DE102005023850B4 - Inverse microscope - Google Patents

Inverse microscope

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Publication number
DE102005023850B4
DE102005023850B4 DE200510023850 DE102005023850A DE102005023850B4 DE 102005023850 B4 DE102005023850 B4 DE 102005023850B4 DE 200510023850 DE200510023850 DE 200510023850 DE 102005023850 A DE102005023850 A DE 102005023850A DE 102005023850 B4 DE102005023850 B4 DE 102005023850B4
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DE
Germany
Prior art keywords
pupil
beam path
modulators
arranged
lens
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.)
Active
Application number
DE200510023850
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German (de)
Other versions
DE102005023850A1 (en
Inventor
Peter Dipl.-Ing. Euteneuer
Ralf Dipl.-Phys. Krüger
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 Wetzlar GmbH
Leica Microsystems CMS GmbH
Original Assignee
Leica Microsystems Wetzlar GmbH
Leica Microsystems CMS GmbH
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Publication date
Application filed by Leica Microsystems Wetzlar GmbH, Leica Microsystems CMS GmbH filed Critical Leica Microsystems Wetzlar GmbH
Priority to DE200510023850 priority Critical patent/DE102005023850B4/en
Publication of DE102005023850A1 publication Critical patent/DE102005023850A1/en
Application granted granted Critical
Publication of DE102005023850B4 publication Critical patent/DE102005023850B4/en
Application status is Active legal-status Critical
Anticipated expiration legal-status Critical

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Classifications

    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS, OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/0004Microscopes specially adapted for specific applications
    • G02B21/0088Inverse microscopes
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS, OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/02Objectives
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS, OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/06Means for illuminating specimens
    • G02B21/08Condensers
    • G02B21/14Condensers affording illumination for phase-contrast observation

Abstract

inverted Microscope (1) for high-contrast imaging of objects (14) with a lens (15) arranged in an imaging beam path with associated Lens pupil (16) or pupil position, a subordinate lens system (17, 19, 22) for generating an intermediate image (23) of the objective pupil and with a at the place of the intermediate image (23) of the objective pupil (16) to be arranged modulator (24) for contrasting an object image, characterized in that different lenses (15) with the same pupil position (30; 31; 32) are provided and the used Lenses (15) a fixed number of such pupil layers (30, 31, 32) and that means (40, 50, 60, 70) are provided is with the for Lenses (15) of different pupil layers (30, 31, 32) suitable, fixedly arranged modulators (24) in the direction of the optical axis to the associated Places of the respective intermediate images (33, 34, 35) of the objective pupils are to be arranged.

Description

  • The The present invention relates to an inverse microscope for high contrast Illustration of objects with one in an imaging beam path arranged lens with associated Lens pupil, a downstream lens system for generating an intermediate image of the objective pupil and with a at the place of Intermediate image of the objective pupil to be arranged modulator for contrasting an object image.
  • inverse Microscopes are characterized in that the to be examined Object is illuminated by transmitted light from above, and that the lenses are mounted under the stage. At inverse Auflichtmikroskopen done both the lighting and the Observation through the lens from below. Such incident light microscopes play an important role in mineralogy and metallurgy, while inverse transmitted light microscopes frequently used for the investigation or manipulation of biological samples become. The main advantage of inverted microscopes is the better accessibility to the object to be examined, since the imaging optics to a large extent below the sample table, ie in the stand, is arranged.
  • Especially Biological samples and small thickness samples may be normal microscopic View as almost transparent. Show such objects usually only different optical thicknesses, while the light amplitude not or over the sample weakened homogeneously becomes. The passage of light through such a sample (phase object) existing optical path differences can be different Contrasting method for make the human eye visible. from the known contrasting methods, like phase contrast, modulation contrast according to Hoffmann, relief contrast, Varial contrast or interference contrast, are only examples here the phase contrast and the modulation contrast as frequent representatives the contrasting explained briefly.
  • At the Phase contrast is an upstream in the illumination beam path Ring orifice from a condenser to infinity. By the ring diaphragm reaching, the sample without bending through penetrating illumination bundle ("zeroth diffraction arrangement") meet in the Focal plane (general pupil) of the lens on a phase ring, that means on an annular, by vapor deposition so tuned layer that against the rays, which penetrate the phase plate adjacent to this layer, reaches a phase difference of λ / 4 becomes. This achieves the following function: While at Amplitude objects the diffracted light over the zeroth order one Phase shift of 180 ° (λ / 2), is with phase objects only 90 ° (λ / 4). By the additional introduced 90 ° shift in the phase ring also result in a total of 180 °, ie equal phase ratios as with an amplitude object. By additional weakening of the Amplitude in the phase ring is the intensity of the zeroth order to the Adjusted diffraction orders. In the intermediate image plane of the microscope results now by interference of the diffraction orders with a an image comparable to an amplitude image. Details with respect to the Environment higher Refractive index appears darker in this image. Of course it is the phase ring tailored to the annular aperture such that the Ring diaphragm is mapped to the phase ring. The phase ring is located yourself in the lens pupil, which is usually inside the lens itself lies. For phase contrast is therefore often used special lenses, in which a phase ring (for example, by vapor deposition on a Lens) is integrated.
  • For the construction A phase contrast layer is usually a combination of used dielectric and metallic layers. The dielectric Layers (for example silicon oxide) serve to adjust the phase shift, the metallic layers (eg chromium) for setting the desired Transmittance.
  • The Choice of suitable phase ring (and thus the associated ring diaphragm) is not only the lens but also the sample to be examined, by the respective transmittance and the respective phase shift is dependent. Furthermore, depending on the desired resolution or depending on the desired Contrast the size of the phase ring to measure. Special lenses with integrated phase ring can open such different requirements do not respond flexibly. This would namely the provision of a wealth of Special lenses require, which requires high effort and high costs. In practice, these are the usual special lenses universally applicable standard solutions, which are often not for special cases the wished Result can deliver.
  • In the so-called modulation contrast is located in a pupil plane of the observation beam path, a plate arranged on which strip-shaped areas of different transparency (usually 0%, 20% and 100%) are located. In this case, since the interference with the diffraction image is not symmetrical to the optical axis of the objective, the phase objects made visible with such a microscope additionally exhibit a relief effect similar to that which occurs when the object is slanted on one side. On the illumination side is conjugated to the modulato ren in the imaging beam path at least one slit-shaped aperture. This is usually mapped onto the strip of the image-side modulator with the medium transmission. Mostly, these slit-shaped diaphragms are located inside a condenser disk, with a special lighting slit being provided for each enlargement.
  • From the EP 0 069 263 is a device for the selective realization of phase contrast and relief contrast in microscopes known. Starting from the problem that the modulators required for the phase and relief contrast are arranged in the objective pupil and thus in the most inaccessible interior of a lens, so that said contrasting methods are generally not compatible with each other, in this document a for both contrasting uniform modulator proposed in the objective pupil. The change between the Kontrastierverfahren is made via a diaphragm change in the illumination beam path. The modulator in the objective pupil consists of a partially translucent plate with two concentric annular layers which influence the amplitude or phase of the light. For the phase contrast observation, an annular aperture is introduced into the illumination beam path in front of the condenser, wherein the annular light-permeable area of the condenser and the objective is imaged onto the phase ring of the modulator located in the objective pupil. For observation in relief contrast, another diaphragm is introduced into the illumination beam path, which has a light-transmissive annular segment-shaped region which is imaged onto the other, only the amplitude-influencing ring of the modulator in the objective pupil. Because of the one-sided oblique angle, under which the light passes through the object plane, then a relief effect occurs, which makes the object appear plastic. Consequently, with this proposed device, it is possible to change between phase contrast and relief contrast solely by changing the diaphragms in the illumination beam path without intervention in the hard-to-reach objective pupil and without exchanging lenses specially made for the particular contrasting method.
  • at this solution mentioned Nevertheless, a special lens is needed, the said modulator consisting of a phase and an amplitude ring, absorbs. Should the size of the phase ring changed or another contrasting method or should be used to work with other lenses, as shown in this solution the initially described limitations in flexibility.
  • Out DE-42 36 803 C2 is a microscope for the examination of amplitude and / or phase objects known. This is in the illumination beam path in front of the condenser a circular sector shutter with a translucent circular sector, the sen tip is located in the optical axis, attached. About condenser and lens, this circular sector is placed on a phase plate, the mounted in the rear focal plane (exit pupil) of the lens is shown. At the location of this figure shows the phase plate a correspondingly formed phase segment, whose tip also in the optical axis, and that is the phase of the transmitted light shifts by λ / 4. The result is a combination in this contrasting process from relief and phase contrast. In this respect, this solution is also regarding their flexibility limited.
  • Finally is from CH-294755 a microscope with a device to achieve known by phase contrast. Here it is proposed by means of a Auxiliary optics an intermediate image of the exit pupil of the lens generate to introduce a phase plate there. This could be on expensive special lenses, which contain a built-in phase plate, be waived. As auxiliary optics here is a telescopic lens system the magnification 1 proposed an intermediate image of the objective exit pupil (hereinafter Intermediate pupil). The phase plate is at the location of the intermediate pupil firmly arranged. When using different lenses with different ones Layers of the intermediate pupils can be the telescopic system for focusing the image of the ring aperture on the Move phase plate along the optical axis. At normal, upright microscopes, it makes sense according to this document, said auxiliary optics and the phase plate alone or jointly displaced to arrange. For ousted (inverse) microscopes, the phase plate is slidable to arrange.
  • By the compulsion, the solution the latter font both the centering as well as the Focusing by means of auxiliary optics with each change of the pupil position It is not necessary to control this change of lens felt as practical.
  • It is therefore an object of the present invention, in an inverse Microscope of the type mentioned without high adjustment different To enable contrasting even with a lens change. The proposed solution in particular, an automated choice of a contrasting method required Modulators suitable for the selected Lens enable.
  • These The object is achieved by an inverted microscope according to the features of the patent claim 1 solved. Find further advantageous embodiments of such a microscope in the subclaims and in the description below.
  • According to the invention are different Lenses with the same pupil position (position of the exit pupil) provided and the lenses used is a fixed number attributable to such pupil positions. Furthermore, a device provided with the for Lenses of different pupil positions suitable in direction the optical axis fixed modulators to the associated locations the respective intermediate images of the objective pupil, in the following Zwischenpupillen, are to be arranged. This allows a quick change the modulator of a particular contrasting depending on from each lens used with associated Zwischenpupille. The Modulators are placed in the freely accessible locations of the intermediate pupils, so that the need for special lenses is eliminated.
  • through the device according to the invention can Modulators not only matched to the lenses used but it is still a simple switch between different contrasting possible. This will be easy at the place of the intermediate pupil the corresponding modulators are exchanged. Furthermore you can to a specific sample type (with certain transmission and phase shift characteristics) trimmed modulators are used. Also different Requirements for resolution and contrast, for example by using different phase rings Size, can Be taken into account. For example, for observation with the human Eye usually desired a high contrast while under observation a camera rather a high resolution stands in the foreground.
  • By Use of the appropriate suitable modulators on the freely accessible Location of the intermediate pupils allows the invention contributes to the flexible realization of different contrasting methods the entire usable range of lenses.
  • Under The term modulator in the present application is a phase plate, a panel or another for understood a corresponding contrasting required modulator, thus So an optical element that the amplitude and / or the phase of the passing light changed and / or the passing light beam spatially limited.
  • A advantageous possible mechanical embodiment of the device for the arrangement of modulators exists in a perpendicular to the imaging beam path displaceable arranged slide, in which in the sliding direction next to each other Modulators are arranged at different heights. By Shift perpendicular to the imaging beam gear then pass different modulators in the imaging beam path. Thereby in that the modulators are related to the direction of the imaging beam path at different heights are arranged, the modulators each arrive at the associated location the intermediate pupil.
  • In another embodiment, the device for arrangement of modulators several, parallel to the imaging beam path arranged Modulators, each of which can be introduced into the imaging beam path are. Here, for example, the device fixed to the microscope be arranged laterally of the imaging beam path. In the facility can the modulators parallel to each other and with predetermined distances from each other be arranged such that a modulator only in the imaging beam path must be pushed to be there at the right place of the intermediate pupil. The Arrangement of the modulators in the device then corresponds to the Arrangement of the intermediate pupils in the imaging beam path.
  • A further advantageous possibility the mechanical configuration of the device according to the invention for the arrangement of modulators is in the nature of a rotatable about an axis Revolvers, as he is known, for example, as an arrangement of lenses is. By rotating the revolver around its axis, the attached to the revolver modulators in the imaging beam path be introduced.
  • It is advantageous if that for generating an intermediate image of a Lens pupil used lens system with respect to the object image Represents afocal system. The Afokalsystem has the advantage that in the Infinity adjusted object imaging beam path interventions (by Mo dulatoren) possible are, without location and size of the object image to influence. An afocal system consists of at least two lenses, the one coming from the infinite ray path to infinity depict.
  • Suitably, the proposed afocal system has a magnification factor of 1, so that the imaging ratios remain unaffected. Due to the afocal system, only the imaging beam path is lengthened, so that the intermediate pupils (intermediate images of the objective pupils) are placed at a suitable location in the tube can, on which a simple external intervention for the arrangement of the modulators is possible. While the afocal system leaves the object image (orthoscopic beam path) unaffected, it images the objective pupil into the respective intermediate pupil planes (conoscopic beam path).
  • With the device according to the invention for arranging modulators it is not only possible for a chosen contrasting method with changing objectives the associated modulators fast and without adjusting effort to the location of the corresponding intermediate pupil but it is also possible between different Change contrasting method. For this purpose, the corresponding Modulators in the imaging beam path and in the illumination beam path arranged. For example, with a fixed lens at the location of his Intermediate pupil once a modulator for phase contrast, the other Time a modulator for Modulation contrast are introduced. For this purpose, for example a slider can be used where the corresponding modulators next to each other at the same height are attached. To be able to consider different lenses, couples can also be used such modulators (ie in each case for phase and modulation contrast) be arranged side by side at different heights. in this connection It should be noted that when changing the contrasting method in the Usually also the corresponding modulator (aperture) in the illumination beam path to change. For the change of the modulators in the illumination beam path are available in general mechanical slide on. The change of contrasting methods lends itself to depending on the preparation the most appropriate method with regard to Resolution, Contrast, object fidelity and examination habits to select can. Furthermore can For several contrasting complementary information about the result in an object to be examined.
  • The Inventive inverse Microscope is preferably suitable for the automation of contrasting methods. Such automation is a great advantage by the operators felt. For this purpose, information about the selected lens (For example, by the position of the nosepiece or by transmission of transponder information of the selected lens) to the device according to the invention transferred to the arrangement of modulators in the imaging beam path (for example, by interposing a logical unit). Correspondingly selected lens (and thus the location of his intermediate pupil) becomes the corresponding Modulator automatically arranged at the location of the intermediate pupil. Such automatic control is in itself for the expert in common Realizable manner, so that details are not mentioned have to.
  • A higher Level of automation can be achieved by depending on selected Contrasting method (and dependent of the chosen Lens) automatically the associated Modulators in the imaging beam path and the illumination beam path be introduced. By transmission the information about the selected Contrasting method and the selected Lens becomes the appropriate modulator at the location of the intermediate pupil arranged. At the same time, the matching modulator (aperture) arranged in the illumination beam path. Details of the realization of a Such automation method are familiar to the expert and Furthermore not the subject of the present invention.
  • Finally, it is possible also within a contrasting method depending on the object structures or even dependent from the viewer (eye or camera) by exchanging modulators For example, the contrast and / or the resolution change. So is with flat object structures often that Main focus on a good contrast, while for thick specimen structures the main focus on the resolution to lay is. Similar is for an observation by the human eye the contrast more important as the resolution, while Vice versa for an observation by a camera applies. For such a design suitable modulators can be used (for one and the same contrasting method) attach to a location of the intermediate pupil and, if necessary, corresponding modulators in the illumination beam path. By change The respective modulators can then images, for example with higher Contrast or higher resolution generate alternately.
  • Next the already mentioned Contrasting methods can be flexibly achieved by the invention in addition to the positive and a negative phase contrast (shift around -λ / 4) or such as spectrally optimized phase or modulation contrast methods or more generally any contrasting method with a modulator in the imaging beam path and any modulator in the illumination beam path realize.
  • Of course it is the invention also for modulators suitable only for the amplitude or the amplitude and the phase of the passing light.
  • in the The following are the invention and its advantages with reference to an in The embodiment illustrated in the following figures will be explained in more detail.
  • 1 shows in a perspective schematic view a typical example of an inverted microscope,
  • 2 shows a schematic view of the conoscopic and the orthoscopic beam path in a preferred embodiment of an inverse microscope according to the invention,
  • 3 represents a section of the conoscopic beam path in an inverted microscope according to 2 showing different positions of objective pupils and corresponding positions of the intermediate pupils,
  • 4 shows a slider for modulators in the imaging beam path in plan view ( 4A ) and in perspective view ( 4B )
  • 5 shows a similar slider, here for modulators of a modulation contrast method, for arrangement in the imaging beam path in a longitudinal section ( 5A ) and in a view from above ( 5B )
  • 6 schematically shows a device for arranging modulators, which are arranged parallel to the imaging beam path and
  • 7 schematically shows again a device for arranging modulators, which is designed as a revolver.
  • An example of an inverse microscope manufactured by the Applicant 1 shows 1 in perspective view. To realize different types of lighting is a transmitted light illumination arm 4 present, which illuminates one on the microscope stage 6 stored object serves from above. For this purpose, a field diaphragm is in the illumination beam path 3 arranged. Below the microscope stage 6 is the nosepiece 5 , by the rotation of different lenses can be introduced into the beam path. To realize a reflected light illumination in this case, two incident light lamp houses 2 provided by a lighting with different types of lamps, so different spectral composition can be implemented. For focusing can be by means of the drive 7 (z-drive) the lens focus in the z-direction move. Alternatively, a z-shift of the microscope stage 6 be realized. Also shown is a documentation port 8th over which a part of the imaging beam path can be extracted for documentation purposes. The tube with the eyepieces is with 11 designated. Also shown is a control panel 10 with associated display 9 ,
  • Clearly visible 1 is the displacement of the imaging beam path in the lower housing part (tripod) of the inverted microscope, so that in space on the microscope stage 6 there is sufficient space for examinations and manipulations of the on the microscope stage 6 make the object. Often, the objects to be examined are in Petri dishes. About the control panel 10 Depending on the microscope type, different microscope settings can be changed, such as, for example, switching the type of illumination, the position of the microscope stage 6 in the three spatial directions, choice of the lens by adjusting the nosepiece 5 , Choice of lens focus on the drive 7 , The corresponding settings can then be shown in the display 9 conveniently visible to the operator below the tube 11 is arranged.
  • With 46 is an opening in the microscope 1 for inserting a slider 40 . 50 (see. 4 and 5 ) with modulators 24 in the imaging beam path (see explanation below).
  • 2 shows a schematic representation of the orthoscopic and conical beam path in a particular embodiment of an inverse microscope according to the invention 1 , Shown is the solid line of the conoscopic, the image of the lens pupil performing beam path and in dashed line the orthoscopic, representing the object image beam path.
  • In 2 is the modulator in the illumination beam path with 12 designated. For the Modulationskontrastverfahren realized here, the modulator 12 a shutter in the form of a lighting slit, which is arranged in the Kondensorpupupille. The condenser is with 13 designated. About this condenser 13 becomes the object 14 illuminated in the object plane. To illuminate the object 14 As a rule, a Köhler type of lighting is chosen, but the critical type of lighting can also be used. Through the lens 15 becomes the object 14 Shown to infinity (orthoscopic beam path), through a tube lens (here second tube lens 25 ) becomes an intermediate image (here second intermediate image 27 ), which can be viewed through one or two eyepieces (shown here only schematically).
  • According to the in 2 illustrated preferred embodiment is an afocal system 28 with magnification factor 1 between lens 15 and (second) tube lens 25 connected. Location and size of the object image remain through the afocal system 28 unaffected. Through the afocal system 28 Once again, rays coming from the infinite are imaged into the infinite (see orthoscopic ray path in 2 ). In the present case, this is afocal 28 from a (first) tube lens 17 , a field lens 19 as well as an achromat 22 , Through the first tube lens 17 and the field lens 19 becomes a first intermediate image 20 of the object 14 generated by the Achromat 22 is again depicted in the infinite. To realize a "U-shaped" beam path are two deflection mirror 18 and 21 intended. It should be noted that instead of the U-shaped beam path and a V-shaped beam path can be realized, in which the beam path from the object 14 directly to the first deflection mirror 18 and from there directly to the second intermediate image 27 runs.
  • Now let us take a closer look at the conoscopic beam path (solid line). The lens 15 the objective pupil is assigned 16 (When imaging parallel beams, the position of the objective pupil corresponds to that of the focal plane of the objective). In the usual today construction of a lens 15 , this in 2 is shown only schematically, is the objective pupil 16 usually in the hard to reach inside of the lens. Through the afocal system 28 becomes the objective pupil 16 in an intermediate image of the objective pupil, in the following the intermediate pupil 23 displayed. According to the invention is at the place of the intermediate pupil 23 a suitable modulator 24 arranged here a modulator suitable for the realization of a modulation contrast method, which consists of three mutually parallel strips of different transmissivity (usually 0%, 20% and 100%). The modulator 24 is firmly in the Zwischenpupille 23 arranged. A displaceable arrangement is not necessary because the intermediate image of the objective pupil 16 in the illustrated construction at the fixed location of the intermediate pupil 23 is produced. Likewise is the Afokalsystem 28 firmly arranged. It forms depending on the selected lens 15 the respective objective pupil 16 into a corresponding intermediate pupil 23 from. Due to the fixed arrangement of the modulators 24 as well as the fixed arrangement of the afocal system 28 omitted in the inverted microscope according to the invention 1 Adjustment and focusing problems.
  • 2 shows the conditions in the realization of a specific contrasting (modulation contrast) with a particular selected lens 15 in a inventions to the invention microscope 1 , From this it is clear that with fixed selected lens 15 a slight change of the contrasting can be done by the corresponding modulators 12 and 24 change. For example, by introducing a ring diaphragm in the illumination beam path as a modulator 12 and by introducing a correspondingly formed phase plate as a modulator 24 in the intermediate pupil 23 a simple transition to a phase contrast method possible. Such a transition can be easily or manually realized by appropriately designed slide. Depending on the selected contrasting method - as already mentioned in the introduction - a phase object 14 in the form of an amplitude image (phase contrast) or in the form of a relief image (modulation contrast). For this purpose, the second tube lens generates 25 a second intermediate picture 27 of the object that can be viewed or viewed by a camera or directly by a viewer (via an eyepiece). For deflection in the inclined tube part is another deflection mirror 26 intended.
  • 3 now clarifies the conditions, as in the usual use of a plurality of lenses 15 available. 3 corresponds to a section of 2 with stronger schematization, where in 3 only the conoscopic beam path is shown. The attention in 3 lies in the different positions of the objective pupils. These pupil layers 30 . 31 and 32 are depending on the lens used 15 set to discrete positions. For example, Applicants' typical objectives have the following pupil positions, defined as the distance between the object and the objective pupil: A: 47.5 mm, W: 40 mm, C: 35 mm, D: 27.5, and E: 20 mm. In this context, it should be mentioned that it is possible by appropriate lens design to assign different lenses of the same class (A, B, C, D or E) of pupil layers. Three representatives of these pupil layers are in 3 With 30 . 31 and 32 designated. By the in 3 shown, based on 2 The lens pupils are discussed in detail 30 . 31 . 32 , in 3 represented by the pupil position 2 (Reference 31 ), with a magnification of 1 in the intermediate pupils 33 . 34 . 35 illustrated, compare intermediate pupil 2 at reference number 34 in 3 , The distance between the pupil layers corresponds to the selected imaging conditions 1 . 2 and 3 that of the intermediate pupils 1 . 2 and 3 , so that, for example, when using the aforementioned lenses of the Applicant a maximum difference of pupil positions or Zwischenpupillenlagen of 27.5 mm results.
  • According to the invention is now in the in 3 used lens 15 the modulator required for the respective contrasting method 24 to the with 34 designated place of the intermediate pupil 2 bring to. When changing the lens 15 with a correspondingly changing objective pupil (for example, pupil position 3 , Reference number 32 ) would then be the corresponding modulator 24 now at the location of the corresponding intermediate pupil (intermediate pupil 3 , Reference number 35 ).
  • Preferably, in a lens change with pupil change of the corresponding modulator 24 automatically at the associated location of the intermediate pupil 33 . 34 or 35 brought in. Possible Means for introducing the modulator 24 at the place of the intermediate pupils 33 . 34 . 35 will be described below. With appropriate training such a device can also be a change of contrasting, z. B. from phase contrast to Modulatonskontrast and vice versa, make. For this purpose, the corresponding modulators 24 be arranged next to each other (see. 3 ).
  • 4 shows a slider 40 for phase contrasting. 4B shows the slider 40 in perspective view. The first opening 41 in this embodiment does not carry a modulator for phase contrast, it is consequently free to pass. The second with 42 termed opening contains a modulator for phase contrast, ie a phase plate, as has been discussed in detail in the introduction to the description. Likewise, the contains with 43 designated third opening in the slide 40 such a phase plate. The phase plates in the openings 42 and 43 are at different locations in the direction of the axis of the respective opening 42 . 43 arranged. The slider 40 can be in an inverse microscope according to the invention 1 in the longitudinal direction of the slider 40 move so that each one of the openings 41 . 42 or 43 is positioned in the imaging beam path. This is in an inverse microscope according to the invention 1 a corresponding opening 46 (see 1 ) for inserting the slider 40 intended. The layers of the phase plates in the openings 42 and 43 of the slider 40 are chosen such that when inserting the slider 40 in the imaging beam path of the inverted microscope 1 the phase plates are each arranged at different Zwischenpupillenlagen.
  • 4A shows the slider 40 in a top view from above. The slider 40 can be adjusted by means of three locking grooves 45 Lock in three different positions on an inverted microscope. The respective position is indicated by the labels 44 clarified. In the position marked "0" is the opening 41 of the slider 40 in the imaging beam path, so that there is no influence on the beam path in this position. In the position marked "5x - 10x" for the corresponding lenses is the opening 42 with the corresponding phase plate in the imaging beam path. These are lenses to which the pupil class A is assigned in the objective program of the Applicant. In the position marked "20x - 63x" for the corresponding lenses is the opening 43 with the corresponding phase plate in the imaging beam path. These are lenses to which the pupil class C is assigned in the objective program of the Applicant. Is the opening located 42 in the imaging beam path, the corresponding phase plate is located at the location of the intermediate pupil of a selected objective. If you choose a different lens of the other pupil class is accordingly the opening 43 in the imaging beam path to place the local phase plate at the correct location of the intermediate pupil of this lens.
  • The in 4 illustrated slide 40 Of course, it is possible to enlarge the slider in the longitudinal direction by attaching further apertures with phase plates, each pupil class being associated with such an aperture. It is also conceivable, instead of such a slider to use several small, which may need to be replaced when changing lenses.
  • 5 shows a slider 50 for modulation contrast method. This slider is used to implement a modulation contrast method instead of the slider 40 in the opening 46 an inverse microscope according to the invention 1 (see 1 ) inserted. Similar to the slider 40 out 4 points the slider 50 three openings 51 . 52 and 53 on, with the first opening 51 represents a free continuous opening, while the openings 52 and 53 corresponding modulators 24 exhibit.
  • As in 5A , in longitudinal section through the slider 50 , shown, are the modulators 24 the respective openings 52 respectively. 53 at different heights (height in relation to the position along the axes 56 and 57 through the openings 52 and 53 ), these layers in turn correspond to the positions of the intermediate pupils in the imaging beam path.
  • 5B shows the view of the slider 50 in plan view from above, which is substantially in accordance with the view 4A equivalent. In this respect, it is related to the corresponding statements 4 directed. In the plan view according to 5B are the modulators 24 to recognize for the modulation contrast
  • The possible embodiments of a slider 50 for modulation contrast (increasing the number of openings or distribution of the openings on several slides) applies above in connection with the slide 40 out 4 Said.
  • 6 shows very schematically a device 60 for the arrangement of modulators 24 in which the modulators 24 par allel to the imaging beam path are arranged and each individually in the imaging beam path can be introduced. The possible speed of movement of the individual modulators 24 with its center on the optical axis 29 is in 6 indicated by the double arrows. With 33 . 34 and 35 are, as in 3 , which designates places of intermediate pupils. At the in 6 represented modulators 24 are phase contrast modulators, in this example a phase contrast modulator 24 for lenses with associated intermediate pupil 34 is introduced into the imaging beam path. The device 60 could be called a drawer. The modulators 24 may be supported in sockets that are to be slid into the imaging beam path.
  • Another possible embodiment of a device for the arrangement of modulators again shows in a schematic view the 7 , This is a revolver 70 with modulators attached 24 , Out 7 it can be seen that the modulators 24 arranged at different heights, which are the positions of the intermediate pupils 33 . 34 . 35 correspond. By turning the revolver 70 around the axis 71 can therefore use different modulators 24 into the intermediate pupils 33 . 34 or 35 be panned. With the modulators 24 in 7 these are in turn phase contrast modulators. In the example shown the 7 is a phase contrast modulator 24 into the intermediate pupil 35 pivoted.
  • 1
    inverse microscope
    2
    Reflected light lamp housing
    3
    Field diaphragm
    4
    Transmitted light illumination arm
    5
    Lens Revolver
    6
    microscope stage
    7
    drive lens focus
    8th
    documentation Sports
    9
    display
    10
    operator
    11
    tube
    12
    modulator in illumination beam path
    13
    condenser
    14
    object
    15
    lens
    16
    objective pupil
    17
    first tube lens
    18
    first deflecting
    19
    field lens
    20
    first intermediate image
    21
    second deflecting
    22
    achromatic
    23
    Intermediate pupil, Intermediate image of the objective pupil
    24
    modulator in intermediate pupil
    25
    second tube lens
    26
    third deflecting
    27
    second intermediate image
    28
    afocal
    29
    optical axis
    30
    pupil position 1
    31
    pupil position 2
    32
    pupil position 3
    33
    intermediate pupil 1
    34
    intermediate pupil 2
    35
    intermediate pupil 3
    40
    pusher for phase contrast
    41 51
    opening
    42 52
    Opening for modulator
    43 53
    Opening for modulator
    44 54
    labeling
    45, 55
    locking grooves
    46
    Opening for slider 40 . 50
    50
    pusher for modulation contrast
    56 57
    Axes through openings 52 . 53
    60
    Facility for arranging modulators
    70
    revolver for modulators
    71
    axis of rotation for revolvers

Claims (10)

  1. Inverted microscope ( 1 ) for contrast-rich imaging of objects ( 14 ) with a lens arranged in an imaging beam path ( 15 ) with associated objective pupil ( 16 ) or pupil position, a subordinate lens system ( 17 . 19 . 22 ) for generating an intermediate image ( 23 ) of the objective pupil and with one at the location of the intermediate image ( 23 ) of the objective pupil ( 16 ) modulator ( 24 ) for contrasting an object image, characterized in that different lenses ( 15 ) with the same pupil position ( 30 ; 31 ; 32 ) and the lenses used ( 15 ) a fixed number of such pupil layers ( 30 . 31 . 32 ), and that a body ( 40 . 50 . 60 . 70 ), which is used for lenses ( 15 ) of different pupil layers ( 30 . 31 . 32 ) suitable, fixed in the direction of the optical axis modulators ( 24 ) to the corresponding locations of the respective intermediate images ( 33 . 34 . 35 ) of the objective pupil are to be arranged.
  2. Inverse microscope according to claim 1, characterized in that the device ( 40 . 50 ) for the arrangement of modulators ( 24 ) a perpendicular to the imaging beam path slidably arranged slide ( 40 . 50 ) in which in the sliding direction next to each other several modulators ( 24 ) are arranged at different heights according to the under different locations of the intermediate images of the objective pupil.
  3. Inverse microscope according to claim 1, characterized in that the device ( 60 ) for the arrangement of a plurality of modulators, in a row parallel to the imaging beam path corresponding to the different locations of the intermediate images of the Ob jective-pupil-arranged modulators ( 24 ), which are each displaceable in the imaging beam path to the corresponding location of the intermediate image of an objective pupil.
  4. Inverse microscope according to claim 1, characterized in that the device ( 70 ) for the arrangement of modulators ( 24 ) one around an axis ( 71 ) rotatable turret ( 70 ), which is arranged such that by rotation of the revolver ( 70 ) modulators mounted therein ( 24 ) are each displaceable in the imaging beam path to the corresponding location of the intermediate image of an objective pupil.
  5. Inverted microscope according to one of claims 1 to 4, characterized in that for generating an intermediate image ( 23 ) of an objective pupil ( 16 ) used lens system ( 17 . 19 . 22 ) with respect to the object image an afocal system ( 28 ).
  6. Inverse microscope according to claim 5, characterized in that the afocal system ( 28 ) is fixedly arranged in the direction of the optical axis.
  7. Inverse microscope according to claim 5 or 6, characterized in that the afocal system ( 28 ) has the magnification factor 1.
  8. Inverse microscope according to one of claims 1 to 7, characterized in that for the realization of different contrasting methods corresponding modulators ( 24 ; 12 ) are to be arranged in the imaging beam path and in the illumination beam path.
  9. Inverse microscope according to one of Claims 1 to 8, characterized in that, depending on the lens selected ( 15 ) automatically the associated modulator ( 24 ) can be introduced into the imaging beam path.
  10. Inverse microscope according to claim 9 and 8 or claim 8, characterized in that, depending on the selected contrasting method, the associated modulators ( 24 ; 12 ) can be introduced into the imaging beam path and into the illumination beam path.
DE200510023850 2005-05-24 2005-05-24 Inverse microscope Active DE102005023850B4 (en)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB0906449D0 (en) 2009-04-15 2009-05-20 Phase Focus Ltd Improvements in imaging
DE102010014501A1 (en) * 2010-04-10 2011-10-13 Carl Zeiss Microimaging Gmbh Inverse microscope
DE102010062341A1 (en) 2010-12-02 2012-06-06 Carl Zeiss Microimaging Gmbh Device for increasing the depth discrimination of optically imaging systems
CN105807413B (en) * 2016-05-18 2018-11-02 麦克奥迪实业集团有限公司 It is a kind of to fall radioglold phase microscope based on light modulation techniques

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CH294755A (en) * 1950-07-21 1953-11-30 Optische Werke C Reichert Akti Microscope with a device for achieving phase contrast and aperture effects.
GB707796A (en) * 1951-02-08 1954-04-21 Leitz Ernst Gmbh Improvements in or relating to arrangements for examining objects by means of phase contrast in colours and/or in polarised light
EP0069263B1 (en) * 1981-07-07 1984-12-19 Firma Carl Zeiss Device for alternately realizing the microscopic observation in phase contrast or in relief
DE8530473U1 (en) * 1985-10-28 1987-02-19 Fa. Carl Zeiss, 7920 Heidenheim, De
DE4036065A1 (en) * 1989-11-15 1991-05-16 Olympus Optical Co Condenser turret for microscopes
DE4236803C2 (en) * 1992-10-30 1996-03-21 Leica Mikroskopie & Syst Microscope for microscopic under investigation amplitude and / or phase objects

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CH294755A (en) * 1950-07-21 1953-11-30 Optische Werke C Reichert Akti Microscope with a device for achieving phase contrast and aperture effects.
GB707796A (en) * 1951-02-08 1954-04-21 Leitz Ernst Gmbh Improvements in or relating to arrangements for examining objects by means of phase contrast in colours and/or in polarised light
EP0069263B1 (en) * 1981-07-07 1984-12-19 Firma Carl Zeiss Device for alternately realizing the microscopic observation in phase contrast or in relief
DE8530473U1 (en) * 1985-10-28 1987-02-19 Fa. Carl Zeiss, 7920 Heidenheim, De
DE4036065A1 (en) * 1989-11-15 1991-05-16 Olympus Optical Co Condenser turret for microscopes
DE4236803C2 (en) * 1992-10-30 1996-03-21 Leica Mikroskopie & Syst Microscope for microscopic under investigation amplitude and / or phase objects

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