DE102007061214A1 - Microscope for observing a sample by means of different contrasting techniques - Google Patents

Abstract

The invention relates to a microscope for observing a sample (13), preferably a biological substance, using various contrast methods comprising a microscope stand (1) and at least partially within the microscope stand (1) extending illumination beam path with optical elements to form a the sample (13) directed beam (11). According to the invention, the microscope is equipped with a device for temporary introduction of a segment diaphragm (14) with a diaphragm opening (16) arranged peripherally to the optical axis (15) of the illumination beam path into the section of the illumination beam path running inside the microscope frame (1).

Description

The The invention relates to a microscope for observing a sample, preferably a biological substance, using various Contrast method comprising a microscope stand and at least one sectionwise within the microscope stand extending illumination beam path with optical elements for forming a sample directed to the sample Beam.

Around at observation of relative thick biological substances by means of a contrast method to get a three-dimensional impression of the sample is a good contrast of vital importance. This is especially true when observed in transmitted light and with a view to a targeted Manipulation on single cells or cell areas. It happens especially on the representation of the deeper in the tissue lying cellular Structures.

To to a certain thickness provides the known differential interference contrast method (DIC) when illuminating the sample with visible or infrared light, results, which clearly outperform the results obtained with other contrasting methods are. However, the differential interference contrast method is also different Details in the tissue increasingly poorer to detect, the thicker the sample is because the light in the tissue is strongly scattered and the degree of polarization decreases dramatically with increasing thickness of the fabric.

Therefore oblique illuminations are used in thicker samples, the sample structures aligned optimally to the incident light also create a three-dimensional impression of the sample to let. Disadvantages of this procedure, however, arise from the inhomogeneity of the small amount of light entering the sample Illumination light, from the preferred representation of only in a certain direction oriented structures of the fabric and therefore mostly insufficient spatial impression.

improvements be introduced by the introduction of a rotatable segment aperture in the Illuminated beam path achieved. The rotation axis of the segment aperture lies in the optical axis of the illumination beam path, while the aperture, for example, in Shape of a ring segment, peripheral to the optical axis of the illumination beam path is arranged. By means of this segment diaphragm is an oblique to the sample directed beam in the form of a Detail taken from a cone of light cone, often as a lightsaber designated. Compared to full beam lighting reduces the lightsaber object-related scattered light, and it is compared to a simple oblique lighting a larger object similarity in the picture achieved. By turning the segment aperture and the associated Change in position of the aperture on one Allow the circle around the optical axis of the illumination beam path structures in all orientations.

Out DE 195 14 358 C2 It is furthermore known to arrange such a segment stop in the light propagation direction for a light-scattering diffuser, for example a diffusing screen.

It Additional modules for microscopes are commercially available, with which the conjugate planes into which the segment aperture and if necessary, the lens in the illumination beam path are to be duplicated. These additional modules have the shape of a tube in which the required for duplication optical elements are housed. The conversion of a Microscope with such a supplementary assembly is done by this outside the microscope stand at the luminaire interface arranged between the lamp attachment and the microscope stand becomes.

functional However, disadvantages arise in this respect, insofar as the additional module not necessarily optimal to the optical properties of the each microscope used is adapted. Furthermore There are disadvantages in ergonomic terms, because the position the luminaire interface for the user of the microscope barely accessible and therefore poorly usable. This Disadvantage is particularly important when the microscope in integrated a measuring stand and additionally surrounded by a Faraday cage.

Furthermore Is it awkward or impossible to be one with one? such additional assembly provided microscope for others Contrasting techniques, such as for the brightfield or differential interference contrast method. This is just when using switching mirrors and a second lighting conceivable.

From that Based on the object of the invention is based on a microscope to propose an uncomplicated and efficient observation the sample using different contrast methods possible is.

According to the invention this is achieved in a microscope of the type mentioned above with a device for temporarily introducing a segment stop with a peripheral to the optical axis of the illumination beam path arranged Blen the opening in the running within the microscope stand illumination beam path.

The Segment aperture should be 360 ° around the optical axis of the Illumination beam path to be rotatably mounted so that the aperture through Turning the segment aperture to arbitrarily aligned sample structures can be adjusted and these are displayed in an optimal contrast can.

In a further advantageous embodiment is provided, in addition to the segment stop a light-scattering element in the within the Microscope tripod extending illuminating beam path, wherein the light-scattering element in the light propagation direction should be located behind the segment panel.

With the subordinate light-diffusing element is better illumination reached the sample, and it becomes the image quality improved, because in the lens pupil additional, for Image formation contributing diffraction orders of the illumination light reach.

The Inserting the segment panel, optionally together with the light-diffusing panel Element, takes place according to the invention advantageously in a first pupil intermediate image plane located in the portion of the illumination beam path is formed, which extends within the microscope stand. For example, this pupil intermediate image layer arises between two lenses L1 and L2.

alternative this can in another embodiment of the invention Microscope structure provided that the segment aperture, If necessary, in turn together with a light-scattering element, is introduced into a second pupil intermediate image plane. These for example, the second pupil intermediate image plane is within one formed in the illumination beam path arranged condenser.

in the The invention also relates to embodiments of microscope assemblies, with which segment apertures with apertures different Size and geometry in the first or second pupil intermediate image plane be introduced temporarily. The so different Segment diaphragms can oppose each other in the illumination beam path be exchanged with a suitable segment aperture one optimal adaptation to the respective observation task to be solved to achieve.

In this regard, The device can be fitted with a bracket for segmental panels equipped, which is displaceable in two end positions, wherein in a first end position, the segment stop or the light-scattering Element is located in the illumination beam path and in the other End position is removed from the illumination beam path.

Conceivable On the other hand, it is also the device for introducing the segment stop or the segment stop and the light-diffusing element as changing device, preferably with a turret wheel, form. The changing device or the turret wheel should be with several brackets for each a segment stop with a specific aperture or for this segment stop and the light-scattering element be equipped. It is advantageous if the changing device a in the illumination beam path to be put empty permanent holder has, so that the illumination light without fading or dimming This holder can pass and interact with the rest optical elements forms a cone of light in the illumination beam path, whose cone tip is directed to the sample and so the observation the sample, for example, in polarization contrast possible is.

Advantageous the device communicates with at least one control element, with the introduction or removal of the segment diaphragm or the Segment aperture and the light-scattering element in or out of the Illumination beam path can be triggered manually.

is the segment stop around the optical axis of the illumination beam path rotatably mounted, the operating element should also trigger a rotational movement of the introduced into the illumination beam path Segment aperture be formed.

Especially user-friendly and clear it is when the device for introducing the segment stop or the segment stop and the light-scattering Element in the Mi kroskopstativ is integrated, the operating element on the other hand is arranged on the outside of the microscope stand.

The Light-scattering element may for example be formed as a diffuser be, and as the light source, a white light source may be present.

optional is a filter wheel when using a white light source for temporary introduction of wavelength filters present to the sample optionally with different colored light to be able to illuminate. It can also be provided, for example, to put an infrared filter in the illumination beam path.

Alternatively to the aforementioned Ausgestal tion variant, it is also conceivable to use as a light source both LEDs that emit white light, as well as LEDs that emit infrared light, with a circuit should be available for alternative switching on the white light LEDs or the infrared light LEDs.

The Invention will be described below with reference to an embodiment be explained in more detail. In the associated Drawings show

1 a view into the interior of a microscope stand from the prior art,

2 the schematic representation of a first embodiment of the microscope assembly according to the invention,

3 the schematic diagram of a second embodiment of the microscope assembly according to the invention.

1 shows in principle the interior of a portion of a microscope stand 1 with a luminaire interface 2 to which a lighting unit 3 is coupled.

The lighting unit 3 has a light source 4 from which light 5 starting, by means of optical elements, such as the lenses 6 to 9 and a condenser 10 , is formed into an illumination beam path, from which finally a beam 11 on a in a sample level 12 located sample 13 is directed. The sample 13 is, for example, a biological substance consisting of several living cells.

The components shown here are used in cooperation with other components of the microscope setup, for example, to observe the sample 13 in the phase contrast transmission method, in the polarization contrast transmission method or in the differential interference contrast transmission method with illumination of the sample 13 , as are common in the prior art.

But also a bundle of rays 11 in the form of a so-called lightsaber can be directed obliquely to the sample, the invention provides the microscope assembly with a device for temporary introduction of a segment stop 14 with a peripheral to the optical axis 15 the illumination beam path arranged aperture 16 equip.

From this institution is in 2 for the sake of clarity, only the segment aperture 14 with the aperture 16 shown, in a position outside the illumination beam path.

Note: In 2 be as well as below in 3 for the components or assemblies already in 1 are shown, the same reference numerals used.

Not shown in 2 a bracket for the segment panel 14 , which is displaceable in two end positions, wherein in a first end position of the holder, the segment stop 14 is held in the illumination beam path, in one between the two lenses 7 and 8th trained first pupil intermediate image plane, while in the other end position of the bracket, the segment aperture 14 is held in a position outside the illumination beam path, so that they have no influence on the formation of the beam 11 Has.

Is the segment aperture 14 outside of the running in the foot of the microscope tripod illumination beam path, can be used with the microscope structure of the contrast method described above. By contrast, by moving the holder from one to the other end position, the segment aperture 14 placed in the illumination beam path, this has, in cooperation with the other optical components with the result that the beam 11 as desired in the form of the already described lightsaber hits the sample.

The constructive possibilities for forming a holder for the segment panel 14 , which is displaceable in two end positions, for example by means of a slide sliding in a straight-line guide, are known from precision engineering and need not be further explained here.

By way of example, the slide can be via mechanical transmission elements 17 with one or more controls 18 connected, which are arranged outside the microscope stand in an ergonomic position. The operating element 18 and the gear members 17 can be designed so that with their operation not only the insertion and removal of the segment aperture 14 in or out of between the lenses 7 and 8th trained pupil intermediate image plane takes place, but also a rotational movement of the segment aperture 14 around the optical axis 15 is triggered to the position of the aperture 16 at the periphery of the optical axis 15 to change and with this change the direction of the light sword striking the sample to vary.

Furthermore, the bracket for the segment panel 14 , the transmission links 17 and the control 18 be designed so that except the segment aperture 14 also a diffuser 19 can be introduced into the illumination beam path, in a position in the vicinity a pupil intermediate image, this position being from the light source 4 seen from the segment aperture 14 should be subordinate.

With the introduction of the lens 19 in addition to the segment panel 14 will illuminate the sample 13 and hence the quality of the sample 13 improved image, since, as already stated, get into the lens pupil additional contributing to the image formation diffraction orders of the illumination light.

3 shows a second embodiment of the microscope assembly according to the invention. The difference to the design after 2 consists essentially in that here the introduction of the segment aperture 14 and optionally also the diffuser 19 not in the first pupil intermediate image plane between the lenses 7 and 8th is provided, but in a pupil intermediate image plane in the condenser 10 is trained.

In this case, instead of the slider integrated into the microscope stand, as above based 2 described, advantageously a changing device can be used with a turret wheel, the multiple mounts for segment panels 14 has, wherein the inserted into the brackets segment panels 14 different apertures 16 have, so that by rotation of the turret wheel a segment stop 14 can be selected and placed in the pupil intermediate image plane whose aperture 16 has a shape with which an optimal adaptation of the sample illumination is achieved to the sample structure or to the currently to be solved observation task.

The operating element 18 is here advantageously at the same position as in the embodiment according to 2 , the transmission links 17 are adapted accordingly constructive.

The operating element 18 and the gear members 17 can also be formed in this embodiment of the microscope assembly according to the invention so that with the operation of the control element 18 both the insertion, the removal and the rotational movement of the segment diaphragm 14 can be triggered to achieve the same effects as they are based on 2 have been described.

The turret wheel should also have an empty holder with no segment stop 14 is placed in the pupil intermediate image plane, but the illumination light can pass through this pupil intermediate image plane without fading or dimming, whereby, as already explained, the observation of the sample in a different contrasting method is possible.

Of course, the turret wheel here also additional holders for lenses 19 each then together with a segment aperture 14 be placed in the illumination beam to the quality of the sample 13 to improve the image obtained.

1

microscope stand

2

Lights interface

3

lighting unit

4

light source

5

light

6 till 9

lenses

10

condenser

11

ray beam

12

sample plane

13

sample

14

sector stop

15

optical axis

16

aperture

17

transmission members

18

controls

19

diffuser

QUOTES INCLUDE IN THE DESCRIPTION

This list The documents listed by the applicant have been automated generated and is solely for better information recorded by the reader. The list is not part of the German Patent or utility model application. The DPMA takes over no liability for any errors or omissions.

Cited patent literature

• - DE 19514358 C2 [0006]

Claims (15)

Microscope for observing a sample ( 13 ) using various contrasting techniques, comprising - a microscope stand ( 1 ), and - at least in sections within the microscope stand ( 1 ) extending illumination beam path with optical elements to form a on the sample ( 13 ) beam ( 11 ) from the light source ( 4 ) radiated light ( 5 ), characterized by - means for temporarily inserting a segment stop ( 14 ) with a peripheral to the optical axis ( 15 ) of the illumination beam path arranged aperture ( 16 ) in the inside of the microscope stand ( 1 ) extending illumination beam path. Microscope according to Claim 1, in which the segment stop ( 14 ) rotatable about the optical axis ( 15 ) of the illumination beam path is mounted. Microscope according to claim 1 or 2, wherein in addition to the introduction of the segment stop ( 14 ) the introduction of a light-scattering element in the inside of the microscope stand ( 1 ) extending illumination beam path is provided, so that the light-scattering element seen in the light propagation direction behind the segment diaphragm ( 14 ) is located. Microscope according to one of the preceding claims, in which - in which within the microscope stand ( 1 ) extending illumination beam path, a first pupil intermediate image plane is formed, and - the introduction of the segment diaphragm ( 14 ) or inserting the segment panel ( 14 ) is provided together with a light-scattering element in this first pupil intermediate image plane. A microscope according to claim 5, wherein the first pupil intermediate image plane is formed between two lenses L1 and L2. Microscope according to one of claims 1 to 3, in which - the insertion of the segment stop ( 14 ) or the segment panel ( 14 ) and the light-diffusing element is provided in a second pupil intermediate image plane, wherein - the second pupil intermediate image plane within a condenser arranged in the illumination beam path ( 10 ) is trained. Microscope according to one of the preceding claims, in which the temporary introduction of segmental diaphragms ( 14 ) with apertures ( 16 ) of different size and geometry is provided in the first or second pupil intermediate image plane. Microscope according to one of the preceding claims, in which - a holder for each segmental aperture ( 14 ) and a light-scattering element is provided, and - the holder is displaceable in two end positions, wherein - in a first end position, the segment diaphragm ( 14 ) or the light-scattering element are located in the illumination beam path and are removed in the second end position of the illumination beam path. Microscope according to one of the preceding claims, in which - the device for inserting the segment stop ( 14 ) or the segment panel ( 14 ) and the light-scattering element is designed as a change device with a turret wheel, wherein - the turret wheel a plurality of holders for each segmental aperture ( 14 ), and wherein - the segment diaphragms ( 14 ) with regard to the geometric shape and / or the size of the apertures ( 16 ). Microscope according to one of the preceding claims, in which - the device for inserting the segment stop ( 14 ) or the segment panel ( 14 ) and the light-scattering element with controls ( 18 ), which - for triggering the insertion or removal of the segment stop, the segment stop ( 14 ) and the light-scattering element in or out of the illumination beam path, or - to trigger a rotational movement of the introduced into the illumination beam path segment aperture ( 14 ) about the optical axis ( 15 ) of the illumination beam path are formed. Microscope according to Claim 10, in which the device for inserting the segment diaphragm ( 14 ) or the segment panel ( 14 ) and the light-scattering element in the microscope stand ( 1 ) and the controls ( 18 ) on the outside of the microscope stand ( 1 ) are arranged. Microscope according to one of the preceding claims, in which the light-scattering element is used as a diffuser ( 19 ) is trained. Microscope according to one of the preceding claims, in which the light source ( 4 ) a white light source is present. Microscope according to claim 13 with a filter wheel for temporary introduction of at least one wavelength filter in the illumination beams gang, preferably an infrared filter. Microscope according to one of claims 1 to 12, in which - the light source both white light radiating LED's as well as infrared emitting LED's, and - A circuit is available as an alternative Switching on the white light LEDs or the infrared LEDs.