Imaging apparatus and imaging method

Abstract

An imaging device (10) is provided with a substrate (1), a first optical system (2), and an imaging unit (3). The object (110) is disposed on the substrate (1). The first optical system (2) irradiates a first emitted light (L1) to an object (110). The imaging unit (3) captures an interference image of the first reflected light (L2) and the second reflected light (L3). The first reflected light (L2) is reflected by the first emitted light (L1) at a first interface (F1) of the outer surface of the matching object (110). The second reflected light (L3) is a light reflected by the first emitted light (L1) at a second interface (F2) between the object (110) and the substrate (1).

Description

Technical Field

[0001] The present invention relates to an imaging device and an imaging method. Background Art

[0002] In the study of cell biology, etc., an imaging device is used to observe the shape of cells based on an interference image obtained using reflected light from the cell surface (for example, see Non-Patent Document 1). In the imaging device, it is required to grasp the three-dimensional shape of the object with high accuracy.

[0003] Prior art literature

[0004] Non-patent literature

[0005] Non-patent document 1: T. Yamauchi et.al., "Low coherent quantitative phasemicroscope for nanometer scale measurement of living cells morphology.", Optics Express 2008, Vol. 16, No. 16, 12227-12238. Summary of the invention

[0006] One aspect of the present invention is an imaging device comprising: a substrate for configuring an object; a first optical system for irradiating a first emitting light toward the object; and an imaging unit for capturing an interference image of a first reflected light and a second reflected light, wherein the first reflected light is light reflected from the first emitting light at a first interface matching an outer surface of the object, and the second reflected light is light reflected from the first emitting light at a second interface between the object and the substrate.

[0007] One aspect of the present invention is an imaging method, which comprises placing an object on a substrate, irradiating the object with a first outgoing light, and capturing an interference image of a first reflected light and a second reflected light, wherein the first reflected light is light reflected by the first outgoing light at a first interface matching an outer surface of the object, and the second reflected light is light reflected by the first outgoing light at a second interface between the object and the substrate. BRIEF DESCRIPTION OF THE DRAWINGS

[0008] Figure 1 It is a schematic diagram showing an imaging device according to an embodiment.

[0009] Figure 2 It is a schematic diagram showing the structure of the gas supply unit.

[0010] Figure 3 Schematic diagram showing a gas supply unit and a substrate.

[0011] Figure 4 It is a schematic diagram showing an enlarged view of a substrate and an object.

[0012] Figure 5 It is a schematic diagram showing a substrate and an object in a further enlarged manner.

[0013] Figure 6 This is an example of an interference pattern.

[0014] Figure 7 It is shown along Figure 6 Plot of the brightness of the interference image of the A-A' line.

[0015] Figure 8 This is an example of an interference image including a transmission image.

[0016] Fig. 9 It is enlarged to show Figure 8 .

[0017] Fig.10 It is an explanatory diagram of the imaging method according to the embodiment.

[0018] Fig.11 It is an explanatory diagram of the imaging method according to the embodiment.

[0019] Fig.12 This is an example of an interference pattern.

[0020] Fig.13 It is shown Fig.12 A graph showing the temporal variation of the brightness of the A-A' line.

[0021] Fig.14 It is shown Fig.13 A graph showing the temporal variation of the brightness of the center of the interference fringe in the BB' line.

[0022] Fig.15 yes Fig.13 Brightness profile of the C-C' line. DETAILED DESCRIPTION

[0023] Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings, as appropriate. In addition, in the accompanying drawings, identical or corresponding reference numerals are used for identical or equivalent parts, and repeated descriptions are omitted. In addition, the dimensional ratios in the drawings are exaggerated for the sake of convenience of description and are not consistent with the actual dimensional ratios.

[0024] [Camera device]

[0025] Figure 1 1 is a schematic diagram showing an imaging device 10 according to the embodiment. Figure 2 It is a schematic diagram showing the structure of the gas supply unit 4 . Figure 2A cross-sectional view along the central axis of the hollow member 41 and a cross-sectional view taken along line II orthogonal to the central axis are shown. Figure 3 It is a schematic diagram showing the hollow member 41 and the base material 1 . Figure 4 It is a schematic diagram showing the bottom plate 11 of the substrate 1 and the object 110 in an enlarged manner. Figure 5 It is a schematic diagram which shows the base plate 11 and the object 110 in a further enlarged manner. The “plan view” means viewing from a direction perpendicular to the upper surface 11 a of the base plate 11 .

[0026] like Figure 1 As shown, the imaging device 10 of the embodiment includes a substrate 1 , a first optical system 2 , an imaging unit 3 , a gas supply unit (substance supply unit) 4 , a second optical system 5 , and a stage 6 .

[0027] Substrate 1 such as Figure 3 The substrate 1 has a bottom plate 11 and a side plate 12. The side plate 12 stands from the periphery of the bottom plate 11. The substrate 1 can store a liquid 100 in an internal space 1a divided by the bottom plate 11 and the side plate 12. The upper surface 11a of the bottom plate 11 is formed flat. The substrate 1 can hold the liquid 100 and the object 110.

[0028] Figure 3 The substrate 1 shown has a bottom plate 11 and side plates 12, but the structure of the substrate is not limited thereto. For example, a flat substrate consisting only of the bottom plate 11 may be used as long as it can hold droplets and objects.

[0029] The bottom plate 11 can transmit the first emitted light L1 (see Figure 4 ). The difference in refractive index between the base plate 11 and the object 110 is 0.1 or more. By making the refractive index of the base plate 11 within this range, the reflectivity at the interface (second interface F2) between the base plate 11 and the object 110 can be increased. The refractive index of the base plate 11 and the refractive index of the object 110 can be higher. The refractive index of the base plate 11 can be, for example, 1.45 to 2.1. As a method for measuring the refractive index, there is JIS K7142:2014 and the like.

[0030] Examples of the material of the substrate 1 include polystyrene, polycarbonate, polymethyl methacrylate (PMMA), polyethylene, polypropylene, poly-4-methylpentene-1, and glass. The refractive index of polystyrene is about 1.5. The refractive index of polycarbonate is about 1.6. The refractive index of PMMA is about 1.5. The refractive index of polyethylene and polypropylene is about 1.5. The refractive index of poly-4-methylpentene-1 is about 1.46. The refractive index of glass is, for example, 1.4 to 2.1.

[0031] The upper surface 11a of the substrate 1 may be subjected to a surface treatment for improving cell adhesion. Examples of the surface treatment include physical hydrophilization treatment such as plasma treatment, and coating with a cell adhesion substance such as gelatin, fibronectin, collagen, or an extracellular matrix.

[0032] The shape of the substrate 1 is not particularly limited. The substrate 1 may be, for example, a culture dish, a well plate with 6 wells, 12 wells, 24 wells, 48 ​​wells, 96 wells, etc., a fluidic device, a slide glass, or the like.

[0033] like Figure 1 As shown, the first optical system 2 includes a first light source 21 , a bandpass filter 22 , an aperture stop 23 , a half mirror 24 , an objective lens 25 , and an aperture control unit 26 .

[0034] As the first light source 21 , for example, a mercury lamp, a halogen lamp, a laser light source, or the like can be used.

[0035] The wavelength of the first emitted light L1 from the first light source 21 only needs to include the near-infrared region. Thus, even when there is a substance that absorbs or scatters in the visible light region inside the object 110, it is possible to suppress the light from being absorbed or scattered by the object 110. Therefore, the size that affects the interference image can be reduced.

[0036] The wavelength in the near-infrared region is, for example, 650 nm to 1.4 μm.

[0037] The wavelength of the first emitted light L1 only needs to be 650 nm or longer. Thus, for example, when the object 110 is a cell, the first reflected light L2 can be obtained without being affected by intracellular substances (organelles, etc.) that absorb and scatter in the visible light region. Therefore, an interference image with less noise can be obtained.

[0038] The wavelength of the first emission light L1 only needs to be, for example, 1.7 μm or less.

[0039] The wavelength width of the first emitted light L1 only needs to be 50 nm or less. By setting the wavelength width to this range, the coherence length can be increased. The interference between the first reflected light and the second reflected light occurs within the range of the coherence length, so the coherence length is substantially the upper limit of the thickness of the object 110 that can be measured. By setting the wavelength width to the above range, the coherence length can be increased, so a good interference image can be obtained even when the thickness of the object 110 is large.

[0040] The wavelength width of the first emission light L1 may be, for example, 0.1 nm or more.

[0041] The wavelength width of the first emitted light L1 only needs to be less than 1 / 10 of the wavelength of the first emitted light L1. The coherence length is determined by the wavelength and the wavelength width, so it is preferable to appropriately define the wavelength width relative to the wavelength. The wavelength width of the first emitted light L1 is less than 1 / 10 of the wavelength, thereby obtaining a good interference image even when the thickness of the object 110 is large.

[0042] The wavelength width of the first emission light L1 only needs to be 1 / 1000 of the wavelength or more.

[0043] The first emitted light L1 may include multiple wavelengths (i.e., multiple wavelength components) with a wavelength difference of less than 20 nm. As a result, the effective central wavelength becomes longer. As long as the thickness of the object 110 converges to one interference fringe, the height information of the outer surface of the object 110 can be obtained based on the brightness of the interference image. The height information of the outer surface of the object 110 can be obtained without complicated analysis of the interference fringes, so the shape of the object 110 can be easily and accurately grasped.

[0044] The first emission light L1 may include a plurality of wavelengths having a wavelength difference of 1 nm or more.

[0045] The bandpass filter 22 selectively transmits light having predetermined characteristics (for example, wavelength, wavelength width, etc.).

[0046] The half mirror 24 reflects the first emission light L1 from the first light source 21 toward the substrate 1 . The half mirror 24 transmits the first reflected light L2 and the second reflected light L3 toward the imaging unit 3 .

[0047] The aperture stop 23 can adjust the numerical aperture of the illumination by the first light source 21 by adjusting the diameter of the light beam of the first outgoing light L1 .

[0048] The aperture control unit 26 controls the diameter of the aperture stop 23. The aperture control unit 26 can reduce the numerical aperture of the illumination, for example, by reducing the diameter of the aperture stop 23. Thus, an interference image with clear interference fringes can be obtained. On the other hand, the illumination light passes through the object lens 25 to illuminate the object 110. Therefore, when the numerical aperture of the object lens 25 is small relative to the numerical aperture of the illumination, the numerical aperture of the object lens 25 becomes the final numerical aperture of the illumination. Therefore, the numerical aperture of the illumination controlled by the aperture stop 23 only needs to be less than the numerical aperture of the object lens 25. The numerical aperture of the object lens 25 only needs to be less than 0.7.

[0049] In addition, the imaging numerical aperture is determined only by the numerical aperture of the objective lens 25. The imaging numerical aperture only needs to be 0.7 or less.

[0050] The first optical system 2 can irradiate the first emitted light L1 from the first light source 21 toward the object 110 from below the substrate 1 through the bottom plate 11 (see Figure 4 ).

[0051] The imaging unit 3 includes a detection unit 31 and an information processing unit 32 .

[0052] The detection unit 31 includes, for example, an imaging element such as a CCD or a CMOS. The detection unit 31 captures an image of the object 110. The imaging element may be an area sensor, a linear sensor, or a point sensor. A detection signal corresponding to the detected light is appropriately A / D-converted by an A / D converter or the like, and is output to the information processing unit 32.

[0053] The information processing unit 32 processes information based on the detection signal from the detection unit 31 .

[0054] like Figure 2 As shown, the gas supply unit 4 includes a hollow member 41 and a control unit 42 .

[0055] The hollow member 41 is formed in a circular tube shape (or cylindrical shape), for example. The hollow member 41 has a flow path 43 through which the gas 40 flows. The hollow member 41 can release the gas 40 from the front end 41a by feeding the gas 40 into the flow path 43. When the front end 41a is located in the liquid 100, bubbles 44 are formed at the front end 41a (see Figure 3 The gas 40 is an example of a “substance”. The gas 40 is, for example, air. The gas 40 may have a refractive index lower than that of the object 110 by 0.1 or more.

[0056] The gas supply unit 4 closes the flow path or the flow path 43 in the gas supply unit 4 so that the gas 40 does not flow back into the hollow member 41, thereby maintaining the bubbles 44. The bubbles 44 can be maintained by adjusting the amount of the gas 40 sucked or supplied from the gas supply unit 4 to control at least one of the pressure and volume of the bubbles 44.

[0057] The hollow member 41 may be configured to be movable in a direction perpendicular to the central axis of the hollow member 41 (horizontal direction) by a driving unit such as a motor (not shown).

[0058] The control unit 42 may include, for example, a pump for supplying the gas 40. The control unit 42 can control the supply amount of the gas 40. Therefore, the control unit 42 can adjust the bubbles 44 (see Figure 3 )’s volume.

[0059] The control unit 42 may include, for example, a pressure sensor for detecting the pressure of the gas 40 (the pressure in the flow path 43). The control unit 42 can control the pressure of the gas 40 by controlling the supply amount of the gas 40 using a pump for supplying the gas 40. Therefore, the control unit 42 can adjust the bubbles 44 (see Figure 3 ) pressure.

[0060] like Figure 1 As shown, the second optical system 5 includes a second light source 51 and a collecting lens 52 .

[0061] The second optical system 5 can irradiate the second outgoing light L4 toward the object 110 from above.

[0062] The stage 6 can place the substrate 1 on its upper surface. The stage 6 can be moved in a direction perpendicular to the central axis of the hollow member 41 (horizontal direction) by a driving unit such as a motor (not shown).

[0063] [Image capture method] (First embodiment)

[0064] like Figure 3 As shown, the object 110 and the liquid 100 are contained in the internal space 1a of the substrate 1. The liquid 100 is, for example, water. The refractive index of water is about 1.33. The liquid 100 may be a liquid culture medium for culturing the object 110. The refractive index of the liquid culture medium is also about 1.33. The object 110 is arranged on the substrate 1 (specifically, on the bottom plate 11).

[0065] The object 110 is disposed in the liquid 100. That is, the object 110 is immersed in the liquid 100. The object 110 is, for example, a cell. The cell may be an animal cell, a plant cell, an insect cell, a bacterium, a yeast, etc. The cell may be an adherent cell or a floating cell. Even if the cell is a floating cell, the substrate 1 may be left still until the cell is attached to the bottom plate 11 and allowed to settle, and the image may be taken. The refractive index of the cell is, for example, approximately 1.35 to 1.60.

[0066] The object 110 is not particularly limited, but when light passes through the object 110, if the refractive index difference at the interface is large, the reflectivity becomes high, and a clear interference image can be obtained, which is preferred. An object with a small absorptivity but a refractive index distribution is called a phase object. In a phase object, if the refractive index difference at the interface is large, the reflectivity becomes high, and clear interference fringes can be obtained.

[0067] The hollow member 41 of the gas supply unit 4 is disposed so that the tip portion 41 a is located in the liquid 100 . The portion of the hollow member 41 including the tip portion 41 a is immersed in the liquid 100 .

[0068] By the control unit 42 (refer to Figure 2) Gas 40 is supplied to the flow path 43 of the hollow member 41, and bubbles 44 are formed at the front end 41a. The refractive index of the gas 40 is different from the refractive index of the liquid 100.

[0069] like Figure 4 as well as Figure 5 As shown in FIG. 1 , the object 110 is in surface contact with the upper surface 11a of the bottom plate 11. Specifically, Figure 5 As shown, the lower surface 110b of the object 110 is in contact with the upper surface 11a of the base plate 11 in all regions. Figure 5 In FIG. 1 , the object 110 has an arc shape formed by a straight line forming a lower surface 110 b and an arc forming an upper surface 110 a. The upper surface 110 a of the object 110 is an outer surface of the object 110 .

[0070] The interface between the base plate 11 and the object 110 is the second interface F2.

[0071] When the hollow member 41 is brought close to the object 110, the bubbles 44 cover the object 110. In a plan view, the bubbles 44 surround the object 110. At this time, a part of the liquid 100 in contact with the object 110 is pressed outward by the bubbles 44.

[0072] A part of the liquid 100 remains in a portion including the peripheral edge of the object 110. The remaining liquid 100 is referred to as a remaining liquid 101. The remaining liquid 101 forms a layer that gradually becomes thinner toward the outside of the object 110 in the radial direction.

[0073] The residual liquid 101 may also be present as a thin layer in the center of the upper surface 110a of the object 110 in a plan view. In this case, the residual liquid 101 exists in a circular region extending around the object 110 in a plan view.

[0074] The remaining liquid 101 may remain only in the annular portion including the peripheral edge of the object 110. In this case, the remaining liquid 101 has an annular shape surrounding the central portion of the object 110 in a plan view.

[0075] When the residual liquid 101 is diffused over the entire circular region centered on the object 110, the interface between the residual liquid 101 and the gas 40 is the first interface F1. When the residual liquid 101 is in a ring shape, the first interface F1 is formed by the ring-shaped interface between the residual liquid 101 and the gas 40 and the circular interface between the central portion of the object 110 and the gas 40.

[0076] At least a portion of the first interface F1 is formed between the remaining liquid 101 and the gas 40 (a gas having a refractive index different from that of the remaining liquid 101 ).

[0077] The remaining liquid 101 is flowable and has surface tension, so the upper surface 101a of the remaining liquid 101 has a smooth shape similar to the upper surface (outer surface) 110a of the matching object 110. Therefore, the first interface F1 has a smooth shape similar to the upper surface (outer surface) 110a of the matching object 110.

[0078] In a state where the first interface F1 is formed by the bubbles 44, the supply amount of the gas 40 is adjusted by the control unit 42 as needed, thereby adjusting the pressure of the gas 40 in the bubbles 44. If the pressure of the gas 40 is too high, the object 110 (cell) may be deformed, but the control unit 42 controls the supply amount of the gas 40 so as not to increase the deformation of the object 110, thereby being able to grasp the three-dimensional shape of the object 110 with high accuracy.

[0079] In a state where the first interface F1 is formed by the bubbles 44, the stage 6 (see Figure 1 ) is moved to a direction (horizontal direction) perpendicular to the central axis of the hollow member 41. Thus, the position of the front end portion 41a relative to the object 110 can be adjusted, and therefore, the shape of the first interface F1 can be adjusted.

[0080] When the first interface F1 is formed by the bubbles 44, the hollow member 41 may be moved in a direction (horizontal direction) perpendicular to the central axis of the hollow member 41 as needed. This allows the position of the tip 41a relative to the object 110 to be adjusted, so that the shape of the first interface F1 can be adjusted.

[0081] like Figure 1 As shown, the first light source 21 emits the first emission light L1. The first emission light L1 is incident on the bandpass filter 22. In the bandpass filter 22, light of predetermined characteristics is selectively transmitted. The first emission light L1 that has passed through the bandpass filter 22 reaches the semi-transparent half-reflecting mirror 24 after the diameter of the light beam is adjusted by the aperture stop 23. The first emission light L1 is reflected by the semi-transparent half-reflecting mirror 24 and directed upward, and is incident on the bottom plate 11 of the substrate 1 through the objective lens 25.

[0082] like Figure 4 As shown, the first outgoing light L1 enters the bottom plate 11 from the lower surface 11 b , passes through the bottom plate 11 , and moves upward to reach the second interface F2 .

[0083] like Figure 5 As shown, the refractive index of the bottom plate 11 (refractive index, for example, about 1.5) and the object 110 (refractive index, for example, about 1.35) are different. Therefore, a part of the first emitted light L1 is reflected at the second interface F2. This reflected light is referred to as "second reflected light L3".

[0084] like Figure 1 As shown, the second reflected light L3 travels downward, passes through the objective lens 25 and the half mirror 24 , and reaches the detection unit 31 of the imaging unit 3 .

[0085] like Figure 5 As shown, the first emitted light L1 ( Figure 4 Reference) toward upward and reaches the first interface F1.

[0086] The refractive index of the residual liquid 101 (refractive index, for example, about 1.33) or the object 110 (refractive index, for example, about 1.35) and the gas 40 (refractive index, for example, 1) are different. Therefore, a part of the first emitted light L1 is reflected at the first interface F1. This reflected light is referred to as "first reflected light L2".

[0087] like Figure 1 As shown, the first reflected light L2 goes downward and passes through the object 110 or the residual liquid 101. The first reflected light L2 further passes through the bottom plate 11 and goes downward, passes through the objective lens 25 and the half mirror 24, and reaches the detection unit 31 of the imaging unit 3.

[0088] The first reflected light L2 and the second reflected light L3 are incident on the detection unit 31. Therefore, the interference image is captured by using the phase difference between the first reflected light L2 and the second reflected light L3 in the imaging unit 3. The detection unit 31 sends the data of the obtained interference image to the information processing unit 32. In the information processing unit 32, a stereoscopic image of the object 110 can be obtained based on the interference image.

[0089] Figure 6 This is an example of an interference pattern. Figure 7 It is shown along Figure 6 The brightness of the interference image of the A-A' line can be based on Figure 7 The shape (height) of the first interface F1 is calculated from the brightness shown. In this way, the three-dimensional shape of the object 110 can be grasped based on the interference image.

[0090] like Figure 5 As shown, there is residual liquid 101 (liquid between the first interface F1 and the second interface F2) near the peripheral portion of the object 110, so in the interference image (refer to Figure 6 ) in which interference fringes are observed in a range wider than the object 110.

[0091] In order to grasp the shape of the object 110 in a top view, the irradiation of the first emitted light L1 can generate heat to evaporate at least a portion of the remaining liquid 101. As a result, the first interface F1 becomes a shape along the outer surface of the object 110, and an interference image matching the object 110 is obtained. Therefore, the shape of the object 110 can be accurately grasped.

[0092] The imaging device 10 of the first embodiment has an imaging unit 3, which captures the interference image of the first reflected light L2 at the first interface F1 and the second reflected light L3 at the second interface F2. The first interface F1 is the interface between the residual liquid 101 (or the object 110) and the gas 40. The first interface F1 is the interface between two substances with a large refractive index difference, so the reflectivity is high. Therefore, a good interference image with clear interference fringes is obtained. Therefore, the three-dimensional shape of the object 110 can be grasped with high precision.

[0093] In the case where the object 110 is a cell, there are intracellular organelles and the like in the cell. The intracellular organelles and the like can have a higher refractive index than the cell nucleus, cytoplasm, and the like (for example, mitochondria: about 1.4, lysosomes: 1.6). Therefore, by utilizing the refractive index difference between the intracellular organelles and the cell nucleus, etc., irregular reflections are generated in the cell, which become the noise component of the interference fringes. On the other hand, the refractive index difference between the cell and the substance (gas) at the first interface F1 is very large. Therefore, the reflectivity of the first interface F1 becomes higher, and the reflected light of the first interface F1 becomes dominant, and the influence of the intracellular organelles and the like on the reflected light can be ignored. Therefore, good interference fringes can be obtained. Therefore, the three-dimensional shape of the object 110 can be grasped with high precision.

[0094] On the other hand, when the object 110 is placed in water, the reflectivity of the first interface (the interface between water and the object 110) tends to be low because the refractive index difference between water and the object 110 is small. Therefore, the reflected light at the first interface becomes weak, and the interference image may not be clear.

[0095] In the imaging method of the first embodiment, the first interface F1 having high reflectivity and being less prone to disturbance of a plane wave is formed, so that a high-quality first reflected light L2 is obtained. Therefore, a clear interference image based on the first reflected light L2 and the second reflected light L3 is obtained. Therefore, the three-dimensional shape of the object 110 can be grasped with high precision.

[0096] [Image capture method] (Second embodiment)

[0097] like Figure 1 As shown, in the imaging method of the second embodiment, similarly to the imaging method of the first embodiment, the first emission light L1 from the first light source 21 is irradiated onto the substrate 1 and the object 110. The first reflected light L2 and the second reflected light L3 are incident on the imaging unit 3.

[0098] In the imaging method of the second embodiment, the second optical system 5 is used. The second light source 51 of the second optical system 5 emits the second emission light L4. The second emission light L4 passes through the object 110 and the bottom plate 11 downward. The second emission light L4 passes through the objective lens 25 and the half mirror 24 and enters the detection unit 31 of the imaging unit 3.

[0099] In the imaging unit 3, in addition to the interference image, a transmission image of the object 110 is obtained using the second emitted light L4. The transmission image is superimposed on the interference image.

[0100] In the imaging method of the second embodiment, an interference image including information based on the transmission image is obtained. Therefore, an interference image to which information on the outer shape of the object 110 is added can be acquired.

[0101] Figure 8 This is an example of an interference image including a transmission image. Fig. 9 It is enlarged to show Figure 8 Figure. Figure 8 as well as Fig. 9 As shown, it can be confirmed that there is a transmission image inside the interference image of the interference fringes.

[0102] [Image capture method] (third embodiment)

[0103] like Figure 4 as well as Figure 5 As shown, in the imaging method of the third embodiment, the hollow member 41 is arranged so that the air bubble 44 covers the object 110 .

[0104] The control unit 42 is used to increase or decrease the amount of gas 40 supplied to the hollow member 41. As a result, the pressure of the gas 40 changes. At this time, the shape of the object 110 grasped from the interference image obtained by the camera unit 3 also changes. In the information processing unit 32, the viscoelastic information and surface tension of the object 110 can be obtained from the pressure change of the gas 40, the shape change of the object 110, the pressure change, and the time required for the shape change.

[0105] The pressure (surface stress) applied to the object 110 can be regarded as the pressure of the gas 40 under certain conditions, so the tension of the first interface F1 does not need to be considered. However, there are cases where the tension inherent to the first interface F1 needs to be considered.

[0106] The surface stress applied by the first interface F1 to the object 110 can be calculated based on the inherent value of the tension of the first interface F1 and the interface shape (deformation of the interface), and therefore can be calculated based on the shape of the first interface F1 grasped from the interference image. For example, it is known that when the first interface F1 is an interface between air and liquid, stress (Lagro pressure) caused by surface tension and the radius of curvature of the interface is generated. The Lagro pressure can be obtained from the shape of the first interface F1 (radius of curvature of the interface) grasped by the surface tension (intrinsic value) and the interference image. In the information processing unit 32 of the camera unit 3, the surface stress applied to the object 110 can be obtained based on the shape information and the physical property values ​​of the interface.

[0107] For example, when the first interface F1 is formed by the bubbles 44 , a first interference image when the pressure of the gas 40 is set to a first pressure and a second interference image when the pressure of the gas 40 is set to a second pressure (pressure different from the first pressure) can be acquired.

[0108] The information processing unit 32 of the imaging unit 3 can obtain viscoelastic information of the object 110 based on the first interference image and the second interference image. At this time, the change time from the first pressure to the second pressure and the shape change of the object can be used to analyze the viscoelastic information.

[0109] [Image capture method] (Fourth embodiment)

[0110] Fig.10 It is an explanatory diagram of the imaging method according to the fourth embodiment.

[0111] The refractive index of the material in contact with the object can be higher than that of the object. Fig.10 As shown, the refractive index of the liquid 200 containing the object 110 is higher than that of the object 110. For example, the refractive index of the liquid 200 may be higher than that of the object 110 by 0.1 or more. The refractive index of the liquid 200 is, for example, 1.45 or more. Examples of the liquid 200 include glycerol aqueous solutions and sugar aqueous solutions. The first interface F1 is an interface between the liquid 200 and the object 110.

[0112] In the present embodiment as well, the three-dimensional shape of the object 110 can be grasped with high accuracy using the interference image between the first reflected light L2 at the first interface F1 and the second reflected light L3 at the second interface F2.

[0113] [Image capture method] (Fifth embodiment)

[0114] Fig.11 It is an explanatory diagram of the imaging method according to the fifth embodiment.

[0115] The material in contact with the object 110 may be a thin film member 300. The thin film member 300 has a different refractive index from that of the object 110, or has a property of reflecting light. The thin film member 300 may have a refractive index higher than that of the object 110 by, for example, 0.1 or more. The refractive index of the thin film member 300 may be, for example, 1.45 or more. The thin film member 300 is formed, for example, by resin. The thin film member 300 overlaps the upper surface 110a of the object 110. The first interface F1 is an interface between the thin film member 300 and the object 110.

[0116] The film member 300 may be formed in a bag shape or a balloon shape that is expanded by introducing gas through the hollow member 41. The film member 300 can be held at the front end portion 41a of the hollow member 41 (see Figure 3 ). By adjusting the position of the front end portion 41a or the pressure inside the hollow member 41, the position and shape of the thin film member 300 can be adjusted. Figure 4 as well as Figure 5 ) Similarly, pressure can be applied to the object 110. In addition, in order to adjust the position and shape of the film member 300, at least one of the pressure and volume of the gas supplied or sucked through the hollow member 41 can be controlled.

[0117] In the present embodiment as well, the three-dimensional shape of the object 110 can be grasped with high accuracy by using the interference image between the first reflected light L2 at the first interface F1 and the second reflected light L3 at the second interface F2 .

[0118] [Image capture method] (Sixth embodiment)

[0119] In order to grasp the three-dimensional image of the object 110, it is preferable to set the position where the first interface F1 reaches the substrate 1 (specifically, the upper surface 11a of the bottom plate 11) as the reference position of the height of the object 110. Then, when it is desired to increase the height of the bubble 44 (see Figure 5 ) exposes the upper surface 11a of the bottom plate 11 due to the pressure of the gas 40, there is a possibility that the object 110 is deformed by the pressure of the gas 40. Therefore, the shape of the object 110 in the state where the pressure of the gas 40 is not applied (original shape: shape before deformation) is effectively grasped by the next method. In addition, the position where the first interface F1 reaches the substrate 1 can be a part of the surface in a state where a small amount of water exists between the first interface F1 and the substrate 1 in a state where the first interface F1 and the substrate 1 are sufficiently close.

[0120] Fig.12 This is an example of an interference pattern. Fig.13 It is shown Fig.12 The brightness of the A-A' line varies with time. Fig.13In FIG. 1 , the horizontal axis shows time and the vertical axis shows brightness on the AA' line. Fig.14 : is a graph showing the temporal change in the brightness of the center of the interference fringes. Fig.15 yes Fig.13 Brightness profile of the C-C' line.

[0121] like Fig.13 As shown, when the pressure of the gas 40 is applied to the object 110 by pressing the bubble 44 to the object 110 (refer to Figure 5 ), interference fringes appear (point B). As time passes until the final time point (point B'), the interference fringes of the interference image change.

[0122] like Fig.14 As shown in FIG. 1 , the brightness of the center of the interference fringes increases and decreases. The change in the interference fringes that appears here shows the deformation process of the object 110. Fig.15 The brightness profile at the final time point (point B′) shown shows the shape of the object 110 at the final time point, and therefore, based on the information of the deformation process and the final shape, the original shape of the object 110 before the pressure of the gas 40 is applied can be predicted and understood.

[0123] On the other hand, the following method is used to control the first interface F1 to reach the substrate 1D while keeping the pressure of the gas 40 at a low level. By adjusting the affinity between the first interface F1 and the substrate 1, the upper surface 11a of the bottom plate 11 can be exposed without increasing the pressure of the gas 40. For example, when the liquid 100 is a liquid culture medium, when the liquid culture medium is replaced with a phosphate buffer having a high surface tension, the affinity of the gas 40 to the substrate 1 becomes higher, and the first interface F1 easily approaches or reaches the upper surface 11a of the bottom plate 11. Similarly, by making the upper surface 11a of the bottom plate 11 water-repellent, the affinity of the gas 40 to the substrate 1 also becomes higher, and the first interface F1 easily approaches or reaches the upper surface 11a of the bottom plate 11.

[0124] As mentioned above, one embodiment of the present invention has been described in detail with reference to the drawings, but the specific configuration is not limited to the above-described embodiment, and various design changes and the like can be made within the scope not departing from the gist of the present invention.

[0125] In the imaging device 10, the first optical system 2 for obtaining an interference image irradiates light from below to the object 110. The second optical system 5 for obtaining a transmission image irradiates light from above to the object 110. The configuration of the imaging device is not limited to this example. The first optical system may irradiate light from below to the object. The second optical system may irradiate light from above to the object.

[0126] Description of Reference Numerals

[0127] 1. Substrate

[0128] 21st optical system

[0129] 3. Camera Department

[0130] 4Gas supply unit (material supply unit)

[0131] 5Second optical system

[0132] 10 Camera device

[0133] 21 No.1 Light Source

[0134] 23 aperture diaphragm

[0135] 26 Aperture control unit

[0136] 31. Inspection Department

[0137] 32 Information Processing Department

[0138] 41Hollow parts

[0139] 43 flow path

[0140] 44 bubbles

[0141] 51 Second Light Source

[0142] F1 1st screen

[0143] F2 2nd interface.

Claims

1. A camera device comprising: A substrate for the object to be arranged; a first optical system for irradiating the object with the first emitted light; and A camera unit that captures interference images of a first reflected light and a second reflected light, wherein the first reflected light is light reflected by the first emitted light at a first interface matching the outer surface of the object, and the second reflected light is light reflected by the first emitted light at a second interface between the object and the substrate.

2. The imaging device according to claim 1, wherein: The difference in refractive index between the substrate and the object is 0.1 or more.

3. The imaging device according to claim 1 or 2, wherein: The substrate is capable of retaining a liquid and the object placed in the liquid.

4. The imaging device according to claim 3, wherein: At least a portion of the first interface is formed between the liquid and a substance having a refractive index different from that of the liquid. The imaging device further includes a substance supply unit configured to supply the substance so that the first interface has a shape matching the outer surface of the object.

5. The imaging device according to claim 4, wherein: The refractive index of the substance is lower than that of the object by at least 0.

1.

6. The imaging device according to claim 4 or 5, wherein: The substance is a gas, The substance supply part is a gas supply part.

7. The imaging device according to claim 6, wherein: The gas supply unit includes a hollow member having a front end disposed in the liquid, and the first interface is formed by bubbles formed and maintained at the front end in the liquid by the gas supplied through a flow path in the hollow member.

8. The imaging device according to any one of claims 1 to 7, wherein: The wavelength of the first emitted light includes a near-infrared region.

9. The imaging device according to claim 8, wherein: The wavelength of the first emitted light is equal to or longer than 650 nm.

10. The imaging device according to any one of claims 1 to 9, wherein: The wavelength width of the first emitted light is 50 nm or less.

11. The imaging device according to any one of claims 1 to 10, wherein: The wavelength width of the first emission light is less than or equal to 1 / 10 of the wavelength of the first emission light.

12. The imaging device according to any one of claims 1 to 11, wherein: The first optical system includes an aperture stop, an aperture control unit for controlling a diameter of the aperture stop, and an objective lens. The aperture control unit adjusts the diameter of the aperture stop so that the illumination numerical aperture is set to be less than the imaging numerical aperture. The imaging numerical aperture is less than 0.

7.

13. The imaging device according to any one of claims 1 to 12, wherein: The first emission light emitted by the first optical system includes a plurality of wavelength components having a wavelength difference of 20 nm or less.

14. The imaging device according to any one of claims 1 to 13, wherein: further comprising a second optical system for irradiating the object with a second light, The imaging unit also captures a transmission image based on the second light transmitted through the object.

15. The imaging device according to any one of claims 1 to 14, wherein: The imaging unit includes a detection unit that captures the interference image, and an information processing unit that acquires a stereoscopic image of the object based on the interference image.

16. The imaging device according to any one of claims 4 to 7, wherein: The camera unit comprises: a detection unit for photographing the interference image; and An information processing unit acquires viscoelastic information of the object from the change in pressure of the substance, the change in shape of the object calculated based on the interference image, the change in pressure, and the time required for the shape change of the object.

17. The imaging device according to claim 4, wherein: The substance has a higher refractive index than the object.

18. The imaging device according to claim 17, wherein: The substance is a liquid containing the object.

19. The imaging device according to claim 17, wherein: The substance is a thin film component.

20. The imaging device according to any one of claims 1 to 19, wherein: The object is a phase object.

21. A method for photographing, wherein: The object is placed on the substrate. irradiating the object with the first emitted light, An interference image of a first reflected light and a second reflected light is captured, wherein the first reflected light is light reflected by the first emitted light at a first interface matching the outer surface of the object, and the second reflected light is light reflected by the first emitted light at a second interface between the object and the substrate.

22. The imaging method according to claim 21, wherein: The object is disposed in a liquid, At least a portion of the first interface is an interface between the object and a gas in contact with the object.

23. The imaging method according to claim 22, wherein: The gas is supplied so that the first interface is formed to match the shape of the outer surface of the object.

24. The imaging method according to claim 22 or 23, wherein: By evaporating at least a portion of the liquid between the first interface and the second interface, the first interface is formed along the object.

25. The imaging method according to claim 22 or 23, wherein: The affinity between the gas or the liquid and the substrate is adjusted so that the first interface is formed along the object.

26. The imaging method according to any one of claims 21 to 25, wherein: A three-dimensional image of the object is acquired from the interference image, using the point where the first interface reaches the substrate as a reference position of the height of the object.

27. The imaging method according to any one of claims 22 to 26, wherein: The front end portion of the hollow member having a flow path is arranged in the liquid, In a state where the first interface is formed by bubbles formed and maintained at the tip portion in the liquid by supplying the gas to the flow path, at least one of the supply amount of the gas and the position of the tip portion is adjusted.

28. The imaging method according to any one of claims 22 to 27, wherein: A first interference image is acquired when the pressure of the gas is set to a first pressure, and a second interference image is acquired when the pressure of the gas is set to a second pressure different from the first pressure.

29. The imaging method according to claim 28, wherein: Viscoelastic information of the object is acquired based on the first interference image, the second interference image, the change in pressure of the gas, the change in shape of the object calculated based on the first interference image and the second interference image, the change in pressure, and the time required for the shape change of the object.

Images