Optical module for a microscope device, microscope device, and use thereof

Abstract

The invention relates to an optical module (10) for a microscope device (100), in particular for observing fluorescent substances to be observed, comprising a filter assembly (35, 35a), which can be rotated about a longitudinal axis (12) and which has a plurality of filters (29a, 30, 30a, 31, 31a, 32, 32a) for light, in particular light of different wavelengths, and an element (28), in particular a disc-shaped element for placing an illumination source (24, 26) and / or image-capturing devices (20, 22), said element (28) being rotatable relative to the filter assembly (35, 35a) about the longitudinal axis (12).

Description

[0001] Description

[0002] Optical module for a microscope setup, microscope setup and its use

[0003] Technical field

[0004] The invention relates to an optical module for a microscope, in particular for observing fluorescent substances, which is characterized in particular by a particularly compact and cost-effective design with high resolution. Furthermore, the invention relates to a microscope using an optical module designed according to the invention, as well as a preferred use of the optical module or the microscope.

[0005] State of the art

[0006] From DE 11 2008 003 363 T5, a microscope device is known which is characterized by a CMOS sensor that interacts with a lens and with a filter arrangement rotatable about an axis, comprising several filters for light of different wavelengths. The individual filters can be alternately aligned with the CMOS sensor, and the object under investigation can be illuminated by means of a light source. Using the microscope device known from the prior art, an observation area of ​​the object can be analyzed for different substances; however, the observation area of ​​the object is relatively small.

[0007] DE 11 2021 001 620 T5 discloses a detector with multiple light-receiving units, which can be arranged in an array. DE 10 2023 009 817 B4 discloses a microscope device for observing the fluorescence of a sample, in which several CMOS sensors are present. However, details regarding the specific arrangement of the CMOS sensors are not provided in this document.

[0008] Disclosure of the invention

[0009] The optical module according to the invention for a microscope device, in particular for observing fluorescent substances, with the features of claim 1, has the advantage that, in connection with the use of several image-taking sensors, it has a structurally relatively simple and compact design.

[0010] The invention is based on the idea of ​​arranging filters for light of preferably different wavelengths and at least one illumination source on two different, mutually adjustable elements and designing the element containing the filters in such a way that illumination of the object to be examined by the illumination source through the element containing the filters is made possible.

[0011] In light of the above explanations, an optical module according to the invention for a microscope device with the features of claim 1 therefore comprises a filter arrangement rotatable about a longitudinal axis, comprising several filters for light, in particular of different wavelengths. Furthermore, a body, in particular disc-shaped, is provided for arranging an illumination source and / or image-receiving devices, wherein the body is rotatable about the longitudinal axis relative to the filter arrangement, and wherein the filter arrangement has at least one through-opening to emit light from the illumination source towards an object, i.e., in particular to allow it to pass between the filters. Preferably, the filter arrangement comprises several through-openings for improved homogeneous illumination of the object. The filter arrangement can also have a body, in particular disc-shaped, wherein the filters and the through-openings are arranged in the body.According to advantageous embodiments, the body comprises the light source and / or one, preferably several, image-capturing devices, in particular cameras.

[0012] Advantageous further developments of the optical module according to the invention for a microscope device, in particular for observing fluorescent substances, are listed in the dependent claims.

[0013] A particularly preferred embodiment of the optical module provides that the optical module comprises image-capturing devices, preferably cameras, preferably CMOS sensors, each interacting with a lens, which can be aligned with the filters of the filter arrangement. The image-capturing devices, preferably together with the lenses, are capable of detecting different observation areas with a common detection area of ​​the object. In particular embodiments, the lenses for the image-capturing devices can also be omitted. This embodiment makes it possible, in particular, to combine the image sections captured by the individual image-capturing devices or CMOS sensors into a relatively extended or large image or observation area of ​​the object. However, the image sections do not necessarily have to be combined into a single overall image.Rather, four individual images of different areas of the object can be generated. In particular, a composite image for each filter can be created if the filter arrangement and the object are rotated completely relative to each other, capturing a section of the image through that filter with each image-taking device and adding it to the composite image. For example, with four filters and four image-taking devices, a total of four composite images for each filter can be generated by just one complete relative rotation of the filter arrangement and the object around their longitudinal axis. It can also be advantageous if the image-taking devices capture areas of different sizes with different resolutions.When using CMOS sensors, these can be, in particular, CMOS sensors known from the mobile communications sector, which are therefore relatively inexpensive and available with high (image) quality. Especially advantageous in the latter design of the optical module is when the lenses, together with the image-taking elements and the body, form a single, integrated unit.

[0014] In another preferred design arrangement, the light source is arranged between the filter arrangement and the lenses when viewed in a longitudinal direction. This makes it easy to attach the lenses together with the light source, for example, to a common element or support.

[0015] In a preferred embodiment of the optical module, it can further be provided that the optical module has four lenses and image-taking devices, each arranged at 90° intervals around the longitudinal axis. Such an arrangement makes it possible to examine a very large section of the object and simultaneously has a centrally located (common) detection area that can be captured by all four image-taking devices or CMOS sensors.

[0016] For example, the filters in the filter arrangement can include at least a blue filter, a green filter, and a red filter. Preferably, these are high-quality interference filters with precisely defined spectral transmission ranges, tailored to the specific application and, in particular, efficiently blocking the excitation light. This enables the detection of three different visible wavelengths of fluorescence using visible light.

[0017] In a preferred further development of the last proposal, the filter arrangement includes an additional pass without a filter or a pass with a clear glass. Such a design makes it possible, in addition to evaluating the individual wavelengths through the different filters, to generate images of the object that depend solely on the wavelength of the light source.

[0018] Furthermore, it is preferred that the light source comprises multiple LEDs. Such LEDs offer the advantage of a particularly compact design as well as high and narrowband light emission. In conjunction with the aforementioned (color) filters, the LEDs are preferably designed as 3-color or RGB LEDs with individually controllable red, green, and blue LEDs. This allows for particularly high image quality when using the appropriate color filter, through a reduction in light scattering, light reflection, and background radiation.

[0019] The prerequisite is always that the emission spectrum of the LEDs used, possibly limited by a bandpass filter, does not overlap with the passband of the camera filter used. In other words, when an image is captured, the area being observed is not illuminated by radiation transmitted by the filter currently in front of the image capture device (camera).

[0020] Alternatively, the lighting source can also comprise one or more other light sources, in particular a laser beam source and optionally optical fibers connected to the laser beam source. The end faces of the optical fibers facing away from the laser beam source are designed to emit light and thereby illuminate the object. According to a particular embodiment, the lighting source can include optical fibers, in particular glass fibers, to which an external light source, in particular a laser light source or an LED, can be connected. In particular, if the lighting source comprises a laser beam source, an optical fiber for connection to a laser beam source, or a monochromatic LED, the filter arrangement can also, according to a particular embodiment, incorporate this lighting source.

[0021] In a particular embodiment, the apertures of the filter arrangement for the passage of light from the illumination source can also include filters, hereinafter referred to as pass-through filters. Particularly when using a white LED or a multi-colored LED as the illumination source, the apertures can incorporate multibandpass filters for selecting several disjoint wavelength ranges. Preferably, these pass-through filters are selected such that the passable wavelength ranges of the pass-through filters do not overlap with passable wavelength ranges of the other filters in the filter arrangement, and preferably at least not with passable wavelength ranges of the filters directly adjacent to the respective pass-through filters. This prevents, or at least reduces, the risk of light passing through the apertures being detected by the image-capturing devices via the other filters.Thus, in the case of fluorescence excitation on the object, the risk of excitation radiation being detected by the imaging devices in addition to fluorescence radiation can be avoided or at least reduced.

[0022] Furthermore, the invention relates to a microscope device with an optical module designed according to the invention as described so far.

[0023] The microscope device preferably includes a control device designed to generate a single image of the object with the respective observation area and to generate a composite image of the object from the individual observation areas by detecting and aligning the common detection area.

[0024] Another particularly preferred design of the microscope device provides that at least the optical module, the filter assembly, and the illumination source are arranged in a common housing. Such a housing, for example a hollow cylindrical one, allows for a compact arrangement of the elements or assemblies required for operating the microscope device. Furthermore, the isolated arrangement of these elements within the housing minimizes disruptive influences from the environment.

[0025] Furthermore, it may be provided that the optical module, together with the lenses and the image-taking devices for focusing the object along the longitudinal axis, is arranged adjustable within the housing.

[0026] Furthermore, it is particularly preferred that the housing be connectable to a slide holder, preferably interchangeably arranged on the housing. The slide serves to hold the sample or object to be examined, allowing it to be prepared outside the actual microscope setup and then attached to the housing. This enables a particularly simple and efficient examination of multiple samples or objects with short changeover times. Moreover, the possibility of a rigid connection or coupling with the other (relevant) optical elements of the setup reduces the influence of external vibrations on the object under investigation. The slide holder can include an additional light source, for example an LED, to illuminate the object from below, thus achieving particularly homogeneous illumination.The slide holder can also be heated to change the temperature of the object depending on the application.

[0027] Finally, the invention also includes the use of an optical module and / or a microscope device according to the invention, in particular for observing fluorescent substances in an object containing the substances in a microfluidic sample arrangement. As already explained above, the microscope device, in particular the optical module, can be used to combine several image sections captured by at least one image-taking device into a complete image.

[0028] Further advantages, features and details of the invention will become apparent from the following description of preferred embodiments of the invention and from the drawings.

[0029] Brief description of the drawings

[0030] Fig. 1 shows a perspective and schematic representation of parts of a microscope device according to the invention,

[0031] Fig. 2 shows a front view of an optical module of the microscope device,

[0032] Fig. 3 shows a perspective view of a light source and a filter arrangement rotatably arranged relative to the light source.

[0033] Fig. 4 is a view of the arrangement according to Fig. 3 in the direction of arrow IV in Fig. 3, Fig. 5 is a side view of a microscope device with a housing and

[0034] Fig. 6 shows a view of the arrangement according to Fig. 3 in the direction of arrow IV in Fig. 3, in an arrangement modified compared to Fig. 4.

[0035] Embodiments of the invention

[0036] Identical elements or elements with the same function are provided with the same reference numbers in the figures.

[0037] The figures show a highly simplified representation of the essential components of a microscope apparatus 100 for observing fluorescent substances on an object O. The microscope apparatus 100 can be, by way of example and without limitation, a component of a larger apparatus. In particular, the microscope apparatus 100 can be used to examine biological samples containing the substances being observed, which are arranged on a chip as object O. The chip represents a microfluidic sample arrangement.

[0038] The microscope device 100 has an optical module 10 comprising several lens modules 14 to 17, in the exemplary embodiment four, each arranged offset from one another by 90° about a longitudinal axis 12, with lenses 18 not individually identifiable. On an end face of each lens module 14 to 17 facing away from the object O, an image-receiving device 20 in the form of a CMOS sensor 22 is arranged, rigidly coupled to the respective lens module 14 to 17. The image-receiving device 20 or the CMOS sensor 22 could, for example, be a CMOS camera from Sony designated ISOCELL JN1.

[0039] On the side of the lens modules 14 to 17 opposite the CMOS sensors 22, a light source 24 is arranged. In the illustrated embodiment, the light source 24 comprises several, in particular five, LEDs 26, which are arranged between the lens modules 14 to 17 and, as shown in Fig. 2, are aligned around the longitudinal axis 12 or at the corners of an imaginary square. Fig. 2 also schematically shows that the optical module 10 comprises two bodies 28, 34, in particular disc-shaped bodies, which are rotatable relative to each other about the longitudinal axis 12. The first body 28 can include the light source 24, 26 and / or the image-receiving devices 20, 22, and the second body 34, as a filter disk, comprises the filter arrangement with the filters and the through-holes.

[0040] The LEDs 26 interact with a single broadband or multiband filter (not shown) to filter the wavelengths of the LEDs 26 and are arranged, for example, as shown in Fig. 3, in the area of ​​a disk-shaped body 28. The body 28 is fixed in position relative to the lens modules 14 to 17 or forms a common assembly with them.

[0041] Viewed in the direction of the longitudinal axis 12, several (light) filters 30 to 32 are connected to the side of the object O facing the LEDs 26 and the lens modules 14 to 17. These filters are arranged in the area of ​​the filter disk 34, which is rotatable about the longitudinal axis 12 in accordance with arrow 33. The filter disk 34 forms a filter arrangement 35, which can be rotated (intermittently) about the longitudinal axis 12 by means of a drive (not shown) such that, as shown in Fig. 4, the circular filters 30 to 32 can be positioned longitudinally with the lens modules 14 to 17.

[0042] Filters 30 to 32 serve to detect the substances being observed and, when using visible light, include, for example, a blue filter, a green filter, and a red filter. Furthermore, the filter arrangement 35 has a passage 36, i.e., not a filter, in the form of a through-opening, in the area of ​​which a clear glass element (not shown) can optionally be arranged. The arrangement of filters 30 to 32 and the passage 36 is such that they are each offset from one another by 90° around the longitudinal axis 12. Through-openings 37 are also formed in the filter disk 34, aligned with the LEDs 26, in order to allow the light from the LEDs 26 to be emitted in the direction of the object O (Fig. 4).According to a particular embodiment, the filter disc 34 comprises only one passage opening 37, preferably, in particular centrally, arranged between the filters 30, 31, 32 and the filterless passage 36, in particular for the passage of light from only one light source 26, in particular an LED 26.

[0043] Figure 1 shows that each of the individual CMOS sensors 22 or lens modules 14 to 17 can detect a (circular) observation area 38 of the object O. In particular, it can also be seen that the arrangement of the lens modules 14 to 17 or the CMOS sensors 22 is such that the (different) observation areas 38 have a common, central detection area as the total set of all observation areas 38, which is detectable by all four CMOS sensors 22. This (common) detection area serves to combine the individual images captured successively by the CMOS sensors 22 and lens modules 14 to 17 into a common image B. For this purpose, the pixels or elements extractable from the common detection area can be processed by software using a control unit 45 to combine the four individual images.To summarize observation areas 38 to the image B with an enlarged image section of object O with high resolution compared to the individual observation areas 38 or partial images.

[0044] Figure 5 shows that the microscope device 100 also has a housing 50, which is designed, for example, in the form of a round hollow cylinder. The housing 50 has a domed area 52 on its upper surface, in which, for example, the drive motors for rotating the filter assembly 35 and the control unit 45 for image processing (not visible in Figure 5) are arranged. Below the area 52 is a lens module area 54 for arranging the lens modules 14 to 16 and a carrier plate 55 that accommodates the CMOS sensors 22. Adjoining the lens module area 54 is an area 56 for accommodating the illumination source 24 and an area 58 for accommodating the filter assembly 35 or the filter disk 34.

[0045] At the lower end of the housing 50, opposite area 52, an interface 59 is provided via which a slide holder 60 can be interchangeably flanged to a slide 61 holding the object O. This allows several objects 0 to be prepared for examination on each slide holder 60, so that the different objects O can be changed or exchanged on the microscope setup 100 with minimal setup time. Slide holders 60 can also be used for differently shaped slides 61.

[0046] Fig. 6 shows another exemplary embodiment with a filter disc 34a and a filter arrangement 35a, in which the feedthroughs, i.e. the through-openings 37, for the light of the illumination source 24, 26 are also fitted with filters 29a, 30a, 31a and 32a. Filters 30 to 32 are for the image-taking devices and have exemplary transmission ranges of 650nm-700nm (filter 30), 450nm-490nm (filter 31) and 540nm-580nm (filter 32). The smaller diameter filters 29a, 30a, 31a and 32a, i.e. the transmission filters described above, for the red / green / blue LEDs 26, have transmission ranges of 510nm-530nm (filter 32a), 620nm-640nm (filter 30a) and 420nm-440nm (filter), or as multibandpass filters, combinations thereof, as with filters 29a and 31a.In this example, filter 31 is flanked by filter 29a with combined transmission ranges of 420-440 nm and 510-530 nm on one side, and filter 31a with transmission ranges of 420-440 nm and 620-640 nm on the other. If only the blue LED 26 is switched on, the area viewed by the camera or image-taking device 20 with filter 31 is illuminated from both sides via filters 29a and 31a with 420 nm-440 nm light. However, this light does not penetrate filter 31, and thus only the excited fluorescence in the area of ​​filter 31 reaches the camera or image-taking device 20. The wavelength ranges mentioned are only examples and do not necessarily have to be explicitly red / green / blue.

[0047] The microscope apparatus 100 described so far can be modified or adapted in a variety of ways without deviating from the inventive concept. For example, it is possible to increase the detection range or resolution by adjusting the distances between the object O under investigation and the focal lengths of the lenses 18. Furthermore, the illumination source 24 can alternatively be implemented using laser light with different wavelengths. This is achieved using a laser beam source and optical fibers connected to the laser beam source, the end faces of which emit light towards the object O. Such a configuration has the advantage that the aforementioned filter can be omitted.

Claims

Claims 1. Optical module (10) for a microscope device (100), in particular for observing fluorescent substances, comprising a filter arrangement (35, 35a) rotatable about a longitudinal axis (12), which has several filters (30, 31, 32) for light, in particular of different wavelengths, and a body (28) in particular shaped like a disc for arranging an illumination source (24, 26) and / or image-taking devices (20, 22), wherein the body (28) is rotatable about the longitudinal axis (12) relative to the filter arrangement (35, 35a), and wherein the filter arrangement (35, 35a) has at least one, preferably several, passage openings (37) to emit light from the illumination source (24) in the direction of an object (O).

2. Optical module according to claim 1, characterized in that this module has image-taking devices (20), preferably each cooperating with a lens (18), in particular cameras or CMOS sensors (22), which can be aligned with the filters (30, 31, 32) of the filter arrangement (35, 35a), wherein different observation areas (38) with a common detection area (40) of the object (O) can be detected by, preferably the lenses (18) and the image-taking devices (20).

3. Optical module according to claim 2, characterized in that the lenses (18) together with the image-receiving devices (20) and the body (28) form a common assembly.

4. Optical module according to claim 2 or 3, characterized in that the light source (24) is arranged between the filter arrangement (35) and the lenses (18) when viewed in the direction of the longitudinal axis (12) and preferably has several LEDs (26).

5. Optical module according to one of claims 2 to 4, characterized in that four lenses (18) and image-receiving devices (20) are provided, each arranged 90° apart from the longitudinal axis (12).

6. Optical module according to one of claims 1 to 5, characterized in that the filters (29a, 30, 30a, 31 , 31 a, 32, 32a) of the filter arrangement (35, 35a) comprise at least one blue filter, one green filter and one red filter.

7. Optical module according to claim 6, characterized in that the filter arrangement (35, 35a) comprises an additional passage (36) without a filter or a passage (36) with a clear glass.

8. Optical module according to one of claims 1 to 7, characterized in that at least one, preferably several of the through-holes (37) have filters (29a, 30a, 31a, 32a), in particular multibandpass filters, wherein preferably passable wavelength ranges of the filters (29a, 30a, 31a, 32a) do not overlap with passable wavelength ranges of the other filters (30, 31, 32) of the filter arrangement (35, 35a), preferably at least not with passable wavelength ranges of the filters (30, 31, 32) directly adjacent to the respective filters (29a, 30a, 31a, 32a) of a through-hole (37).

9. Optical module according to one of claims 1 to 8, characterized in that the illumination source (24) comprises a laser beam source and optical fibers connected to the laser beam source.

10. Microscope device (100) comprising an optical module (10) configured according to any one of claims 1 to 9.

11. Microscope apparatus according to claim 10, wherein a control device (45) is provided which is configured to generate a single image of the object (O) with the respective observation area (38) and to generate a composite image (B) of the object (O) by detecting and aligning the common detection area (40) of the individual observation areas (38).

12. Microscope apparatus according to claim 10 or 11, wherein at least the optical module (10), the filter arrangement (35, 35a) and the illumination source (24) are arranged in a common housing (50).

13. Microscope device according to claim 12, characterized in that the optical module (10) together with the lenses (18) and the image-receiving devices (20) for focusing the object (O) along the longitudinal axis (12) is adjustably arranged within the housing (50).

14. Microscope device according to claim 12 or 13, characterized in that the housing (50) can be connected to a slide holder (60) preferably arranged interchangeably on the housing (50) for receiving at least one slide (61) for an object (O).

15. Use of an optical module (10) according to one of claims 1 to 9 and / or a microscope device according to one of claims 10 to 14, in particular for observing fluorescent substances of an object (O) containing the substances in a microfluidic sample arrangement.

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