TW201732366A - Imaging arrangement determination method for imaging device and imaging device - Google Patents

Abstract

Provided is a method for simply determining an imaging arrangement that makes it possible to efficiently obtain an image with good quality in imaging using an imaging device having two illumination optical systems. First, an effective field of view for when imaging is carried out using the imaging device is determined (step S210). Then, a plurality of imaging positions at which imaging using the first illumination optical system is to be carried out are arranged along the wall surface of a sample accommodation part (steps S220 and S230). Subsequently, a plurality of imaging positions at which imaging using the second illumination optical system is to be carried out are arranged in an area outside of the effective fields of view obtained through the imaging at the plurality of imaging positions determined in steps S220 and S230 so that a given position in a sample container is included in the effective field of view obtained through imaging at one or more of the imaging positions (step S240).

Description

Imaging configuration determination method and imaging device in imaging device

The present invention relates to a method of determining an imaging arrangement (arrangement of a plurality of imaging positions) when an imaging device that generates an entire image by combining a plurality of captured images obtained by imaging from a plurality of imaging positions is imaged.

In the fields of medical treatment, drug development, and the like, cells cultured in a sample container called "aperture plate" or "microplate" have been observed as samples. In the sample container, a plurality of sample storage portions called concave holes are formed. Generally, the sample is injected into the holes together with the liquid medium. In recent years, an image pickup apparatus such as a CCD (Charge Coupled Device) camera is mounted to image an image of the sample, and the image data obtained by the imaging is used to observe the sample. For example, in the drug development research for cancer, observation or analysis of cancer cells is performed by imaging an cancer cell into a blood cell together with a liquid (culture solution) as a medium by an imaging device. In such an image pickup apparatus, when the illumination light is emitted from the upper side of the hole toward the hole, the illumination light is refracted by the meniscus of the surface of the liquid (the liquid as the medium) formed in the hole, thereby being in the hole The brightness of the peripheral image is insufficient. Therefore, in the image pickup apparatus disclosed in Japanese Laid-Open Patent Publication No. 2015-118036, the imaging optical system can be configured to have a characteristic of the object-focusing telecentricity, and the traveling direction can be deviated from the optical axis due to the refraction. The curved light in the direction is efficiently concentrated. [Prior Art Document] [Patent Document] [Patent Document 1] Japanese Patent Laid-Open No. 2015-118036

[Problems to be Solved by the Invention] When it is necessary to observe a sample at a high magnification, it is possible to image one hole from a plurality of imaging positions. In this case, an image of the entire aperture is generated by combining a plurality of captured images obtained by imaging from a plurality of imaging positions. Moreover, in this case, the imaging field of view (the imaging range when imaging is performed from one imaging position) is smaller than the area of the hole. If the imaging field of view is smaller than the area of the hole, it may occur in a region where the imaging field is included in the region affected by the meniscus, and in a region where the imaging field of view does not include the region affected by the meniscus. According to the image pickup apparatus disclosed in Japanese Laid-Open Patent Publication No. 2015-118036, the light is efficiently collected in the region affected by the meniscus as described above, so that the image obtained by the imaging has sufficient brightness. However, in the case where the region affected by the meniscus is not included in the imaging field of view, the brightness at the peripheral portion of the imaging field of view is insufficient. Therefore, two illumination optical systems are prepared, and imaging is performed while switching the two illumination optical systems in accordance with the imaging position. In this case, it is necessary to determine a position (a plurality of imaging positions) for imaging using one illumination optical system and a position (a plurality of imaging positions) for imaging using another illumination optical system before imaging by the imaging device. That is, it is necessary to determine the imaging configuration (the configuration of a plurality of imaging positions). However, if a large number of positions are set as the imaging position, the imaging becomes inefficient and waste occurs. Conversely, when the position set as the imaging position is small, an area where the brightness is insufficient may be generated in the entire image obtained by the combination of the plurality of captured images. In this way, a better imaging configuration cannot be easily obtained. Accordingly, it is an object of the present invention to provide a method of easily determining an imaging configuration in which an image of a high-quality image can be efficiently obtained by imaging an imaging device having two illumination optical systems. [Means for Solving the Problems] The first aspect of the present invention is a method for determining an imaging arrangement, wherein the image capturing device has a plurality of imaging positions, and the imaging device includes a first illumination optical system and In the second illumination optical system, the illumination optics used to switch between the first illumination optical system and the second illumination optical system are based on the type of the sample container having one or more sample storage portions and the imaging position. The system performs imaging, and the imaging arrangement determining method includes: an effective visual field determining step of determining an effective visual field region when imaging by the imaging device; and a first imaging arrangement determining step of walling the sample storage portion a plurality of imaging positions for imaging using the first illumination optical system; and a second imaging arrangement determining step for including at least one of all the imaging positions by an arbitrary position in the sample storage unit The method of determining the effective field of view area obtained by the imaging is determined by using the first imaging configuration described above. A region other than the effective field of view of the imaging area on the imaging position of the plurality of decision obtained, the configuration should be conducted using a plurality of imaging positions of the second imaging illumination optical systems. According to a second aspect of the present invention, in the effective field of view region determining step, the effective field of view region when the first illumination optical system is used for imaging is determined, and the first aspect is used. (2) an effective field of view area when the illumination optical system performs imaging, and each of the effective fields of view obtained by imaging at two imaging positions adjacent to each other along the wall surface of the sample storage unit in the first imaging arrangement determining step In a manner in which one of the regions overlaps each other, the arrangement of the plurality of imaging positions using the imaging of the first illumination optical system is determined, and in the second imaging arrangement determining step, the first imaging arrangement determining step is determined. In a region other than the effective field of view obtained by the imaging at the plurality of imaging positions, one of the effective field of view regions obtained by the imaging at the two imaging positions adjacent to each other overlaps with each other. The arrangement of the plurality of imaging positions of the imaging by the second illumination optical system is used. According to a second aspect of the present invention, in the second aspect of the present invention, the shape of the bottom surface of the sample storage portion is circular, and the first imaging arrangement determining step includes: a reference position determining step of determining a reference position which is any one of a plurality of imaging positions of the imaging using the first illumination optical system; and an imaging position order determining step of connecting the center of the sample storage unit to the reference position The line segment is rotated at a specific angle by the center of the sample storage portion as a center of rotation, and each time the rotation is performed, the end of the two end points of the rotated line segment and the center of the sample storage portion are The position of the end point different in the point is selected as the imaging position at which the imaging of the first illumination optical system is to be performed. According to a fourth aspect of the present invention, in the third aspect of the present invention, in the reference position determining step, imaging using the first illumination optical system is performed while gradually moving the imaging position. The brightness at the position of the wall surface of the sample storage unit is selected to be substantially the same as the brightness of the center of the sample storage unit when the image is captured by the second illumination optical system at the center of the sample storage unit. It is the above reference position. According to a fifth aspect of the present invention, in the second imaging arrangement determining step, the imaging is performed at all imaging positions by the imaging device. The number of scanning times required is the smallest, and it is determined that the arrangement of the plurality of imaging positions using the imaging of the second illumination optical system is performed. According to a sixth aspect of the present invention, in the second aspect of the present invention, in the second imaging arrangement determining step, the imaging number of the imaging device is minimized. The arrangement of a plurality of imaging positions using the imaging of the second illumination optical system described above is performed. According to a seventh aspect of the present invention, in the first aspect of the present invention, in the first imaging arrangement determining step, the image is detected by the imaging device with respect to the main scanning The imaging interval in the direction perpendicular to the direction is close to the equal interval, and fine adjustment of a plurality of imaging positions to be imaged using the first illumination optical system is performed. According to an eighth aspect of the present invention, in the first aspect of the present invention, in the effective field of view region determining step, the type of the sample container and the injection into the sample storage are considered. The culture conditions of the imaging target in the section determine the effective visual field area. According to a ninth aspect of the present invention, the first aspect of the present invention, characterized in that the incident state of the chief ray toward the bottom surface of the sample storage portion is in the first illumination optical system The second illumination optical system is different in the above. According to a ninth aspect of the present invention, the first illumination optical system is configured such that the principal ray is incident on the bottom surface of the sample storage portion in a state in which the principal ray is parallel. The sample accommodating portion emits light, and the second illuminating optical system emits light toward the sample accommodating portion so that the principal ray incident on the bottom surface of the sample accommodating portion has a component away from the optical axis. An imaging optical system including a light that receives a component whose principal ray has a direction away from the optical axis. According to an eleventh aspect of the present invention, in an image pickup apparatus, an image pickup object that is held in a sample storage portion having a light-transmitting property on a bottom surface together with a liquid is provided, and a container holding portion is provided. a sample container having one or more sample storage portions; an illumination unit that emits light to an imaging target held by the sample storage unit; and an imaging unit that holds an imaging target held in the sample storage unit a driving unit that integrally moves the imaging unit and the illumination unit according to an imaging position; and a control unit that controls an operation of the illumination unit, the imaging unit, and the driving unit; the illumination unit includes an orientation a first illumination optical system and a second illumination optical system in which incident directions of principal rays of the bottom surface of the sample storage portion are different from each other, wherein the control portion is a plurality of imaging positions arranged along a wall surface of the sample storage portion a plurality of imaging positions of the first imaging position group and an area other than the effective field of view area obtained by the imaging of the first imaging position group (2) a method of imaging on the imaging position group, controlling the imaging unit and the driving unit, and controlling the light emitted from the first illumination optical system when imaging is performed at an imaging position included in the first imaging position group The illumination unit controls the illumination unit such that light is emitted from the second illumination optical system when imaging is performed at an imaging position included in the second imaging position group. According to a twelfth aspect of the present invention, the control unit includes an imaging position adjustment unit that is based on a position of the sample container on the container holding portion and In the orientation, the imaging position from the external direction is corrected, and the imaging position when the imaging unit actually performs imaging is obtained. According to a thirteenth aspect of the invention, the first illumination optical system is characterized in that the first illumination optical system is incident on a bottom surface of the sample storage portion in a state where the principal ray is parallel. The second illumination optical system emits light toward the sample storage unit such that the principal ray incident on the bottom surface of the sample storage portion has a component away from the optical axis. The imaging unit includes an imaging optical system configured to receive light having a principal ray having a component away from the optical axis. [Effects of the Invention] According to the first aspect of the present invention, when an image is taken by an image pickup apparatus provided with two illumination optical systems (the first illumination optical system and the second illumination optical system), when the effective field of view area is selected A plurality of imaging apparatuses that use the first illumination optical system (for example, an illumination optical system suitable for imaging in a region affected by the meniscus) are disposed so as to arrange a plurality of imaging positions along the wall surface of the sample storage portion. The configuration of the camera position. When considering the arrangement of a plurality of imaging positions along the wall surface of the sample storage unit, if one imaging position near the wall surface is selected, the imaging position should be performed for all imaging positions to be imaged using the first illumination optical system. The distance between the wall surfaces of the accommodating portion is selected. Further, the distance between the two imaging positions adjacent to each other may be determined in consideration of the effective visual field area. According to the above, it is relatively easy to determine the arrangement of a plurality of imaging positions to be imaged using the first illumination optical system. In addition, when the arrangement of the imaging position of imaging using the second illumination optical system (for example, an illumination optical system suitable for imaging in a region not affected by the meniscus) is performed, the first illumination optical system is used based on the configuration. The effective field of view of the plurality of imaging positions of the camera has been selected. Therefore, the breadth of the remaining area and the breadth of the effective visual field area when the second illumination optical system is used can be considered, and the arrangement of the imaging position at which the imaging using the second illumination optical system is to be performed can be relatively easily determined. As described above, it is possible to easily determine the arrangement of the imaging arrangement (a plurality of imaging positions) when imaging is performed using an imaging device having two illumination optical systems. According to the second aspect of the present invention, since the imaging arrangement is determined such that one of the effective field of view regions obtained by the imaging at the two imaging positions adjacent to each other overlaps each other, it is possible to surely prevent the plurality of An area where the brightness is insufficient is generated in the entire image obtained by the combination of the captured images. According to the third aspect of the present invention, the plurality of imaging positions to be imaged using the first illumination optical system can determine the remaining imaging positions relatively easily by determining the reference position. According to the fourth aspect of the present invention, since the brightness of the image of the image of the wall surface portion of the sample storage portion is equal to the brightness of the image of the image of the center portion of the sample storage portion, a high-quality overall image can be obtained. . According to the fifth aspect of the present invention, since the number of scans is small, the time required until the end of imaging at all the imaging positions is shortened. According to the sixth aspect of the present invention, since the number of images is small, resources can be effectively utilized. According to the seventh aspect of the present invention, a plurality of imaging positions are efficiently arranged in an area where imaging by the second illumination optical system is to be performed. According to the eighth aspect of the present invention, since the effective field of view area is selected in consideration of the type of the sample container or the culture condition, a plurality of imaging positions can be more efficiently arranged, and synthesis by a plurality of captured images can be suppressed. An area where the brightness is insufficient is generated in the overall image obtained. According to the ninth aspect of the present invention, the same effects as those of any of the first to eighth aspects of the present invention can be obtained. According to the tenth aspect of the present invention, for example, it is possible to easily determine an imaging arrangement for imaging by an imaging device that switches the illumination optical system between an area affected by the meniscus and an area that is not affected by the meniscus. According to the eleventh aspect of the present invention, when imaging is performed, a different illumination optical system is used in a region in the vicinity of the wall surface of the sample storage portion and in other regions. Therefore, even if a meniscus is formed on the surface of the liquid injected into the sample storage unit, it is possible to suppress an area where the brightness is insufficient in the entire image obtained by the combination of the plurality of captured images. Further, since the imaging is performed at a plurality of imaging positions that are efficiently arranged, the imaging processing can be performed efficiently. According to the twelfth aspect of the present invention, even if a design error of the sample container or a positional shift when the sample container is set occurs, a desired image pickup image can be obtained. Further, since there is no need to reset the sample container when there is a positional shift, it is possible to obtain a desired captured image without causing damage to the sample (cell or the like). According to the thirteenth aspect of the present invention, for example, in an image pickup apparatus that switches an illumination optical system in an area affected by a meniscus and an area that is not affected by a meniscus, the plurality of captured images can be combined to be generated. The image processing is performed efficiently in the entire image.

Hereinafter, an embodiment of the present invention will be described with reference to the accompanying drawings. <1. (Configuration of Image Pickup Apparatus) Fig. 1 is a view showing a schematic configuration of an image pickup apparatus 1 according to an embodiment of the present invention. The imaging device 1 is for detecting a cell, a cell population, a bacterium, or the like (hereinafter referred to as "cell, etc.") which is cultured in a liquid injected into a hole W formed on the upper surface of the orifice plate WP. Sample device. The orifice plate WP has a flat shape. In the orifice plate WP, a plurality of (for example, six, 24, 96, 384, etc.) holes W as the sample storage portions having openings on the upper surface side and having a transparent bottom surface on the lower surface side are arranged. In the above, the example in which the orifice plate WP is used as the sample container will be described. However, the present invention is not limited thereto, and a container called a dish (a container having only one sample storage portion) may be used. Used as a sample container. Regarding the shape of the hole W, the cross section is typically circular and the bottom surface is flat. However, the shape of the cross section and the bottom surface of the hole W is not limited to this. The diameter and depth of the hole W are generally from several mm to several 10 mm. A specific amount of a liquid (culture liquid) as a medium M for providing a growth environment for cells or the like is injected into each of the wells W. The amount of the liquid injected into each of the holes W is generally about 50 to 200 μl. In the present embodiment, cells or the like which are cultured under the specific culture conditions in the liquid become an imaging target. As shown in FIG. 1, the imaging device 1 includes an illumination unit 10 that emits light for imaging, a holder 12 that holds the aperture disk WP, an imaging unit 13 that images an image (cell or the like) in the hole W, and a control illumination unit. The control unit 14 that operates the imaging unit 13 and the drive unit 15 that moves the illumination unit 10 and the imaging unit 13 during imaging. The illumination unit 10 is disposed on the upper portion of the imaging device 1. The holder 12 is disposed below the illumination unit 10, and the imaging unit 13 is disposed below the holder 12. In the following, a region which is affected by the meniscus when included in the imaging field in the region in the hole W is referred to as a "hole edge portion". Further, when imaging the edge portion of the hole, there is a case where the imaging position is located further outward than the edge end (wall surface) of the hole W. Therefore, a region including the region closer to the outer side than the side end portion and the region closer to the inner side than the side end portion is referred to as a "hole peripheral region". The edge portion of the hole is a region in the peripheral portion of the hole that is further inside than the edge portion. Further, a region (not strictly speaking, a region where the influence of the meniscus is sufficiently small) which is not affected by the meniscus when included in the imaging field in the region in the hole W is referred to as a "hole center region". <1. 1 Illumination unit> The illumination unit 10 includes two light sources (a first light source 101 and a second light source 111) such as a white LED (Light Emitting Diode), two mirrors 102 and 105, and two light collecting lenses 103. 112, the beam splitter 104, and the collecting lens 106. The light emitted from the first light source 101 is folded back by the mirror 102, and then incident on the beam splitter 104 via the collecting lens 103. The light emitted from the second light source 111 is incident on the beam splitter 104 via the collecting lens 112. The light emitted from the dichroic mirror 104 is oriented in the (-Z) direction by the mirror 105, that is, the vertical direction. Then, the light whose traveling direction is in the downward direction is emitted downward from the illumination unit 10 via the condensing lens 106. The light emitted from the illumination unit 10 is incident on at least one hole W from above the orifice plate WP supported by the holder 12, and illuminates the object to be imaged in the hole W. As described above, the illumination unit 10 of the present embodiment includes an illumination optical system (hereinafter referred to as a "first illumination optical system") 100 that uses the first light source 101 as a light source, and illumination optical with the second light source 111 as a light source. The system (hereinafter referred to as "second illumination optical system") 110. The first illumination optical system 100 includes a first light source 101, a mirror 102, a collecting lens 103, a beam splitter 104, a mirror 105, and a collecting lens 106. The second illumination optical system 110 includes a second light source 111, a collecting lens 112, a beam splitter 104, a mirror 105, and a collecting lens 106. Further, the beam splitter 104, the mirror 105, and the collecting lens 106 are shared by the first illumination optical system 100 and the second illumination optical system 110. The first light source 101 and the second light source 111 are selectively turned on based on a control signal supplied from the light source control unit 146 in the control unit 14. Therefore, the illumination unit 10 can emit light emitted from the first illumination optical system 100 (hereinafter referred to as "first illumination light") and light emitted from the second illumination optical system 110 (hereinafter referred to as "second illumination light". ") Selectively incident into the hole W. The first illumination light and the second illumination light are combined by the beam splitter 104, and these can be emitted coaxially. In other words, the central axes of the first illumination light and the second illumination light emitted from the condenser lens 106 match each other. 2 is a ray diagram of the first illumination optical system 100 and the second illumination optical system 110. In FIG. 2, the first illumination optical system 100 and the second illumination optical system 110 are separately described in order to clearly show the optical path. Moreover, for convenience of explanation, the optical axis which is actually bent by the mirrors 102 and 105 and the beam splitter 104 is indicated by a straight line. Therefore, the illustration of the mirrors 102 and 105 and the beam splitter 104 having the function of bending the optical axis is omitted. In the first illumination optical system 100, the light emitted from the first light source 101 is condensed by the collecting lens 103. The light that has been collected by the condensing lens 106 is emitted toward the sample surface on which the cells to be imaged or the like are present. Usually, the sample surface is the bottom surface of the hole W. The collecting lens 103 images the image of the first light source 101 between the collecting lens 103 and the collecting lens 106. That is, the conjugate point C1 of the first light source 101 exists between the collecting lens 103 and the collecting lens 106. Further, the collecting lens 103 and the collecting lens 106 are configured such that the chief ray from the collecting lens 106 toward the sample surface is parallel to the optical axis. That is, the first illumination optical system 100 forms telecentric illumination. The light exit surface of the first light source 101 is an angular range defining the light incident on the collecting lens 103, and an aperture stop 107 is provided as needed. The NA (numerical aperture) of the illumination can be adjusted by the aperture stop 107. Further, a field stop 108 is provided as needed on the rear side of the collecting lens 103 and on the front side of the conjugate point C1. Thereby, only the range required for imaging is illuminated, so that glare in the imaging optical system can be prevented from occurring. In the second illumination optical system 110, the light emitted from the second light source 111 is condensed by the collecting lens 112. The condensed light is emitted toward the sample surface via the condensing lens 106. The position of the condensing point C2 to which the second light source 111 is applied in the collecting lens 112 is closer to the rear side than the condensing lens 106, and the refractive index is further forward than the sample surface. The light exit surface of the second light source 111 is an angular range defining the light incident on the collecting lens 112, and an aperture stop 113 is provided as needed. The aperture of the aperture stop 113 is set such that the NA of the illumination light emitted from the condenser lens 106 becomes NA or more of the objective lens 131. Thereby, it is possible to prevent the resolution of the imaging optical system from being restricted by illumination. Further, between the collecting lens 112 and the collecting lens 106, a field stop 114 is provided as needed. Thereby, only the range required for imaging is illuminated, so that glare in the imaging optical system can be prevented from occurring. The first illumination optical system 100 and the second illumination optical system 110 share the condensing lens 106. A beam splitter 104 for realizing this sharing is disposed between each of the collecting lenses 103, 112 and the collecting lens 106. More specifically, in the first illumination optical system 100, the rear side of the collecting lens 103 (the field stop 108 when the field stop 108 is provided) is further rearward than the collecting lens 106. The position of the second illumination optical system 110 is further rearward than the collecting lens 112 (the field stop 114 when the field stop 114 is provided) and is further forward than the collecting lens 106. Position, the beam splitter 104 is set. <1. 2 Bracket> When imaging is performed by the imaging device 1, the orifice disk WP including the plurality of holes W holding the sample and the medium M is held in the holder 12. The bracket 12 abuts against the peripheral portion of the lower surface of the orifice plate WP to maintain the orifice plate WP in a substantially horizontal posture. <1. 3 Imaging Unit> The imaging unit 13 includes an objective lens 131, a low-magnification afocal system 132, a high-magnification afocal system 133, a mirror 134, an imaging lens 135, and an imaging element 136. The objective lens 131 is disposed at a position directly below the orifice plate WP. The optical axis of the objective lens 131 is oriented in the vertical direction, and is coaxial with the optical axes of the first illumination optical system 100 and the second illumination optical system 110. The light emitted from the illumination unit 10 and incident on the liquid (medium medium M) from above the hole W illuminates the imaging target, and the light transmitted downward from the bottom surface of the hole W is incident on the objective lens 131. Below the objective lens 131, a low-magnification afocal system 132 and a high-magnification afocal system 133 are switchably provided. Here, the switching between the low-magnification afocal system 132 and the high-magnification afocal system 133 will be described. The low-magnification non-focus system 132 and the high-magnification non-focus system 133 can be integrally moved in the horizontal direction by a drive mechanism (not shown), and one of the two is selectively disposed in the objective lens 131 at the time of imaging. Directly below. As shown by the solid line in FIG. 1, the high-magnification-based afocal system 133 is disposed at a position directly below the objective lens 131, and constitutes an imaging optical system including a high magnification of the objective lens 131 and the high-magnification afocal system 133. At this time, the relatively narrow range of the imaging target is imaged at a high magnification. On the other hand, as shown by the broken line in FIG. 1, the low-magnification non-focus system 132 is disposed at a position directly below the objective lens 131, and the low-magnification imaging including the objective lens 131 and the low-magnification afocal system 132 is included. Optical system. At this time, the range in which the object to be imaged is relatively wide is imaged at a low magnification. The light emitted from the afocal system (the low-magnification non-focus system 132 or the high-magnification non-focus system 133) is folded back by the mirror 134, and then incident on the image pickup element 136 via the imaging lens 135. As described below, the imaging optical system including the objective lens 131, the low-magnification afocal system 132, and the imaging lens 135 has an optical characteristic of an object-focusing telecentric. On the other hand, the imaging optical system including the objective lens 131, the high-magnification afocal system 133, and the imaging lens 135 has an optical property of an object telecentric. The imaging element 136 is an area image sensor having a two-dimensional light receiving surface. As the imaging element 136, a CCD sensor, a CMOS (Complementary Metal Oxide Semiconductor) sensor, or the like can be used. The image of the imaging target image formed on the light receiving surface of the imaging element 136 by the imaging lens 135 is imaged by the imaging element 136. The imaging element 136 converts the optical image obtained by the light into an electrical signal, and outputs the electrical signal as an image signal. According to such an imaging method, it is possible to image a cell or the like as an imaging target in a non-contact, non-destructive, and non-invasive manner, thereby suppressing damage to cells and the like caused by imaging. Further, the operation of each unit of the imaging unit 13 is controlled by the imaging control unit 143 provided in the control unit 14. <1. 4 control unit> The control unit 14 includes a CPU (Central Processing Unit) 141, an interface (IF) unit 142, an imaging control unit 143, and an AD converter (A/D (analog to digital) 144. The mechanical control unit 145, the light source control unit 146, the image memory 147, and the memory 148. The CPU 141 performs control of various operations of the components in the control unit 14 or various arithmetic processing. The interface 142 has a function of accepting a function input from a user, a function of displaying information such as a result of processing by the user, and a function of communicating data with another device via a communication line. Further, the interface 142 is connected to an input accepting unit (a keyboard or a mouse) that accepts an operation input, a display unit that performs information display, a communication line, and the like. The imaging control unit 143 controls the operation of the imaging unit 13 so as to image the imaging target in accordance with the scanning movement recipe described below. The AD converter (A/D) 144 receives an image signal (analog data) output from the image pickup element 136, and converts the image signal into digital image data. Based on the digital image data, the CPU 141 performs appropriate image processing. The machine control unit 145 moves the imaging unit 13 in the horizontal direction or the vertical direction by actuating the drive mechanism 15. The imaging unit 13 is moved in the horizontal direction with respect to the hole W by moving the imaging unit 13 in the horizontal direction. Further, focus adjustment is performed by moving the imaging unit 13 in the vertical direction. Further, the machine control unit 145 moves the illumination unit 10 in the horizontal direction by actuating the drive mechanism 15. The light source control unit 146 selectively lights the first light source 101 and the second light source 111 in accordance with the imaging position. The image memory 147 holds digital image data. The memory 148 holds a program to be executed by the CPU 141 or data generated by the CPU 141. Furthermore, the image memory 147 and the memory 148 may also be integrated. Moreover, the image memory 147 and the memory 148 can also be realized by an appropriate combination of mass storage and semiconductor memory. <1. 5 Drive Mechanism> The drive mechanism 15 moves the illumination unit 10 in the horizontal direction. Further, the drive mechanism 15 moves the imaging unit 13 in the horizontal direction or the vertical direction. In the imaging device 1, the positional relationship between the illumination unit 10 and the imaging unit 13 is selected such that the center of the emitted light from the illumination unit 10 substantially coincides with the optical axis of the objective lens 131. Therefore, when the imaging unit 13 moves the imaging unit 13 in the horizontal direction, the illumination unit 10 moves the illumination unit 10 integrally with the imaging unit 13. Thereby, a good illumination state can be maintained regardless of which of the holes W is imaged. Further, in Fig. 1, the Z direction indicates the vertical direction, the Y direction indicates the main scanning direction, and the X direction indicates the sub-scanning direction. <2. Illumination Optical System> FIG. 3 is a view showing the first illumination light L1 emitted from the first illumination optical system 100. As shown in FIG. 3, the first illumination light L1 emitted from the condensing lens 106 in the first illumination optical system 100 is incident on a state in which the chief ray is parallel to the sample bottom surface Wb which is distributed on the imaging target. . That is, the first illumination optical system 100 forms a telecentric illumination in which the exit pupil position is at infinity. FIG. 4 is a view showing the second illumination light L2 emitted from the second illumination optical system 110. As shown in FIG. 4, the second illumination light L2 emitted from the condensing lens 106 in the second illumination optical system 110 travels close to the optical axis of the second illumination optical system 110, and is closer to the hole bottom surface Wb. The upper position (the position viewed from the illumination optical system closer to the front side than the hole bottom surface Wb) intersects the optical axis. In other words, when the image of the second light source 111 (more strictly, the image of the aperture stop 113) is imaged on the optical path of the second illumination light L2, the emission pupil position Pp is observed from the second illumination optical system 110. It is located closer to the hole bottom surface Wb than the sample surface on which the imaging object is distributed. More specifically, in the illumination of the second illumination optical system 110, the image of the second light source 111 is performed at a position between the output end of the condensing lens 106 from which the second illumination light L2 is emitted and the objective lens 131 of the imaging optical system. Imaging. That is, there is a conjugate point for the second light source 111 at this position. Further, the holder 12 holds the orifice plate WP such that the hole bottom surface Wb is located between the conjugate point and the objective lens 131. Therefore, the chief ray of the second illumination light L2 incident on the hole bottom surface Wb has a directional component in a direction away from the optical axes of the second illumination optical system 110 and the objective lens 131. As described above, the first illumination optical system 100 and the second illumination optical system 110 are different from each other in the pupil position. The first illumination optical system 100 and the second illumination optical system 110 are switched and used as described below. Furthermore, the flasher illumination is used during imaging. In other words, the illumination light is emitted only for a short time when the imaging unit 13 performs imaging. Therefore, the light source control unit 146 can switch between the illumination light by selecting which one of the two light sources (the first light source 101 and the second light source 111) is to be lit. Further, as described above, a liquid as the medium M is injected into each of the holes W. Therefore, the illumination light incident from above the hole W is incident on the hole bottom surface Wb (sample surface) via the liquid surface of the medium M. Here, the liquid surface in the hole W forms a concave meniscus. Therefore, the traveling line of the illumination light is bent outward from the center of the hole W due to the refraction. The refractive index is small near the center of the hole W, and the edge portion (wall surface) closer to the hole W becomes larger. In the present embodiment, the imaging optical system forming object-focusing telecentric optical system including the objective lens 131 efficiently condenses the light thus curved outward and guides it to the image-capturing element 136. That is, the obliquely incident light can be imaged in the imaging element 136 at a position away from the optical axis of the lens. <3. Separate Use of Illumination Optical System> Next, the separate use of the first illumination optical system 100 and the second illumination optical system 110 will be described. Here, attention is paid to the case where the area to be imaged (the entire area of the hole) is wider than the imaging field of view. When the area to be imaged is wider than the imaging field of view, the area is divided into a plurality of images for imaging. Then, a plurality of captured images obtained by imaging are synthesized by image processing, whereby an image indicating the entire area to be imaged is generated. FIG. 5 is a view showing a state of imaging when the first illumination optical system 100 is temporarily used when the hole edge portion WR is not included in the imaging field of view. When the imaging field of view V includes only the central region of the hole away from the edge end of the hole W, the influence of the meniscus on the optical path is sufficiently small. Further, as described above, the imaging optical system including the objective lens 131 forms a fixed-focus telecentric optical system. In other words, the imaging optical system is configured to receive light that is obliquely inclined toward the refraction of the meniscus at a position away from the optical axis (light shown by a broken line in FIG. 5 in the first illumination light L1). However, the light passing through the hole W does not linearly retreat by the refraction of the meniscus, so the inclination of the chief ray in the incident light does not coincide with the inclination of the chief ray on the light receiving side. When the telecentric illumination is used as described above, the light incident on the optical axis of the objective lens 131 is condensed and incident on the imaging element 136, but the difference in inclination between the incident light and the chief ray of the optical system occurs at a position away from the optical axis. And the resulting mismatch. As a result, deterioration of image quality is caused particularly in the peripheral portion of the imaging field of view V. On the other hand, when the hole edge portion WR is included in the imaging field of view V, when the first illumination optical system 100 is used, as shown in FIG. 6, the inclination of the chief ray of the light refracted by the meniscus at the hole edge portion WR is as shown in FIG. The light is substantially uniformly aligned with the inclination of the chief ray on the light receiving side, so that the light is efficiently collected. As described above, when the hole edge portion WR is included in the imaging field of view V, imaging is performed using the first illumination optical system 100. FIG. 7 is a view showing a state of imaging when the second illumination optical system 110 is used when the hole edge portion WR is not included in the imaging field of view V. As described above, the second illumination light L2 emitted from the condensing lens 106 in the second illumination optical system 110 travels close to the optical axis. Therefore, when there is no influence of the meniscus, the chief ray of the second illumination light L2 incident on the hole bottom surface Wb does not become parallel to each other. In the present embodiment, the exit pupil position Pp (observed from the illumination optical system) of the second illumination optical system 110 is located closer to the front side than the hole bottom surface Wb, so that the second illumination light L2 incident on the hole bottom surface Wb is The chief ray becomes a broader one from the optical axis of the objective lens 131. Here, when the imaging optical system is configured such that the inclination of the chief ray of the incident light to the objective lens 131 coincides with the inclination of the chief ray on the objective lens 131 side, as shown in FIG. 7, the light passing through the bottom surface Wb of the aperture is provided by the objective lens 131. The light is concentrated and eventually directed to the imaging element 136. When the hole edge portion WR is not included in the imaging field of view V, the second illumination optical system 110 performs imaging. As described above, in the imaging device 1 of the present embodiment, the first illumination optical system 100 that is suitable for imaging of a region affected by the meniscus, and the imaging device that is suitable for an area that is not affected by the meniscus are provided. 2 Illumination optical system 110 as an illumination optical system combined with an imaging optical system having a telecentric telecentric characteristic. Further, when the hole edge portion WR is included in the imaging field of view (that is, when the region affected by the meniscus is included in the imaging field of view) and the hole edge portion WR is not included in the imaging field (that is, the region affected by the meniscus) These two illumination optical systems are used separately when they are not included in the imaging field of view. Thereby, it is possible to obtain an image of a high quality image as a whole of the area to be imaged. <4. Overall Process Flow> Next, a schematic flow of the overall process when the image pickup apparatus 1 performs image pickup will be described. In connection with this, first, the matter to be described before the image pickup apparatus 1 will be described. In the image pickup apparatus 1, the area in which the first illumination optical system 100 is imaged and the area in which the second illumination optical system 110 is imaged are set in advance for each type of the sample container. As described above, the first illumination optical system 100 is suitable for imaging of an area affected by the meniscus, and the second illumination optical system 110 is suitable for imaging of an area not affected by the meniscus. Therefore, for example, when focusing on one hole W of a certain type of sample container (hole plate WP), the area indicated by reference numeral 51 in FIG. 8 is set as an area where imaging is performed using the first illumination optical system 100. The area indicated by symbol 52 in FIG. 8 is set as an area where imaging is performed using the second illumination optical system 110. Such information relating to the setting of the area is previously written in the image pickup apparatus 1 to, for example, a setting file of a specific format. The area indicated by reference numeral 51 in Fig. 8 corresponds to the peripheral region of the hole, and the area indicated by reference numeral 52 in Fig. 8 corresponds to the central portion of the hole. The imaging device 1 is configured to perform imaging at the plurality of imaging positions in accordance with a scanning movement process recipe indicating a plurality of imaging positions and the imaging sequence thereof. In the present specification, the "imaging position" refers to a position corresponding to the center of the imaging field of view at the time of imaging (this position coincides with the position of the optical axis of the objective lens 131). For example, when the imaging configuration (the configuration of the plurality of imaging positions) is as shown in FIG. 9 (the positions indicated by the symbols P1 to P23 indicate the imaging positions), the processing recipe is moved in accordance with the scanning, for example, as indicated by symbol 53 in FIG. Scanning for imaging is performed like an arrow. Further, the orifice plate WP includes a plurality of holes W, but in FIGS. 9 and 10, one hole W is focused on for the sake of convenience. Further, when the imaging device 1 performs imaging in accordance with the scanning movement recipe recipe, based on the above-described information relating to the setting of the region (see FIG. 8), two illumination optical systems (the first illumination optical system 100 and the second image are used depending on the imaging position). One of the illumination optical systems 110). FIG. 11 is a flow chart showing the overall processing flow when the imaging device 1 performs imaging. First, information on the type of the sample container (for example, the manufacturer name and model number) of the imaging target and the imaging conditions such as the amount of the liquid used as the medium M are obtained (step S10). Next, the type of the sample container and the imaging conditions are considered, and the imaging arrangement as shown in FIG. 9 is determined (step S20). Further, a detailed description of a method of determining an imaging configuration will be described below. After the imaging configuration decision, a scan moving recipe recipe is created based on the imaging configuration (step S30). Thereafter, the scanning movement process recipe is given to the image pickup apparatus 1, and the type of the sample container of the image pickup target is designated (selected) by the operator in the image pickup apparatus 1 (step S40). Thereby, the imaging apparatus 1 performs imaging based on the scanning movement process recipe (step S50). <5. Method of determining the imaging configuration> Next, the method of determining the imaging configuration will be described in detail. Fig. 12 is a flow chart showing the procedure for determining the imaging configuration. <5. (1) Determination of the effective field of view area. First, an effective field of view area which is an area which is considered to be an image in which a sufficient amount of light is obtained when a sufficient amount of light is imaged during imaging is determined (step S210). The image in only the effective field of view determined in step S210 in the captured image constitutes an image that is ultimately presented to the user. In other words, when one hole W is imaged at a plurality of imaging positions, an image representing only one hole W is created by combining images in only the effective field of view of each of the plurality of captured images. The image. In the imaging device 1 of the present embodiment, the first illumination optical system 100 is used when imaging is performed in the peripheral region of the aperture, and the second illumination optical system 110 is used when imaging is performed in the central region of the aperture. Therefore, in step S210, the effective field of view area when the first illumination optical system 100 is imaged and the effective field of view when the second illumination optical system 110 is used for imaging are determined. A specific example of each of the effective field of view area when the first illumination optical system 100 is imaged and the effective field of view area when the second illumination optical system 110 is used for imaging will be described with reference to FIGS. 13 to 17 . As shown by reference numeral 54 in Fig. 13, when the peripheral edge region of the hole is included in the imaging field of view, imaging is performed using the first illumination optical system 100. At this time, a captured image as shown in FIG. 14 is obtained, for example. From Fig. 14, it can be grasped that there is an area where the brightness is insufficient in the imaging field of view. Therefore, for example, "the area where the brightness is 50% or more when the minimum brightness is 0% and the maximum brightness is 100%" is selected as the effective field of view when the first illumination optical system 100 is used for imaging. In this example, the effective field of view area becomes the area surrounded by the thick line of the symbol 57 in FIG. As shown by reference numeral 55 in Fig. 13, when the peripheral edge region of the hole is not included in the imaging field of view, imaging is performed using the second illumination optical system 110. At this time, a captured image shown in, for example, FIG. 16 is obtained. From Fig. 16, it can be understood that the brightness of the peripheral portion of the imaging field of view is insufficient. Therefore, for example, a rectangular region of 70% of the entire imaging field of view (the center of the rectangular region is aligned with the center of the imaging field of view) is selected as the effective field of view when the second illumination optical system 110 is used for imaging. In this example, the effective field of view area becomes the area surrounded by the thick line of the symbol 58 in FIG. In particular, regarding the imaging on the peripheral region of the hole, the region where sufficient image quality is obtained varies depending on the culture conditions. For example, the area where sufficient image quality is obtained varies depending on the amount of liquid used as the medium M. For example, when a certain amount of liquid is used as the medium M, even if the area indicated by the hatching of the symbol 61 in Fig. 18 obtains sufficient image quality, the case where different amounts of the same liquid are used as the medium M is used. At the time, there is a case where only the area indicated by the hatching of the symbol 62 in Fig. 18 obtains sufficient image quality. Since the area where the sufficient image quality is obtained is changed by the culture conditions as described above, the determination of the effective visual field area (step S210) is preferably carried out in consideration of the culture conditions. <5. 2. Determination of the imaging position in the peripheral region of one hole> After the effective visual field region is determined as described above, one of the plurality of imaging positions in the peripheral region of the hole is determined (step S220). Hereinafter, the imaging position determined in step S220 will be referred to as a "reference position". Here, when the hole W is viewed in a certain direction, an example in which the reference position BP is placed above the hole W as shown in FIG. 19 will be described. In addition, the area indicated by the hatching of the symbol 63 in FIG. 19 indicates the effective field of view area at the reference position BP (the effective field of view area when the reference position BP is the center of the imaging field of view). When the reference position BP is determined, the brightness of the edge portion (wall surface) WE of the hole W is observed, and the imaging field of view V is gradually moved as shown in FIG. More specifically, the center of the imaging field of view V is located on the straight line 65 passing through the center WC of the hole W and the one end portion WE of the hole W, and the area in the hole W included in the imaging field of view V is gradually enlarged. The imaging field of view V is gradually moved while observing the brightness of the edge portion WE. Then, the luminance obtained when the center WC of the aperture W of the second illumination optical system 110 is used (the luminance of the center WC of the aperture W) and the luminance of the edge portion WE obtained when the peripheral edge region of the imaging aperture of the first illumination optical system 100 is used are used. The substantially consistent position is selected as the reference position BP. However, there is also a case where the range (area) at which sufficient brightness is obtained is largely changed by simply moving the imaging field of view V by a small amount. Therefore, when the brightness of the edge portion WE is substantially the same as the brightness of the center WC of the hole W, it is preferable to select the reference position BP so that the range (area) of sufficient brightness is as large as possible. As can be grasped from Fig. 21, the distance LE between the reference position BP and the edge end WE of the hole W is selected by selecting the reference position BP. In other words, the distance between the imaging position at the time of imaging using the first illumination optical system 100 and the edge portion WE of the hole W is selected. After the determination of the reference position BP, the remaining imaging position (the imaging position other than the reference position BP) in the peripheral region of the hole is determined (step S230). In step S230, first, as shown in FIG. 22, the straight line 70 of the center WC of the connection hole W and the reference position BP is rotated about the center WC of the hole W, and the imaging position 73 adjacent to the reference position BP is set. The imaging position 73 adjacent to the reference position BP is determined such that the upper effective visual field area 72 partially overlaps the effective visual field area 71 on the reference position BP. At this time, the distance from the edge portion WE of the hole W to the imaging position 73 is equal to the distance LE from the edge portion WE of the hole W to the reference position BP (see FIG. 21). In other words, the line segment of the center WC of the connection hole W and the reference position BP is rotated about the center WC of the hole W, and the end point of the line segment after the rotation is different from the end point of the position at the center WC of the hole W. The position of the other end point is selected as the imaging position 73 adjacent to the reference position BP. After the imaging position 73 adjacent to the reference position BP is determined in this manner, the imaging position adjacent to the imaging position 73 (adjacent to the side opposite to the reference position BP) is determined in the same manner. Repeat the above process. At this time, the overlapping portion of the effective view region at the two imaging positions adjacent to each other is fixed in size (the center WC of the connection hole W is fixed at an angle formed by the two line segments of the two imaging positions adjacent to each other). In this way, as shown in FIG. 23, all the imaging positions that should be disposed in the peripheral region of the hole are determined. Further, as shown in FIG. 24, there is also a case where the imaging position is disposed outside the edge portion WE of the hole W. As described above, in the present embodiment, first, a plurality of imaging positions are arranged along the wall surface of the hole W as the sample storage portion, and the hole peripheral region is determined (the imaging using the first illumination optical system 100 should be performed). The configuration of a plurality of imaging positions in the area). <5. 2. Determination of the imaging position in the central region of the second hole> Thereafter, the imaging position (usually a plurality of imaging positions) in the central region of the hole (the region other than the peripheral edge region of the hole) is determined (step S240). In this step S240, the imaging position is determined such that the region other than the effective visual field region based on the imaging position in the peripheral region of the hole is efficiently filled as the effective visual field region based on the imaging position in the central region of the aperture. In the present embodiment, specifically, the imaging position is determined such that the number of scans required for imaging by the imaging device 1 at all imaging positions is minimized. This will be described with reference to FIGS. 25 and 26. An example of an imaging arrangement in a partial area is shown in FIGS. 25 and 26. In Fig. 25, the imaging positions of the peripheral regions of the holes are indicated by symbols P31 and P32, and the imaging positions of the central regions of the holes are indicated by symbols P41 to P44. In Fig. 26, the imaging positions of the peripheral regions of the holes are indicated by symbols P51 and P52, and the imaging positions of the central regions of the holes are indicated by symbols P61 to P64. In addition, in FIGS. 25 and 26, the outer edge of the effective field of view area at each imaging position is indicated by a thick line. If it is assumed that the imaging configuration in a certain area is as shown in FIG. 25, it is necessary to obtain the captured image of the area, and the imaging unit 13 is reciprocated once in the main scanning direction. That is, the scanning of the imaging position in the peripheral region of the hole is different from the scanning of the imaging position in the central portion of the hole. On the other hand, if the imaging arrangement in a part of the area is as shown in FIG. 26, the imaging image of the area can be obtained by causing the imaging unit 13 to perform only one-way movement in the main scanning direction (that is, one scan). In this way, it is possible to grasp the influence of the arrangement method of a plurality of imaging positions on the efficiency of imaging. Therefore, as described above, in the present embodiment, the imaging position in the central portion of the hole is determined such that the number of scans required for imaging by the imaging device 1 at all imaging positions is minimized. In the above manner, the imaging position in each of the hole peripheral region and the hole central region is determined. Thereby, for example, the imaging configuration is determined as shown in FIG. Furthermore, in the present embodiment, the effective field of view region determining step is implemented by the above-described step S210, the first imaging arrangement determining step is implemented by the steps S220 and S230, and the second imaging configuration is realized by the above step S240. Decide the steps. Further, the reference position determining step is realized by the above-described step S220, and the imaging position order determining step (see FIG. 12) is realized by the above-described step S230. <6. [Effects] According to the present embodiment, the first illumination optical system 100 provided with an image suitable for the region affected by the meniscus and the second illumination optical system 110 adapted to image the region not affected by the meniscus are used as the second illumination optical system 110. When the imaging device 1 of the illumination optical system performs imaging, when the effective field of view area in the case where each illumination optical system is used is selected, a plurality of imaging positions are arranged along the edge end (wall surface) WE of the hole W, and the determination should be made. The arrangement of the plurality of imaging positions of the imaging of the first illumination optical system 100 is used. When an arrangement of a plurality of imaging positions along the edge portion WE is considered, if one imaging position (the above-described reference position BP) near the edge portion is selected, all imaging positions using the imaging of the first illumination optical system 100 are performed correspondingly. The distance from the edge end WE is selected. Further, the distance between the two imaging positions adjacent to each other may be determined by considering the effective visual field area. According to the above, the arrangement of the plurality of imaging positions to be imaged using the first illumination optical system 100 can be relatively easily determined. Moreover, when the arrangement of the imaging position using the imaging of the second illumination optical system 110 is to be considered, the effective visual field area based on the plurality of imaging positions to be imaged using the first illumination optical system 100 is selected. Therefore, the breadth of the remaining area and the breadth of the effective field of view when the second illumination optical system 110 is used can be considered, and the arrangement of the imaging position at which the imaging using the second illumination optical system 110 is to be performed can be relatively easily determined. In addition, the imaging position at which the imaging using the second illumination optical system 110 is performed is determined such that the number of scans required for imaging by the imaging device 1 at all imaging positions is minimized. Therefore, efficient imaging can be performed. Further, since the effective field of view area when each illumination optical system is used is selected in consideration of the type or culture condition of the sample container, a plurality of imaging positions can be disposed more efficiently, and a plurality of captured images can be suppressed. An area where the brightness is insufficient in the overall image obtained by the synthesis. As described above, according to the present embodiment, it is possible to easily determine an imaging configuration in which an image of a high quality can be efficiently obtained by imaging of an imaging device having two illumination optical systems. <7. Variation Example Hereinafter, a modification of the above embodiment will be described. <7. (1) Variations relating to the determination of the imaging position on the central portion of the hole. In the above embodiment, the imaging of the central portion of the hole is determined such that the number of scans required for imaging by the imaging device 1 at all imaging positions is minimized. position. However, the present invention is not limited to this, and it is also possible to determine the imaging position in the central portion of the hole in such a manner that the number of imaging sheets is minimized (that is, the number of imaging positions is minimized). In this way, resources can be used effectively. <7. (2) Variations relating to the determination of the imaging position on the peripheral edge of the hole. In the above embodiment, focusing on the imaging position in the peripheral region of the hole, the portion of the effective imaging region at the two imaging positions adjacent to each other overlaps. The size is fixed (the angle between the center WC of the connection hole W and the two line segments of the two imaging positions adjacent to each other is fixed). In this case, if the X coordinate of the imaging position in the central portion of the hole is aligned with the X coordinate of the imaging position in the peripheral region of the hole to reduce the number of scanning times for imaging, as shown in FIG. 27, relative to the main scanning. In the direction perpendicular to the direction (sub-scanning direction), as the center WC of the hole W approaches the edge end portion WE, the interval between the imaging positions is narrowed. If the area where the interval of the imaging position is dense is generated in this way, the imaging arrangement may be inefficient in reverse. Therefore, when it is assumed that the X coordinate of the center WC of the hole W is 0, as shown in FIG. 28, if the absolute value of the X coordinate of the imaging position is larger, the centers WC of the respective connection holes W may be adjacent to each other. The angle formed by the two line segments of the two imaging positions is larger. In Fig. 28, K1 to K3 indicate angles, and "K1>K2>K3" holds. In the same manner, the imaging interval in the direction perpendicular to the main scanning direction when the imaging device 1 is imaged is close to the equal interval, and the plurality of imaging positions to be imaged using the first illumination optical system 100 are performed. The adjustment is performed to efficiently arrange a plurality of imaging positions in the central portion of the hole. <7. (3) Variations relating to Determination of Effective Field of View Area In the above embodiment, the type of the sample container and the amount of the liquid as the medium M are considered when determining the effective field of view area. However, the invention is not limited thereto. When determining the effective field of view, for example, the state of the surface processing of the sample container, the material of the sample container (the reflectance varies depending on the material), and the physical properties (for example, viscosity, transmittance) of the culture solution (medium) may be considered. <7. 4. Variations relating to the configuration of the imaging device> When imaging is performed by the imaging device 1, the operator places the sample container (the orifice plate WP in the above embodiment) at a specific position on the holder 12. However, there is a case where a desired image of the image cannot be obtained due to a design error of the sample container or a positional shift when the sample container is set. Therefore, in order to obtain the desired captured image even in the presence of such a design error or positional shift, the alignment process can be performed at the time of imaging. Therefore, in the present modification, the alignment processing unit (imaging position adjustment unit) is provided in the control unit 14 of the imaging device 1. For example, although the orifice plate WP should originally be placed on the bracket 12 as shown in Fig. 29, it is assumed that the orifice plate WP is placed on the bracket 12 in a state of being inclined in a plan view as shown in Fig. 30. In this case, the offset between the current position and the original position can be determined for each hole W based on the center position of the plurality of holes W in the orifice plate WP. Based on the offset obtained in this way, the alignment processing unit corrects the imaging position based on the scanning movement process recipe, and obtains the imaging position when the imaging is actually performed. By performing such processing, the desired imaged image can be obtained regardless of the design error of the sample container or the positional deviation when the sample container is set. Further, when the positional shift is present, if the orifice plate WP is newly set, damage to cells or the like may occur. In this respect, if the alignment processing as described above is employed, the re-arrangement of the orifice plate WP is not required, so that the desired captured image can be obtained without causing damage to cells or the like.

1‧‧‧ camera
10‧‧‧Lighting Department
12‧‧‧ bracket
13‧‧‧Photography Department
14‧‧‧Control Department
15‧‧‧Drive mechanism
51‧‧‧ hole peripheral area
52‧‧‧Central area of the hole
53‧‧‧ arrow
54‧‧‧Video field of view
55‧‧‧Video field of view
57‧‧‧ thick line
58‧‧‧ Thick line
61‧‧‧ Shadow
62‧‧‧ Shadow
63‧‧‧ Shadow
65‧‧‧ Straight line
70‧‧‧ Straight line
71‧‧‧effective field of view
72‧‧‧effective field of view
73‧‧‧Photography location
100‧‧‧1st illumination optical system
101‧‧‧1st light source
103‧‧‧ collecting lens
106‧‧‧Concentrating lens
107‧‧‧Aperture aperture
108‧‧‧ Field of view
110‧‧‧2nd illumination optical system
111‧‧‧2nd light source
112‧‧‧ collecting lens
113‧‧‧ aperture diaphragm
114‧‧‧ Field of view
131‧‧‧ Objective lens
BP‧‧‧ reference position
C1‧‧‧ conjugate point
C2‧‧‧ conjugate point
K1‧‧‧ angle
K2‧‧‧ angle
K3‧‧‧ angle
L1‧‧‧1st illumination
L2‧‧‧2nd illumination light
LE‧‧‧ distance
M‧‧‧ medium
Pp‧‧‧ shot out of the position
P1‧‧‧ Camera location
P2‧‧‧ Camera location
P3‧‧‧Photography location
P4‧‧‧Photography location
P5‧‧‧Photography location
P6‧‧‧Photography location
P7‧‧‧ Camera location
P8‧‧‧Photography location
P9‧‧‧Photography location
P10‧‧‧Photography location
P11‧‧‧Photography location
P12‧‧‧ Camera location
P13‧‧‧Photography location
P14‧‧‧Photography location
P15‧‧‧Photography location
P16‧‧‧Photography location
P17‧‧‧Photography location
P18‧‧‧Photography location
P19‧‧‧Photography location
P20‧‧‧Photography location
P21‧‧‧Photography location
P22‧‧‧Photography location
P23‧‧‧Photography location
P31‧‧‧Photography location in the area around the hole
The location of the camera in the area around P32‧‧
P41‧‧‧ Camera location in the central area of the hole
P42‧‧‧ Camera location in the central area of the hole
P43‧‧‧ Camera location in the central area of the hole
P44‧‧‧ Camera location in the central area of the hole
P51‧‧‧Photography location around the hole
P52‧‧‧Photography location in the area around the hole
P61‧‧‧ Camera location in the central area of the hole
P62‧‧‧ Camera location in the central area of the hole
P63‧‧‧ Camera location in the central area of the hole
P64‧‧‧ Camera location in the central area of the hole
V‧‧‧ camera field of view
W‧‧‧ hole
Wb‧‧‧ hole bottom
WC‧‧‧ Well Center
WE‧‧‧ edge of the hole
WP‧‧‧ hole plate
WR‧‧‧ hole edge

Fig. 1 is a view showing a schematic configuration of an image pickup apparatus according to an embodiment of the present invention. Fig. 2 is a ray diagram of the first illumination optical system and the second illumination optical system of the above embodiment. Fig. 3 is a view showing the first illumination light emitted from the first illumination optical system in the above embodiment. Fig. 4 is a view showing a second illumination light emitted from the second illumination optical system in the above embodiment. FIG. 5 is a view showing a state of imaging when the first illumination optical system is temporarily used when the peripheral edge region of the hole is not included in the imaging field of view. FIG. 6 is a view showing a state of imaging when the first illumination optical system is used when the peripheral edge region of the hole is included in the imaging field of view. FIG. 7 is a view showing a state of imaging when the second illumination optical system is used when the peripheral edge region of the hole is not included in the imaging field of view. FIG. 8 is a view showing an example of setting of a region in which imaging is performed using the first illumination optical system and a region in which imaging is performed using the second illumination optical system in the embodiment. Fig. 9 is a view showing an example of an imaging arrangement in the above embodiment. Fig. 10 is a view for explaining scanning for imaging in the above embodiment. Fig. 11 is a flow chart showing the overall processing flow when imaging is performed by the imaging device in the above embodiment. Fig. 12 is a flow chart showing the procedure for determining the imaging arrangement in the above embodiment. Fig. 13 is a view for explaining a method of determining an effective field of view area in the above embodiment. FIG. 14 is a view showing an example of a captured image obtained when imaging is performed using the first illumination optical system in the peripheral region of the hole in the embodiment. Fig. 15 is a view showing an effective field of view area in the case where imaging is performed using the first illumination optical system in the above embodiment. Fig. 16 is a view showing an example of a captured image obtained when imaging is performed using the second illumination optical system in the central portion of the hole in the above embodiment. Fig. 17 is a view showing an effective field of view area in the case where imaging is performed using the second illumination optical system in the above embodiment. Fig. 18 is a view for explaining the difference in the effective visual field area due to the difference in culture conditions in the above embodiment. Fig. 19 is a view for explaining the determination of the reference position among a plurality of imaging positions in the peripheral region of the hole in the above embodiment. Fig. 20 is a view for explaining the determination of the reference position among a plurality of imaging positions in the peripheral region of the hole in the above embodiment. Fig. 21 is a view for explaining the distance between the reference position and the edge end portion of the hole in the above embodiment. Fig. 22 is a view for explaining the determination of the imaging position adjacent to the reference position in the above embodiment. Fig. 23 is a view showing an arrangement example of a plurality of imaging positions in the peripheral region of the hole in the embodiment; Fig. 24 is a view showing an arrangement example of a plurality of imaging positions in the peripheral region of the hole in the embodiment; Fig. 25 is a view for explaining the determination of the imaging position in the central portion of the hole in the above embodiment. Fig. 26 is a view for explaining the determination of the imaging position in the central portion of the hole in the above embodiment. Fig. 27 is a view for explaining a variation of the determination of the imaging position in the peripheral region of the hole. Fig. 28 is a view for explaining a variation of the determination of the imaging position on the peripheral region of the hole. Fig. 29 is a view for explaining an alignment process in a modification of the above embodiment. Fig. 30 is a view for explaining an alignment process in a modification of the above embodiment.

S210~S240‧‧‧Steps

Claims (13)

A method of determining an imaging configuration, wherein the imaging device is configured to determine a plurality of imaging positions, and the imaging device includes a first illumination optical system and a second illumination optical system, and has one or more tests. The type of the sample container and the imaging position of the sample storage unit are imaged while switching the illumination optical system used between the first illumination optical system and the second illumination optical system, and the imaging arrangement determination method includes: an effective field of view a region determining step of determining an effective field of view region when the imaging device performs imaging; and a first imaging device determining step of arranging a plurality of imagings using the first illumination optical system along a wall surface of the sample storage portion And a second imaging arrangement determining step of utilizing the effective field of view area obtained by imaging at at least one of all imaging positions in an arbitrary position in the sample storage unit Obtained by a plurality of imaging positions determined in the first imaging configuration determining step In the area other than the effective field of view area, a plurality of imaging positions using the imaging of the second illumination optical system are arranged. The imaging configuration determining method according to claim 1, wherein in the effective field of view region determining step, an effective field of view region when the image is captured by the first illumination optical system and an effective field of view when the image is captured by using the second illumination optical system are determined. In the first imaging arrangement determining step, the region is determined such that one of the effective field of view regions obtained by imaging at two imaging positions adjacent to each other along the wall surface of the sample storage portion overlaps with each other. The arrangement of the plurality of imaging positions using the imaging of the first illumination optical system is performed in the second imaging arrangement determining step by the imaging at the plurality of imaging positions determined in the first imaging arrangement determining step. The area other than the effective field of view area is determined such that one of the respective effective field of view areas obtained by imaging at two imaging positions adjacent to each other overlaps each other, and the plural image capturing using the second illumination optical system is determined. The configuration of the camera position. The imaging arrangement determining method according to claim 2, wherein the shape of the bottom surface of the sample storage unit is circular, and the first imaging arrangement determining step includes: a reference position determining step of determining to use the first illumination optical system And an imaging position order determining step of the line segment connecting the center of the sample storage unit and the reference position to the center of the sample storage unit as a reference position of any one of the plurality of imaging positions; The center of rotation is rotated in units of a specific angle, and each time the position of the end point of the line segment after the rotation is different from the end point of the center position of the sample storage portion is selected as An imaging position using imaging of the first illumination optical system described above is performed. In the imaging position determining method of claim 3, in the reference position determining step, while the imaging position is gradually moved, the imaging using the first illumination optical system is performed, and the wall surface of the sample storage portion is placed. The imaging position at which the brightness is substantially the same as the brightness at the center of the sample storage unit when the imaging is performed using the second illumination optical system at the center of the sample storage unit is selected as the reference position. The imaging configuration determining method according to any one of claims 1 to 4, wherein in the second imaging configuration determining step, the number of scannings required for imaging at all imaging positions performed by the imaging device is minimized. It is determined that the arrangement of the plurality of imaging positions using the imaging of the second illumination optical system described above should be performed. The imaging arrangement determining method according to any one of claims 1 to 4, wherein in the second imaging arrangement determining step, the second illumination optical system is determined to be used so that the number of imaging times of the imaging device is the smallest The configuration of multiple camera positions for camera recording. In the first imaging configuration determining step of the first aspect, the imaging interval in the direction perpendicular to the main scanning direction at the time of imaging by the imaging device is close to the equal interval. Fine adjustment of a plurality of imaging positions using the imaging of the first illumination optical system described above is performed. In the method of determining the imaging configuration of claim 1, in the effective field of view region determining step, the effective visual field region is determined in consideration of the type of the sample container and the culture condition of the imaging target injected into the sample storage unit. In the imaging arrangement determining method of claim 1, the incident state of the chief ray toward the bottom surface of the sample storage portion is different between the first illumination optical system and the second illumination optical system. The imaging arrangement determining method according to claim 9, wherein the first illumination optical system emits light toward the sample storage unit such that the principal ray is incident on the bottom surface of the sample storage unit in a state in which the principal ray is parallel, the second The illumination optical system emits light toward the sample storage unit such that a principal ray incident on a bottom surface of the sample storage unit has a component away from the optical axis, and the imaging device includes a receiving main ray having a distance away from the optical axis. An imaging optical system composed of the light of the component of the direction. An imaging device that images an imaging target that is held in a sample storage unit that is transparent to a bottom surface together with a liquid, and that has a container holding portion that holds one or more of the above-described objects. a sample container of the sample storage unit; an illumination unit that emits light to an imaging target held by the sample storage unit; and an imaging unit that captures an imaging target held by the sample storage unit; and a driving unit; The imaging unit and the illumination unit are integrally moved according to the imaging position, and the control unit controls the operation of the illumination unit, the imaging unit, and the driving unit. The illumination unit includes a bottom surface facing the sample storage unit. The first illumination optical system and the second illumination optical system in which the incident states of the chief ray are different from each other, wherein the control unit is a first imaging position group that is a plurality of imaging positions arranged along a wall surface of the sample storage unit, and a plurality of imaging positions, that is, a second imaging position group, which are disposed in an area other than the effective field of view area obtained by the imaging of the first imaging position group The imaging unit and the driving unit control the imaging unit and the driving unit, and control the illumination unit to emit light from the first illumination optical system when imaging is performed at an imaging position included in the first imaging position group. When imaging is performed at an imaging position included in the second imaging position group, the illumination unit is controlled such that light is emitted from the second illumination optical system. The imaging device according to claim 11, wherein the control unit includes an imaging position adjustment unit that corrects an imaging position from an external instruction based on a position and an orientation of the sample container on the container holding portion. The imaging position at the time of imaging is actually performed by the imaging unit. The imaging device according to claim 11 or 12, wherein the first illumination optical system emits light toward the sample storage unit such that the chief ray is incident on the bottom surface of the sample storage unit in a state in which the principal ray is parallel, the second The illumination optical system emits light toward the sample storage unit such that a principal ray incident on a bottom surface of the sample storage unit has a component away from the optical axis, and the imaging unit includes a receiving main ray having a distance from the optical axis. An imaging optical system composed of a light of a component of a direction.