APPARATUS FOR ESTABLISHING THE EFFECT OF ACTIVE INGREDIENTS ON NEMATODES AND OTHER ORGANISMS IN AQUEOUS TESTS

Abstract

The invention relates to an apparatus (1) for establishing the effect of active ingredients on nematodes and other organisms in aqueous tests, comprising a holder (40) for receiving a cell culture plate (50) with wells (57), in which the nematodes with the active ingredients can be filled, wherein the cell culture plate (50) has a bottom side (51), a top side (52) and a plurality of side faces, which extend between the bottom side (51) and top side (52), a camera (20) that serves to record images of preferably the bottom side (51) of the cell culture plate (50), and an illumination device (30) for illuminating the cell culture plate (50), wherein the illumination device (30) comprises a plurality of light sources. According to the invention, in the use position of the cell culture plate (50), provision is made for the light of a first light source (31) to enter the cell culture plate (50) from the outside through a first side face (53), the light of a second light source (32) to enter the cell culture plate (50) from the outside through a second side face (54), the light of a third light source (33) to enter the cell culture plate (50) from the outside through a third side face (55) and the light of a fourth light source (34) to enter the cell culture plate (50) from the outside through a fourth side face (56). Moreover, the invention relates to a method for adjusting the individual light sources.

Claims

1. Apparatus for recording one or more whole-area images of a cell culture plate with one or more wells, comprising a holder for receiving the cell culture plate with wells, wherein the cell culture plate has a bottom side, a top side and a plurality of side faces, which extend between the bottom side and top side, a camera that serves to record images of the bottom side of the cell culture plate within a predefined depth of field, an illumination device for illuminating the cell culture plate, wherein the illumination device comprises a plurality of light sources for producing light beams wherein, in a use position of the cell culture plate, light of a first light source enters the cell culture plate from the outside through the entire length of a first side face, light of a second light source enters the cell culture plate from the outside through the entire length of a second side face, light of a third light source enters the cell culture plate from the outside through the entire length of a third side face, and light of a fourth light source enters the cell culture plate from the outside through the entire length of a fourth side face, wherein the intensity of the produced light beams, the vertical height of the produced light beams in relation to the cell culture plate, the horizontal distance of the light sources from the respective side face and the inclination angle of the produced light beams about a longitudinal axis are individually adjustable for each light source by way of means for adjusting the illumination device.

2. Apparatus according to claim 1, wherein the first light source has a Fresnel lens arrangement in order to produce a uniform light over an effective length of the first light source.

3. Apparatus according to claim 1, wherein each light source and/or a holder for fastening the light source is connected to one or more controllers for adjusting the illumination device.

4. Apparatus according to claim 1, wherein the controller for adjusting the illumination device is configured to carry out a method for adjusting the illumination device.

5. Apparatus according to claim 1, wherein the first light source is arranged laterally next to the first side face.

6. Apparatus according to claim 1, wherein the intensity of the produced light beams, the vertical height of the produced light beams in relation to the cell culture plate, the horizontal distance of the light sources from the respective side face and the inclination angle of the produced light beams about a longitudinal axis are set for each light source in such a way that the cell culture plate is illuminated from the bottom to a predefined depth of field.

7. Apparatus according to claim 1, wherein a vertical height of a light beam emerging from the first light source is 2 to 6 mm.

8. Apparatus according to claim 1, wherein the holder has a plurality of receiving corners, wherein a first receiving corner serves to receive one end of the first side face and a second receiving corner serves to receive an opposite end of the first side face, and wherein a distance between the first receiving corner and the second receiving corner is greater than a length of a row or column of wells, which extends along the first side face.

9. Apparatus according to claim 1, wherein the camera comprises a telecentric lens.

10. Apparatus according to claim 1, wherein the camera comprises an image sensor, and the cell culture plate and/or the image sensor are adjustable in relation to one another in all directions such that the cell culture plate is adjusted in all directions parallel to the image sensor.

11. Apparatus according to claim 1, wherein, for purposes of establishing an effect of one or more active ingredients on nematodes and other organisms in aqueous tests, the wells of the cell culture plate are filled with one or more nematodes and/or other organisms in aqueous solutions with and without active ingredients.

12. Method for adjusting the illumination device of the apparatus according to any one of claim 1, wherein, in relation to the cell culture plate, the relative height of each light source, the inclination angle of each light source and a light intensity of each light source are set individually in order to obtain a good illumination of the individual wells of the cell culture plate and a uniform illumination of the cell culture plate.

13. Method according to claim 12, wherein the camera is used to create an image of a monochrome sheet that is situated in a plane above the illuminated cell culture plate and in that the adjustment is implemented on the basis of an evaluation of the image.

14. Method according to claim 13, wherein a histogram of the light intensity is produced from the image and adjustment by way of controlling the means for adjusting the illumination device is implemented on the basis of said histogram until the histogram has a minimum width.

Description

[0035] The invention should be explained in more detail on the basis of an exemplary embodiment illustrated in the figures. In the figures:

[0036] FIG. 1 schematically shows the structure of an apparatus according to the invention;

[0037] FIG. 2 schematically shows a cell culture plate and an illumination device with four light sources; and

[0038] FIG. 3 shows a section along the line III-III in FIG. 2.

[0039] FIG. 1 schematically shows an apparatus for establishing the effect of active ingredients on dirofilaria and other organisms in aqueous tests. The apparatus, which is denoted by 1 in the totality thereof, comprises a housing 10 with a top separating plate 11 and a middle separating plate 12. The separating plates 11, 12 divide an interior surrounded by the housing 10 into a top region 13, a middle region 14 and a bottom region 15. A camera 20 is assembled in the bottom region 15. The camera 20 has a telecentric lens 21, the end 22 of which facing away from the camera 20 protrudes into the middle region 14 of the housing 10. The middle separating plate 12 has a circular opening 16, through which the lens 21 passes. Here, a conical-frustum-shaped portion 23 of the lens 21 is supported on the circular edge of the opening 16.

[0040] An illumination device 30 and a holder 40 for a cell culture plate 50 are housed in the top region 13 of the housing 10. The illumination device 30, the holder 40 and the cell culture plate 50 will still be described in more detail below on the basis of FIGS. 2 and 3.

[0041] In addition to the camera 20, the bottom region 15 of the housing 10 also holds a controller 60 for the camera 20 and the illumination device 30. Here, the controller 60 can consist of separate control units for the camera 20 and the illumination device 30. The controller 60 can be connected to a computer.

[0042] Using the apparatus 1 according to the invention, it is possible to produce images of a bottom side 51 of the cell culture plate 50. Therefore, a glass panel 17 is incorporated in the separation plate 11.

[0043] FIG. 2 shows the illumination device 30 from above, said illumination device having a first light source 31, a second light source 32, a third light source 33 and a fourth light source 34. The first light source 31 comprises a plurality of light-emitting diodes (not illustrated), which are arranged next to one another in a row and which emit a strip-shaped light beam 35 over virtually the entire length of the first light source 31. The light beam is illustrated by the parallel arrows 35. Over virtually the entire length thereof, the remaining light sources 32, 33, 34 also each emit a strip-shaped light beam in the direction of the cell culture plate 50. However, corresponding arrows for labelling the respective light beams are not illustrated in FIG. 2 for reasons of clarity.

[0044] In addition to the bottom side 51 already mentioned above, the cell culture plate 50 has a top side 52 and a first side face 53, a second side face 54, a third side face 55 and a fourth side face 56. On account of the rectangular basic form of the cell culture plate 50, the opposing side faces 53, 54 extend perpendicular to the other pair of side faces 55, 56 extending in parallel.

[0045] The cell culture plate 50 has a multiplicity of wells 57, which are arranged in 8 rows with in each case 12 wells. As can be gathered from FIG. 3, each individual well 57 has a U-shaped form in the longitudinal section. Here, each well 57 can be filled with an aqueous solution from above, i.e. from the top side 52, the dirofilaria and an active ingredient being situated in said aqueous solution. Once filling is complete, the wells can be covered by way of a cover film.

[0046] As becomes clear from the overview of FIGS. 2 and 3, the individual light sources 31, 32, 33, 34 each emit light into the cell culture plate 50 through the side faces 53, 54, 55, 56. Thus, light from the first light source passes into the cell culture plate 50 from the outside through the first side face 53. The same applies analogously to light sources 32, 33, 34. In order to achieve the best possible illumination of the individual wells 57 of the cell culture plate 50, the position of the light source 31 can be modified in relation to the cell culture plate 50.

[0047] The double-headed arrow 36 indicates that a distance between the first light source 31 and the first side face 53 of the cell culture plate 50 can be increased or reduced. The double-headed arrow 37 (see FIG. 3) indicates that the first light source 31 can be moved upwards or downwards in relation to the cell culture plate 50. Moreover, the first light source 31 can be tilted about a longitudinal axis 38, which extends parallel to the first side face 53 of the cell culture plate 50. The corresponding adjustability about the longitudinal axis 38 or about an axis extending parallel thereto is indicated by the bent arrow 39. Moreover, the light intensity of the first light source 31 can be set by way of the controller 60. The horizontal distance from the respective side face, the vertical position, the inclination angle and the light intensity of the respective light source can be set individually by the corresponding setting parameters of the remaining light sources 32, 33, 34. Consequently, it is possible, for example, to adjust the illumination device in such a way that the light intensity of the first light source 31 deviates not only from the light intensities of the light sources 33, 34 arranged transversely thereto, but also from the light intensity of the opposing light source 32.

[0048] The holder 40 has a first receiving corner 41, a second receiving corner 42, a third receiving corner 43 and a fourth receiving corner 44. Preferably, three of the four receiving corners 41, 42, 43, 44 are embodied as stationary stops while the remaining fourth receiving corner is embodied as a sprung stop. A distance between the first receiving corner and the second receiving corner, for example, is dimensioned in such a way that the light beam 35 of the first light source 31 can strike unhindered the region of the first side face in which the wells 57 are situated. Expressed differently, a distance between the receiving corners 41, 42 is greater than a length of the first column of wells, which extends along the first side face 53.

[0049] On account of the different lengths of the longer side faces 55, 56 and the shorter side faces 53, 54, the light sources 33, 34 on the one hand and the light sources 31, 32 on the other hand have different lengths. Here, the width of a light beam from a light source is always greater than the length of the region of the associated side face in which the wells are situated.

[0050] All regions of the side faces of the cell culture plate 50 in which the wells 57 are situated are impinged by light from the light sources as a result of the illumination device 30 and the special arrangement of the individual receiving corners of the holder 40. As a result of the individual adjustability of the individual light sources, the illumination device 30 can be adjusted in such a way that an almost optimal illumination of the wells 57 is achieved. A preferred monitoring criterion for the illumination that is as optimal as possible has already been described above. An optimal illumination can be assumed when the well-specific results in a cell culture plate whose individual wells have been filled with the same active ingredient in each case do not deviate from one another in respect of the effectiveness of the same active ingredient in each case or have a standard deviation that approximately corresponds to the estimated standard deviation on account of the biological organisms.

LIST OF REFERENCE SIGNS

[0051] 1 Apparatus [0052] 10 Housing [0053] 11 Top separating plate [0054] 12 Middle separating plate [0055] 13 Top region [0056] 14 Middle region [0057] 15 Bottom region [0058] 16 Opening [0059] 17 Glass plate [0060] 20 Camera [0061] 21 Lens [0062] 22 End [0063] 23 Conical-frustum-shaped region [0064] 30 Illumination device [0065] 31 First light source [0066] 32 Second light source [0067] 33 Third light source [0068] 34 Fourth light source [0069] 35 Arrow (light beam) [0070] 36 Double-headed arrow [0071] 37 Double-headed arrow [0072] 38 Arrow [0073] 40 Holder [0074] 41 First receiving corner [0075] 42 Second receiving corner [0076] 43 Third receiving corner [0077] 44 Fourth receiving corner [0078] 50 Cell culture plate [0079] 51 Bottom side [0080] 52 Top side [0081] 53 First side face [0082] 54 Second side face [0083] 55 Third side face [0084] 56 Fourth side face [0085] 57 Well [0086] 60 Controller