EP2064578A1 - Procédé pour examiner un objet avec un microscope et microscope - Google Patents

Procédé pour examiner un objet avec un microscope et microscope

Info

Publication number
EP2064578A1
EP2064578A1 EP07788340A EP07788340A EP2064578A1 EP 2064578 A1 EP2064578 A1 EP 2064578A1 EP 07788340 A EP07788340 A EP 07788340A EP 07788340 A EP07788340 A EP 07788340A EP 2064578 A1 EP2064578 A1 EP 2064578A1
Authority
EP
European Patent Office
Prior art keywords
scan
slide
examined
objects
plane
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP07788340A
Other languages
German (de)
English (en)
Inventor
Frank Sieckmann
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Leica Microsystems CMS GmbH
Original Assignee
Leica Microsystems CMS GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Leica Microsystems CMS GmbH filed Critical Leica Microsystems CMS GmbH
Priority to EP11173796A priority Critical patent/EP2381287A1/fr
Publication of EP2064578A1 publication Critical patent/EP2064578A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
    • G02B21/365—Control or image processing arrangements for digital or video microscopes
    • G02B21/367—Control or image processing arrangements for digital or video microscopes providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • G01N1/31—Apparatus therefor
    • G01N1/312—Apparatus therefor for samples mounted on planar substrates
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0052—Optical details of the image generation
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/24—Base structure
    • G02B21/241—Devices for focusing
    • G02B21/244—Devices for focusing using image analysis techniques

Definitions

  • the present invention relates to a method for examining an object with a microscope, in particular a confocal scanning microscope, and to a microscope for carrying out the method.
  • confocal laser scanning microscopes are used, which can be used in the field of fluorescence microscopy.
  • the present invention is therefore based on the object to provide a method and a microscope of the type mentioned, after which a particularly fast object examination is achieved with the smallest possible volume of data.
  • the above object is achieved by a method having the features of claim 1. Thereafter, a selection of an object to be examined or a plurality of objects to be examined containing scan area in a predeterminable spatial direction. It must be ensured that the selected scan area contains the object (s) to be examined. In the next step, taking a series of images in the scan area along the Direction of space, so that the individual images are uniquely assignable to individual levels in the spatial direction.
  • the use of a suitable image analysis method makes it possible to reliably determine at least one preferred scan plane, which, for example, contains as much object information as possible.
  • the originally selected scan area extending in the spatial direction can be limited. In most cases, a large number of layers originally contained in the scan area can be separated out and ignored if the scan is repeated in the spatial direction. This saves considerable time, especially in the case of long-term measurements, wherein, in addition, significantly smaller amounts of data have to be processed, for example, stored and analyzed.
  • Data volume could be applied to the image analysis method or another image analysis method for determining boundary levels in the spatial direction, so that for at least one object defined by the boundary planes scan area containing object information levels in the spatial direction is definable, thus the respective object contains completely, but has no layers without object information.
  • boundary planes which still contain object information, with planes adjacent to these boundary planes no longer having object information.
  • a stack or stack of levels can be determined, which is sufficient for the complete examination of the object under consideration. Other originally selected layers do not need to be parsed.
  • Objects assigned limited scan areas are determined. In other words, each object to be examined can be assigned to a different limited scan area. In an original
  • Scan area could therefore be several different limited
  • the spatial direction is the Z direction, i. the direction in which the object to be examined is usually viewed by a user.
  • the method could be carried out in a further or even along all spatial directions, in order in the latter case at least one of limited scan areas of the respective
  • the at least one criterion of the image analysis method could include geometry, shape, color, texture, colocalization, environment, timing, or a number of objects or combinations thereof.
  • the criterion with respect to the geometry could be based on surfaces, circumference and roundness.
  • the shape could be directed to a search for, for example, oval shapes, smooth edges, irregular shaped edges, and holes.
  • the texture criterion could be used for differences in stripe patterns, textures, and porosities.
  • the colocalization criterion may indicate that a predetermined number of small objects must be present in each large object to result in selection by image analysis.
  • the criterion of the environment refers to the fact that, for example, every large object must be surrounded by a certain number of small objects.
  • Time behavior can also serve as a criterion in image analysis, since the size or shape of an object, for example in the case of living cells, can be time-dependent.
  • the criterion regarding the number of objects can be used in such a way that the image analysis method could detect when there are a certain predetermined number of objects in the image to be examined or in the plane to be examined. All criteria can be used in any combination.
  • the respective image analysis method could be selected by the user depending on individual requirements. Such a selection of the image analysis method could also be carried out automatically according to predefinable criteria.
  • Object examination be performed.
  • a usual scan of a scan volume in which the object or objects exist in any case could be performed, with captured data stored and analyzed with respect to the location of desired objects in accordance with the invention.
  • a narrowing of the scan area and / or scan volume could already be done to save the recording and management of amounts of data that contain no object information. This is especially beneficial when many scan repeats are required in long-term studies.
  • the examination procedure is therefore almost capable of learning and can considerably reduce examination times.
  • the method according to the invention in no case requires additional time for a conventional object examination.
  • a map may be generated from location coordinates of preferred levels.
  • scan repetitions can be oriented in order to save examination time and data volumes.
  • the selection of a scan area containing a object to be examined or several objects to be examined in a predeterminable spatial direction as one proceeds from a first location coordinate to a second location coordinate could take into account the position of the preferred plane at the first location coordinate. This is based on the consideration that a second location coordinate of a neighboring preferred level does not differ significantly from a first
  • Location coordinate of a first preferred level will differ if there is a substantially uniform arrangement of objects. Thus, a successive construction of a map takes place without the need for an extensive scan along a spatial direction when generating an adjacent location coordinate.
  • the first method step of the method according to the invention consists in selecting a scan area which contains an object to be examined or several objects to be examined. This first selection is usually done by means of an estimate. In order to carry out this estimation as efficiently as possible, the selection of a scan area containing an object to be examined or several objects to be examined could take place in a predeterminable spatial direction taking into account the surface shape and / or structure of a slide. This is based on the consideration that an object to be examined arranged on a slide usually and inevitably is positioned very close to the surface of the slide.
  • the estimate of the scan area can be tuned to the known surface position. In this case, one could select a scan region which has the position of the surface of the object carrier and deviates from this position by a predefinable distance in the plus direction and the minus direction of the spatial direction. There is a high probability that the object will be within the estimated scan area. It is therefore particularly advantageous to know the surface shape and / or structure of a slide precisely in order to be able to automatically determine limited scan areas and scan volumes along preferably flat examination areas on the slide. The entire successive construction of a map of location coordinates of preferred levels and / or limited scan areas and / or scan volumes is achieved by considering the
  • the surface of the slide could be triangulated from at least four known points.
  • the surface of the slide could be approximated by n-corner or polygon.
  • the surface of the object carrier could be approximated from n known points by a polynomial fit.
  • the approach to the surface shape and / or structure of the slide could be based on manually determined points.
  • individual points could be measured manually by the user.
  • the known points could also be determined automatically by the microscope.
  • the known points starting from the
  • Surface of the slide can be divided into multiple scan fields and, after determining the position of a first scan field in the Determining the position of an adjacent scan field, the position of the first scan field are taken into account. As a result, a successive determination of the surface of the object carrier could be carried out.
  • a regular or irregular pattern could be introduced or introduced onto the slide on one of its surfaces or into the interior of the slide.
  • Such a pattern greatly facilitates the presentation and determination of the surface or shape and structure of the slide.
  • the pattern could be black and white or colored and / or have fluorescent dots or areas. In any case, make sure that the pattern can be detected safely by the microscope.
  • the fluorescent dots or areas could react to a predeterminable wavelength. This allows optical separation of the fluorescent spots or areas of a fluorescent object to be examined.
  • the pattern could be used to calculate, triangularise or approximate the surface of the slide.
  • the pattern could be applied to a substantially planar slide.
  • the pattern could be applied to a microtiter plate.
  • the pattern could be applied in a particularly simple manner between the pots and / or in the pots of a microtiter plate.
  • a different wavelength of the illumination light could be used in the method for determining the preferred plane than in the object examination. This also applies to the determination of limited scan areas and scan volumes.
  • the wavelength used in the object inspection could be shorter than the wavelength used to determine the preferred plane and the limited scan area or scan volume. It is considered that illumination light with a longer wavelength usually causes less damage to the object than illumination light with a shorter wavelength.
  • the method could be repeated after specifiable and / or ascertainable time intervals.
  • time-dependent temperature drifts could be counteracted.
  • this behavior could be used to define time-dependent scan areas and scan volumes. In other words, at different times, a long-term measurement could be different
  • Scan areas and scan volumes are scanned. Such a requirement could, for example, be present in time-varying objects. This would allow further optimization of the examination procedure.
  • the initially formulated object is further solved by a microscope for carrying out the method according to the invention.
  • a microscope for carrying out the method according to the invention.
  • the present invention can be used in particular in confocal laser scanning microscopes.
  • the invention is used for preferably automatically finding and setting a preferred, characterized by selectable features image plane.
  • a use with both normal lenses and with immersion lens lenses is possible.
  • both a preferred focal plane and an optimized scan region in a spatial direction can be determined in order to completely detect a three-dimensional object.
  • the field of application of the invention covers all imaging methods in which an optical cutting is made possible, which thus produce a series of images in which no or only a few fuzzy regions are contained.
  • an adaptive autofocus takes place.
  • a scan range module can be coupled to a conventional microscope in order to enable the method.
  • the method can also be used with microscopes that use structured illumination or use the spinning disc technique.
  • the method according to the invention does not search for the sharpest image plane in the concrete, but for the image plane with the most information regarding certain selectable features.
  • a feature-related detector has advantages.
  • a jump prediction is possible, as it were, in which the focus is searched for independently of wavelength in an adjustable probability range.
  • the method is adaptable to different image contents, wherein such an adjustment can be done manually or automatically.
  • the entire method is capable of learning due to the consideration of previously determined data.
  • a focus map can be created for surface scans. Furthermore, a survey of the
  • Fig. 1 in a sketch a Z-stack of images by spherical
  • Fig. 5 is a schematic representation of the determination of a limited
  • Fig. 6 shows the sequence of a refinement of the scan areas of on a
  • Fig. 8 is a schematic representation of a calculated skew
  • Scan areas starting from the estimated plane 10 shows in a flowchart the determination of a plane
  • 1 1 is a schematic representation of a triangularization
  • 13 is a flowchart of a method sequence for
  • 17 is a schematic representation of a to the points of FIG.
  • 19 is a schematic representation of a scan area divided into a curved scan area
  • 21 is a schematic representation of the jump prediction in the scan of the first three fields of a matrix
  • FIG. 22 is a schematic plan view and a side view of a scanned slide and FIG Fig. 23 in a schematic plan view and in a schematic
  • FIG. 1 is a schematic representation of a Z-stack or z-stack of images taken through spherical objects.
  • the Z-stack consists of six cutting planes - slices - which are identified by the letters A, B, C, D, E and F. Each slice - has a different amount of objects.
  • the objects in the sectional images differ in shape, size and number.
  • the following table characterizes the individual sections in terms of shape, size and number.
  • Fig. 2 shows a z-stack of images through spherical and pyramidal objects.
  • the z-stack consists of six cutting planes - slices - marked with the letters A to F.
  • Each cut contains a different amount of objects.
  • the objects in the sectional images differ in shape - cuts through spheres are circular, by pyramids square, size and number.
  • SOI preferred levels
  • Slice D could also be considered the best plane for the pyramidal object.
  • slice D or slice E is detected as the best level.
  • classifiers on the other hand, can be better evaluated in pure number form, for example the number of objects.
  • a combination of both types of classification can be used, namely fuzzy classification and conventional classification.
  • feature extractions such as "if more than 20 relatively small objects were found, then choose that plane" are possible.
  • FIG 3 shows a schematic representation of two scan stacks in the Z direction.
  • the left stack shows an originally selected scan area and the right stack shows the limited or optimized scan area.
  • the level is of greatest informational content and the start area for the z-scan and the stop area for the z-scan area are shown.
  • the start area and the stop area can be determined automatically by the method according to the invention. All outside the optimized Scan Scheme lying scan sections have no or no relevant object information.
  • FIG. 4 shows an exemplary embodiment of the method according to the invention in a flowchart.
  • An optimal scan volume is characterized in that it encloses the object to be examined as closely as possible. All object information lies within the optimal scan volume, while there is no object information outside of the optimal scan volume.
  • FIG. 5 shows this in a schematic representation, wherein an adapted limited scan volume is formed after variation of the Z, X and Y scans. The individual scans are performed with the proviso to automatically determine the layer with the most object information and the optimal scan area.
  • Vopt V (Dx, Dy, Dz)
  • the point of maximum information Pbest is to the searched object.
  • Pbest does not have to be in the center of Vopt, but always within Vopt.
  • the method described above is described for a single scan point, the method being summarized as follows:
  • Fig. 6 shows the situation before the estimation and after clarification in this regard.
  • the scanning area with the objects to be examined is shown.
  • the objects are distributed over the scan area at different X-Y-Z positions.
  • the middle picture shows a view from the side on the scan surface.
  • the objects to be examined are not equidistant from the support surface, which is present, for example, in the form of a slide. They are in different Z positions. Since the exact location of the objects has not yet been determined, an estimate of the scan area is started, which captures the objects with certainty.
  • the scan range of the estimate is always greater than or equal to the optimal scan range. The highest information position is always within the scan area. Therefore, before the specification:
  • FIG. 7 shows three points in the case of a confocal microscope, the Z stacks showing the three image stacks recorded at the three points of the plane.
  • the plane shown in Fig. 8 has been clearly described with reference to the three points. In order to determine the point that lies exactly on the plane - slide - two variants have been realized.
  • Variant 1 comprises a manual setting, whereby the operator searches for an object and manually sets the highest information position by adjusting the Z-plane.
  • an automatic adjustment can take place, wherein the system automatically searches for the position of the highest information using the search method described above.
  • Fig. 10 is a flowchart showing the scheme of the implementation of the previous embodiment.
  • Fig. 1 1 shows schematically in Fig. 1 1, wherein in the upper part of a flat surface is adopted as an approximation.
  • the surface was triangularized according to the lower part of FIG. 11, ie subdivided into a plurality of triangular planes, which allow a better adaptation to a curved surface than a single surface.
  • a division into two individual planes takes place, which are at a favorable angle to each other.
  • the number of triangularizing triangles can be increased and an ever better approximation of the "estimation surface" to the actual, generally curved surface of the scan object, preferably a slide, can be achieved. Then, this estimate, previously based on the assumption of a plane and now piecewise adaptable to a curved surface by triangularization, is followed by a precision scan in which, as described above, the exact highest order Z positions and the optimum scan range for each point This basic procedure is set forth in the flow chart shown in Fig. 13. Unlike the previous embodiment, more than three interpolation points are now taught in. The xyz values of the interpolation points are then used to intersect the surface
  • a triangle is placed between three points - subset of the n scanned points. Subsequently, as before, a precision scan is performed. In addition to triangles, you can approximate a surface by other surface types such as n-corner or polygons.
  • Fig. 14 shows examples with squares.
  • n set points P (x "y ,, z y )
  • P P (x "y ,, z y )
  • the square of the distance of a curve point from the corresponding measuring point is minimized.
  • FIG. 15 shows a two-dimensional curve fit. The line represents a B-spline fit through the measurement points.
  • an approximation of a surface course is carried out in such a way that the surface represents the surface of an object, preferably a slide.
  • a set of points within a given scan area is measured. Since the scan surface can be curved, a plane equation is not suitable as a basis for the determination of all surface values from the measurement of a few points, preferably three.
  • the surface is approximated by the measurement of some points and by a subsequent three-dimensional fit with a function that places a curved surface through the measurement points such that the pitch of the corresponding points becomes minimum.
  • a family of measured points is shown. Each point has an xyz coordinate and lies on the surface of the slide or object whose surface shape is to be estimated.
  • the curved surface of the function that has been fitted is the new treasure plane, that is, the plane where objects, preferably cells, lie approximately.
  • the new treasure plane that is, the plane where objects, preferably cells, lie approximately.
  • Fig. 18 shows a flow chart for carrying out the last-described embodiment.
  • FIG. 19 shows scan fields which lie adjacent approximately in the same Z position. If one thus knows the exact Z position of the slide surface in a scan field, one can make a prediction of its neighboring positions with respect to the Z direction for its neighboring scan fields.
  • the current scan field is the start field whose exact surface position in the Z direction is known.
  • the three adjacent adjacent scan fields must therefore have approximately the same surface position in the Z direction. So, if you move the slide table to the next scan field to look for the surface position in the Z direction, then you already know where to look, which saves a lot of time and fully exploits its advantage when the scan field is very large, for example having 386 fields
  • the scan follows the surface to a certain extent, using already acquired scan data for a jump prediction.
  • FIG. 20 shows, according to FIGS. A to e, a method sequence in which Z-planes of highest information about all points of a scan matrix are found by means of jump prediction.
  • image a in a first step, the first field is scanned over a large area to find the highest information plane.
  • this plane position is stored for use in the next scan field.
  • the Z position of the measurement of the first field is transmitted to the neighboring field in the sense of a jump prediction. It is assumed that the Z-position of highest information of a field does not change appreciably when the table is moved to the immediate neighboring field and when the neighboring field is not far away.
  • the Z position of the measurement of this field is transmitted to the neighboring field similarly as described in the step according to c in the sense of a jump prediction and then proceeded with this field as described in the step according to d.
  • each jump prediction may be applied in the scan direction to every three neighbors.
  • FIG. 21 shows a corresponding scheme for this purpose, this being a schematic representation of the jump prediction during the scan of the first three fields of a matrix.
  • the advantage of a method by jump prediction lies essentially in its speed. In this case, it is possible to dispense with determining an estimated value with subsequent clarification of the actual position. Particularly in the case of biological objects to be examined, photochemical reactions can occur in the illumination, which can lead to a fading of the fluorescent dyes used. Such bleaching is also called bleaching. The bleaching results in a decrease in the luminous intensity of the dye and therefore requires an increase in the laser beam intensity in order to obtain enough light for an observation.
  • This effect depends on the wavelength used and more energetic radiation - blue, UV - more pronounced than red wavelengths or IR radiation, where this effect is weaker or not at all.
  • Noise ratio the load on the sample during the focus scan can be further reduced.
  • the analysis of a cell reveals a variety of detectable features. These features are usually stained with fluorescent labels and thus detectable.
  • the objects within the cell are thus different in shape, number, color, spatial and temporal distribution, wherein a temporal distribution change, for example, occurs when the cell divides.
  • Many objects are also colocalized, i. For example, they are always within a cell.
  • the nucleus is close to the slide" or "in the cell division phase enlarges the cell and the nucleus has a greater distance to the slide".
  • the required hardware for carrying out the method according to the invention could be a confocal-operating microscope unit and a precisely operating Z-adjustment device.
  • the adjustment can also work manually.
  • the precision may drop.
  • the image acquisition can take place via a photomultiplier or a CCD, which in conjunction with, for example, a structured illumination or "spinning
  • the z-stack should contain the object or objects that the system should focus on.
  • an image analysis algorithm is selected manually or automatically with which the Z plane and the required scan area can be determined.
  • the image analysis algorithm is adapted by an expert - human or machine - to the respective problem.
  • the features are linked so that the desired Z-plane - the one with the most information about the searched objects - can be found by comparing the measured feature values with preset limits.
  • the start and end points of the scan area are determined, which is characterized in that it completely captures the searched object or objects, but does not scan additional slices in which there is no information.
  • the algorithm for this does not have to match the algorithm used in 4..
  • Fig. 22 shows a slide having a regular or irregular pattern on one of its surfaces or in its interior.
  • the pattern can be colored or black and white. It can also consist of fluorescent dots. In the case of fluorescence, it is possible in particular to choose an embodiment of the specimen slide in which the fluorescent pattern reacts to a specific wavelength.
  • FIG. 22 shows a slide 1 with an applied spot pattern 2 for focusing in plan view.
  • the slide 1 with the applied dot pattern 2 is shown in a side view.
  • objects 3 are arranged to be examined. These are, for example, cells.
  • the dot pattern 2 can be applied to a slide 1, for example by embossing. However, a bedding or projection of the pattern is also conceivable. This pattern can also be focused.
  • the dot pattern could be periodic to be filtered out of the image by Fourier filtering. Alternatively, the dot pattern may be statistically available to be statistically filtered from the image signal. Dot patterns can be made from high contrast colors or from
  • the focusing method may use an artificially applied dot pattern instead of the present cells.
  • Fig. 23 shows a microtiter plate 1 which can be used instead of a glass or plastic slide.
  • the microtiter plate has a dot pattern between the individual preparation pots, wherein the spaces between the pots of the microtiter plate 1 can be used for focusing.
  • the applied dot pattern is designated by the reference numeral 2.
  • the pots of the microtiter plate are with the
  • the lower part of FIG. 23 shows the microtiter plate 1 in a side view.
  • the space between the individual pots 3 of the microtiter plate 1 is identified by the reference numeral 4.
  • the interspace bottom of the microtiter plate 1 with the dot pattern 2 is identified by the reference numeral 5.
  • the reference numeral 7 shows the bottom of a potty 3 of the microtiter plate 1 without dot pattern.
  • the bottom of the microtiter plate 1 could also have a dot pattern.
  • the microtiter plate 1 has in the empty spaces between the individual pots 3 in their bottom area an applied dot pattern for the purpose of focusing on.
  • the bottom regions of the pots through which they are microscopically also may have dot patterns, which preferably fluoresce at a particular wavelength and can thus be focused upon using an appropriate excitation wavelength.
  • the point patterns are either periodic or random, in order to be filtered out in a suitable manner.

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Abstract

Procédé pour examiner un objet avec un microscope, notamment un microscope à balayage à foyer commun, présentant les étapes consistant à : sélectionner une zone à balayer contenant un objet à examiner ou plusieurs objets à examiner dans une direction spatiale pouvant être prédéfinie; enregistrer une série d'images dans la zone à balayer le long de la direction spatiale de sorte que les images individuelles puissent être associées explicitement à des plans individuels dans la direction spatiale; sélectionner un procédé d'analyse d'image avec au moins un critère pouvant être prédéfini concernant l'objet à examiner ou les objets à examiner, et finalement appliquer le procédé d'analyse d'image à chacun des plans afin de déterminer un plan préférentiel qui satisfait au moins largement au critère ou aux critères pouvant être prédéfinis. L'invention concerne également un microscope destiné à mettre en œuvre le procédé.
EP07788340A 2006-09-06 2007-08-09 Procédé pour examiner un objet avec un microscope et microscope Withdrawn EP2064578A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP11173796A EP2381287A1 (fr) 2006-09-06 2007-08-09 Procédé d'examen d'un objet à l'aide d'un microscope et un microscope

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE200610042242 DE102006042242A1 (de) 2006-09-06 2006-09-06 Verfahren zur Untersuchung eines Objekts mit einem Mikroskop und ein Mikroskop
PCT/EP2007/058278 WO2008028745A1 (fr) 2006-09-06 2007-08-09 Procédé pour examiner un objet avec un microscope et microscope

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EP2064578A1 true EP2064578A1 (fr) 2009-06-03

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EP07788340A Withdrawn EP2064578A1 (fr) 2006-09-06 2007-08-09 Procédé pour examiner un objet avec un microscope et microscope

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JP4954800B2 (ja) 2007-06-06 2012-06-20 オリンパス株式会社 顕微鏡撮像システム
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