JP2015029461A - Imaging device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To image a biological sample with a simple constitution in an environment suitable for culture.SOLUTION: An imaging device 100 comprises a culture dish 15, a light source 3 and an imaging element 4. A biological sample 14 is arranged in the culture dish 15, and optical observation of the biological sample 14 can be allowed from the outside of the culture dish 15. The light source 3 can illuminate the culture dish 15. The imaging element 4 is arranged opposite to the light source 3 through the culture dish 15. The imaging element 4 images the biological sample 14 illuminated by the light source 3 at a magnification of 3 times or less.

Description

  The present invention relates to an imaging apparatus.

  In fields such as recent biological research, drug discovery research, and future clinical applications of ES (Embryonic Stem) cells and iPS cells, it is important to observe and record constantly changing cells and biological tissues for a long time. It is becoming. For example, it is unpredictable when and how cellular responses to various drugs occur. In addition, in order to control the differentiation of ES cells and iPS cells, it is necessary to finely change the culture conditions according to the state of the cells that change over time. Therefore, it is indispensable to observe cells and living tissues over time.

  In general, an optical microscope is used to observe such cells and biological tissues over time. However, an ordinary optical microscope cannot be observed while maintaining cell activity. Therefore, in order to maintain the activity of cells and biological tissues, a technique of observing in combination with an optical microscope and a culture apparatus capable of maintaining the culture conditions of the cells and biological tissues is used.

  As such a technique, a technique has been proposed in which a small culture incubator is installed on the stage of an optical microscope, and a cell culture dish is fixed therein (Patent Document 1). This technique allows observation while maintaining the activity of cells and living tissue in the stage cell culture dish. There has also been proposed a method of installing the entire optical microscope system in a place (in an incubator) suitable for cell culture (Non-patent Document 1).

  In addition, a method of performing high-magnification observation of about 250 to 450 times using a lens array has been proposed (Non-Patent Document 2). In this method, a plurality of illumination light emitting diodes are provided so as to correspond to the respective lenses of the lens array. In this method, illumination is turned on / off so that images from the lenses of the lens array do not overlap, and the imaging optical system is optimized so as to obtain an image optimally, and the image is guided to the image sensor.

  In addition, a technique has been proposed in which images acquired using a microlens array are collectively captured by a single image sensor (Non-patent Document 3). In this method, it is possible to observe a wide range or a plurality of samples by moving the sample placed on the stage.

JP 2008-259430 A

“Incubator Fluorescence Microscope LCV110”, [online], Olympus Corporation, [searched on June 27, 2013], Internet <URL: http://www.olympus.co.jp/jp/lisg/bio-micro/ product / lcv110 /> Brian McCall et al., “Toward a low-cost compact array microscopy platform for detection of tuberculosis”, 2011, Elsevier Ltd., Tuberculosis 91, pp.S54-S60 Antony Orth et al., “Gigapixel fluorescence microscopy with a water immersion microlens array”, 28 January 2013, OPTICS EXPRESS, Vol. 21, No. 2, PP.2361-2368 Wahbe Bishara et al., “Lensfree on-chip microscopy over a wide fieldof-view using pixel super-resolution”, 24 May 2010, OPTICS EXPRESS, Vol. 18, No. 11, PP.11181-11191 Guoan Zheng et al., “The ePetri dish, an on-chip cell imaging platform based on subpixel perspective sweeping microscopy (SPSM)”, October 11, 2011, PNAS, vol. 108, No. 41, pp.16889-16894

  However, the inventor has found that the above-described method has the following problems. In the method of Patent Document 1, it is necessary to install one culture incubator one by one on the observation stage of the optical microscope. Therefore, when observing, the culture incubator must be taken out of an environment suitable for culture, which adversely affects the cells in the culture incubator.

  On the other hand, the method of Non-Patent Document 1 can solve the problem of Patent Document 1 because the entire optical microscope system is placed in an environment suitable for cell culture for culturing. However, it is necessary to prepare a large thermostat or the like that can contain the optical microscope system, and the system is too large for the size of the sample that can be observed. Therefore, this system requires a large operating cost.

  The present invention has been made in view of the above circumstances, and an object of the present invention is to image a biological sample with a simple configuration in an environment suitable for culture.

  The imaging apparatus according to the first aspect of the present invention includes a sample holding unit configured to allow a biological sample to be disposed inside and optically observing the biological sample from the outside, a light source that illuminates the sample holding unit, An imaging unit that is arranged to face the light source via the sample holding unit and that images the biological sample illuminated by the light source at a magnification of 3 times or less. Thereby, imaging of a biological sample can be performed while maintaining the culture environment of the biological sample with a simple configuration.

  An imaging apparatus according to a second aspect of the present invention is the above-described imaging apparatus, wherein the plurality of imaging units are arranged on the first surface, and the plurality of light sources are provided to each of the plurality of imaging units. It is arranged so as to correspond. Thereby, it is possible to collectively perform imaging of a plurality of biological samples while maintaining the culture environment of the plurality of biological samples with the configuration.

  An imaging apparatus according to a third aspect of the present invention is the imaging apparatus described above, wherein the one or more sample holding means are formed with a microchannel configured as a flow cell through which the biological sample flows, All or a part of the plurality of the imaging means is arranged at a position where a plurality of locations of the microchannel can be observed, thereby imaging the biological sample flowing in the flow cell while maintaining the culture environment of the biological sample. Can be done continuously.

  An imaging apparatus according to a fourth aspect of the present invention is the imaging apparatus described above, wherein a distance between the biological sample in the sample holding unit and the imaging unit is 1 mm or less, and the one imaging unit When the first light source and the second light source, which are the light sources, are provided, and an image acquired by the one imaging unit when illuminated by the first light source is illuminated by the second light source In addition, the position of the image acquired by the one image pickup unit is different by about half of the pixels of the one image pickup unit. Thereby, the biological sample can be imaged without a lens while maintaining the culture environment of the biological sample.

  An imaging apparatus according to a fifth aspect of the present invention is the imaging apparatus described above, wherein the optical apparatus is disposed between the sample holding unit and the imaging unit, and forms an image of the biological sample on the imaging unit. A system is further provided. Thereby, the image of the biological sample can be formed on the imaging means at a predetermined magnification.

  An imaging apparatus according to a sixth aspect of the present invention is the imaging apparatus described above, wherein the imaging means is a CCD or CMOS image sensor. Thereby, imaging of a biological sample can be performed.

  According to the present invention, a biological sample can be imaged with a simple configuration in an environment suitable for culture.

  The above and other objects, features and advantages of the present invention will be more fully understood from the following detailed description and the accompanying drawings. The accompanying drawings are presented for purposes of illustration only and are not intended to limit the present invention.

1 is a perspective view schematically showing an example of the appearance of an imaging apparatus 100 according to a first embodiment. It is a perspective view showing typically an example of imaging device 100 when upper case 1 and lower case 2 are separated. It is sectional drawing which shows typically the example of the cross-sectional structure of the imaging device 100 in the III-III line | wire of FIG. FIG. 2 is a diagram schematically illustrating an example of how the imaging apparatus 100 according to the first embodiment is used. FIG. 6 is a cross-sectional view schematically illustrating an example of a cross-sectional configuration of an imaging apparatus 200 according to a second embodiment. FIG. 10 is a cross-sectional view schematically illustrating an example of a cross-sectional configuration of an imaging apparatus 201 that is a modification of the imaging apparatus 200. FIG. 6 is a perspective view schematically illustrating an example of an appearance of an imaging apparatus 300 according to a third embodiment. FIG. 10 is a perspective view illustrating an example when the upper housing 31 of the imaging apparatus 300 according to the third embodiment is removed. It is sectional drawing which shows typically the example of the cross-sectional structure of the imaging device 300 in the VIII-VIII line of FIG. FIG. 6 is a perspective view schematically illustrating an example of an appearance of an imaging apparatus according to a fourth embodiment. FIG. 9 is a perspective view illustrating an example in which an upper housing 41 of an imaging apparatus 400 according to a fourth embodiment is removed. It is sectional drawing which shows typically the example of the cross-sectional structure of the imaging device 400 in the XI-XI line | wire of FIG. FIG. 9 is a cross-sectional view schematically illustrating an example of a cross-sectional configuration of an imaging apparatus 500 according to a fifth embodiment. It is sectional drawing which shows typically the example of the cross-sectional structure of the imaging device 501 which is the modification which made the light source of the imaging device 500 into multiple.

  Embodiments of the present invention will be described below with reference to the drawings. In the drawings, the same elements are denoted by the same reference numerals, and redundant description is omitted as necessary.

Embodiment 1
First, the imaging apparatus 100 according to the first embodiment will be described. FIG. 1 is a perspective view schematically illustrating an example of the appearance of the imaging apparatus 100 according to the first embodiment. The imaging apparatus 100 includes a biological sample and devices necessary for imaging the biological sample in a box formed by the upper housing 1 and the lower housing 2. Further, hereinafter, for convenience of explanation, a direction parallel to the front surface of the imaging apparatus 100 is defined as an X direction. A direction perpendicular to the front of the imaging apparatus 100 (that is, a direction perpendicular to the X direction) is defined as a Y direction. A direction perpendicular to the upper surface and the lower surface of the imaging apparatus 100 (that is, a direction perpendicular to the X direction and the Y direction) is defined as a Z direction.

  FIG. 2 is a perspective view schematically showing an example of the imaging device 100 when the upper housing 1 and the lower housing 2 are separated. The imaging device 100 is configured to be able to store a culture dish 15 containing cells and biological tissue. The culture dish 15 is a sample holding means that holds an internal biological sample and can optically observe the biological sample from the outside. The sample holding means is not limited to the culture dish as long as the biological sample held inside can be optically observed from the outside.

  FIG. 3 is a cross-sectional view schematically showing an example of a cross-sectional configuration of the imaging apparatus 100 taken along line III-III in FIG. The imaging apparatus 100 includes an upper casing 1, a lower casing 2, a light source 3, an imaging element 4, a lens 5, and a control unit 6.

  The upper housing 1 is disposed on the lower housing 2 and forms a hollow box integrally with the lower housing 2. The upper housing 1 and the lower housing 2 can be attached and detached. The upper housing 1 and the lower housing 2 can be made of a material such as resin or metal. The upper housing 1 and the lower housing 2 are provided with vent holes 8 for supplying air into the imaging device 100. In addition, the position and the number of the vent holes 8 are merely examples, and are not limited to this example.

  A light source 3 is provided on the ceiling surface 11 inside the upper housing 1. As the light source 3, for example, a light emitting diode or a small lamp can be used. The light source 3 is desirably arranged at the center of the ceiling surface 11, but the arrangement location is not limited thereto.

  For example, a notch 13 is provided on the upper portion of the lower housing 2 and can support a culture dish 15 for culturing the biological sample 14.

  On the inner bottom surface 12 of the lower housing 2, the image sensor 4 is provided as an imaging unit. For example, a high-resolution CMOS (Complementary Metal-Oxide Semiconductor) image sensor (for example, a pixel size of about 1 μm and a number of pixels of about 16 million) can be used as the imaging device 4. The biological sample 14 includes one or a plurality of cells, and the size of these cells is about 10 to 20 μm. Therefore, an image of a cell can be obtained by using the image sensor 4 having a pixel size of about 1 μm. Here, a CMOS image sensor is exemplified as the image pickup means, but other image pickup elements in which a large number of pixels are arranged on a two-dimensional plane such as a CCD (Charge Coupled Device) can also be used.

  The lens 5 is fixed to the lower housing 2 so as to be positioned between the imaging device 4 and the culture dish 15. The lens 5 forms an image of the biological sample 14 in the culture dish 15 on the image sensor 4. The lens 5 is a low magnification lens having a magnification of about 1 to 3 times.

  The control unit 6 controls the light source 3 and the image sensor 4. The control unit 6 transmits image or video data captured by the image sensor 4 to the outside of the imaging device 100 by wired communication or wireless communication. Further, the control unit 6 can control the image sensor 4 and turn on / off the light source 3 in accordance with an external command received by wired communication or wireless communication.

  Note that the light source 3, the imaging element 4, and the control unit 6 require a power source. However, the imaging device 100 may be provided with a battery or may be provided outside the imaging device 100.

  In this configuration, since the imaging device 4 is brought close to the culture dish 15 and imaging is performed at a low magnification, there is no need to use a large and complicated optical system such as a normal optical microscope. Therefore, according to this configuration, the size of the imaging device can be reduced.

  From the viewpoint of the small size of the imaging device 100, it is suitable that the imaging magnification of the imaging device 4 is 3 times or less. Thereby, it is not necessary to use a large imaging optical system provided in an optical microscope for observing at a high magnification of several tens to several hundred times, and downsizing of the imaging device can be realized.

  As described above, since the imaging device 100 can be downsized, the imaging device 100 can be used in a specific manner. FIG. 4 is a diagram schematically illustrating a usage example of the imaging apparatus 100 according to the first embodiment. As shown in FIG. 4, the imaging device 100 can be placed in a thermostatic chamber 1001 that can maintain an environment suitable for culturing a biological sample containing cells. Even if the thermostat 1001 is sealed with the imaging device 100 placed inside the thermostat 1001, the control unit 6 of the imaging device 100 can communicate with the external control computer 1002 by the wireless communication means 1003, for example. Therefore, even in this case, the imaging apparatus 100 can receive a command from the control computer 1002 or send imaging data to the control computer 1002.

  In this manner, the imaging device 100 can be arranged inside a thermostat or the like that can be normally used by utilizing the fact that the size can be reduced. Therefore, it is not necessary to take the biological sample out of the environment suitable for culture every time it is observed. As a result, it is possible to observe a biological sample while maintaining an environment suitable for culture.

  Furthermore, the image acquired by the imaging apparatus 100 can be subjected to desired image processing by, for example, an external computer (for example, the control computer 1002 in FIG. 4). Thereby, even if there is an influence of aberration or the like on the captured image, the aberration can be corrected by image processing. This means that aberrations that can be corrected by image processing can be allowed in pursuit of miniaturization of the imaging apparatus. In other words, within the correctable range, it is possible to reduce the size of the image sensor without worrying about the influence of aberration.

Embodiment 2
Next, the imaging apparatus 200 according to the second embodiment will be described. The external appearance of the imaging apparatus 200 is the same as the external appearance of the imaging apparatus 100 shown in FIG. FIG. 5A is a cross-sectional view schematically illustrating an example of a cross-sectional configuration of the imaging apparatus 200 according to the second embodiment. FIG. 5A shows a cross section of the imaging device 200 at the same position as the cross section of the imaging device 100 of FIG. The imaging apparatus 200 has a configuration in which an actuator 7 is added to the imaging apparatus 100 according to the first embodiment.

  The actuator 7 is disposed between the lower housing 2 and the image sensor 4. The actuator 7 can change the distance (distance in the Z direction) between the image sensor 4 and the culture dish 15 by driving the image sensor 4.

  According to this configuration, the focal position at the time of imaging with the imaging element 4 can be changed in the depth direction (Z direction). Therefore, a clearer image can be obtained by performing imaging a plurality of times while changing the focal position and synthesizing the image with the control unit 6 or an external computer.

  Furthermore, if the imaging magnification can be made 1, the configuration can be more advantageous. FIG. 5B is a cross-sectional view schematically illustrating an example of a cross-sectional configuration of an imaging apparatus 201 that is a modification of the imaging apparatus 200. The imaging device 201 has a configuration in which the lens 5 of the imaging device 200 is replaced with a 1 × lens array 9. Since the other configuration of the imaging apparatus 201 is the same as that of the imaging apparatus 200, description thereof is omitted.

  The lens array 9 has a configuration in which a plurality of 1 × lenses are arranged in one or both of the X direction and the Y direction. FIG. 5B shows an example in which three 1 × lenses 9a, 9b and 9c are arranged in the X direction. Note that the number of lenses arranged is not limited to this example. In this way, when a lens array is configured by using a plurality of small lenses, a wider viewing angle can be secured as compared with the case of using the lens 5 (FIG. 5A). In addition, since a lens array can be configured using a lens having a small numerical aperture, it is more advantageous from the viewpoint of downsizing the imaging device.

Embodiment 3
Next, the imaging apparatus 300 according to the third embodiment will be described. The imaging apparatus 300 has a configuration in which a plurality of imaging units are arranged in parallel. FIG. 6 is a perspective view schematically illustrating an example of the appearance of the imaging apparatus 300 according to the third embodiment. The imaging apparatus 300 includes a biological sample and equipment necessary for imaging the biological sample in a box formed by the upper housing 31 and the lower housing 32.

  FIG. 7 is a perspective view illustrating an example in which the upper housing 31 of the imaging apparatus 300 according to the third embodiment is removed. A so-called well plate 16 (also referred to as a microplate or a microwell plate) that contains cells and biological tissue is placed on the lower housing 32 of the imaging apparatus 300. The well plate 16 is provided with a partition inside, and a region for arranging the sample is arranged on the matrix. Here, as an example, it is assumed that a total of 36 sample arrangement regions of 6 rows and 6 columns are formed on the well plate 16.

  FIG. 8 is a cross-sectional view schematically showing an example of a cross-sectional configuration of the imaging apparatus 300 along the line VIII-VIII in FIG. In the imaging apparatus 300, a plurality of imaging cells 301 are formed in a matrix. The imaging cell 301 has the same configuration as that of the imaging device 100 according to the first embodiment. That is, the upper housing 1 of the plurality of imaging cells 301 is integrated to form the upper housing 31. The lower housing 2 of the plurality of imaging cells 301 is integrated to form the lower housing 32. Each of the imaging cells 301 is provided with one microplate well. In other words, the imaging device 300 can be understood as having a configuration in which a plurality of imaging devices 100 are integrated.

  The control unit 6 collectively manages video data captured by the plurality of imaging cells 301.

  Since this configuration is a configuration in which small imaging devices 100 are bundled, the entire imaging device 300 can be placed in a constant temperature bath or the like by selecting the number of imaging cells 301 according to the application. Thereby, it becomes possible to observe a plurality of biological samples simultaneously.

  In the present embodiment, an example using a well plate has been described, but a large culture dish without a partition may be placed.

Embodiment 4
Next, an imaging apparatus 400 according to the fourth embodiment will be described. An imaging device 400 is a modification of the imaging device 300 according to the third embodiment. FIG. 9 is a perspective view schematically illustrating an example of the appearance of the imaging apparatus 400 according to the fourth embodiment. The imaging device 400 has a configuration in which the upper housing 31 and the lower housing 32 of the imaging device 300 are replaced with an upper housing 41 and a lower housing 42, respectively. In addition, the imaging apparatus 400 is provided with an inlet 47 and an outlet 48 described later. The discharge port 48 is not shown in FIG.

  FIG. 10 is a perspective view illustrating an example in which the upper housing 41 of the imaging apparatus 400 according to the fourth embodiment is removed. In addition, inside the imaging device 400, a flow cell 45 in which a micro flow path 46 (not shown in FIG. 10) used for flow cytometry or the like is formed is disposed. The flow cell 45 is an example of a sample holding unit. The biological sample flows in the microchannel 46 (not shown in FIG. 10). A biological sample is introduced into the flow cell 45 from the outside of the imaging device 400 through the inlet 47. In addition, the biological sample is discharged from the flow cell 45 to the outside of the imaging device 400 via the discharge port 48. Since the other configuration of the imaging apparatus 400 is the same as that of the imaging apparatus 300, description thereof is omitted.

  FIG. 11 is a cross-sectional view schematically showing an example of a cross-sectional configuration of the imaging apparatus 400 taken along the line XI-XI in FIG. As shown in FIG. 11, a plurality of imaging cells 301 are arranged in the extending direction (X direction) of the micro flow path 46. Therefore, it is possible to continuously observe the biological sample 49 flowing in the microchannel in synchronization with the flow velocity.

Embodiment 5
Next, an imaging apparatus 500 according to the fifth embodiment will be described. An imaging apparatus 500 is a modification of the imaging apparatus 100 according to the first embodiment. The external appearance of the imaging apparatus 500 is the same as the external appearance of the imaging apparatus 100, and thus description thereof is omitted. FIG. 12 is a cross-sectional view schematically illustrating an example of a cross-sectional configuration of the imaging apparatus 500 according to the fifth embodiment. FIG. 12 shows a cross section of the imaging device 500 at the same position as the cross section of the imaging device 100 of FIG.

  In the imaging apparatus 500, the notch 13 is provided below the imaging apparatus 100. For this reason, the culture dish 15 is disposed in the vicinity of the image sensor 4. Therefore, the biological sample 14 and the image sensor 4 are close to each other. For example, the distance W2 between the biological sample 14 and the image sensor 4 is 1 mm or less. On the other hand, the biological sample 14 and the light source 3 are separated from each other as compared with the imaging device 100.

  In this configuration, by bringing the biological sample 14 and the image sensor 4 close to each other, an image of the biological sample 14 can be formed on the image sensor 4 without the lens 5. Thereby, so-called lensless imaging can be realized. As a result, it is advantageous from the viewpoint of reducing the size and cost of the imaging apparatus by removing the lens.

  In addition, a plurality of light sources of the imaging device 500 can be used. FIG. 13 is a cross-sectional view schematically showing an example of a cross-sectional configuration of an imaging apparatus 501 that is a modified example in which a plurality of light sources of the imaging apparatus 500 are used. The plurality of light sources are, for example, a horizontal direction (X direction in FIG. 13) or a direction perpendicular to the paper surface (a direction perpendicular to the X direction and the Z direction in FIG. 13, that is, a direction connecting the front side and the back side of the paper. Are arranged side by side in the Y direction). The plurality of light sources may be arranged in an array in the X direction and the Y direction. For the plurality of light sources, for example, an LED array or an optical fiber array can be used. FIG. 13 shows an example in which the light sources 3a and 3b are arranged side by side in the X direction.

  The light source 3a and the light source 3b are alternatively turned on. The image acquired by the image sensor 4 when illuminated by the light source 3b is displaced in the X direction by about half of the pixels of the image sensor 4 with respect to the image acquired by the image sensor 4 when illuminated by the light source 3a. Thus, the light source 3a and the light source 3b are arranged. Accordingly, high-resolution lensless imaging can be performed under so-called subpixel imaging optical axis control (Non-Patent Documents 4 and 5).

Other Embodiments The present invention is not limited to the above-described embodiments, and can be appropriately changed without departing from the spirit of the present invention.

  The actuator 7 according to the second embodiment can be incorporated in the imaging apparatuses according to the third and fourth embodiments. Also, the imaging apparatus according to the third and fourth embodiments described above can be configured to perform lensless imaging similarly to the imaging apparatus according to the fifth embodiment.

  In the above-described embodiment, the example in which the imaging apparatus images a biological sample has been described, but this is merely an example. That is, the imaging target of the imaging device according to the above-described embodiment is not limited to a biological sample, and does not hinder imaging of other samples. In addition, imaging of various samples including biological samples can acquire one or both of a still image and a moving image. As an observation method for various materials including a biological sample, various observation methods used in optical observation such as a scattered image, a reflected image, a bright field image, a dark field image, a phase difference observation, and a fluorescence observation can be applied. Furthermore, these various observation methods can be applied singly or in combination.

  In the above-described embodiment, a culture environment (eg, temperature, humidity, carbon dioxide concentration, type and concentration of a drug, type of culture solution, etc.) in a sample holding means including a thermostat or a culture dish in which an imaging device is placed. It is also possible to perform imaging of a biological sample while changing. This makes it possible to continuously observe changes in the biological sample accompanying changes in the culture environment. In addition, a large number of samples can be observed at the same time or at any timing while being placed in the same environment.

DESCRIPTION OF SYMBOLS 1, 31, 41 Upper housing | casing 2, 32, 42 Lower housing | casing 3 Light source 4 Image pick-up element 5, 9a, 9b, 9c Lens 6 Control part 7 Actuator 8 Vent 9 Lens array 11 Ceiling surface 12 Bottom surface 13 Notch 14 , 49 Biological sample 15 Culture dish 16 Well plate 45 Flow cell 46 Micro flow path 47 Inlet port 48 Outlet port 100, 200, 300, 400, 500, 501 Imaging device 301 Imaging cell 1001 Constant temperature bath 1002 Control computer 1003 Wireless communication means

Claims (6)

A sample holding means configured so that a biological sample is disposed inside and optical observation of the biological sample is possible from the outside;
A light source for illuminating the sample holding means;
An imaging unit that is arranged to face the light source via the sample holding unit and that images the biological sample illuminated by the light source at a magnification of 3 times or less,
Imaging device. A plurality of the imaging means are disposed on the first surface;
The plurality of light sources are arranged to correspond to each of the plurality of imaging means.
The imaging device according to claim 1. One or two or more sample holding means are formed with a microchannel configured as a flow cell through which the biological sample flows,
All or a part of the plurality of the imaging means is arranged at a position where a plurality of locations of the microchannel can be observed.
The imaging device according to claim 3. The distance between the biological sample in the sample holding means and the imaging means is 1 mm or less,
A first light source and a second light source, which are the light sources, are provided for one imaging means;
The image acquired by the one imaging unit when illuminated by the first light source is compared with the image acquired by the one imaging unit when illuminated by the second light source. The position is different by about half of the pixels,
The imaging device according to any one of claims 1 to 3. An optical system that is disposed between the sample holding unit and the imaging unit and forms an image of the biological sample on the imaging unit;
The imaging device according to any one of claims 1 to 4. The imaging means is a CCD or CMOS image sensor.
The imaging device according to any one of claims 1 to 5.

Images