DE102006044214B4 - Microscope, in particular a polarization and / or a fluorescence microscope - Google Patents

Abstract

A microscope (1) having an illumination beam path (15) and an imaging beam path (16) extending from an object to a detector and / or a tube (122), and in which first and second deflection means (23, 24) and wherein the imaging beam path (16) is in the form of a wide-field imaging beam path, wherein the first deflection means (23) deflects a first part of the imaging light in a first direction (25), the second deflection means (16). 24) deflects a second portion of the imaging light in a second direction (26) and both deflection means (23, 24) pass a third portion of the imaging light for visual viewing by an operator of the microscope (1) and the first and second deflection means (23, 23; 24) deflect the first and the second part of the imaging light toward a documentation exit (30) of the microscope (1), to which at least one detection device (27) is attached wherein the imaging light deflected by the two deflection means (23, 24) is detectable simultaneously with the detection device (27), wherein the imaging optics (11) generates at least one intermediate image (12) of the object in the beam path of the third part of the imaging light and wherein the two deflection means (23, 24) and the at least one detection device (27) no means are provided which generate a further intermediate image.

Description

The present invention relates to a microscope and in particular a polarization and / or a fluorescence microscope. The microscope comprises an illumination beam path and an imaging beam path. The imaging beam path extends from an object to a detector and / or to a tube. The imaging beam path has an imaging optical unit imaging an object and is designed in the form of a wide-field imaging beam path. In the imaging beam path, a first and a second deflection means are provided. With the first deflection means, a part of the imaging light can be deflected in a first direction. With the second deflection means, a further part of the imaging light can be deflected in a second direction.

A microscope of the type mentioned is, for example, from US 5,926,283 A is known and is used in particular in the life science sector, ie in particular in basic bio-medical research. There are applications in which dynamic cellular processes occur, which are microscopically observed. These processes should also be recorded or recorded or documented, but this causes problems. As a rule, such dynamic processes take place relatively quickly, and it is therefore necessary to record and store many images of the object per time unit in order to appropriately document the temporal change of the object and thus such dynamic processes.

Especially in fluorescence microscopy, individual object areas are specifically marked with several different fluorescence markers. The fluorescent markers are, in particular, molecule building blocks provided with fluorescent dyes which specifically attach to complementary object areas and thus mark these object areas with the corresponding fluorescent dye. Accordingly, to make the specifically marked object areas visible, the fluorescent dyes must be illuminated with light of a suitable wavelength, and the fluorescent light must preferably be supplied to the detection means without the excitation light. If individual object areas with several different fluorescence markers or fluorescent dyes are to be specifically marked and these are to be simultaneously recorded and stored for documentation of the dynamic process, this is currently possible with a fluorescence microscope in conjunction with a color camera, if it is for the different fluorescent dyes used there is a single suitable beam splitter filter which is capable of transmitting only the fluorescent light of the different fluorescent dyes to the detection device and not allowing the respective excitation light to reach the detection device. With such a so-called dual or triple-band filter, however, one is limited to a few fluorescent dyes on the bio-medical application side, so that some experiments are not possible at all due to this limitation. The beam paths of these individual fluorescence channels are coaxial and are detected with one and the same active detector surface.

Therefore, the optical structure according to the US 5,926,283 A the approach of providing a microscope with a comparable unit to a spectrometer, with which an image field-wise object detection of multiple fluorescence channels is simultaneously possible.

An image field-wise object detection in a microscope in this context and in particular in the sense of the present invention is to be understood that the imaging beam path is in the form of a wide-field imaging beam path. In other words, two-dimensional images are generated and / or detected by the object.

The from the US 5,926,283 A known optical beam path has a consisting of a plurality of optical components image-reversing module, which is adaptable to a documentation port of a microscope and which is an intermediate image of the microscope with a further imaging stage to double or quadruple in a position. This is expensive, since many optical components provide and adapt to each other or to adjust. Furthermore, high demands have to be placed on the imaging quality of the further imaging stage, in particular on the monochromatic and chromatic imaging properties, in order to avoid color aberrations in the image for the most high-resolution applications. Transmission losses, caused by the additionally introduced interfaces of the optics used, generate additional false light due to multiple reflections with the consequence of a further reduction in contrast, which is extremely disadvantageous, in particular in the case of weak fluorescence signals.

US 5,926,283 A discloses an achromatic lens spectrometer for displaying a two-dimensional intermediate image at an internal field stop. The light emanating from this intermediate image is collimated with further lenses and subsequently spectrally split. The wavelength-split light is imaged onto a detector.

From the EP 1 424 579 A1 For example, a microscope illumination apparatus is known which comprises a semitransparent mirror which splits the light from a light source into two beams. The two beams are each supplied to an excitation filter and then brought together again by means of a further semitransparent mirror.

DE 10 2004 044 629 A1 discloses an apparatus and a method for the simultaneous detection of multiple spectral regions of a light beam, in particular for the simultaneous detection of multiple spectral regions of a detection light beam in a preferably confocal scanning microscope.

The present invention is therefore based on the object of specifying and developing a microscope of the type mentioned above, with which a simultaneous detection of dynamic processes of an object examined by the microscope with simpler means and improved detection properties is possible.

The inventive microscope of the type mentioned solves the above problem by the features of claim 1.

According to the invention, it has first been recognized that the dynamic processes mentioned at the outset can be documented in a relatively simple and inexpensive manner if the deflection means required for this purpose are integrated directly into the beam path of the microscope and do not provide any further optical imaging. As a result, fewer optical components can be provided in the imaging beam path in a particularly advantageous manner, which on the one hand entails a reduction of the manufacturing costs and, on the other hand, a reduced adjustment effort, since fewer components have to be adjusted to one another. Furthermore, due to the reduced number of optical components, fewer light losses, aberrations and / or (multiple) reflections occur at interfaces in the imaging beam path, so that increased detection efficiency can be achieved with improved imaging properties. This is of great advantage, in particular with normally weak fluorescent light signals. In accordance with the invention, therefore, no additional optical imaging is provided in order to detect the light deflected by the two deflection means with the detection device. By avoiding further or additional imaging in the sense of the present invention, it is to be understood in particular that under certain circumstances the light deflected by a deflection means or that deflected by the two deflection means can be shaped with beam-shaping optical means (eg lenses). However, no optical means is provided which, starting from the conventional or existing beam path of the microscope, generates a further real image or intermediate image in order to detect the light deflected by the two deflection means with the detection device. Therefore, no means for additional optical imaging are preferably provided between the deflection means and the at least one detection device or from the point of the beam path, from which the imaging beam path only serves to detect the light deflected by the deflection means.

In a very preferred embodiment, the first direction, under which part of the imaging light is deflected out of the imaging beam path with the first deflection means, deviates from the second direction, under which a further part of the imaging light is deflected with the second deflection means out of the imaging beam path. Specifically, the two deflection means could deflect the light in the first and second directions substantially in a spatial direction, but the two directions have an angle with each other which is a few degrees, e.g. B. 0.5 degrees to 5 degrees.

Depending on the particular bio-medical application, it may be necessary to simultaneously detect more than just two parts of the imaging light. Therefore, in a preferred embodiment, at least one further deflection means is provided with which a further part of the imaging light can be deflected from the imaging beam path in another direction.

In a preferred embodiment, the light deflected by the deflection means is detected by a detection device. In principle, it is possible to separately detect the light deflected by the two deflection means, in each case with a detection unit of the detection device, for example with a camera which has two CCD chips arranged next to one another. However, the light deflected by the deflecting means is preferably only detected by a detection device. This could be done with a chip formed in the form of a CCD camera detection device.

The imaging light deflected by the deflection means is in each case spatially separated from each other detectable by the detection device according to a preferred embodiment. Thus, the light deflected by the first deflection means is detected with the detection device two-dimensionally or two-dimensionally. The light deflected by the second deflection means is also detected with the detection device in a planar or two-dimensional manner, wherein the two area areas are not or at most only insignificantly (eg on a Border area) spatially overlap and can adjoin one another.

The detection device could comprise a CCD camera or a CMOS camera. The camera could have an active detection area with a diagonal of 1 inch to 1/4 inch, and the active detection area could be as small as 0.5 inch to 0.125 inch since the CCD chip may have other electronics at the periphery, but none there having active detection area.

The first and / or the second deflection means could comprise a beam splitter. Depending on the desired microscopy method, a polarization beam splitter or a wavelength-selective beam splitter could be provided as a beam splitter. If a polarization microscopic application is performed, the beam splitter could comprise a polarization beam splitter.

If a fluorescence microscopic application is carried out, the wavelength-selective beam splitter could be designed in such a way that at least one predeterminable wavelength range of the light is reflectable and a further, specifiable, further wavelength range of the light can be transmitted. The wavelength-selective beam splitter could conventionally comprise a coated glass plate, with the aim of minimizing so-called cross-talk. The cross-talk is then minimized when the excitation light is almost completely faded out or blocked out of the imaging beam path and the fluorescent light is conducted almost completely to the detection unit. Also, the excitation and fluorescent light of other fluorescent dyes should also be hidden from the wavelength-selective beam splitter from the imaging beam path, so that only fluorescent or imaging light of the desired fluorescent dye is detected with the detection device.

In a specific embodiment, the first deflection means could be arranged on a prism, for example in the form of a boundary layer of the prism provided with a coating. The second deflection means could be arranged on a wedge plate or on another prism, wherein the wedge plate could be arranged between the first deflection means and the detection device. In order for the first direction to be different from the second direction, the wedge plate or the further prism could have non-parallel surfaces. On one surface of the wedge plate, the second deflection means could be provided, namely also in the form of a surface provided with a coating. Alternatively, the first and the second deflection means could be formed solely by a wedge plate or a corresponding prism, wherein the two surfaces arranged in the beam path are each provided with a specific coating.

The first and / or the second deflection means could comprise a reflector. In particular, if the first deflection means is designed as a fluorescence beam splitter, it may be expedient for the second deflection means to have a reflector. If both deflection means are then provided in the imaging beam path of the microscope, the object can not be observed with the eyepiece during detection with the detection device as a function of the entire beam path of the microscope.

Such a configuration could also be advantageous in polarization applications. Then, the first deflection means could comprise a polarization beam splitter and the second deflection means could comprise a reflector. The first deflection means could in this case be arranged between the second deflection means and the detection device.

In a very particularly preferred embodiment, the two deflection means are arranged spaced apart from an intermediate image plane in the imaging beam path. Thus, the two deflection means could be arranged in a region of the imaging beam path in which the beam path is collimated or converging or divergent. Although in principle the two deflection means could also be arranged in the immediate vicinity of an intermediate image plane of the imaging beam path, this is not preferred.

In a very particularly preferred embodiment, the first and / or the second deflection means are arranged in a region of the imaging beam path in which a beam splitter of the microscope is usually arranged. Such an area of the imaging beam path could be where usually a neutral, fluorescence, polarization beam splitter or a reflector is arranged. In commercially available microscopes, such beam splitters or reflectors are arranged in filter slides or filter wheels. Thus, at such a point, a reflector or a neutral beam splitter can be reversibly introduced into the beam path, namely the imaging light completely or in a corresponding proportion (depending on the reflection or transmission properties of the neutral beam splitter) in the direction of the documentation of the microscope with the use of a suitable neutral beam splitter, a portion is also directed to the eyepiece and thus for visual observation by a microscope operator. Therefore, the two deflection means can be integrated in a filter slide or in a filter wheel and - Design of the filter slide or filter wheel - are reversibly introduced into the imaging beam path. As a result, in a particularly advantageous manner, a plurality of wide field images can be switched between a conventional microscope mode and a microscope mode with simultaneous detection, as a result of which the microscope according to the invention can be used in a versatile manner for the special life-care applications.

It is therefore very particularly preferable for the light deflected by the first and / or the second deflection means to be deflected to a documentation exit or documentation port of the microscope. At this documentation port with a generally standardized attachment interface, a CCD camera could be adapted, with which simultaneously two or more wide-field images of the same object can be detected.

As already indicated, the first and / or the second deflection means could be arranged on a filter slide or on a filter holder wheel. The filter slide or filter wheel could be used either in the imaging beam path, either mechanically or manually, for the components fitted therein. If at least one of the two deflection means can be introduced into the imaging beam path by motor, depending on the degree of motorisation of the microscope, further components could be motor-activated or deactivated, namely with a correspondingly operator-controlled selection of the required and correct beam splitters, fluorescence filters and / or polarization filters to position the beam path of the microscope.

As already indicated, the first and / or the second deflection means could in principle be arranged in a collimated or in a converging or diverging region of the imaging beam path. If the first and / or the second deflection means are arranged in a collimated region of the imaging beam path, detection of the light deflected by the deflection means or devices is advantageously relatively simple. In this case, in most cases, no additional beam-shaping means are required because even after the deflection of the light by a deflection means is still a collimated beam path. Under certain circumstances, it may be necessary to provide a beam diameter-adjusting optical means or a focusing device, with which the deflected by the deflection means light can be focused on the detection device. This could be done, for example, with a tube lens arranged between the two deflection means and the detection device. In a comparable manner this applies to the case in which the imaging beam path has at least partially an infinity beam path in which the first and / or the second deflection means are arranged.

In the other case, the first and / or the second deflection means could be arranged in a converging or divergent region of the imaging beam path. If the first and / or the second deflection means is reversibly introduced into the imaging beam path, for example with the aid of a corresponding filter slide, a tube lens or another focusing means to be arranged between the deflection means and the detection device would also be reversible and preferably with the two deflection means in FIGS To introduce imaging beam path.

According to a preferred embodiment, at least one beam-shaping means is provided, which is arranged before and / or downstream of a deflection means in the imaging beam path. With the beam-shaping means, the beam path can be converted from a converging or divergent to a collimated beam path, provided it has corresponding properties. It is also conceivable to use the beam-shaping means to convert the beam path from a collimated to a converging or diverging beam path, if this is necessary for the specific design of the imaging beam path. With the beam-shaping means, however, no optical imaging is effected in the sense that a real image or intermediate image is thereby produced.

Most preferably, a diaphragm or a field diaphragm or a hatch is provided, which is arranged in the illumination beam path or in an intermediate image plane in the imaging beam path. The diaphragm could have a rectangular, square or round aperture or passage and impress a corresponding shape on the corresponding beam path. If the diaphragm is provided in the illumination beam path, it is expedient to arrange it at a location where there is a plane corresponding to the focal plane of the objective. Should the diaphragm be provided in the imaging beam path, this could be arranged, for example, at an intermediate image plane, wherein the intermediate image plane likewise corresponds to the focal plane of the objective. The aim of the diaphragm is, in particular, to limit the image field-side size of an image, which can be generated on the detection device due to the deflection of the first and / or the second deflection means. Thus, it is possible, for example, the images generated by the two deflection means side by side on a CCD chip in the form of a CCD camera Detection device side by side and without mapping overlapping image areas.

According to a particularly preferred embodiment, the detection device is designed such that with it within a short time many object images can be recorded and stored. Thus, time-critical observations of events on the object can be documented. Accordingly, one must select the detection device which is able to detect and read or store image data on the one hand in the required temporal resolution and on the other hand has a suitable sensitivity, so that the detected image data also have a sufficient dynamic range, preferably 8 to 16 bits per pixel.

In a preferred embodiment, the microscope according to the invention has an operating state in which the imaging light can be detected by the two deflection means. It also has a further operating state in which the imaging light can be detected in a conventional manner. As already indicated above, it is possible to switch back and forth between the two operating states if the two deflection means can be reversibly introduced into the imaging beam path, for example with the aid of a filter slide. Here it is also conceivable that both operating states can be applied simultaneously, namely, if the two deflecting deflect only a portion of the imaging light in the direction of the detection device and the remaining part of the eyepiece and thus for visual observation by an operator passes. In this case, on the one hand, the operator can observe the experiment in the eyepiece and, if necessary, adjust settings on the microscope (for example, slightly focusing or moving the object laterally with the microscope stage). On the other hand, the course of the experiment is detected or documented with the detection device.

The microscope can basically have an upright or inverse microscope construction. Since micromanipulators and / or microinjectors are usually used in the abovementioned life science experiments, an inverted microscope setup for this area of application seems more appropriate. In any case, the microscope according to the invention may be a microscope with a conventional tripod, wherein a light source suitable for the respective application can be coupled into the microscope or adapted to the microscope stand.

Usually, the imaging optics will have at least one microscope objective. In general, a tube lens could be provided, which is adapted with regard to the imaging properties of the imaging properties of the microscope objective. This is the case in particular in the case of a beam path of a research microscope currently available on the market with a so-called infinity beam path, where the objective has a collimated beam path running substantially parallel to the image. With the tube lens downstream of the microscope objective in the imaging beam path, an intermediate image is generated, which can be viewed with an eyepiece.

An optical arrangement, which can be introduced into a beam path of a microscope, in particular a polarization or fluorescence microscope, comprises a first deflection means and a second deflection means arranged fixedly relative to the first deflection means. With the first deflection means, a part of the imaging light can be deflected in a first direction and in the direction of a detection device of the microscope. With the second deflection means, a further part of the imaging light can be deflected in a second direction and in the direction of the detection device of the microscope. The first direction differs from or differs from the second direction. The first direction and the second direction are selected such that the light deflected by the two deflection means is detectable spatially separated from one another by the detection device.

In a particularly advantageous manner, therefore, a microscope already installed in a laboratory can be upgraded to a microscope according to the invention, namely if an optical arrangement can be integrated or adapted into the microscope. Under certain circumstances, it is even possible to detect with an already provided on the conventional microscope CCD camera (which then performs the function of the detection device) and the images that can be generated by the deflection with the two deflection means. In that regard, the optical arrangement represents a cost-effective upgrade kit or option with which a conventional microscope can be retrofitted to a microscope according to any one of claims 1 to 23 according to the invention. In that regard, to avoid repetition in respect of the components required for this purpose, namely in particular of the first and the second deflection means, reference is made to the preceding part of the description.

There are now various possibilities for designing and developing the teaching of the present invention in an advantageous manner. For this purpose, on the one hand to the claims subordinate to claim 1 and on the other hand to refer to the following explanation of the preferred embodiments of the invention with reference to the drawings. In conjunction with the explanation of the preferred embodiments of the invention with reference The drawing also generally preferred embodiments and developments of the teaching are explained. In the drawing, in each case in a schematic representation

1 a beam path of a microscope according to the invention,

2 in a detailed view of the part of the beam path 1 in which the two deflection means are arranged and

3 in a further detail view of the part of the beam path 2 to which a conventional beam splitter is provided.

Identical or similar components are identified in the figures with the same reference numerals. In 1 is the beam path of a microscope according to the invention 1 which differs from the beam path of a conventional microscope by a modified arrangement, which in the dashed areas 2 and 19 to 22 could be arranged. Specifically, it is the microscope 1 around a fluorescence microscope, though the microscope 1 alternatively or additionally could also be designed in the form of a polarizing microscope or could have a phase and / or interference contrast mode.

First, briefly on the beam path of the microscope 1 received. With the light source 3 is about a Kondensoroptik 4 one in the object plane 5 arranged, not an extra drawn object illuminated. With the lens 6 the light coming from the object is picked up and with the lens 7 after a reflection on the mirror 8th on a first intermediate picture 9 displayed. With the lens 10 and the tube lens 11 becomes the first intermediate image 9 on a second intermediate picture 12 pictured, which is a user of the microscope 1 can look at what with a symbol 13 is schematically indicated for an eye of an operator. Between the first intermediate picture 9 and the lens 10 is the mirror 14 arranged.

Accordingly, the illumination beam path extends 15 from the light source 3 to the object level 5 or to the object. The imaging beam path 16 extends from the object plane 5 to the tube shown only schematically 122 or to the eye 13 of the operator. The imaging beam path 16 is formed in the form of a wide-field imaging beam path. In other words, a two-dimensional image of the object is generated for the operator. The same applies to a detection device, which will be discussed later. At the in 1 shown beam path of the microscope 1 is the orthoscopic beam path 17 with solid lines and the conoscopic beam path 18 dashed lines.

In the imaging beam path 16 out 1 a first and a second deflection means (in 1 not shown) are introduced at locations or areas, which with dash-dotted squares with the reference numerals 2 . 19 to 22 are indicated. Only schematically is in 2 indicated that a first and a second deflection means 23 and 24 at the point 20 out 1 in the imaging beam path 16 can be introduced. In 2 is therefore only schematically the object plane 5 , the objective 6 and the lens designed as a tube lens 7 shown. Alternatively, the in 2 with the reference number 20 edged components also in the reference numeral 2 marked area of the imaging beam path 16 out 1 be introduced. The illumination beam path is in 2 Not shown. From that in the object plane 5 lying object diverges the object light and is from the lens 6 collected and converted into a collimated, infinite beam path. With the lens designed as a tube lens 7 the infinite beam path is focused on one in 2 not shown tube of the microscope. In the in 2 shown representation of the imaging beam path 16 is both the first deflector 23 as well as the second deflection means 24 in the imaging beam path 16 brought in. With the first deflection 23 is a part of the imaging light in a first direction 25 distractible. The beam path of this imaging light is shown by solid lines. With the second deflection means 24 is another part of the imaging light in a second direction 26 distractible. The beam path of this imaging light is shown with dashed lines.

According to the invention that of the deflection 23 . 24 deflected imaging light simultaneously with a detection device 27 detectable without providing an additional optical image for this purpose. Accordingly, between the deflection means 23 . 24 and the at least one detection device 27 no means for additional optical imaging provided. The first direction 25 deviates from the second direction 26 from. In the concrete is the first direction 25 to the second direction 26 arranged at an angle α, which of the angle, the two surfaces of the wedge plate 28 include, depends. In concrete terms, this is the first means of deflection 23 formed in the form of a prism, which has a triangular base. On one side of the prism is a wedge plate 28 appropriate. The wedge plate 28 facing side of the prism is provided with a coating which light of shorter wavelength, in particular green light, in the first direction 25 reflected and which light of longer wavelength in the direction 29 transmitted. The the Prism facing away from the wedge plate 28 is also provided with a coating which light of longer wavelength, in particular red light, in the second direction 26 reflected. The side of the wedge plate facing away from the prism 28 reflected light of longer wavelength passes unimpeded the coated side of the prism. In that regard, the deflection means 23 . 24 out 2 adapted for fluorescence microscopy applications. The deflection means 23 . 24 are accordingly in the form of wavelength-selective beam splitters.

Although in 2 not explicitly shown, could be the first and / or second deflection means 23 . 24 alternatively have a polarization beam splitter, which is similar to the wavelength-selective beam splitter according to 2 is trained. So could also a prism 23 be provided with a triangular base, on which also a wedge plate 28 adapted. Between the prism 23 and the wedge plate 28 could be arranged a linear polarization beam splitter, which linearly polarized light with a in the plane of the 2 lying vibration direction in the direction 25 reflected. On the prism 23 opposite side of the wedge plate 28 could be arranged another linear polarization beam splitter, which linearly polarized light with a perpendicular to the plane of the 2 lying vibration direction in the direction 26 reflected.

Now that could be the second deflecting means 24 have a reflector, with which the entire of the lens 6 coming light in the direction of the detection device 27 is reflected. In this case, no light is going in the direction 29 to the tube of the microscope 1 and thus transmitted to an operator. If, however, to a detection with the detection device 27 at the same time a part of the lens 6 coming light to the tube of the microscope 1 is to be directed and thus an operator simultaneously a microscope image is to be made available, it is expedient, the second deflection means 24 in the form of a beam splitter, which forms a part of the imaging light in the direction 29 pass through.

Although in 2 not shown so it would be conceivable that a further wedge plate and thus a further deflection means is provided, with which a further part of the imaging light from the imaging beam path 16 is deflectable in another direction. This further part of the imaging light could also be detected by the detection device 27 be detected.

2 is removable, that of the deflection means 23 . 24 deflected light from only the one detection device 27 which is detected by the deflection means 23 . 24 deflected imaging light from the detection device 27 each spatially separated from each other. The detection device 27 is formed in the form of a CCD camera.

1 is removable, that the bodies 2 and 19 to 22 of the imaging beam path to which the two deflection means 23 . 24 can be arranged, both from the first intermediate image 9 as well as the second intermediate picture 12 spaced apart.

Most preferably, the first and the second deflection means 23 . 24 in one area 2 of the imaging beam path 16 arranged in which usually the in 1 not shown fluorescence beam splitter and the filter wheel of the microscope 1 is arranged.

In 2 is indicated that of the first and the second deflection means 23 . 24 deflected light to a documentation exit 30 of the microscope 1 is distracted. At this documentation exit 30 is the detection device 27 cultivated, what in the schematic representation of the 2 not shown.

Only schematically is by the reference numeral 31 hinted that in the square 20 drawn optical components, in particular the first and the second deflection means 23 . 24 is arranged on a filter holder not shown, so that in the square 20 provided optical components reversible in the imaging beam path 16 motor or manually be moved.

In 2 are the first and second deflection means 23 . 24 in a converging region of the imaging beam path 16 arranged. They are first and second jet-forming agents 32 . 33 provided, which are formed in the form of lenses. The first jet-forming agent 32 is with respect to the propagation direction of the imaging light (from the object plane 5 to the detection device 27 ) in front of the first deflection means 23 in the imaging beam path 16 arranged. With the first jet-forming agent 32 the convergent beam path is converted to a collimated beam path. Between the two deflection means 23 . 24 and the detection device 27 is the second beam-forming agent 33 arranged. It is thus with respect to the propagation direction of the imaging light, the two deflection means 23 . 24 downstream. With the second jet-forming agent 33 the collimated beam path in the prism is converted to a convergent beam path, wherein the refractive power of the two lenses or of the two beam-shaping means 32 . 33 is dimensioned such that the imaging length is identical to a conventional beam splitter (such as in 3 shown). This will do so in both directions 25 . 26 deflected imaging light in the same way to the detection device 27 imaged or focused, as would be the case with a conventional beam splitter which can likewise be introduced at the location.

With the reference numerals 19 and 22 out 1 are, for example, the locations of the imaging beam path 16 in which the first and / or the second deflection means 23 . 24 in a collimated region of the imaging beam path 16 could be arranged. in these areas 19 and 22 has the imaging beam path 16 an infinite beam path. Should at these points the first and / or the second deflection means 23 . 24 in the imaging beam path 16 can be arranged, it may be appropriate, between the deflection 23 . 24 and the detection device 27 to arrange a tube lens. The first and the second direction 25 . 26 are from the drawing level of 1 out, preferably substantially perpendicular thereto, aligned.

In the first intermediate image plane 9 of the imaging beam path 16 (for the sake of clarity, put something in this regard 1 drawn) is an aperture 34 intended. The aperture 34 has a rectangular aperture. Thus edge light areas of the imaging beam path 16 hidden, so that the simultaneous on the detection device 27 imaged images do not overlap in the border area.

3 shows a section of the imaging beam path 16 which is comparable to the one in 2 shown section of the imaging beam path 16 is. In place of the prism and the wedge plate 28 out 2 is a neutral beam splitter 36 in the imaging beam path 16 introduced, which is also formed in the form of a prism with a triangular base. This will be 50% of the imaging light in the direction of the detection device 27 reflected. The remaining 50% of the imaging light will be in the direction 29 transmitted for visual viewing by the operator.

In 2 is an operating state of the microscope 1 shown in which the imaging light with the two deflection means 23 . 24 is detectable. Accordingly, the microscope is located 1 in 2 in an "operating mode according to the invention". As well in 1 as well as in 3 is another operating state of the microscope 1 in which the imaging light is visually in the conventional manner with the operator's eye 13 is detectable.

In 1 An inverse microscope construction is shown, although in principle also that of an upright microscope is suitable for a microscope according to the invention.

2 shows an embodiment of an optical arrangement 35 moving into an imaging beam path 16 a microscope 1 can be introduced. The microscope 1 is designed in the form of a fluorescence microscope. The optical arrangement 35 has a first deflection means 23 and a relative to the first deflection means 23 fixedly arranged second deflection means 24 on. With the first deflection 23 is a part of the imaging light in a first direction 25 and to a detection device 27 of the microscope 1 distractible. With the second deflection means 24 is another part of the imaging light from the imaging beam path 16 in a second direction 26 and to the detection device 27 of the microscope 1 distractible. The first direction 25 points from the second direction 26 from. The first direction 25 and the second direction 26 are chosen such that the of the two deflection 23 . 24 deflected light each spatially separated from each other with the detection device 27 is detectable.

LIST OF REFERENCE NUMBERS

1

microscope

2

Area of the beam path with respect to a conventional microscope modified optical arrangement

3

light source

4

condenser

5

object level

6

lens

7

lens

8th

mirror

9

first intermediate picture

10

lens

11

tube lens

12

second intermediate picture

122

tube

13

Eye of a server

14

mirror

15

Illumination beam path

16

Imaging beam path

17

orthoscopic beam path

18

conoscopic beam path

19-22

Places or areas of ( 16 ), on which ( 23 ) and ( 24 ) in ( 16 ) can be introduced

23

first deflection means

24

second deflection means

25

first direction

26

second direction

27

detection device

α

Angle between ( 25 ) and ( 26 )

28

wedge plate

29

Direction to the tube of ( 1 )

30

documentation finish

31

filter wheel

32

first jet-forming agent

33

second jet-forming agent

34

cover

35

optical arrangement

36

Neutral beam splitter

Claims (23)

Microscope ( 1 ) with an illumination beam path ( 15 ) and an imaging beam path ( 16 ) extending from an object to a detector and / or a tube ( 122 ) and in which a first and a second deflection means ( 23 . 24 ) and an object imaging imaging optics ( 6 ) are arranged, wherein the imaging beam path ( 16 ) in the form of a wide-field imaging beam path, wherein the first deflection means ( 23 ) a first part of the imaging light in a first direction ( 25 ), the second deflection means ( 24 ) a second part of the imaging light in a second direction ( 26 ) deflects and both deflection means ( 23 . 24 ) a third part of the imaging light for visual observation by an operator of the microscope ( 1 ) and the first and the second deflection means ( 23 . 24 ) the first and second parts of the imaging light towards a documentation exit ( 30 ) of the microscope ( 1 ) at which at least one detection device ( 27 ) is attached, whereby that of the two deflection means ( 23 . 24 ) deflected imaging light simultaneously with the detection device ( 27 ) is detectable, wherein the imaging optics ( 11 ) in the beam path of the third part of the imaging light at least one intermediate image ( 12 ) of the object and wherein between the two deflection means ( 23 . 24 ) and the at least one detection device ( 27 ) means are provided which generate a further intermediate image. Microscope according to claim 1, characterized in that the first direction ( 25 ) from the second direction ( 26 ) deviates. Microscope according to claim 1 or 2, characterized in that at least one further deflection means is provided, with which a further part of the imaging light can be deflected in a further direction. Microscope according to one of claims 1 to 3, characterized in that of the deflection means ( 23 . 24 ) deflected light only by a detection device ( 27 ) is detectable. Microscope according to one of claims 1 to 4, characterized in that the deflection means ( 23 . 24 ) deflected imaging light from the detection device ( 27 ) is each spatially separated from each other detectable. Microscope according to one of claims 1 to 5, characterized in that the detection device ( 27 ) comprises a CCD camera or a CMOS camera, which preferably has an active detection range with a diagonal of 1 inch to 1/4 inch. Microscope according to one of claims 1 to 6, characterized in that the first and / or the second deflection means ( 23 . 24 ) have a beam splitter, preferably a polarization or wavelength-selective beam splitter. Microscope according to claim 7, characterized in that the beam splitter is a wavelength-selective beam splitter and is formed such that at least one predeterminable wavelength range of the imaging light is reflectable and a different, predeterminable further wavelength range of the imaging light is transmissible. Microscope according to claim 7 or 8, characterized in that the first deflection means ( 23 ) is arranged on a prism and that the second deflection means ( 24 ) on a wedge plate ( 28 ) is arranged that the wedge plate ( 28 ) has non-parallel surfaces and that on one surface of the wedge plate ( 28 ) the second deflection means ( 24 ) is provided. Microscope according to one of claims 1 to 9, characterized in that the first and / or the second deflecting means ( 23 . 24 ) have a reflector. Microscope according to one of claims 1 to 10, characterized in that the first deflection means ( 23 ) a polarization beam splitter and that the second deflection means ( 24 ) has a reflector. Microscope according to one of claims 1 to 11, characterized in that the two deflection means ( 23 . 24 ) spaced from an intermediate image plane ( 9 . 12 ) in the imaging beam path ( 16 ) are arranged. Microscope according to one of claims 1 to 12, characterized in that the first and / or the second deflection means ( 23 . 24 ) in a region of the imaging beam path ( 16 ) are arranged, in which usually a beam splitter or a reflector of the microscope ( 1 ), for example a neutral, fluorescent or polarization beam splitter. Microscope according to claim 12 or 13, characterized in that the first and / or the second deflection means ( 23 . 24 ) on a filter slide or on a filter holder wheel ( 31 ) are arranged, which in the imaging beam path ( 16 ) are motor or manually be moved. Microscope according to one of claims 1 to 14, characterized in that the first and / or the second deflection means ( 23 . 24 ) in a collimated region of the imaging beam path ( 16 ) are arranged or that the imaging beam path ( 16 ) at least partially has an infinity beam path in which the first and / or the second deflection means ( 23 . 24 ) and that between the two deflection means ( 23 . 24 ) and the detection device ( 27 ) A tube lens is arranged. Microscope according to one of claims 1 to 14, characterized in that the first and / or the second deflecting means ( 23 . 24 ) in a converging or diverging region of the imaging beam path ( 16 ) are arranged. Microscope according to one of claims 1 to 16, characterized in that at least one beam-shaping agent ( 32 . 33 ), which is a deflection means ( 23 . 24 ) in the imaging beam path ( 16 ) and with which the beam path from a converging or diverging to a collimated beam path or with which the beam path from a collimated to a converging or diverging beam path is convertible. Microscope according to one of claims 1 to 17, characterized in that a diaphragm ( 34 ) is provided, which in the illumination beam path ( 15 ) or in an intermediate image plane ( 9 . 12 ) in the imaging beam path ( 16 ) and which has a rectangular, square or round aperture. Microscope according to one of claims 1 to 18, characterized in that the detection device ( 27 ) is designed such that within a short time many object images can be recorded and stored, so that time-critical observations of events can be documented on the object. Microscope according to one of claims 1 to 19, characterized by an operating state in which the imaging light with the two deflection means ( 23 . 24 ) is detectable, and by another operating condition in which the imaging light is detectable in a conventional manner. Microscope according to one of claims 1 to 20, characterized by an upright or inverse microscope construction. Microscope according to one of claims 1 to 21, characterized in that the imaging optics a microscope objective ( 6 ) and preferably a tube lens ( 11 ) having. Microscope according to one of the preceding claims, characterized in that it is a polarization and / or a fluorescence microscope.

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