Observation system, observation device

Abstract

To enable a sample in a container to be observed with high contrast in a wide range of the container.SOLUTION: A system 100 comprises: a placement table on which a sample container is placed; a planar light source that is located in one of two regions divided by the placement table and has an emission plane; an observation optical system that is located in the other of two regions; a conveyance mechanism that moves the observation optical system in a direction orthogonal to an optical axis of the observation optical system and changes an observation position where an accommodation unit bottom face of the sample container and the optical axis intersect; and a control device 120 that controls an emission patten that is defined by an emission region on an emission plane where light is emitted. The control device 120 executes first emission pattern control by which the emission pattern is changed in accordance with the observation position, or second emission pattern control by which the emission pattern is switched among a plurality of cyclic emission patterns differing in a phase from each other.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] This disclosure relates to observation systems and observation devices. [Background technology]

[0002] For example, there is an observation device for observing cells contained in a culture vessel, as described in Patent Document 1. With this observation device, it is possible to photograph and observe the cells in the vessel simply by placing the vessel on the stage of the observation device. [Prior art documents] [Patent Documents]

[0003] [Patent Document 1] International Publication No. 2016 / 158782 [Overview of the Initiative] [Problems that the invention aims to solve]

[0004] To observe cells, which are phase objects, with high contrast, it is desirable to use polarized illumination. However, polarized illumination usually only works in a relatively narrow area relative to the overall size of the container. Therefore, when moving the observation optical system to observe various regions within the container, the observation position easily extends beyond the area where polarized illumination is effective.

[0005] One possible solution to these problems is to move the illumination optical system in conjunction with the observation optical system, thereby shifting the region in which polarized illumination is achieved. However, in this approach, if polarized illumination is achieved using transmitted illumination, the mechanical structure required to synchronize the movement of the observation optical system and the illumination optical system, which are located on opposite sides of the container, becomes complex, leading to other problems such as the need for a larger observation device.

[0006] Based on the circumstances described above, one aspect of the present invention is to provide a technology that enables high-contrast observation of a sample inside a container over a wide area of ​​the container. [Means for solving the problem]

[0007] An observation system according to one aspect of the present invention comprises: a mounting platform on which a sample container is placed; a surface light source having a light-emitting plane, disposed in one of two regions separated by the mounting platform; an observation optical system disposed in the other of the two regions; a transport mechanism that moves the observation optical system in a direction perpendicular to the optical axis of the observation optical system to change the observation position at which the bottom surface of the storage portion of the sample container intersects with the optical axis; and a control unit that controls a light emission pattern defined by a light-emitting region, which is a light-emitting region on the light-emitting plane. The control unit performs a first light emission pattern control that changes the light emission pattern according to the observation position, or a second light emission pattern control that switches the light emission pattern between a plurality of periodic light emission patterns with different phases. The control unit then performs the first light emission pattern control to change at least one of the position or width of the light emission region according to the observation position, and when the portion of the edge of the light emission region closest to the optical axis is located in the positive region, the following conditional expression of the light emission region is satisfied.

number

number

[0008] An observation apparatus according to one aspect of the present invention comprises: a mounting stage on which a sample container is placed; a light-emitting unit disposed in one of two regions separated by the mounting stage, the light-emitting unit forming a light-emitting region consisting of a plurality of fringe regions periodically aligned in a first direction on the emission surface; an observation optical system disposed in the other of the two regions; and a transport mechanism that moves the observation optical system in a direction perpendicular to the optical axis of the observation optical system within the other of the two regions to change the observation position at which the bottom surface of the housing of the sample container intersects with the optical axis. The light-emitting unit comprises a surface light source having a light-emitting plane, and a lenticular lens disposed between the mounting stage and the surface light source, the lenticular lens having a plurality of cylindrical lens elements aligned in the first direction with the same period as the plurality of fringe regions. The observation apparatus satisfies the following conditional expression.

number

[0009] However, Y1 is the shortest distance in the first direction between the central axis of the curved surface of the cylindrical lens element and the fringe region closest to the cylindrical lens element. Y2 is the maximum distance in the first direction between the central axis of the curved surface of the cylindrical lens element and the fringe region closest to the cylindrical lens element. NA is the numerical aperture on the object side of the observation optical system. P is the pitch of the plurality of fringe regions. D is the air-equivalent length of the distance between the exit surface and the lenticular lens. Fl is the focal length of the lenticular lens. [Effects of the Invention]

[0010] According to the above embodiment, it is possible to observe the sample inside the container with high contrast over a wide area of ​​the container. [Brief explanation of the drawing]

[0011] [Figure 1] This diagram illustrates the configuration of the system according to the first embodiment. [Figure 2] This figure illustrates the configuration of an observation device according to the first embodiment. [Figure 3] This is a diagram illustrating an example of a light emission pattern. [Figure 4] This diagram illustrates the changes in the illumination state of the pupil and the resulting image, depending on the observation position. [Figure 5] This diagram illustrates the movement of the luminescent region depending on the observation position. [Figure 6] This diagram illustrates the switching of light emission patterns according to the observation position. [Figure 7] This diagram illustrates images acquired at each observation location. [Figure 8] This is a diagram to explain the parameters. [Figure 9] This is a diagram illustrating the arrangement of observation positions. [Figure 10] This diagram shows specific examples of the position of the luminescent region depending on the observation position. [Figure 11] Figure 10 is a table showing the system parameter values ​​and whether or not they meet the predetermined conditions for each light emission pattern. [Figure 12] This figure shows another specific example of the position of the luminescent region depending on the observation position. [Figure 13] Figure 12 is a table showing the system parameter values ​​and whether or not they meet the predetermined conditions for each light emission pattern. [Figure 14] This figure shows yet another specific example of the position of the luminescent region depending on the observation position. [Figure 15] This figure shows the light emission pattern corresponding to Figure 14. [Figure 16] Figure 14 is a table showing the system parameter values ​​and whether or not they meet the predetermined conditions for each light emission pattern. [Figure 17] This figure illustrates the light emission control in the system according to the second embodiment. [Figure 18] This diagram illustrates the switching between multiple periodic emission patterns. [Figure 19] This is a diagram to explain the parameters. [Figure 20]This diagram illustrates the relationships between multiple periodic light emission patterns. [Figure 21] This is a flowchart of the multi-point imaging process performed by the system. [Figure 22] This is a flowchart of the image processing performed by the system. [Figure 23] Figure 22 shows an example of a biased illumination image generated by the image processing shown. [Figure 24] This figure illustrates the XY coordinates in the observation device according to the second embodiment. [Figure 25] This is a diagram illustrating the arrangement of observation positions according to the second embodiment. [Figure 26] This figure shows specific examples of multiple periodic emission patterns. [Figure 27] This is a table showing the system's parameter values. [Figure 28] Figure 27 is a table showing the details of the multiple periodic emission patterns. [Figure 29] Figure 27 is a graph illustrating the illumination state on the specimen surface for each of the multiple periodic emission patterns shown. [Figure 30] This is a table that shows whether or not the system meets the specified conditions. [Figure 31] This figure illustrates the switching of multiple periodic light emission patterns according to the third embodiment. [Figure 32] This is a diagram illustrating the parameters of the first periodic emission pattern. [Figure 33] This is a diagram illustrating the relationship between the first periodic emission patterns. [Figure 34] This is a diagram illustrating the parameters of the second periodic emission pattern. [Figure 35] This figure illustrates a biased illumination image generated by image processing. [Figure 36] This figure shows specific examples of multiple periodic emission patterns. [Figure 37] This is a table showing the system's parameter values. [Figure 38] This table shows the details of several first periodic emission patterns among the multiple periodic emission patterns shown in Figure 36. [Figure 39] This table shows the details of several second periodic emission patterns among the multiple periodic emission patterns shown in Figure 36. [Figure 40] Figure 36 is a graph illustrating the illumination state on the specimen surface for each of the multiple first periodic emission patterns shown. [Figure 41] This is a table showing whether or not a predetermined condition is met regarding the first periodic light emission pattern of the system. [Figure 42] This is a table showing whether or not a predetermined condition is met regarding the second periodic light emission pattern of the system. [Figure 43] This is a diagram illustrating the container used in the fourth embodiment. [Figure 44] This is a cross-sectional view of the observation optical system, container, and light source along the optical axis of the observation optical system. [Figure 45] This is a ray diagram of light incident on the well from the fringed emission region of the second periodic emission pattern. [Figure 46] This is a ray diagram of light incident on the well from the fringed emission region of the second periodic emission pattern. [Figure 47] This graph illustrates the illumination state on the specimen surface for each of the multiple second periodic emission patterns. [Figure 48] This graph illustrates the illumination state on the specimen surface for each of the multiple first periodic emission patterns. [Figure 49] This figure illustrates the contrast of the polarized illumination images generated for each position within the well. [Figure 50] This is a diagram illustrating the configuration of the observation device according to the fifth embodiment. [Figure 51] This is a diagram illustrating the configuration of the light-emitting section. [Figure 52] This is a diagram illustrating the structure of a slit plate. [Figure 53]This is a diagram illustrating the structure of a lenticular lens. [Figure 54] This is a diagram illustrating the parameters of the light-emitting section. [Figure 55] This diagram illustrates the relationship between the focal length and emitted light of a lenticular lens. [Figure 56] This is a diagram illustrating the arrangement of the container and the lenticular lens. [Figure 57] This diagram shows the specific parameter values ​​for the light-emitting section. [Figure 58] This is a table showing whether or not the observation device meets the specified conditions. [Figure 59] This is a diagram illustrating the configuration of the light-emitting section of the observation device according to the sixth embodiment. [Figure 60] This diagram illustrates the change in the state of emitted light depending on the position of the striped region. [Figure 61] This is a diagram illustrating the parameters of the light-emitting section. [Figure 62] This diagram shows the specific parameter values ​​for the light-emitting section. [Figure 63] This figure shows specific examples of multiple periodic emission patterns. [Figure 64] This is a table showing whether or not a system meets certain conditions in a periodic light emission pattern. [Figure 65] This figure illustrates the configuration of the fly-eye lens included in the light-emitting section of the observation device according to the seventh embodiment. [Figure 66] This is a diagram illustrating the parameters of the light-emitting section. [Figure 67] This figure shows specific examples of multiple periodic emission patterns. [Figure 68] This is a table showing whether or not the system meets the predetermined conditions in the first periodic light emission pattern. [Figure 69] This is a table showing whether or not the system meets the predetermined conditions in the second periodic emission pattern. [Figure 70]This diagram illustrates the hardware configuration of a computer used to implement a control device. [Modes for carrying out the invention]

[0012] (First Embodiment) Figure 1 is a diagram illustrating the configuration of system 100 according to this embodiment. Figure 2 is a diagram illustrating the configuration of observation device 1 according to this embodiment. Figure 3 is a diagram illustrating an example of a light emission pattern. Figure 4 is a diagram illustrating the illumination state of the pupil surface and the image that changes according to the observation position. Figure 5 is a diagram illustrating the movement of the light emission area according to the observation position. Figure 6 is a diagram illustrating the switching of the light emission pattern according to the observation position. Figure 7 is a diagram illustrating images acquired for each observation position. Hereinafter, system 100 will be described with reference to Figures 1 to 7.

[0013] The system 100 shown in Figure 1 is an observation system for observing a sample contained in container 2 while culturing it. The sample to be observed is a transparent phase object, such as any cultured cells. Container 2 can be any container for containing the sample (hereinafter also referred to as the sample container), such as a Petri dish, but other culture vessels such as flasks or multiwell plates may also be used.

[0014] The system 100 comprises one or more observation devices 1 that acquire images of a sample cultured in a container 2, and a control device 120 that controls the observation devices 1. Each observation device 1 and the control device 120 only need to be able to exchange data with each other. Therefore, each observation device 1 and the control device 120 may be connected via a wired connection as shown in Figure 1, or they may be connected via a wireless connection.

[0015] Observation device 1 is an imaging device that images a sample contained in container 2 from below container 2. In order to image the sample without removing it from incubator 110, observation device 1 is used while positioned inside incubator 110, for example, as shown in Figure 1. More specifically, as shown in Figure 2, observation device 1 is positioned inside incubator 110 with container 2 placed on the mounting stage 11 of observation device 1, and acquires an image of the sample contained in container 2 according to instructions from control device 120.

[0016] As shown in Figure 2, the observation device 1 comprises a box-shaped housing 10 with a transparent mounting platform 11 on which the container 2 is placed on top, and a positioning member 12 for positioning the container 2 to a predetermined position relative to the observation device 1 on the mounting platform 11. The observation device 1 further comprises an observation unit 40 including an observation optical system 20 and an image sensor 30, a transport mechanism 50, and a surface light source 60.

[0017] The observation unit 40 and the transport mechanism 50 are located inside the housing 10. In contrast, the surface light source 60 is located outside the housing 10. Specifically, the surface light source 60 is positioned in one of the two regions separated by the mounting platform 11, while the observation optical system 20 included in the observation unit 40 is positioned in the other of the two regions.

[0018] The observation unit 40 is a unit that integrates the observation optical system 20 and the image sensor 30. The observation optical system 20 is an optical system that focuses light from the bottom surface 3 of the housing part of the container 2 onto the image sensor 30, and is equipped with an aperture diaphragm 21 at the pupil position. Furthermore, the observation optical system 20 has a telecentric optical system on the object side so that the magnification does not change even if the focus position changes. The image sensor 30 is an image sensor, and is not particularly limited, but examples include a CCD (Charge-Coupled Device) image sensor and a CMOS (Complementary MOS) image sensor.

[0019] The transport mechanism 50 is a device that moves the observation unit 40 within the housing 10, and the observation unit 40 is fixed to the transport mechanism 50. The transport mechanism 50 moves the observation unit 40 relative to the container 2 in a direction perpendicular to the optical axis AX of the observation optical system 20. By changing the relative position of the observation unit 40 with respect to the container 2, the transport mechanism 50 changes the position where the bottom surface 3 of the housing part of the container 2 and the optical axis of the observation optical system 20 intersect (hereinafter referred to as the observation position).

[0020] The observation unit 40 is movable in a direction perpendicular to the optical axis AX of the observation optical system 20, more specifically in the X and Y directions which are parallel to and perpendicular to the mounting stage 11, but it may also be movable in the Z direction which is perpendicular to both the X and Y directions.

[0021] The surface light source 60 is a surface light source having an emitting plane 61. The surface light source 60 is, for example, a display device having multiple pixels, and various light emission patterns are formed by controlling the light emission from each of the multiple pixels.

[0022] Observation device 1 acquires an image of a sample by operating each part under the control of control device 120. Specifically, the surface light source 60 emits light according to instructions from control device 120, illuminating the sample on the bottom surface 3 of the container 2 from above. The light transmitted through the sample is focused onto the image sensor 30 by the observation optical system 20. The image sensor 30 images the sample according to instructions from control device 120 and acquires an image of the sample. The image acquired by observation device 1 is output to control device 120.

[0023] The control device 120 is a device that controls the observation device 1. The observation device 1 only needs to include one or more processors and one or more non-temporary computer-readable media, and may be, for example, a general-purpose computer. Each of the one or more processors is an electrical circuit consisting of hardware such as a CPU (Central Processing Unit), GPU (Graphics Processing Unit), DSP (Digital Signal Processor), etc., and performs programmed processing by executing programs stored in one or more non-temporary computer-readable media. In addition, one or more processors may include ASICs (Application Specific Integrated Circuits), FPGAs (Field-Programmable Gate Arrays), etc.

[0024] The control device 120, configured as described above, transmits an image acquisition command to the observation device 1 placed inside the incubator 110 and receives the image acquired by the observation device 1. The control device 120 may also display the image acquired by the observation device 1 on the display device provided by the control device 120, thereby allowing the system 100 to function as an observation system for users to observe a sample during cultivation.

[0025] The control device 120 may communicate with the client terminals shown in Figure 1 (client terminal 130, client terminal 140), and may display the images acquired by the observation device 1 on the display devices provided by the client terminals. The client terminals only need to have a display unit and may be, for example, desktop or notebook computers, tablets, or smartphones.

[0026] Incidentally, in order to visualize and recognize a sample, such as a cell or other phase object, with high contrast, it is desirable to focus the light incident on the sample at an appropriate angle to form an image of the sample, that is, to achieve polarized illumination. However, generally, polarized illumination is only possible within a relatively narrow range. Therefore, under certain lighting conditions (for example, a specific emission pattern), if the observation position is changed, the new observation position easily deviates from the range in which polarized illumination is possible. Consequently, it is not possible to observe a wide area inside the container with good contrast.

[0027] For example, let's consider a case where the control device 120 controls the surface light source 60 to emit light in a light emission pattern P0 having a rectangular light emission region 62 approximately in the center of the light emission plane 61, as shown in Figure 3. The light emission pattern is a spatial pattern defined by the light emission region 62, which is the area emitting light on the light emission plane 61. The light emission pattern is controlled by the control device 120.

[0028] In this case, as shown in Figure 4, when the observation unit 40 (i.e., the observation position) is located directly below the light-emitting region 62, an illumination region 22 is formed in the center of the pupil on the pupil plane where the aperture diaphragm 21 is placed, resulting in bright-field illumination. As a result, no shading is produced in the image of the sample, which is a phase object, and the sample cannot be observed with good contrast. By shifting the observation unit 40 (observation position) to a certain extent from directly below the light-emitting region 62, polarized illumination is achieved, and shading is produced in the image of the sample, allowing the sample to be observed. However, if the observation unit 40 (observation position) is shifted too far from directly below the light-emitting region 62, light will not enter the pupil plane, resulting in dark-field illumination, and the sample will no longer be observable.

[0029] Therefore, in system 100, the control device 120 performs light emission pattern control to change the light emission pattern according to the observation position so that biased illumination is achieved at the observation position determined by the position of the observation unit 40. Specifically, the control device 120 performs light emission pattern control to change at least one of the position or width of the light emission area 62 of the surface light source 60 according to the observation position. Hereafter, the light emission pattern control that controls the light emission pattern according to the observation position will be referred to as the first light emission pattern control, and will be distinguished from the light emission pattern control that does not depend on the observation position (second light emission pattern control) which will be described later.

[0030] To illustrate with the example shown in Figure 5, when the observation unit 40 moves to the first position from the left, the control device 120 changes the position of the light-emitting area 62 to position A shown in Figure 5, thereby changing the light emission pattern of the surface light source 60 to light emission pattern P1 shown in Figure 6(a). Similarly, when the observation unit 40 moves to the second, third, fourth, and fifth positions from the left, the control device 120 changes the position of the light-emitting area 62 to positions B, C, D, and E shown in Figure 5, thereby changing the light emission pattern of the surface light source 60 to light emission patterns P2, P3, P4, and P5 shown in Figures 6(b) through 6(e).

[0031] As a result, in system 100, polarized illumination is achieved at each observation position, allowing for the acquisition of high-contrast images such as images M1 to M5 shown in Figures 7(a) to 7(e). Images M1 to M5 are images of a sample acquired at the 1st to 5th positions from the left, respectively. The sample is, for example, a circular cell placed on the optical axis, and the shape of the cell is recognizable by the creation of bright areas 32 and dark areas 33 against the background 31. Images M1, M2, and M4 are images obtained with polarized illumination from the positive side, while images M3 and M5 are images obtained with polarized illumination from the negative side.

[0032] Figure 8 is a diagram illustrating the parameters. The following describes the desirable conditions that the system 100 should satisfy in the first light emission pattern control, with reference to Figure 8.

[0033] First, the definition of parameters related to the first light emission pattern control will be described. NA is the numerical aperture on the object side of the observation optical system 20. H is the air-equivalent length of the distance between the light emission plane 61 and the observation position. Y s is the Y coordinate of the observation position. Y min , Y max are the Y coordinates of both ends of the light emission region 62, respectively, and Y min <Y max is. That is, Y max is the Y coordinate of the other end of the light emission region 62 located on the positive side of one end of the light emission region 62 corresponding to Y min , and is the Y coordinate of the positive side end of the light emission region 62. Y min is the Y coordinate of the negative side end of the light emission region 62. Note that the Y coordinate is, in this example, the coordinate in the Y direction in which the observation position changes.

[0034] S min is the sine of the incident angle of the illumination light from one end (negative side end) of the light emission region 62 corresponding to Y min to the observation position. S max is the sine of the incident angle of the illumination light from the other end (positive side end) of the light emission region 62 corresponding to Y max to the observation position.

[0035] When performing oblique illumination from the positive side with respect to the observation position, it is desirable for the system 100 to satisfy the following conditional expression.

Equation

[0036] As a result, since the illumination region 22 is formed at a position shifted from the center of the pupil, oblique illumination can be established at the observation position. And in this case, it is desirable for the system 100 to further satisfy the following conditional expression.

Equation

[0037] As a result, the illumination area 22 reaches the edge of the pupil, allowing the observation optical system 20 to illuminate the sample at the maximum possible angle of incidence. Therefore, the sample can be visualized with high contrast.

[0038] Furthermore, when performing biased illumination from the negative side relative to the observation position, it is desirable that system 100 satisfies the following condition.

number

[0039] As a result, the illumination area 22 is formed at a position offset from the center of the pupil, thereby enabling biased illumination at the observation position. Furthermore, in this case, it is desirable that the system 100 also satisfies the following condition.

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[0040] As a result, the illumination area 22 reaches the edge of the pupil, allowing the observation optical system 20 to illuminate the sample at the maximum possible angle of incidence. Therefore, the sample can be visualized with high contrast.

[0041] Note that the angle of incidence is expressed as a positive value when the illumination light is incident from the positive side relative to the optical axis, and as a negative value when the illumination light is incident from the negative side relative to the optical axis. Therefore, S min S max It is defined by the following formula:

number

[0042] Figure 9 is a diagram illustrating the arrangement of observation positions. Container 2 shown in Figure 9 is a rectangular Petri dish with dimensions of 87 mm × 87 mm. Three specific examples of the first emission pattern control performed by system 100 are shown for acquiring images of a sample at a total of 20 observation positions (sp11-sp15, sp21-sp25, sp31-sp35, sp41-sp45) in a 4 × 5 arrangement within container 2. In all of these examples, H is 60 mm, NA is 0.25, and the actual field of view is 2.8 mm × 2.1 mm.

[0043] Figure 10 shows a specific example of the position of the light-emitting region according to the observation position. Figure 11 is a table T1 showing the system parameter values ​​and whether or not a predetermined condition is met for each light-emitting pattern shown in Figure 10. Figures 10 and 11 are the first specific example, showing an example of changing the position of the light-emitting region 62.

[0044] Positions A, B, C, and D of the light-emitting region 62 shown in Figure 10 correspond to bright area positions 1, 2, 3, and 4 shown in Figure 11. As shown in Figure 11, in the first specific example, when observing the observation positions (sp11~sp15, sp21~sp25, sp31~sp35), system 100 satisfies conditions (1) and (2), and polarized illumination is achieved from the positive side. Also, when observing the observation positions (sp41~sp45), system 100 satisfies conditions (3) and (4), and polarized illumination is achieved from the negative side.

[0045] Figure 12 shows another specific example of the position of the light-emitting region according to the observation position. Figure 13 is Table T2, which shows the system parameter values ​​and whether or not predetermined conditions are met for each light-emitting pattern shown in Figure 12. Figures 12 and 13 are a second specific example, showing an example of changing the position of the light-emitting region 62. The second specific example differs from the first specific example in that it prioritizes biased illumination from the negative side.

[0046] The positions A, B, C, and D of the light-emitting region 62 shown in Figure 12 correspond to the bright area positions 1, 2, 3, and 4 shown in Figure 13. As shown in Figure 13, in the second specific example, when observing the observation positions (sp11~sp15), the system 100 satisfies conditions (1) and (2), and polarized illumination is achieved from the positive side. Furthermore, when observing the observation positions (sp21~sp25, sp31~sp35, sp41~sp45), the system 100 satisfies conditions (3) and (4), and polarized illumination is achieved from the negative side.

[0047] Figure 14 shows yet another specific example of the position of the light-emitting region according to the observation position. Figure 15 shows the light-emitting pattern corresponding to Figure 14. Figure 16 is Table T3 showing the system parameter values ​​and whether or not predetermined conditions are met for each light-emitting pattern shown in Figure 14. Figures 14 to 16 are a third specific example, showing an example of changing the position and width of the light-emitting region 62. Note that the third specific example differs from the first specific example in that control of illumination light vignetted by the pupil is omitted as much as possible.

[0048] Positions A, B, C, and D of the light-emitting region 62 shown in Figure 14 correspond to light-emitting patterns P6, P7, P8, and P9 shown in Figure 15, and to bright area positions 1, 2, 3, and 4 shown in Figure 16. As shown in Figure 16, in the third specific example, similar to the first specific example, when observing the observation positions (sp11~sp15, sp21~sp25, sp31~sp35), the system 100 satisfies conditions (1) and (2), and polarized illumination is achieved from the positive side. Also, when observing the observation positions (sp41~sp45), the system 100 satisfies conditions (3) and (4), and polarized illumination is achieved from the negative side.

[0049] In all three specific examples described above, polarized illumination is established from the positive side at observation positions (sp11~sp15) and from the negative side at observation positions (sp41~sp45). This is to prevent vignetting of the illumination light at the walls of container 2. Therefore, as long as the illumination light is not vignetted at the walls, polarized illumination may be performed from either the positive or negative side.

[0050] According to the system 100 of the first embodiment, since biased illumination is achieved at least at the observation position, regardless of the observation position, the sample can be observed with high contrast over a wide area of ​​the container.

[0051] (Second embodiment) The system according to this embodiment (hereinafter also simply referred to as "this system") is the same as system 100 in that it includes an observation device 1 and a control device 120. This system differs from system 100 in that, instead of the control device 120 performing emission pattern control (first emission pattern control) according to the observation position, it performs emission pattern control (hereinafter referred to as second emission pattern control) which switches the emission pattern between a plurality of periodic emission patterns with different phases from each other.

[0052] Figure 17 is a diagram illustrating the light emission control in the system according to this embodiment. Figure 18 is a diagram illustrating the switching of multiple periodic light emission patterns. The second light emission pattern control will be described below with reference to Figures 17 and 18.

[0053] In the system according to this embodiment, the control device 120 performs a second light emission pattern control at each observation position to sequentially switch the light emission pattern formed on the light emission plane 61 to a plurality of periodic light emission patterns. This second light emission pattern control is performed by the control device 120 regardless of the observation position. A periodic light emission pattern is a spatial pattern defined by a light emission region 63 consisting of a plurality of striped regions 64 that are periodically aligned in one direction (for example, the Y direction), as shown in Figures 17 and 18.

[0054] As shown in Figure 18, the control device 120 can obtain images of multiple samples acquired by the observation device 1 under different illumination conditions by switching between multiple periodic emission patterns (emission patterns P10 to P15) that have different phases. Here, different phases mean that the reference positions of the fringe regions 64 (for example, the position of the first fringe) are different.

[0055] As will be described later, by appropriately designing multiple periodic emission patterns to be switched, the acquired images will include at least one image taken under conditions of polarized illumination, regardless of the observation position. Therefore, this system does not require different control for each observation position; high-contrast images can be obtained simply by performing a constant emission pattern control (i.e., a second emission pattern control) regardless of the observation position.

[0056] Figure 19 is a diagram illustrating the parameters. Figure 20 is a diagram illustrating the relationships between multiple periodic emission patterns. Below, with reference to Figures 19 and 20, the conditions that this system should ideally satisfy in the second emission pattern control will be described.

[0057] First, let's explain the definitions of the parameters related to the control of the second emission pattern. NA is the numerical aperture on the object side of the observation optical system 20. P is the pitch of the multiple fringe regions 64. d is the width of each of the multiple fringe regions 64. H is the air-equivalent length of the distance between the emission plane 61 and the observation position. δ is the distance on the emission plane 61 corresponding to the phase difference between the multiple periodic emission patterns. In other words, δ corresponds to the amount of movement of the fringe regions 64 that occurs when the periodic emission patterns are switched.

[0058] In this case, the Y coordinate of the negative end of each fringe region 64 of the periodic light emission pattern (light emission pattern P16) shown in Figure 19 is Y M,k,min and the Y coordinate of the positive end Y M,k,max It is defined by the following formula:

number

[0059] Here, M is the number of the striped region, which identifies which of the multiple striped regions 64 included in the periodic emission pattern. k is the number of the periodic emission pattern, which identifies which of the multiple periodic emission patterns (for example, emission patterns P17 to P19 shown in Figure 20) it is. δ0 is the Y coordinate of the negative end of the first striped region of the first periodic emission pattern. dmin , Y dmax These are the minimum and maximum Y coordinates of the light-emitting plane 61, and Y dmin <Y M,k,min <Y dmax , Y dmin <Y M,k,max <Y dmax This is true.

[0060] This system should ideally satisfy the following conditions.

number

[0061] This ensures that the spacing (Pd) between the striped regions 64 is set appropriately without becoming too narrow. Therefore, it is possible to avoid the observation position being simultaneously illuminated from both the positive and negative sides by illumination from two adjacent striped regions 64. This allows for the acquisition of high-contrast images with polarized illumination. However, if polarized illumination occurs from both the positive and negative sides, the shadows cancel each other out, making it difficult to obtain high-contrast images.

[0062] Furthermore, it is desirable that this system also satisfies the following conditions.

number

[0063] This limits the amount of movement of the striped region 64 caused by switching the periodic emission pattern, allowing it to be set appropriately. As a result, polarized illumination is always achieved in at least one of the multiple periodic emission patterns. However, if the amount of movement of the striped region 64 is too large, polarized illumination may not be achieved in any of the periodic emission patterns, making it difficult to obtain a high-contrast image.

[0064] Furthermore, it is desirable that this system also satisfies the following conditions.

number

[0065] This ensures that the width of the striped region 64 is set appropriately without becoming too wide. As a result, the illumination conditions at the observation position change due to the switching of multiple periodic emission patterns, thus avoiding unnecessary shooting such as acquiring the same image multiple times.

[0066] Furthermore, it is desirable that this system satisfies the following condition: N is an integer greater than or equal to 1.

number

[0067] As a result, as shown in Figure 20, the same periodic emission pattern occurs every N+1 times (every 4 times in the example in Figure 20). Therefore, by switching at least N times (3 times in the example in Figure 20), it is possible to acquire an image of the observation position under the same conditions regardless of which periodic emission pattern is started with.

[0068] Figure 21 is a flowchart of the multi-point imaging process performed by the system. Figure 22 is a flowchart of the image processing performed by the system. Figure 23 is an example of a biased illumination image generated by the image processing shown in Figure 22. The multi-point imaging process performed by this system will be explained below with reference to Figures 21 to 23.

[0069] In this system, when the multi-point imaging process shown in Figure 21 is started, the control device 120 first acquires the project settings (step S1). Here, the project is a plan of when, where, and under what conditions imaging will be performed, and the project settings include at least information on the imaging points (observation positions) for multi-point imaging. In addition to the information on the imaging points (observation positions), the project settings may also include information on the scheduled time of imaging.

[0070] The control device 120 moves the observation optical system 20 (step S2). Here, the observation position is changed by moving the observation optical system 20 to the position specified in the project settings acquired in step S1.

[0071] Subsequently, the control device 120 causes the surface light source 60 to emit light in a periodic emission pattern (step S3), images the sample with the image sensor 30 (step S4), and then saves the acquired image (step S5). The process from step S3 to step S5 is repeated for the number of periodic emission patterns. When the repetition process for the number of periodic emission patterns is completed (step S6YES), the control device 120 performs the image processing shown in Figure 22 (step S7).

[0072] In the image processing shown in Figure 22, the control device 120 outputs a biased illumination image based on multiple images acquired with multiple periodic emission patterns through the iterative processing described above. A biased illumination image is an image in which a phase object, such as a sample, is visualized with good contrast, and may be the image itself acquired under conditions of biased illumination. It may also be an image processed to resemble an image acquired under conditions of biased illumination, or it may be a new image generated based on multiple images, for example, one of which includes at least one image acquired under conditions of biased illumination.

[0073] Specifically, as shown in Figure 22, the control device 120 performs the following processes: removing lighting unevenness (step S11), normalizing the image intensity (step S12), and combining the images (step S13).

[0074] In step S11, the control device 120 performs a filter process on each of the multiple images to cut out the low-frequency cut component. This filter process is performed on the entire image area. As a result, the multiple images obtained by shooting (hereinafter also referred to as multiple original images) are converted into multiple images from which the illumination unevenness has been removed (hereinafter referred to as multiple filtered images). In the following steps, the intensity of each pixel in the multiple original images is set to I o,k,x,y Refer to and take the intensity of each pixel in multiple filtered images as I LCF,k,x,y Refer to it as follows: k is the image number, x is the pixel row number, and y is the pixel column number.

[0075] In step S12, the control device 120 normalizes the multiple filtered images and converts them into multiple normalized images with equal average intensity. Specifically, the control device 120 may generate multiple normalized images such that each pixel of the multiple normalized images has an intensity calculated by the following formula.

number

[0076] Here, I nml,k,x,y This is the intensity of each pixel in multiple normalized images. ave n is the average pixel intensity of multiple normalized images. n is the number of pixels in the filtered image.

[0077] In step S13, the control device 120 synthesizes multiple normalized images to generate a polarized illumination image. Specifically, the control device 120 should generate the polarized illumination image such that each pixel of the polarized illumination image has an intensity calculated by the following formula. In other words, the polarized illumination image is generated such that each pixel has an intensity that is different from the background intensity by a value proportional to the sum of the differences between the pixel intensity and the pixel average intensity of the multiple normalized images.

number

[0078] Here, I dm,x,yis the intensity of each pixel in the polarized illumination image. bg is the background intensity. A is the contrast adjustment value. N is the number of normalized images.

[0079] By performing the image processing shown in Figure 22, the contrasts present in multiple original images are effectively added together, making it possible to generate a polarized illumination image with strong contrast, such as image M6 in Figure 23(a) and image M7 in Figure 23(b). When A > 0, the polarized illumination image becomes a positive contrast image in which the sample is represented by bright areas 32, as in image M6. On the other hand, when A < 0, the polarized illumination image becomes a negative contrast image in which the sample is represented by dark areas 33, as in image M7.

[0080] Figure 24 is a diagram illustrating the XY coordinates in the observation device according to this embodiment. Figure 25 is a diagram illustrating the arrangement of observation positions according to this embodiment. As shown in Figure 24, a coordinate system is defined with the origin at the corner of the container 2 that is in contact with the positioning member 12. The container 2 shown in Figure 25 is a rectangular Petri dish with dimensions of 87 mm × 87 mm. Therefore, the coordinates of the corner furthest from the origin of the container 2 are (87, 87).

[0081] The following describes a specific example of the second emission pattern control performed by this system when acquiring images of a sample at a total of 20 observation positions (sp11-sp15, sp21-sp25, sp31-sp35, sp41-sp45) in a 4x5 arrangement within container 2 shown in Figure 25, using the coordinate system shown in Figure 24. In all of these examples, H is 50 mm, NA is 0.25, and the actual field of view size is 2.8 mm x 2.1 mm.

[0082] Figure 26 is a diagram showing specific examples of multiple periodic emission patterns. Figure 27 is Table T4 showing the system parameter values. Figure 28 is Table T5 showing details of the multiple periodic emission patterns shown in Figure 27. Figure 29 is a graph explaining the illumination state on the sample surface for each of the multiple periodic emission patterns shown in Figure 27. Figure 30 is Table T6 showing whether the system meets the predetermined conditions.

[0083] As shown in Figure 26, the system according to this embodiment acquires images at each observation position by switching between six periodic emission patterns (emission patterns P21 to P26). Each periodic emission pattern has an emission region 63 consisting of six striped regions 64, as shown in Figure 26. The detailed parameters of this system are shown in Figure 27. The Y coordinates of the striped regions 64 of each periodic emission pattern are shown in Figure 28. Furthermore, as shown in Figure 30, this system satisfies all of the conditions from (9) to (12).

[0084] Under the above conditions, by executing a second emission pattern control that switches between each periodic emission pattern, as shown in Figures 29(a) to 29(f), the area where polarized illumination is achieved moves in the Y direction with each switch of the periodic emission pattern, and by executing all periodic emission patterns, the entire container is covered as an area where polarized illumination is achieved. As a result, any observation position can be observed with good contrast.

[0085] In Figure 29, the horizontal axis of the graph represents the observation position (Y-coordinate), and the vertical axis represents the sine of the angle of incidence of the illumination light at the observation position. The solid line represents S min The dashed line indicates S max This shows the range of incident angles of illumination light. The shaded areas enclosed by solid and dashed lines indicate the range of observation positions where polarized illumination from the positive side is achieved, and the dashed arrows indicate the range of observation positions where polarized illumination from the negative side is achieved.

[0086] Similar to the system 100 according to the first embodiment, this system also achieves biased illumination regardless of the observation position, allowing for high-contrast observation of the sample over a wide area of ​​the container. Furthermore, with this system, the control device 120 does not need to change its control settings for each observation position, and a high-contrast image can be obtained at any observation position. Moreover, since adjustments for each observation position are unnecessary with this system, high robustness can be achieved.

[0087] In the above, the image processing in step S7 was exemplified as including image synthesis as shown in Figure 22, but the content of the image processing is not limited to the example shown in Figure 22. The control device 120 may also include the process of selecting a biased illumination image from multiple images.

[0088] In that case, in step S12, the control device 120 normalizes the multiple filtered images and converts them into multiple normalized images with equal average intensity. Specifically, the control device 120 may generate multiple normalized images such that each pixel of the multiple normalized images has an intensity calculated by the following formula: I' nml,k,x,y This represents the intensity of each pixel in multiple normalized images.

number

[0089] Subsequently, the control device 120 calculates the variance of the intensity of each normalized image using the following formula: 2 k This is the variance of the intensity of the normalized image.

number

[0090] Finally, the control device 120 selects a biased illumination image based on the calculated variance. Specifically, it should select the image with the highest variance as the biased illumination image. The biased illumination image may be the normalized image with the highest variance itself, a filtered image corresponding to the normalized image with the highest variance, or the original image corresponding to the normalized image with the highest variance. For example, when analyzing cell number or cell density, a filtered image corresponding to the normalized image with the highest variance is most desirable from the standpoint of stability of the analysis results.

[0091] In this system, the second emission pattern control ensures that polarized illumination is achieved at the observation position in at least one of the multiple periodic emission patterns. Therefore, even when image processing including the image selection described above is performed, it is possible to obtain a polarized illumination image, and an effect similar to that obtained when image synthesis, as shown in Figure 22, is performed.

[0092] (Third embodiment) Figure 31 is a diagram illustrating the switching of multiple periodic light emission patterns according to this embodiment. The system according to this embodiment (hereinafter simply referred to as "this system") is the same as the system according to the second embodiment in that the control device 120 performs second light emission pattern control. This system differs from the system according to the second embodiment in that the multiple periodic light emission patterns include, as shown in Figure 31, a plurality of first periodic light emission patterns (light emission patterns P27 to P31) with different phases from each other, and a plurality of second periodic light emission patterns (light emission patterns P32 to P36) with different phases from each other.

[0093] Each of the multiple first periodic emission patterns corresponds to an emission region 63 consisting of multiple first fringe regions (fringe regions 64) periodically aligned in a first direction (e.g., the X direction), whereas each of the multiple second periodic emission patterns corresponds to an emission region 63 consisting of multiple second fringe regions (fringe regions 64) periodically aligned in a second direction (e.g., the Y direction) different from the first direction. In this example, the first and second directions are orthogonal.

[0094] Figure 32 illustrates the parameters of the first periodic emission pattern. Figure 33 illustrates the relationships between the first periodic emission patterns. Figure 34 illustrates the parameters of the second periodic emission pattern. Figure 35 illustrates an example of a biased illumination image generated by image processing. Hereinafter, referring to Figures 32 to 35, the desirable conditions that the system should satisfy in controlling the second emission pattern will be described.

[0095] First, we will explain the definition of the parameters related to the first periodic emission pattern. NA is the numerical aperture on the object side of the observation optical system 20. P x d is the pitch of multiple fringe regions 64 included in the first periodic emission pattern. x δ is the width of each of the multiple fringe regions 64 included in the first periodic emission pattern. H is the air-equivalent length of the distance between the emission plane 61 and the observation position. x δ is the distance on the emission plane 61 corresponding to the phase difference between multiple first periodic emission patterns. x This corresponds to the amount of movement of the striped region 64 that occurs when the first periodic emission pattern is switched.

[0096] In this case, the X coordinate of the negative end of each fringe region 64 of the first periodic light emission pattern (light emission pattern P37) shown in Figure 32 is X L,kx,min and the X coordinate of the positive end L,kx,max It is defined by the following formula:

number

[0097] Here, L is the number of the striped region, which identifies which of the multiple striped regions 64 included in the first periodic emission pattern. kx is the number of the first periodic emission pattern, which identifies which of the multiple first periodic emission patterns (for example, emission patterns P38 to P41 shown in Figure 33) it is. δ x0 This is the X-coordinate of the negative end of the first fringe region of the first periodic emission pattern. dmin , X dmax These are the minimum X coordinate and maximum Y coordinate of the light-emitting plane 61, and X dmin <X L,kx,min <X dmax , X dmin <X L,kx,max <X dmax This is true.

[0098] This system should ideally satisfy the following conditions.

number

[0099] Conditional expressions (19) through (22) correspond to conditional expressions (9) through (12) mentioned above, and their meanings are the same. Conditional expression (23) is derived from conditional expressions (20) and (22).

[0100] Next, we will explain the definition of the parameters related to the second periodic emission pattern. P y d is the pitch of multiple fringe regions 64 included in the second periodic emission pattern. y δ is the width of each of the multiple fringe regions 64 included in the second periodic emission pattern. y This is the distance on the emission plane 61 corresponding to the phase difference between multiple second periodic emission patterns. That is, δ y This corresponds to the amount of movement of the striped region 64 that occurs when the second periodic emission pattern is switched.

[0101] In this case, the Y coordinate of the negative end of each fringe region 64 of the second periodic light emission pattern (light emission pattern P42) shown in Figure 34 is Y M,ky,min and the Y coordinate of the positive end Y M,ky,max It is defined by the following formula:

number

[0102] Here, M is the number of the striped region 64, which is one of the multiple striped regions 64 included in the second periodic emission pattern. ky is the number of the second periodic emission pattern, which is one of the multiple second periodic emission patterns. δ y0 This is the Y coordinate of the negative end of the first fringe region of the first second periodic emission pattern. dmin , Y dmax These are the minimum and maximum Y coordinates of the light-emitting plane 61, and Y dmin <Y M,ky,min <Y dmax , Y dmin <Y M,ky,min <Y dmax This is true.

[0103] This system should ideally satisfy the following conditions.

number

[0104] Conditional expressions (26) to (29) correspond to conditional expressions (9) to (12) mentioned above, and their meanings are the same. Conditional expression (30) is derived from conditional expressions (27) and (29).

[0105] In this embodiment as well, the control device 120 performs the same steps as in the second embodiment, as shown in Figure 22: a process to remove lighting unevenness (step S11), a process to normalize the image intensity (step S12), and a process to combine the images (step S13).

[0106] In step S11, the control device 120 performs a filter process to cut out the low-frequency cut component from each of the multiple first images acquired with multiple first periodic light emission patterns and the multiple second images acquired with multiple second periodic light emission patterns. This filter process is performed on the entire image area. As a result, the multiple first images (hereinafter also referred to as multiple first original images) and the multiple second images (hereinafter also referred to as multiple second original images) are converted into multiple first images and multiple second images (hereinafter referred to as multiple first filtered images and multiple second filtered images) from which the illumination unevenness has been removed.

[0107] Furthermore, from here on, the intensity of each pixel in multiple first original images will be set to I o,kx,x,y Refer to and take the intensity of each pixel in multiple first filtered images as I LCF,kx,x,y Refer to the following. Also, the intensity of each pixel in multiple second original images is I o,ky,x,y Refer to and take the intensity of each pixel in multiple second filtered images as I LCF,ky,x,y Refer to the following: kx is the number of the first image, ky is the number of the second image, x is the row number of the pixel, and y is the column number of the pixel.

[0108] In step S12, the control device 120 normalizes the multiple first filtered images and the multiple second filtered images to convert them into multiple first normalized images and multiple second normalized images with equal average intensity.

[0109] Specifically, the control device 120 may generate multiple first normalized images such that each pixel of the multiple first normalized images has an intensity calculated by the following formula.

number

[0110] Here, I nml,kx,x,y is the intensity of each pixel in multiple first normalized images. aven is the average pixel intensity of multiple first normalized images (second normalized images). n is the number of pixels in the first filtered image (second filtered image).

[0111] Furthermore, the control device 120 may generate multiple second normalized images such that each pixel of the multiple second normalized images has an intensity calculated by the following formula.

number

[0112] Here, I nml,ky,x,y is the intensity of each pixel in multiple second normalized images.

[0113] In step S13, the control device 120 generates a polarized illumination image by combining a plurality of first normalized images and a plurality of second normalized images. Specifically, the control device 120 may generate the polarized illumination image such that each pixel of the polarized illumination image has an intensity calculated by the following formula. In other words, the polarized illumination image is generated such that each pixel has an intensity that is different from the background intensity by a value proportional to the sum of the differences between the pixel intensity and the pixel average intensity of the plurality of normalized images (the first normalized image and the second normalized image).

number

[0114] Here, I dm,x,y is the intensity of each pixel in the polarized illumination image. bg is the background intensity. A is the contrast adjustment value. N x N is the number of first normalized images. y This is the number of second-standardized images.

[0115] By performing the image processing described above, the contrasts present in multiple first original images and multiple second original images are effectively added together. Therefore, in this embodiment as well, it is possible to generate a biased illumination image with strong contrast, such as image M8 in Figure 35(a) and image M9 in Figure 35(b). Furthermore, in this embodiment, by combining an image obtained with illumination from the X direction and an image obtained with illumination from the Y direction, a biased illumination image with improved symmetry compared to the second embodiment can be obtained. When A > 0, the biased illumination image becomes a positive contrast image in which the sample is represented by bright areas 32, as in image M8. On the other hand, when A < 0, the biased illumination image becomes a negative contrast image in which the sample is represented by dark areas 33, as in image M9.

[0116] Figure 36 is a diagram showing specific examples of multiple periodic emission patterns. Figure 37 is Table T7 showing the system parameter values. Figure 38 is Table T8 showing details of multiple first periodic emission patterns among the multiple periodic emission patterns shown in Figure 36. Figure 39 is Table T9 showing details of multiple second periodic emission patterns among the multiple periodic emission patterns shown in Figure 36. Figure 40 is a graph explaining the illumination state on the sample surface for each of the multiple first periodic emission patterns shown in Figure 36. Figure 41 is Table T10 showing whether or not the system meets predetermined conditions regarding the first periodic emission pattern. Figure 42 is Table T11 showing whether or not the system meets predetermined conditions regarding the second periodic emission pattern.

[0117] As shown in Figure 36, the system according to this embodiment acquires images at each observation position by switching between 12 periodic emission patterns (emission patterns P43 to P54). The 12 periodic emission patterns include six first periodic emission patterns (emission patterns P43 to P48) that are periodic in the X direction and six second periodic emission patterns (emission patterns P49 to P54) that are periodic in the Y direction. The first periodic emission patterns have an emission region 63 consisting of three fringe regions 64. The second periodic emission patterns have an emission region 63 consisting of six fringe regions 64.

[0118] The detailed parameters of this system are shown in Figure 37. The X coordinates of the fringe regions 64 of each first periodic emission pattern are shown in Figure 38, and the Y coordinates of the fringe regions 64 of each second periodic emission pattern are shown in Figure 39. Furthermore, as shown in Figures 41 and 42, this system satisfies all of the conditions (19) through (21), (23), (26) through (28), and (30).

[0119] Under the above conditions, by executing the second light emission pattern control, the area where polarized illumination is achieved moves in the X direction each time the first periodic light emission pattern is switched, and by executing all of the first periodic light emission patterns, the entire container is covered as an area where polarized illumination is achieved. Furthermore, each time the second periodic light emission pattern is switched, the area where polarized illumination is achieved moves in the Y direction, and by executing all of the second periodic light emission patterns, the entire container is covered as an area where polarized illumination is achieved. Figures 40(a) to 40(f) show this movement in the X direction. The way to interpret the graph in Figure 40 is the same as the graph in Figure 29. This allows observation of any observation position with good contrast.

[0120] Similar to the system according to the embodiment described above, this system also achieves biased illumination regardless of the observation position, allowing for high-contrast observation of the sample over a wide area of ​​the container. Furthermore, this system is similar to the system according to the second embodiment in that it does not require changes to the control settings for each observation position and achieves high robustness. Moreover, the system according to this embodiment can obtain images with higher symmetry than the system according to the embodiment described above.

[0121] (Fourth embodiment) Figure 43 is a diagram illustrating the container used in this embodiment. Figure 44 is a cross-sectional view of the observation optical system, container, and light source along the optical axis of the observation optical system. The system according to this embodiment (hereinafter simply referred to as "this system") differs from the system according to the third embodiment in that it uses a container 4, which is a microplate, as shown in Figures 43 and 44, instead of a container 2, which is a Petri dish. Other aspects, including the control of the illumination pattern, are the same as those of the system according to the third embodiment. The container 4 is a 96-well microplate, with a well top diameter of 7 mm, a well bottom diameter of 6.45 mm, and a well center-to-center distance of 9 mm.

[0122] Figures 45 and 46 are ray diagrams of light incident on the well from the fringed emission region of the second periodic emission pattern. More specifically, they show the observation position with the well center coordinates (X, Y) = (38.1, 59.3). Figure 47 is a graph illustrating the illumination state on the specimen surface for each of the multiple second periodic emission patterns. Figure 48 is a graph illustrating the illumination state on the specimen surface for each of the multiple first periodic emission patterns. Figure 49 is a diagram illustrating the contrast of the biased illumination image generated for each position in the well.

[0123] Referring to Figures 45 and 46, it can be seen that in a 96-well microplate, the illumination light is vignetted by the well walls (shown by dashed lines), and the illumination light is significantly bent at the surface of the culture medium due to surface tension. As a result, as shown in Figure 47, the illumination area 22 (i.e., S) at the pupil surface is affected. min and S max The first periodic emission pattern does not move regularly in response to the switching of the second periodic emission pattern, but rather widens and narrows. Therefore, the region in which polarized illumination is achieved does not change regularly with the switching of the second periodic emission pattern. However, despite this disruption of regularity, it is possible to achieve polarized illumination in the entire region from the negative end to the positive end of the well by switching between multiple second periodic emission patterns. Furthermore, in the X direction, as shown in Figure 48, it is possible to achieve polarized illumination in the entire region from the negative end to the positive end of the well by switching between multiple first periodic emission patterns.

[0124] Therefore, unlike conventional systems, this system allows for high-contrast observation of any observation position without creating areas that cannot be observed, even in containers with narrow storage areas such as 96-well microplates. However, near the well edges (around the wells), the illumination light from the well walls is disrupted and biased in the illumination direction due to vignetting and curvature of the liquid surface. As a result, as shown in Figure 49, the images acquired around the wells (images M13 to M19) have a limited range of contrast compared to the images acquired in the central area (images M10 to M12). Nevertheless, the ability to acquire images with high contrast around the entire circumference of the wells is a significant advantage compared to conventional systems.

[0125] (Fifth embodiment) Figure 50 is a diagram illustrating the configuration of the observation device according to this embodiment. Figure 51 is a diagram illustrating the configuration of the light-emitting unit. Figure 52 is a diagram illustrating the configuration of the slit plate. Figure 53 is a diagram illustrating the configuration of the lenticular lens. Hereinafter, the configuration of the system according to this embodiment will be described with reference to Figures 50 to 53. Note that the system according to this embodiment differs from the systems according to the other embodiments described above in that it includes the observation device 5 shown in Figure 50 instead of the observation device 1, and the control device 120 does not control the spatial light emission pattern.

[0126] Observation device 5 differs from observation device 1 in that, as shown in Figure 50, it is equipped with a light-emitting unit 80 instead of a surface light source 60. In other respects, it is the same as observation device 1. Specifically, the observation unit 40 and a transport mechanism 50 (not shown) are arranged in one of the two regions separated by the mounting table 11, and the light-emitting unit 80 is arranged in the other region.

[0127] As shown in Figure 51, the light-emitting unit 80 comprises a surface light source 81, a slit plate 82, and a lenticular lens 83. The slit plate 82 is positioned between the surface light source 81 and the lenticular lens 83, and the lenticular lens 83 is positioned between the surface light source 81 and the mounting base 11. Figure 51 shows the configuration of the area indicated by the dashed line in Figure 50.

[0128] The surface light source 81 is any light source having an emitting plane, but like the surface light source 60 described above, it may be, for example, a display device having multiple pixels. The slit plate 82 has a rectangular opening 82a periodically in a certain direction (first direction, for example, the Y direction), as shown in Figure 52. The surface light source 81 and the slit plate 82 included in the light-emitting unit 80 constitute a periodic light emission pattern forming unit that forms an emitting region consisting of multiple fringe regions periodically aligned in the first direction on the emission surface. The lenticular lens 83 has multiple cylindrical lens elements 84 aligned in the first direction with the same period as the multiple fringe regions formed by the periodic light emission pattern forming unit, as shown in Figure 53.

[0129] In the observation device 5, by appropriately setting multiple striped regions (apertures 82a) and cylindrical lens elements 84, as shown in Figure 51, the illumination light emitted from the aperture 82a of the slit plate 82 is emitted at approximately the same angle by the lenticular lens 83 at each position within the aperture 82a. The light ray L1 shown in Figure 51 represents the illumination light emitted from the negative end of the aperture 82a, and the light ray L2 represents the illumination light emitted from the positive end of the aperture 82a.

[0130] Therefore, the observation device 5 can establish biased illumination under almost the same conditions over a wide area of ​​the container, and high-contrast images can be obtained regardless of the observation position. Furthermore, the observation device 5 establishes biased illumination under almost the same conditions at various sample heights. Therefore, even when using various containers with different base heights 3, high-contrast images can be obtained regardless of the observation position.

[0131] Figure 54 is a diagram illustrating the parameters of the light-emitting section. Figure 55 is a diagram illustrating the relationship between the focal length of the lenticular lens and the emitted light. The following describes the conditions that the observation device 5 should preferably satisfy, with reference to Figures 54 and 55.

[0132] First, the definitions of the parameters of the observation device 5 will be explained. NA is the numerical aperture on the object side of the observation optical system 20. D is the air-equivalent length of the distance between the exit surface of the slit plate 82 and the lenticular lens 83. More specifically, it is the air-equivalent length of the distance from the exit surface to the top of the cylindrical lens element 84. P is the pitch of the multiple apertures 82a, and also the pitch of the multiple cylindrical lens elements 84. That is, it is the pitch of the multiple fringe regions. Y1 is the shortest distance in the first direction (in this case, the Y direction) between the center axis CX of the curved surface of the cylindrical lens element 84 and the fringe region closest to the cylindrical lens element 84. Y2 is the maximum distance in the first direction between the center axis CX of the curved surface of the cylindrical lens element 84 and the fringe region closest to the cylindrical lens element 84.

[0133] In this case, the Y coordinate of the center axis CX of the curved surface of each cylindrical lens element 84 of the lenticular lens 83 is Y ax,N And the effective range in the Y direction of each cylindrical lens element 84 ef,N Y is defined by the following formula, where N is any integer. ax0 This is the Y-coordinate of the central axis C of the curved surface of the reference cylindrical lens element 84.

number

[0134] Furthermore, for both the case where polarized illumination is performed from the positive side and the case where polarized illumination is performed from the negative side, the range in the Y direction of the Nth fringe region is expressed by the following formula. (Polarized illumination from the negative side)

number

number

[0135] Observation device 5 should preferably satisfy the following condition.

number

[0136] This ensures that illumination light from the striped region enters at least the pupil plane. However, if the absolute value of Y1 exceeds the upper limit, the illumination light will not enter the pupil of the observation optical system 20 and will all be vignetted, preventing the establishment of polarized illumination. Conversely, if the absolute value of Y1 falls below the lower limit, the effect of polarized illumination will decrease, and the image contrast will decline.

[0137] Furthermore, it is desirable that this system also satisfies the following conditions.

number

[0138] As a result, the illumination light from the fringed region is distributed across the outer edge of the pupil. However, if the absolute value of Y2 exceeds the upper limit, the illumination light from the adjacent fringed region (aperture 82a) enters the pupil of the observation optical system 20. In this case, the direction of incidence of the illumination light is reversed from the normal direction, which reduces the contrast of the image. Also, if the absolute value of Y2 falls below the lower limit, the entire illumination light from the fringed region is contained within the pupil of the observation optical system 20. Therefore, good biased illumination is not achieved, and the image will not have sufficient contrast.

[0139] Furthermore, it is desirable that the observation device 5 satisfies the following condition. Here, F l This is the focal length of the lenticular lens.

number

[0140] The focal length of the lenticular lens 83 is defined by the following formula, where R is the radius of curvature of the cylindrical surface of the cylindrical lens element 84, and n is the refractive index of the lenticular lens 83.

number

[0141] This prevents excessive convergence and divergence of the luminous beam from the fringed region. As shown in Figure 55(a), D / F l If the value exceeds the upper limit, the power of the lenticular lens 83 becomes too strong, causing the light beam to converge and resulting in variations in illumination depending on the observation position. Also, as shown in Figure 55(d), D / F l If the value falls below the lower limit, the power of the lenticular lens 83 becomes too weak, causing the light beam to diverge. In this case, the illumination condition also varies depending on the observation position. In contrast, as shown in Figures 55(b) and 55(c), by satisfying the conditional equation, excessive convergence and divergence of the illumination light beam can be avoided, thus achieving a stable illumination condition regardless of the observation position.

[0142] FIG. 56 is a diagram for explaining the positional relationship between the container and the lenticular lens. FIG. 57 is a diagram showing specific parameter values of the light emitting unit. FIG. 58 is a table T12 showing the non of predetermined conditions of the observation device 5.

[0143] As shown in FIG. 56, the observation device 5 according to the present embodiment is provided with a lenticular lens 83 over a range wider than the region where the container 2 is disposed. Further, the detailed parameters of the observation device 5 are as follows. Here, β is the magnification of the observation optical system 20. L is the distance from the apex of the surface of the lenticular lens 83 to the lower plane. NA = 0.25, β = 2.2, H = 70 mm, D = 0.6 mm, P = 0.5 mm, Y1 = 0.1 mm, Y2 = 0.2 mm, R = 0.5 mm, L = 2 mm, n = 1.493, F l = 1.014 mm

[0144] Further, as shown in FIG. 58, the observation device 5 satisfies all of the conditional expressions (38) to (40). Therefore, substantially the same deflected illumination is realized regardless of the observation position. Therefore, in the observation device 5 and the present system as well, similar to the system according to the above-described embodiment, deflected illumination is established regardless of the observation position, so that the sample can be observed with high contrast over a wide range of the container. Further, according to the observation device 5 and the present system, an image with high contrast can be obtained at an arbitrary observation position without the control device 120 controlling the light emission pattern.

[0145] (Sixth Embodiment) FIG. 59 is a diagram for explaining the configuration of the light emitting unit of the observation device according to the present embodiment. FIG. 60 is a diagram for explaining the change in the state of the emitted light according to the position of the stripe region. The system according to the present embodiment (hereinafter simply referred to as the present system) is different from the system according to the fifth embodiment in that, as shown in FIG. 59, the light emitting unit 80 of the observation device 5 includes a surface light source 60 instead of the surface light source 81 and the slit plate 82, and the control device 120 executes second light emission pattern control.

[0146] In this system as well, similar to the system according to the fifth embodiment, a light-emitting region composed of a plurality of stripe regions 64 periodically aligned in the first direction (Y direction) is formed on the front side of the lenticular lens 83. This system is different from the system according to the fifth embodiment in that it performs second light emission pattern control by moving the stripe regions 64 little by little in the first direction.

[0147] When the stripe regions 64 are moved on the front side of the lenticular lens 83, as shown in FIG. 60, the illumination state changes depending on the position of the stripe regions 64. Specifically, a dark-field illumination state as shown in FIG. 60(a), a negative-side deflected illumination state as shown in FIG. 60(b), a bright-field illumination state as shown in FIG. 60(c), and a positive-side deflected illumination state as shown in FIG. 60(d) occur in order. Therefore, by performing image processing similar to that in the second embodiment using a plurality of images acquired in a plurality of illumination states, a deflected illumination image with high contrast can be obtained.

[0148] FIG. 61 is a diagram for explaining the parameters of the light-emitting unit. Hereinafter, the conditions that this system preferably satisfies will be described while referring to FIG. 61.

[0149] First, the definition of the parameters of this system will be described. In this system, instead of Y1 and Y2, Y0 and d are defined. Y0 is the distance from the curved surface center axis CX of the cylindrical lens element 84 to the center of the stripe region. Note that when Y0 is on the positive side with respect to the curved surface center axis CX, it is particularly denoted as Y 0+ and when it is on the negative side, it is particularly denoted as Y 0- d is the width of each of the plurality of stripe regions.

[0150] In this case, the Y coordinate Y ax,N of the curved surface center axis CX of each cylindrical lens element 84 of the lenticular lens 83 and the effective range Y ef,N in the Y direction of each cylindrical lens element 84 are as described in the fifth embodiment.

[0151] And for each of the cases of performing deflected illumination from the positive side and from the negative side, the range in the Y direction of the N-th stripe region is represented by the following formula. (Deflected illumination from the positive side)

Equation

Equation

[0152] It is further desirable for this system to satisfy the following conditional expression.

Equation

[0153] Thereby, by appropriately setting Y0, deflected illumination can be realized while preventing light from an adjacent stripe region from entering the pupil of the observation optical system 20. Note that if d exceeds the upper limit value, even if deflected illumination is established by the illumination light from the stripe region, the illumination light from an adjacent stripe region will enter the pupil of the observation optical system 20. Since the incident direction of the illumination light from an adjacent stripe region is opposite to that of the normal one, the contrast of the image will be reduced. If d is below the lower limit value, no illumination light will be generated.

[0154] It is further desirable for this system to satisfy the following conditional expressions for each of the cases of performing deflected illumination from the positive side and from the negative side.

Equation

[0155] Thereby, deflected illumination can be realized. When Y 0+ exceeds the upper limit value, or when Y 0- is below the lower limit value, the illumination light does not enter the pupil and deflected illumination is not established. Also, when Y 0+ is below the lower limit value, or when Y 0-When it exceeds the upper limit value, all the illumination light from the stripe region is contained within the pupil, so the effect of the deflected illumination decreases and the contrast of the image becomes low.

[0156] It is desirable for this system to further satisfy the following conditional expressions when performing deflected illumination from the positive side and when performing deflected illumination from the negative side.

Equation

[0157] This can avoid the illumination light from adjacent stripe regions from entering the pupil. Y 0+ When it exceeds the upper limit value, or when Y 0- When it is below the lower limit value, the illumination light from adjacent stripe regions enters the pupil. Since the incident direction of the illumination light from adjacent stripe regions is opposite to the normal one, the contrast of the image is decreased. Also, when Y 0+ When it is below the lower limit value, or when Y 0- When it exceeds the upper limit value, the illumination light spreads across the pupil center of the observation optical system 20, so the deflected illumination does not hold and the image does not have contrast.

[0158] It is desirable for this system to further satisfy the following conditional expression. The meaning of this expression is the same as that of the fifth embodiment.

Equation

[0159] FIG. 62 is a diagram showing specific parameter values of the light emitting unit. FIG. 63 is a diagram showing a specific example of a plurality of periodic light emission patterns. FIG. 64 is a table T13 showing the non - satisfaction of predetermined conditions of the system in the periodic light emission pattern.

[0160] The detailed parameters of the system according to this embodiment are as follows. Note that β is the magnification of the observation optical system 20. NA=0.25, β=2.2, H=70mm, D=6mm, P=4.8mm, d=1mm, R=5mm, L=6mm, n=1.493, F l = 1.014 mm

[0161] As shown in Figure 63, the system according to this embodiment acquires images at each observation position by switching between six periodic emission patterns (emission patterns P55 to P60). Each periodic emission pattern has an emission region 63 consisting of three striped regions 64. Y0(Y 0,k The following applies: Y 0,1 = -1.6 mm, Y 0,2 = -0.8 mm, Y 0,3 = 0.0 mm Y 0,4 =0.8mm, Y 0,5 =1.6mm, Y 0,6 = 2.4 mm

[0162] Furthermore, as shown in Figure 64, in the first periodic emission pattern (emission pattern P55), conditions (44), (46), (48), and (49) are satisfied, and polarized illumination from the negative side is achieved. Also, in the fifth periodic emission pattern (emission pattern P59), conditions (44), (45), (47), and (49) are satisfied, and polarized illumination from the positive side is achieved. Therefore, in this system as well as in the system according to the above embodiment, polarized illumination is achieved regardless of the observation position, so the sample can be observed with high contrast over a wide area of ​​the container. Also, similar to the second embodiment, the control device 120 does not need to change the control content for each observation position, and a high-contrast image can be obtained at any observation position. Furthermore, since adjustment for each observation position is unnecessary with this system, high robustness can be achieved.

[0163] (Seventh Embodiment) Figure 65 is a diagram illustrating the configuration of the fly-eye lens included in the light-emitting section of the observation device according to this embodiment. The system according to this embodiment (hereinafter simply referred to as "this system") differs from the system according to the sixth embodiment in that the light-emitting section 80 of the observation device 5 is equipped with a fly-eye lens 93 as shown in Figure 65 instead of a lenticular lens 83, and the multiple periodic light emission patterns switched by the control device 120 include multiple first periodic light emission patterns and multiple second periodic light emission patterns.

[0164] The fly-eye lens 93 has a plurality of lens elements 94 that are aligned in a first direction with the same period as a plurality of first fringe regions included in a first periodic emission pattern, and that are aligned in a second direction with the same period as a plurality of second fringe regions included in a second periodic emission pattern.

[0165] Figure 66 is a diagram illustrating the parameters of the light-emitting section. The following explanation will describe the desirable conditions that this system should satisfy, with reference to Figure 66.

[0166] First, let's explain the definition of the parameters of this system. This system differs from the sixth embodiment in that, in addition to Y0, X0 is also defined. X0 and Y0 are the distances in the X and Y directions from the center axis CX of the curved surface of the lens element 94 to the center of the fringe region, respectively. Note that when X0 is on the positive side with respect to the center axis CX of the curved surface, it is particularly X 0+ It is noted that when it is on the negative side, it is especially X 0- It is noted that Y0 is particularly Y when it is on the positive side with respect to the central axis CX of the curved surface. 0+ It is noted that when it is on the negative side, Y 0- It is written as follows.

[0167] In this case, the X coordinate of the curved central axis CX of each lens element 94 of the fly-eye lens 93 is X ax,M Y coordinate Y ax,N And the effective range X in the X direction of each lens element 94 ef,M Effective range in the Y direction ef,N X is defined by the following equation, where M and N are arbitrary integers. ax0 , Yax0 These are the X and Y coordinates of the central axis C of the curved surface of the reference lens element 94.

number

[0168] Then, for both the case where polarized illumination is performed from the positive side and the case where polarized illumination is performed from the negative side, the range X in the X direction of the M-th fringe region of the first periodic emission pattern is defined. iLL,M , range Y in the Y direction of the Nth fringe region of the second periodic emission pattern iLL,N It can be expressed by the following formula. (Positive illumination from the positive side)

number

number

[0169] Furthermore, it is desirable that this system also satisfies the following conditions.

number

[0170] This allows for selective illumination to be achieved by appropriately setting Y0, while preventing light from adjacent fringe regions from entering the pupil of the observation optical system 20. However, if d exceeds the upper limit, even if selective illumination is achieved by illumination light from the fringe region, illumination light from adjacent fringe regions will enter the pupil of the observation optical system 20. Since the direction of incidence of illumination light from adjacent fringe regions is opposite to that of normal, it will reduce the contrast of the image. If d falls below the lower limit, no illumination light is generated.

[0171] Note that conditional expressions (58) and (67) are the same as conditional expressions (44) and (49) mentioned above, and their meanings are also the same. lThis is the focal length of the fly-eye lens 93. Note that the conditions (59) to (62) for the X direction and (63) to (66) for the Y direction correspond to the conditions (45) to (48) mentioned above, and their meanings are also the same.

[0172] Figure 67 is a diagram showing specific examples of multiple periodic light emission patterns. Figure 68 is a table T14 showing whether the system meets the predetermined conditions in the first periodic light emission pattern. Figure 69 is a table T15 showing whether the system meets the predetermined conditions in the second periodic light emission pattern.

[0173] The detailed parameters of the system according to this embodiment are as follows. Note that β is the magnification of the observation optical system 20. NA=0.25, β=2.2, H=70mm, D=6mm, P=4.8mm, d=1mm, R=5mm, L=6mm, n=1.493, F l = 1.014 mm

[0174] As shown in Figure 67, the system according to this embodiment acquires images at each observation position by switching between 12 periodic emission patterns (emission patterns P61 to P72). The 12 periodic emission patterns include six first periodic emission patterns (emission patterns P61 to P66) that are periodic in the X direction and six second periodic emission patterns (emission patterns P67 to P72) that are periodic in the Y direction. The first periodic emission patterns have an emission region 63 consisting of three fringe regions 64. The second periodic emission patterns have an emission region 63 consisting of six fringe regions 64. X0(X) of each of the emission patterns P61 to P66 0,k ) is as follows. Also, the respective Y0(Y) values ​​for emission patterns P67 to P72 are as follows. 0,k The following applies: X 0,1 = -1.6 mm, X 0,2 = -0.8mm, X 0,3 = 0.0 mm X 0,4 =0.8mm, X0,5 =1.6mm, X 0,6 = 2.4 mm Y 0,1 = -1.6 mm, Y 0,2 = -0.8 mm, Y 0,3 = 0.0 mm Y 0,4 =0.8mm, Y 0,5 =1.6mm, Y 0,6 = 2.4 mm

[0175] Furthermore, as shown in Figure 68, the system according to this embodiment satisfies conditions (58), (61), (62), and (67) in the first periodic emission pattern (emission pattern P61), and biased illumination from the negative side is achieved. Also, in the fifth first periodic emission pattern (emission pattern P65), conditions (58), (59), (60), and (67) are satisfied, and biased illumination from the positive side is achieved. Furthermore, as shown in Figure 69, the system according to this embodiment satisfies conditions (58), (65), (66), and (67) in the first second periodic emission pattern (emission pattern P67), and biased illumination from the negative side is achieved. Furthermore, in the fifth second periodic emission pattern (emission pattern P71), conditions (58), (63), (64), and (67) are satisfied, and polarized illumination from the positive side is achieved. Therefore, in this system, as with the system according to the embodiment described above, polarized illumination is achieved regardless of the observation position, so that the sample can be observed with high contrast over a wide area of ​​the container. Also, similar to the third embodiment, the control device 120 does not need to change the control content for each observation position, and a high-contrast image can be obtained at any observation position. Moreover, since adjustment for each observation position is unnecessary with this system, high robustness can be achieved.

[0176] The embodiments described above are specific examples provided to facilitate understanding of the invention, and the present invention is not limited to these embodiments. Modified forms of the embodiments described above and alternative forms that replace the embodiments described above may be included. In other words, each embodiment can be modified in terms of its components without departing from its spirit and scope. Furthermore, new embodiments can be implemented by appropriately combining multiple components disclosed in one or more embodiments. In addition, some components may be deleted from the components shown in each embodiment, or some components may be added to the components shown in an embodiment. Moreover, the processing procedures shown in each embodiment may be performed in a different order, as long as they do not contradict each other. That is, the observation system and observation device of the present invention can be modified in various ways without departing from the scope of the claims.

[0177] In the embodiments described above, the observation device and the control device are shown as separate devices, but they may be configured as a single device. That is, the observation device itself may operate as the control device, or it may function as a control unit that performs the light emission control described above, or as an image processing unit that performs image processing. Furthermore, the control device, which is a separate device from the observation device, may implement only one of the functions of the control unit that performs light emission control or the image processing unit that performs image processing, while the observation device may implement the other function. In other words, the observation device alone may operate as the observation system described above.

[0178] Figure 70 illustrates a computer hardware configuration for implementing a control device. The hardware configuration shown in Figure 70 includes, for example, a processor 1001, memory 1002, storage device 1003, reader 1004, communication interface 1006, and input / output interface 1007. The processor 1001, memory 1002, storage device 1003, reader 1004, communication interface 1006, and input / output interface 1007 are connected to each other, for example, via a bus 1008.

[0179] The processor 1001 is any electrical circuit and may be, for example, a single processor, a multi-processor, or a multi-core processor. The processor 1001 may perform the light emission control and image processing described above by reading and executing a program stored in the memory device 1003.

[0180] Memory 1002 is, for example, a semiconductor memory and may include a RAM area and a ROM area. Storage device 1003 is, for example, a semiconductor memory such as a hard disk or flash memory, or an external storage device.

[0181] The reader 1004 accesses the storage medium 1005, for example, according to instructions from the processor 1001. The storage medium 1005 can be implemented by, for example, a semiconductor device, a medium through which information is input / output by magnetic action, or a medium through which information is input / output by optical action. A semiconductor device is, for example, a USB (Universal Serial Bus) memory. A medium through which information is input / output by magnetic action is, for example, a magnetic disk. A medium through which information is input / output by optical action is, for example, a CD (Compact Disc)-ROM, a DVD (Digital Versatile Disk), a Blu-ray Disc, etc. (Blu-ray is a registered trademark).

[0182] The communication interface 1006 communicates with other devices, for example, according to instructions from the processor 1001. The input / output interface 1007 is, for example, an interface between an input device and an output device. The input device may be, for example, a device such as a keyboard, mouse, or touch panel that receives instructions from the user. The output device may be, for example, a display device such as a display, and an audio device such as a speaker.

[0183] The program executed by processor 1001 is provided to computer 1000 in the following form, for example. (1) It is pre-installed on the storage device 1003. (2) Provided by the storage medium 1005. (3) Provided from a server such as a program server.

[0184] The hardware configuration of the computer 1000 for realizing the control device described with reference to Figure 70 is illustrative, and the embodiment is not limited thereto. For example, some of the above configuration may be deleted, or new configurations may be added. In another embodiment, for example, some or all of the functions of the above electrical circuit may be implemented as hardware such as an FPGA (Field Programmable Gate Array), SoC (System-on-a-Chip), ASIC (Application Specific Integrated Circuit), and PLD (Programmable Logic Device).

[0185] In this specification, the expression "based on A" does not mean "based solely on A," but rather "based on at least A," and furthermore, "based at least partially on A." That is, "based on A" may also mean based on B in addition to A, or based on a part of A. [Explanation of Symbols]

[0186] 1, 5: Observation device, 2, 4: Container, 3: Bottom surface, 10: Housing, 11: Mounting platform, 12: Positioning member, 20: Observation optical system, 21: Aperture diaphragm, 22: Illumination area, 30: Image sensor, 31: Background, 32: Bright area, 33: Dark area, 40: Observation unit, 50: Transport mechanism, 60, 81: Surface light source, 61: Light-emitting plane, 62, 63: Light-emitting area, 64: Striped area, 80: Light-emitting part, 82: Slit plate, 82a: Aperture, 83: Lenticular lens 84: Cylindrical lens element, 93: Fly-eye lens, 94: Lens element, 100: System, 110: Incubator, 120: Control device, 130, 140: Client terminal, 1000: Computer, 1001: Processor, 1002: Memory, 1003: Storage device, 1004: Reader, 1005: Storage medium, 1006: Communication interface, 1007: Input / Output interface, 1008: Bus

Claims

1. A platform on which a sample container is placed, A surface light source having a light-emitting plane is positioned in one of the two regions separated by the aforementioned mounting platform, An observation optical system arranged in the other of the two regions, A transport mechanism that moves the observation optical system in a direction perpendicular to the optical axis of the observation optical system to change the observation position at which the bottom surface of the storage section of the sample container intersects with the optical axis, The system comprises a control unit that controls a light emission pattern defined by a light emission region, which is a light-emitting region on the light-emitting plane, The control unit, A first light emission pattern control that changes the light emission pattern according to the observation position, or A second light emission pattern control is performed to switch the aforementioned light emission pattern between a plurality of periodic light emission patterns with different phases. The control unit performs the first light emission pattern control to change at least one of the position or width of the light emission region according to the observation position. When the portion of the edge of the light-emitting region closest to the optical axis is located in the positive region, the following conditional expression for the light-emitting region is satisfied, [Math 1] When the portion of the edge of the light-emitting region closest to the optical axis is located in the negative region, the following conditional expression for the light-emitting region is satisfied. [Math 2] An observation system characterized by the following: However, Smin is the sine of the angle of incidence of the illumination light from one end of the light-emitting region to the observation position. Smax is the sine of the angle of incidence of the illumination light from the other end of the light-emitting region, which is located on the positive side of the one end, to the observation position. NA is the numerical aperture on the object side of the observation optical system.

2. In the observation system described in claim 1, When the portion of the edge of the light-emitting region closest to the optical axis is located in the positive region, the following conditional expression for the light-emitting region is satisfied, [Math 3] When the portion of the edge of the light-emitting region closest to the optical axis is located in the negative region, the following conditional expression for the light-emitting region is satisfied. [Math 4] An observation system characterized by the following:

3. A platform on which a sample container is placed, A surface light source having a light-emitting plane is positioned in one of the two regions separated by the aforementioned mounting platform, An observation optical system arranged in the other of the two regions, A transport mechanism that moves the observation optical system in a direction perpendicular to the optical axis of the observation optical system to change the observation position at which the bottom surface of the storage section of the sample container intersects with the optical axis, The system comprises a control unit that controls a light emission pattern defined by a light emission region, which is a light-emitting region on the light-emitting plane, The control unit, A first light emission pattern control that changes the light emission pattern according to the observation position, or A second light emission pattern control is performed to switch the aforementioned light emission pattern between a plurality of periodic light emission patterns with different phases. Each of the aforementioned plurality of periodic emission patterns corresponds to a emission region consisting of a plurality of striped regions periodically aligned in a first direction. The control unit performs the second light emission pattern control to sequentially switch the light emission pattern to the plurality of periodic light emission patterns. An observation system characterized by the following:

4. In the observation system described in claim 3, An observation system characterized by satisfying the following conditional equation. [Math 5] However, NA is the numerical aperture on the object side of the observation optical system. P is the pitch of the plurality of fringe regions. d is the width of each of the plurality of fringe regions. H is the air-equivalent length of the distance between the light-emitting plane and the observation position. δ is the distance on the light-emitting plane corresponding to the phase difference between the plurality of periodic light-emitting patterns.

5. In the observation system described in claim 4, An observation system characterized by satisfying the following conditional equation. [Math 6]

6. In the observation system described in claim 5, An observation system characterized by satisfying the following conditional equation. [Number 7] However, N is an integer greater than or equal to 1.

7. In the observation system described in claim 3, further, A lenticular lens disposed between the mounting base and the surface light source, comprising a plurality of cylindrical lens elements aligned in the first direction with the same period as the plurality of stripe regions. An observation system characterized by the following:

8. In the observation system described in claim 7, An observation system characterized by satisfying the following conditional equation. [Number 8] However, NA is the numerical aperture on the object side of the observation optical system. P is the pitch of the plurality of fringe regions. d is the width of each of the plurality of fringe regions. D is the air-equivalent distance between the light-emitting plane and the lenticular lens. Fl is the focal length of the lenticular lens.

9. A platform on which a sample container is placed, A surface light source having a light-emitting plane is positioned in one of the two regions separated by the aforementioned mounting platform, An observation optical system arranged in the other of the two regions, A transport mechanism that moves the observation optical system in a direction perpendicular to the optical axis of the observation optical system to change the observation position at which the bottom surface of the storage section of the sample container intersects with the optical axis, The system comprises a control unit that controls a light emission pattern defined by a light emission region, which is a light-emitting region on the light-emitting plane, The control unit, A first light emission pattern control that changes the light emission pattern according to the observation position, or A second light emission pattern control is performed to switch the aforementioned light emission pattern between a plurality of periodic light emission patterns with different phases. The plurality of periodic light emission patterns include a plurality of first periodic light emission patterns having different phases from each other, and a plurality of second periodic light emission patterns having different phases from each other. Each of the plurality of first periodic emission patterns corresponds to the emission region, which consists of a plurality of first striped regions periodically aligned in a first direction. Each of the plurality of second periodic emission patterns corresponds to a emission region consisting of a plurality of second fringe regions periodically aligned in a second direction different from the first direction. The control unit performs the second light emission pattern control to sequentially switch the light emission pattern to the plurality of periodic light emission patterns. An observation system characterized by the following:

10. In the observation system described in claim 9, An observation system characterized by satisfying the following conditional equation. [Number 9] However, NA is the numerical aperture on the object side of the observation optical system. Px is the pitch of the plurality of first fringe regions. Py is the pitch of the plurality of second fringe regions. dx is the width of each of the plurality of first fringe regions. dy is the width of each of the plurality of second fringe regions. H is the air-equivalent length of the distance between the light-emitting plane and the observation position. δx is the distance on the light-emitting plane corresponding to the phase difference between the plurality of periodic light-emitting patterns. δy is the distance on the light-emitting plane corresponding to the phase difference between the plurality of periodic light-emitting patterns.

11. In the observation system according to claim 10, An observation system characterized by satisfying the following conditional equation. [Number 10]

12. In the observation system according to claim 11, An observation system characterized by satisfying the following conditional equation. [Math 11] However, Nx is an integer greater than or equal to 1. Ny is an integer greater than or equal to 1.

13. In the observation system described in claim 9, further, The fly-eye lens is positioned between the mounting base and the surface light source and has a plurality of lens elements that are aligned in the first direction with the same period as the plurality of first fringe regions and aligned in the second direction with the same period as the plurality of second fringe regions. An observation system characterized by the following:

14. In the observation system described in claim 13, An observation system characterized by satisfying the following conditional equation. [Math 12] However, NA is the numerical aperture on the object side of the observation optical system. P is the pitch of the plurality of fringe regions. d is the width of each of the plurality of fringe regions. D is the air-equivalent length of the distance between the light-emitting plane and the fly-eye lens. Fl is the focal length of the fly-eye lens.

15. In the observation system according to any one of claims 3 to 14, further, The image sensor into which the light collected by the aforementioned observation optical system is incident, The system comprises an image processing unit that processes images acquired by the image sensor, The image processing unit outputs a biased illumination image based on the multiple images acquired with the multiple periodic emission patterns. An observation system characterized by the following:

16. In the observation system according to claim 15, The image processing unit synthesizes a plurality of normalized images, each with a normalized intensity, to generate the biased illumination image. An observation system characterized by the following:

17. In the observation system according to claim 15, The image processing unit selects the biased illumination image from the plurality of images based on the variance of the intensity of the plurality of images. An observation system characterized by the following:

18. A platform on which the sample container is placed, A light-emitting unit located in one of the two regions separated by the aforementioned mounting base, the light-emitting unit forming a light-emitting region consisting of a plurality of striped regions periodically aligned in a first direction on the emission surface, An observation optical system arranged in the other of the two regions, The transport mechanism includes moving the observation optical system within the other of the two regions in a direction perpendicular to the optical axis of the observation optical system to change the observation position where the bottom surface of the sample container housing intersects with the optical axis. The light-emitting part is, A surface light source having an emitting plane, A lenticular lens disposed between the mounting base and the surface light source, having a plurality of cylindrical lens elements aligned in the first direction with the same period as the plurality of stripe regions, An observation device characterized by satisfying the following conditional equation. [Number 13] However, Y1 is the shortest distance in the first direction between the central axis of the curved surface of the cylindrical lens element and the fringe region closest to the cylindrical lens element. Y2 is the maximum distance in the first direction between the central axis of the curved surface of the cylindrical lens element and the fringe region closest to the cylindrical lens element. NA is the numerical aperture on the object side of the observation optical system. P is the pitch of the plurality of fringe regions. D is the air-equivalent length of the distance between the exit surface and the lenticular lens. Fl is the focal length of the lenticular lens.

Images