid of corresponding graphics tools, which is then automatically followed by the laser beam with the aid of a computer, in order to cut out the object thus selected. In a similar manner, a desired object can also be marked on the screen or on the video image for catapulting out, the separate laser pulse or laser shot then being set at the desired location.

Although in the previously described known laser microdissection systems there is already basically computer-aided and automated processing of the material on the carrier, the processing of a large number of biological objects, which in particular can be of different types, is relatively complex since the objects are either can only be edited individually or in their entirety. The present invention is therefore based on the object of providing a laser microdissection system of the type described above in which the user-friendliness and variety of functions are improved.

This object is achieved according to the invention by a laser microdissection system with the features of claim 1. The subclaims each define preferred and advantageous embodiments of the present invention.

The present invention is described below with reference to the cutting and / or catapulting of biological objects. However, the invention is also applicable to non-biological objects (inanimate matter), e.g. can be microscopic objects made of glass, silica, plastic etc. or artificially produced vesicles etc. in a biological mass. The present invention is also applicable to non-biological masses or materials, e.g. Polymer masses or the like can be used, from which microscopic objects are to be removed using the laser microdissection system.

The laser microdissection system according to the invention comprises a laser light source for generating a laser beam which is to be directed onto the material to be processed located on a corresponding carrier. Furthermore, an image recording device is provided, for example with a CCD camera, which generates a video or image of the material located on the carrier and displays it on a display device, for example a screen, of the laser microdissection system. This image is overlaid with a user interface of the laser microdissection system in order to select the desired objects to be processed with the laser beam and to be able to assign them to the object groups. The laser Microdissection system comprises control means for evaluating the user selection thus carried out and for creating a list in which the selected objects are included with an indication or description of the respectively assigned object group in such a way that a further selection of the objects and / or an object group-specific selection is based on the object group specification for subsequent processing with the laser beam. These control means are implemented in particular in a PC computer or computer of the laser microdissection system.

For the selection and assignment of the desired objects to be processed with the laser beam, different types of marking are made available to the user, so that the user on the display device determines the desired object on the display device, for example by corresponding selection of the marking, and at the same time assigns it to the corresponding object group , with a different marker assigned to each object group. For example, the user can be offered different colors for marking the desired objects, the user using the appropriate graphic aids, e.g. can draw a cutting line for a biological object to be cut out on the display device in the desired color. In this way it is possible, for example, to mark healthy cells in a first color and tumor cells in a second color on the display device, the individual objects being sorted and grouped by color in the list created by the control means. By selecting an object group

(Which of the control means is preferably displayed on the display device with the corresponding color), it is then possible to process all objects belonging to this object group automatically, that is to say with the aid of a computer, the corresponding objects being approached one after the other and with the aid of the laser beam, for example, are cut out and / or catapulted out. Of course, instead of marking using different colors, any other type of distinctive marking is also possible. For example, a section line drawn on the video image around a desired object can be overlaid with a unique identifier that identifies the desired object group, etc.

The selection of the objects to be summarized in an object group can, as described, be done both manually by the operator and automatically and computer-aided by digital image processing / image evaluation, in the second case, for example, in a manner known per se by means of fluorescence evaluation or the like between healthy cells or tumor cells, etc. a distinction can be made and an assignment to a corresponding object group can take place automatically. The term selection means used here thus includes both a manual selection and a software or computer-controlled automatic selection of the desired objects.

The procedure described above makes it possible to process the previously selected and marked objects with the laser beam in a group-specific manner. This means, for example, that initially all objects assigned to a first object group can be processed with the laser beam, while all objects assigned to a second object group can then be processed. This enables all objects of the first object group to be collected in a first collecting container and all objects of the second object group in a second collecting container after the cutting and / or catapulting process has been carried out. The objects in the first group can - as already mentioned - be tumor cells, for example, while it is the objects of the second group can be healthy cells, for example. The separation of the desired objects is significantly simplified and accelerated for the user in this way.

As a result of the group-specific laser processing, a different type of laser processing can advantageously be selected and set for each object group. For this purpose, the laser microdissection system provides several different laser functions, and in particular the number of repetitions of the preset laser processing can also be selected separately for each object group.

The list generated by the control means of the laser microdissection system, in which the objects previously selected by the user are summarized in groups, preferably includes for each group an indication of the number of objects contained therein and the total area of the objects contained therein. For this purpose, the laser microdissection system is equipped with an automatic area calculation function, which enables the area of the thus selected object to be automatically calculated after drawing a cutting line around a desired object. In this way, the user is provided with important additional information about the processed objects - divided into the respective object groups.

In addition to the list section explained above, the control means preferably also generates and displays on the display device a list section which contains information about each individual object to be processed, about the respective object type, about the respective object area and / or about the respectively assigned object group can. With regard to the object type, a distinction can be made, for example, whether it is an object defined by a cutting line or only an object defined by a catapult point, etc. Within this list section, each individual object or any selection of the displayed objects can be marked by the to be able to process the correspondingly marked objects together with the desired laser function.

The list generated by the control means can optionally also only have the first-mentioned list section, in which the selected objects are grouped together, or only the second-mentioned list section, in which the individual selected objects are contained. It is important, however, that an object group specification is provided in each case in order to enable further selection based on the object group specification.

The list created by the control means or the object information contained therein can be stored in a suitable storage medium or in the working memory of the laser microdissection system. It is advantageous to define a reference position on the corresponding carrier for the selected objects, to which the respective object positions of the selected objects relate, so that when the corresponding carrier is subsequently processed and when the stored object information is loaded, a reference position is used simple approaching and locating the object positions stored for this purpose is possible.

The combination of the selected objects in object groups also enables the positions of the corresponding objects on the slide to be repeated one after the other after laser processing of the objects of a specific object group. drive to check the correct cutting or catapulting of the corresponding objects, this being done both by the operator visually by checking the cutting or catapulting limits on the screen and automatically by computer-assisted evaluation of the cutting or catapulting limits by means of digital image processing or image analysis can.

A pulsed UV laser is preferably used as the laser source of the laser microdissection system. A glass slide can be used as the carrier, which can preferably be coated with a carrier film consisting of a UV-absorbing polymer film with a thickness between, for example, 5 μm and 15 μm, the absorption behavior of the carrier film at the wavelength of the UV laser is adapted and thus preferably has an absorption maximum in the vicinity of the laser wave length. Likewise, support foils stretched on frames or support membrane or Teflon membrane in the form of so-called pet bowls etc. can be used as supports. A collecting substrate, which can be in the form of a film or plate or also in the form of a pot-shaped container, can be used as the collecting device for collecting or picking up objects detached from the material to be processed. In particular, microcentrifuge containers are recommended as collecting devices, as they are used in molecular biology, or the caps thereof, in particular several such collecting containers can be arranged next to one another in order to be able to request different objects in different collecting containers one after the other. Likewise, the use of a microtiter plate with a plurality of wells is possible as a collecting device, so that several objects can be collected one after the other from different wells. The collecting device can be provided with an adhesive layer sem, so that detached objects can be fixed by the adhesive layer. A computer-controlled adjustment device is preferably provided for the carrier and / or the collecting device in order to enable these units to be positioned automatically.

The function of the control means according to the invention explained above is implemented in software in particular in the form of a corresponding control program for the laser microdissection system. The present invention thus relates not only to the laser microdissection system as such, but also to the design of the corresponding control program or the computer-readable storage medium which stores this control program.

The present invention is explained in more detail below with reference to the accompanying drawing based on preferred exemplary embodiments.

FIG. 1 shows the structure of a laser microdissection system according to a preferred exemplary embodiment of the present invention,

FIG. 2 shows the representation of an exemplary image screen image of the laser microdissection system shown in FIG. 1 for the selection and marking of desired biological objects, and

FIG. 3 shows an example of a screen display of the laser microdissection system shown in FIG. 1 with a list which contains biological objects previously selected and marked by a user.

The laser microdissection system shown in FIG. 1 comprises a laser device 4 in which a laser light source for detecting Generation of a laser beam is housed. Furthermore, an optical system 6 is accommodated in the laser device 4, which is required in order to couple the laser beam into a microscope 1 and to match the laser focus in the object plane to the optical focus of the microscope 1. In the present case, it is, for example, a pulsed UV nitrogen laser with a wavelength of 337 nm, a pulse energy of 270 μj, a pulse duration of 3 ms and a pulse frequency of 1-30 pulses / second.

For precise adjustment of the laser energy, a quartz filter 5 is arranged perpendicular to the laser beam path, which can be adjusted manually or automatically via a control panel (not shown) for the appropriate adjustment of the laser energy. In addition to setting the laser energy, the laser focus can also be set independently of the microscope focus, i.e. the focal point of the laser can be shifted in the m z direction relative to the object plane of the microscope 1, the lenses 6 shown in FIG. 1 being able to be moved for this purpose via a stepper motor. This adjustment can also be done both manually and automatically.

The laser beam is coupled into the microscope 1 via a plurality of coated beam splitters and deflected to a 12 μm lens. The diameter of the laser beam impinging on the object plane is largely dependent on the numerical aperture of the objective 12. A lens with a relatively high numerical aperture enables laser beam diameters of less than 1 μm. In addition, care should be taken to ensure that the lens 12 used in each case has a high transmittance for the respective laser wavelength in order to minimize energy losses.

The laser beam emitted via the objective 12 finally strikes a motorized and computer-controlled one Microscope or carrier table 3, on which a carrier with a biological material to be processed is arranged. Above the support table 3 there is a manually operable or preferably likewise motorized and computer-controlled manipulator 2. The components 2 and 3 enable exact object positioning with high precision and the automatic execution of μ manipulation procedures.

The motorized support table 3 can be moved along two linear axes (x / y direction). For example, a needle or micropipette for microinjection can be attached to the motorized manipulator 2. In the context of the present invention, however, it is assumed that a collecting device is attached to the manipulator 2 in order to collect biological objects catapulted away from the carrier. The motorized manipulator 2 can be moved both in the m x / y direction and in the z direction.

The microscope 1 can be any microscope. In particular, the use of an inverse as well as an upright microscope or a laser microscope is conceivable. The laser microdissection system shown in FIG. 1 is an inverse structure in which the laser beam strikes the carrier from below in order to catapult biological objects located thereon upward to the collecting device. In the case of an upright construction, on the other hand, the laser beam strikes the carrier from above, so that objects extracted from the biological material, depending on the laser energy, fall down or are catapulted onto the collecting device located below the carrier.

The microscope 1 is equipped with a video camera, in particular one

CCD video camera ("Charge Coupled Device") equipped, which records the area of the carrier 3 above the lens 12. The video signal of this video camera is fed to a commercially available computer (“personal computer”) 7 and processed there, so that the corresponding video image can be displayed in real time on the screen or monitor 8 of the computer 7. A storage of individual video images on a suitable one is also possible Storage medium of the computer 7 possible. Furthermore, an analog or digital video recorder for recording the video images supplied by the video camera can also be coupled to the computer 7.

As described in more detail below, smd on the computer

7 or implemented various functions by the software running on it, which both a computer-based, i.e. enable automatic control of the laser device 4 and the microscope 1, so that, for example, the laser is automatically activated and the manipulator 2 or the carrier table 3 can be moved automatically. These computer-aided functions also enable a particularly user-friendly selection and processing of desired biological objects of the biological material on the carrier. Conventional input means, such as a keyboard 9 or a computer mouse 10, are provided for setting or selecting these functions. Furthermore, a foot switch 11 is assigned to the laser device 4, by means of which the laser can be activated manually.

The functions provided in the laser microdissection system shown in FIG. 1 will be explained in more detail below.

After turning on the computer 7 will appear on the screen

8 shows the microscope image currently recorded by the video camera, as shown for example in FIG. 2. The laser target point 13 is shown in FIG Shown cross. In addition to this microscope image, software-implemented setting options for setting the laser energy, the laser focus, the laser function, the magnification of the objective lens 12 used, for storing the microscope image, etc., or for calling up further menu windows are shown, which are not described in more detail here to be received. The control program is essentially controlled via the computer mouse 10 shown in FIG. 1, although the essential functions can also be called up by corresponding key combinations on the keyboard 9.

A distinction can essentially be made between two different operating modes.

In the so-called cursor mode, 10 menus can be opened with the mouse, corresponding menu functions selected and so-called buttons clicked. In contrast, in the travel mode, movements of the mouse 10 are converted directly into corresponding adjustment signals and thus corresponding mechanical movements of the support table 3 or the manipulator 2. A status window can be displayed below the microscope image. indicates whether the control is currently in cursor mode or in traversing mode. this status window can also display the X and Y coordinates, which indicate the absolute position (in μm) of the microscope or Define support table 3 in relation to a zero position.

In addition to the functions explained above, so-called graphics tools are shown on the screen 8 in addition to the microscope image, with the aid of which freehand lines or predetermined figures, such as rectangles, circles, straight lines or ellipses, are drawn on the screen 8 or on the microscope image shown can, so that Microscope image with these graphic elements superimposed on the screen 8 is displayed. In addition, a color palette can be displayed on the screen 8, so that the desired color that appears on the screen 8 can be selected for each element drawn. The color selected in each case is stored as a default for all elements of the currently selected element type. In this way it is possible to use different biological objects of the biological material 14 on the carrier 3, which form of the video or Microscope image is displayed on the screen 8 by selecting and marking it with different colors.

In the example shown in FIG. 2, it is assumed that the user has placed a freehand cut line 16 m of a first color, which is indicated by a square in FIG. 2, around two biological objects with the aid of the graphic tools explained above. In a second color, which is indicated by a circle in FIG. 2, two further objects were marked with the aid of corresponding freehand cut lines. Three other objects were selected or marked in an analogous manner by means of a freehand cut in a third color, which is indicated by a triangle. In a fourth color, which is indicated by an asterisk, a further freehand cut film was finally placed around another biological object, but in contrast to the freehand cut film explained above, this freehand cut film is not completely closed. The cutting lines shown in FIG. 2 serve not only for marking and selection of the corresponding biological objects, but also for the subsequent laser processing of the biological material 14 located on the support table 3 as a specification for the automatic relative movement between the support table 3 and the laser beam of the laser device 4 to the laser beam along the to guide the specified cutting line and thus to free the selected biological object from the surrounding biological mass.

In the context of the present exemplary embodiment, it is assumed that the different biological objects 15 are marked in different colors for the subsequent laser processing, all objects marked in one and the same color forming a corresponding object group. Of course, apart from the use of different colors, a different way of marking the individual biological objects on the screen 8 is also possible. For example, as shown in FIG. 2, a different type of graphic representation (FIG. 2 with a rectangle, circle, triangle or star) can be selected for each cutting line. This is only relevant with regard to the fact that the marking is chosen in such a way that the individual groups, in which the biological objects with the same marking are combined, can be distinguished as clearly as possible.

In the example shown in FIG. 2, the graphics tools explained above are set in such a way that each biological object marked or selected with a freehand cutting line 16 is provided with a consecutive number, which is also shown on the screen 8. The biological objects with the numbers "1" and "2", the biological objects with the numbers "3" and "4", the biological objects with the numbers "5" - "7" and the biological object with the number "8 "thus each form an object group. The display of these numbers on the screen 8 can optionally also be deactivated. In addition, the software of the laser microdissection system includes a function with which the distance between two points selected on the microscope image can be measured for example on the Microscope image a starting point can be selected, wherein the measurement of the distance to this starting point takes place during the movement of the mouse 10 when a corresponding mouse button on the mouse 10 is held down. Likewise, the software can have a function for automatically calculating the area of a biological object selected or marked in the previously explained manner, which is delimited by a previously drawn intersection line 16. The area of the respective biological object is then displayed, for example, μm ² on the screen 8 of the laser microdissection system. As a further function, the graphics tools of the laser microdissection system can also have a “radio rubber” function in order to delete graphic elements previously drawn on the screen 8. Likewise, a desired function can be selected at any point on the video image displayed on the screen 8 Text comment can be inserted.

The previously described marking or selection of the desired biological objects 15 serves to select the biological objects desired for the subsequent laser treatment, i.e. those biological objects are determined which are later to be automatically freed and / or catapulted to the collecting device. For the biological objects selected in this way, an entry is generated in a list, which is also shown on the screen 8. The structure of this list will be explained in more detail below with reference to FIG. 3.

In an upper list section of FIG. 3 there is an entry or an element for each previously selected object. In a column A, the individual selected or marked objects are numbered in the order of their marking, analogously to FIG. 2. In a column B is for each Element indicated what type it is, a distinction being made in particular between a type “line” for a cutting line and a type “dot” for a single catapulting point, which can also be determined on the video image using the specified graphics tools. In the example shown in FIG. 3, all elements are pre-drawn lines. Furthermore, the area enclosed by the corresponding section line (preferably in μm ² ) is given in a column C for each selected biological object. In column D, a comment can optionally be added for each entry.

As can be seen from FIG. 3, the marking or color selected in the selection of the respective object is indicated in a further column 18 for each element or for each object.

The list is continuously updated. That during the marking of further desired biological objects by the user on the screen 8, a new entry m in the list is automatically generated for each additionally marked biological object.

In a lower list section 29, a summary is shown in which the objects are sorted and summarized according to object groups or markings / colors. For each marking / color, the number of the objects contained in each column F and the total area of the objects assigned to this object group are specified in a column G. A final line of this list section 29 provides information about the total number of objects of all object groups, ie the total number of all marked objects, and their total area. In the example shown in FIG. 3, a selection field 19 is activated, but when it is activated only list entries of the “Line” type are displayed. However, entries of the “Dot ^x” or text entries etc. type are activated when the selection field 19 is activated not shown.

The list shown can be saved and closed by clicking a button 20. By clicking a button 21, on the other hand, the list can be closed without being saved and thus discarded. By clicking a button 22, all elements of the list as well as their properties (e.g. marking / color, number or type etc.) as well as the summarizing values can be exported to a file.

A button 23 enables all elements of the displayed list to be saved in a file, the position m in relation to a previously selected reference position of the respective biological material being stored in particular for each element or for each biological object. Before saving, this reference position of the wearer or of the biological material located thereon must be determined by computer. This reference position is necessary in order to ensure, when a slide is used again with a biological material that has already been examined, that the biological objects selected and marked there can be approached or positioned correctly. A different reference position can be determined for each slide or for each sample. When the slide is inserted, the desired biological objects are then approached with reference to the previously determined reference position, ie the position data stored for each element or for each selected biological object are relative position data which relate to the previously determined reference position. The user can use an appropriate software function when setting the reference reference position on the microscope image shown, these are also marked with the laser in the biological material, so that later it is easy to find the reference position.

The activation of a button 24 enables the deletion of all elements of the displayed list, while a button 28 only allows the deletion of one or more selected elements of the list. By clicking a button 25, the remaining elements of the list can then be renumbered.

With the help of a button 26, each individual biological object can be approached in such a way that it appears centered on the screen 8. For the centered representation of the object number “5”, only the corresponding line in the upper list section of FIG. 3 has to be marked with the mouse 10 and the button 26 then activated.

Each individual element or each individual biological object can be processed separately with the laser beam. If, for example, the biological object No. “5” is to be processed, only the corresponding entry in the upper list section has to be marked with the mouse and then a laser start button 30 has to be activated. The laser beam is then computer-aided by a suitable relative movement between the laser beam and the carrier table 3 positioned on the desired cutting line of the respective biological object as shown in Figure 2 and moved along the predetermined cutting line in order to prepare the biological object freely. In an analogous manner, several of the selected biological objects can also be marked and then activated by activating the laser start button 30 processed one after the other. It is of particular advantage, however, that a corresponding selection of one or more object groups can also be made in the lower list section 29, so that after the laser start button 30 is subsequently activated, only the biological objects belonging to the selected object group or the selected object groups are processed.

If, for example, tumor cells are to be placed in a collecting container and healthy cells are placed in a further collecting container, it is advisable to mark the desired cells during the movement of the microscope image shown on the screen 3 according to FIG. 2 in such a way that the cut lines of the tumor cells are in a first color and the cut lines of the healthy cells are drawn in a second color. For example, suppose that the color blue is used for marking the tumor cells, while the color yellow is used for marking the healthy cells. A line with a summary of the "blue" tumor cells and a line with the summary of the "yellow" healthy cells would then appear in the lower list section 29. By marking the “blue” line (“high-light mg”), correspondingly positioning the collecting container above the laser beam and then activating the laser star button 30, all the “blue” tumor cells are then processed with the laser beam and - depending on the selection field 32 set laser function - catapulted into the collecting container. Then the "yellow" line can be marked with the mouse 10, a new collecting container positioned over the laser beam and the laser star button 30 activated again so that all "yellow" in the second collecting container healthy cells are caught.

In this regard, the laser microdissection system is advantageously designed in such a way that the different collecting containers for the individual object groups and / or objects objects are automatically positioned and thus moved into the catch position, ie before processing the objects of a selected object group, an appropriate catch container is automatically moved into the catch position, so that the objects em and the same object group are collected in one and the same catch container, which facilitates the storage and subsequent analysis of these objects.

In addition to column 18, a further column 17 is provided in the upper list section, which shows the processing status for each element or for each biological object. As soon as a biological object has been processed with the laser beam after activation of the laser start button 30, the “check box” corresponding to this biological object or this list entry is marked.

For each activation of the laser start button 30, the number of repetitions of the laser treatment can be entered into a further selection window 31 by entering a corresponding value. In the example shown in FIG. 3, only a simple implementation of the laser function set according to the selection window 32 is provided.

As has already been briefly explained above, the selection window 32 can be used to select a specific one of a plurality of predetermined laser functions for each laser processing. With the laser function “RoboLPC” preset according to FIG. 3, when the laser start button 30 is activated, the predetermined cutting line of the respective biological object is automatically traversed to a predetermined residual web and then a separate laser shot is placed on the center of this residual web to expose the desired biological object of the surrounding biological mass to catapult into the collecting container Section line - as in the biological object shown in FIG. 2 with the number “8” - the section line being left open more than would have been the width of this residual web preset in the system, this oversized gap is automatically made by a straight line to the specified one Width of the remaining web reduced.

Another laser function "LPC" can be provided, for example, for setting separate catapulting laser shots, ie without prior preparation, e laser shot is set at the desired location in order to catapult out the corresponding biological object. In certain preparations, such as, for example, cytocentric cells, this can separately set laser shots are sufficient for catapulting out. A further laser function "Cut" can only be provided for cutting along the predetermined cutting line without a subsequent catapulting laser shot being set. The laser shot can then be placed separately at a desired location on the freely prepared biological object using the laser function explained above. Even with this pure cutting function, the respective biological object is preferably not completely exposed, but a narrow remaining web of a predetermined width is left. A further provided laser function "CloseCut" can correspond to the previously explained cutting function, however the drawn cutting line is completely scanned with the laser beam in order to prepare the respective biological object completely freely. If the cutting line was not completely closed by the user, the start and the end point of the cutting line are connected by a straight line from the laser microdissection system in order to obtain a closed cutting line. Another laser function “AutoLPC” that can be set via the selection window 32 can be used to remove a surface or a previously marked area on the microscope image. nes previously marked biological object. When this laser function is selected, the area within the pre-drawn line is removed by a large number of successive laser shots and catapulted into the corresponding collecting container. The number of laser shots per unit area can be set via a corresponding menu in the laser microdissection system. Another laser function "CloseCut &AutoLPC" finally provides a combination of the two laser functions explained above, ie the desired biological object is first separated from the surrounding biological mass with the aid of a completely closed cutting line and then removed by a large number of successive laser shots This procedure is particularly useful if the user wants to rule out any risk of contamination of the biological material to be removed by neighboring biological material.

For each laser processing operation, i.e. for each activation of the laser start button 30, the laser function can thus be set via the selection window 32 and the number of repetitions of this laser function via the selection window 31. On the other hand, each activation of the laser star button 30 can be selected by appropriately marking the desired elements or objects or object groups in the list shown, whether the corresponding process should extend to individual objects or entire object groups.

As has already been mentioned previously, the corresponding “check box” is set for each processed object in column 17 as soon as the processing for this object has been completed. At the same time, the number of cutting and catapulting operations carried out for the corresponding object are recorded and displayed.

The above-described design of the laser microdissection system enables, for example, the different object groups, which are combined biological objects, to be processed with the laser in different ways. For example, only the cutting laser function can be set for the objects of a first object group, while the "RoboLPC" laser function preset in FIG. 3 is provided for the objects of a second object group. In this way, the greatest possible degree of flexibility is achieved.

Claims

1. Laser microdissection system for processing a material (14) located on a support (3), with a laser light source (4) for generating a laser beam to be directed onto the material (14) to be processed in order to carry the material (14) with it to process the laser beam, with an image recording device (1) for generating an image of at least a portion of the material on the carrier (3), with a display device (8) for displaying the image generated by the image recording device (1), with selection means (9, 10) for selecting objects (15) of the material (14) on the carrier (3) to be processed with the laser beam and for assigning the respectively selected object to a corresponding object group, and with control means (7) for Evaluation of the selection of the objects (15) to be processed using the selection means (9, 10) and to create a list of the selected objects and / or the object group n with one

The corresponding object group contains smd, whereby any object group in the list can be selected via the selection means (9, 10) and the control means (7) are designed such that they process the objects (15) assigned to the selected object group. Arrange group-specific with the laser beam.

2. Laser microdissection system according to claim 1, characterized in that an object to be processed (15) can be selected by appropriately marking the object on the image of the material on the carrier (3) represented by the display device (8), and that the control means (7) are designed in such a way that they provide different types of marking, each type of marking being assigned to an object group, so that all objects (15) marked and selected with a specific type of marking form a corresponding object group.

3. Laser microdissection system according to claim 1 or 2, characterized in that the control means (7) are designed such that they create the list with a list section (29) in which the selected objects (15) are summarized the corresponding object groups.

4. Laser microdissection system according to claim 3, characterized in that the list with the list section (29) from the control means (7) is displayed on the display device (8), and that via the selection means (9, 10) the object group in the List section (29) can be selected and the control means (7) are designed such that they cause the objects (15) assigned to the selected object group to be processed with the laser beam.

5. Laser microdissection system according to claim 2 and claim 3 or 4, characterized in that the control means (7) are designed such that in the list section (29) of the list for each object group used in the selection of the corresponding objects (15) Marking is specified.

6. Laser microdissection system according to one of claims 3-5, characterized in that the control means (7) are designed such that the list section (29) of the list contains the number of objects assigned to this object group for each object group

7. Laser microdissection system according to one of claims 3-6, characterized in that the control means (7) comprise an area calculation function for calculating the area of an object (15) selected via the selection means (9, 10), and in that the control means (7 ) is designed in such a way that the list section (29) of the list for each object group contains the total area of all objects assigned to the corresponding object group.

8. Laser microdissection system according to one of the preceding claims, characterized in that the control means (7) are designed such that they select the list with a list entry with object information about the individual via the selection means (9, 10) and with the laser beam generate processing objects (15).

9. Laser microdissection system according to claim 8, characterized in that an object to be processed (15) by marking the corresponding object via the selection means (9, 10) on the image of the display device (8) of the image on the carrier (3) material can be selected so that the control means (7) for marking the objects (15) to be processed provide different types of marking and that the control means (7) are designed such that each list entry of a selected object to be processed with the laser beam ( 15) a representation of the marking of the respective object contains the type of marking used.

10. Laser microdissection system according to claim 8 or 9, characterized in that the control means (7) comprise an area calculation function for calculating the area of an object (15) selected via the selection means (9, 10) and to be processed with the laser beam, and in that the control means (7) are designed in such a way that they generate the list entries with object information which indicate the respective object area for the selected objects (15).

11. Laser microdissection system according to one of claims 8-10, characterized in that the control means (7) are designed in such a way that each list entry contains an indication of whether the corresponding object (15) has already been processed with the laser beam or not.

12. Laser microdissection system according to one of claims 8-

11, characterized in that the control means (7) are designed such that the list entries of the list generated by the control means (7) contain an indication of the type of the corresponding object (15).

13. Laser microdissection system according to one of claims 8-12, characterized in that the control means (7) represent the list on the display device (8), that an arbitrary list entry from the list shown on the display device (8) can be selected via the selection means (9, 10), and that the control means (7) are designed such that they process the objects (15) corresponding to the selected list entries by the Initiate laser beam.

14. Laser microdissection system according to one of the preceding claims, characterized in that the control means (7) in the presence of a corresponding selection via the selection means (9, 10) processing objects selected via the selection means (9, 10) in the list ( 15) or cause object groups with the laser beam in accordance with a laser function that can be selected using the selection means (9, 10).

15. Laser microdissection system according to claim 14, characterized in that via the selection means (9, 10) the desired one

Laser function can be selected from a group of predefined laser functions, the group of laser functions having at least a first laser function for cutting with the laser beam along a cutting line specified by the selection means (9, 10) and a second laser function for setting a laser shot on one of the Selection means (9, 10) predetermined position on the material (14) located on the carrier (3).

16. Laser microdissection system according to claim 14 or 15, characterized in that via the selection means (9, 10) the number of repetitions of processing the objects (15) or object groups selected via the selection means (9, 10) with the laser beam is adjustable.

17. Laser microdissection system according to one of the preceding claims, characterized in that the control means (7) comprise an area calculation function for calculating the area of an object (15) selected via the selection means (9, 10), and in that the control means (7) are designed such that they generate the list with an indication of the total number of all objects selected via the selection means (9, 10) and with an indication of the total area of the objects selected via the selection means (9, 10).

18. Laser microdissection system according to one of the preceding claims, characterized in that via the selection means (9, 10) an object to be processed with the laser beam (15) by corresponding marking of the object to be processed (15) on the display - Device (8) shown image of the material located on the carrier (3) is selected such that the control means (7) are designed such that they provide different types of marking for marking the objects (15) to be processed and all via the selection means (9, 10) assign objects (15) marked with the same type of marking to a corresponding object group, so that each object group includes objects marked with the same type of marking.

19. Laser microdissection system according to claim 18, characterized in that the different types of marking different marking colors with which the objects to be processed (15) on the displayed on the display device (8) Image of the material on the carrier (3) are marked.

20. Laser microdissection system according to one of the preceding claims, characterized in that the list m storage means can be stored.

21. Laser microdissection system according to claim 20, characterized in that the control means (7) comprise a function for determining a reference position on the material located on the carrier (3), and in that the control means (7) are designed such that they are used for Saving the list in the memory means storing positions of the selected objects with the material on the carrier (3) in relation to the reference position.

22. The laser microdissection system as claimed in claim 21, characterized in that the control means (7) are designed such that they comprise a function (26) for the targeted approach to a selected object (15) contained in the list, the control means (7) are designed such that they determine the position of the corresponding object (15) in relation to the reference position and, based on the reference position, cause the carrier (3) to be adjusted such that the corresponding object (15) on the carrier (3) located material appears at a certain point in the image shown on the display device (8).

23. Computer-readable storage medium in which a control program for computer-aided control of a laser microdissection system for processing a material (14) located on a carrier (3) is stored, the laser microdissection system comprising:

- a laser light source (4) for generating a laser beam to be directed onto the material to be processed (14) in order to process the material (14) with the laser beam, - an image recording device (1) for generating an image of at least a portion of the on the carrier (3) material (14),

- a display device (8) for displaying the image generated by the image recording device (1), and - selection means (9, 10) for selecting objects (15) to be processed with the laser beam of the material (14) located on the carrier (3) and for assigning the respectively selected object to a corresponding object group, the control program being designed such that, when executed in the computer-controlled laser microdissection system, the selection of the objects (15) to be processed carried out with the aid of the selection means (9, 10) evaluates, creates a list which can be displayed on the display device (8) and which contains the selected objects and / or the object groups with a description of the respective corresponding object group, and one when selecting any object group from the list via the selection means (9, 10) Processing of the objects (15) assigned to the selected object group in a group-specific manner using the laser beam causes.

24. Computer-readable storage medium according to claim 23, characterized in that the control program is set up in such a way that it executes the function of the control means (7) of the laser microdissection system according to one of claims 1-22 when executed in the computer-controlled laser microdissection system.