JP2013025023A - Virtual slide device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain more accurate spectral data in shorter time.SOLUTION: An imaging section 108 images an image according to light made incident on from a sample 101. A color sensor 110 captures spectral data of light made incident from the sample 101. A light source control part 105 turns on a light source 104 in a sampling period at regular times and turns off the light source 104 in a sampling period at dark times. Based on spectral data obtained by the color sensor 110 in the sampling period at the regular times and spectral data obtained by the color sensor 110 in the sampling period at the dark times, a correcting part 111 corrects the color of an image imaged by the imaging part 108. A stage control part 103 moves the relative position of a stage 102, objective lens 106, and half mirror 107 in the sampling period at the dark times.

Description

  The present invention relates to a virtual slide device.

  A virtual slide device that digitally images a slide image of a pathological specimen is known (see, for example, Patent Document 1).

  FIG. 7 is a schematic diagram showing a configuration of a conventionally known virtual slide device. In the illustrated example, the virtual slide apparatus 1000 includes a stage 1002 on which the sample 1001 is placed, and a stage control unit 1003 that drives the stage 1002 in the horizontal direction and the optical axis direction. The virtual slide apparatus 1000 includes a light source 1004 that illuminates the sample 1001, an objective lens 1005 that includes a plurality of lenses so as to face the sample 1001, and an imaging unit 1006 that captures an image of the sample 1001. Yes. In addition, when the virtual slide device 1000 generates a single image by combining a plurality of images, the virtual slide device 1000 includes information that specifies adjacent images and information that specifies an area to be overlapped with the adjacent images. A composite information generation unit 1007 that generates information, an image file generation unit 1008 that stores a plurality of images and composite information in one file, and an imaging control unit that controls the stage control unit 1003 and the imaging unit 1006 1009.

  Next, a method for imaging the sample 1001 using the virtual slide apparatus 1000 will be described. Light from the light source 1004 passes through a predetermined region of the sample 1001 placed on the stage 1002 and the objective lens 1005 and enters the imaging unit 1006. The imaging unit 1006 photoelectrically converts incident light and captures a high-magnification image of a predetermined area of the sample 1001. Subsequently, the stage control unit 1003 has a margin 100 that can capture a high-magnification image of an area adjacent to the predetermined area and has an area that overlaps the image of the adjacent predetermined area. The sample 1001) is moved in the horizontal direction. Note that the imaging control unit 1009 controls the imaging unit 1006 and the stage control unit 1003. The imaging unit 1006 and the stage control unit 1003 repeatedly perform the above processing under the control of the imaging control unit 1009, so that the imaging unit 1006 captures a high-magnification image having a margin for each predetermined region of the sample 1001.

  After the imaging unit 1006 has captured high-magnification images of all predetermined areas of the sample 1001, the bonding information generation unit 1007 recognizes and recognizes adjacent high-magnification images and margins of each high-magnification image. Bonding information is generated based on the result. Subsequently, the image file generation unit 1008 stores the combination information generated by the combination information generation unit 1007 and a plurality of high-magnification images captured by the imaging unit 1006 in one file. A playback unit (not shown) combines a plurality of high-magnification images stored in one file based on the bonding information, generates a single high-magnification image showing the entire sample 1001, and displays it on a liquid crystal display or the like. To do. By performing the above-described processing, the virtual slide apparatus 1000 can generate a high-magnification image of the entire sample 1001.

  Also, a method is known in which a spectrum of an object is measured at multiple points using a small spectrometer attached to an RGB camera, and the color reproducibility from an RGB image is improved using the information (for example, see Non-Patent Document 1). ). If a conventionally known method for improving color reproducibility is applied to a conventionally known virtual slide apparatus 1000, a high-magnification image of the entire sample 1001 can be generated while improving color reproducibility.

JP 2006-292999 A

Kunihiko Ietomi, Yuri Murakami, Masahiro Yamaguchi, Nagaaki Oyama, “Experimental Evaluation of Color Estimation Method for Color Images Using Multi-point Measurement Spectral Information”, Proceedings of the 54th Joint Conference on Applied Physics, p. 1071

  Generally, the visual field range of the color sensor is smaller than the image area (imaging range) of the RGB image sensor. FIG. 8 is a schematic diagram showing a range of a field-of-view range 801 of a conventionally known color sensor and an image area 802 of an RGB image sensor. In the illustrated example, the visual field range 801 of the color sensor is φ400 μm, and the image area 802 of the RGB image sensor is 20 mm × 20 mm. In this example, the visual field range 801 of the color sensor is a very small area of about 0.13% of the image area 802 of the RGB image sensor. Therefore, the amount of light incident on the color sensor is less than the amount of light incident on the RGB image sensor. Therefore, the output signal of the spectrum data output from the color sensor is a finer signal than the output signal of the RGB image sensor.

  One of the methods for improving the accuracy of spectral data based on a minute output signal is to use dark output that is spectral data acquired by the color sensor in a state where light is not incident on the color sensor (dark state). Correction of dark output for correcting the spectral data of a subject is known. Since the dark output varies depending on the integration time and temperature fluctuation of the color sensor, the accuracy of the corrected spectrum data deteriorates unless the color sensor is acquired in the dark state as frequently as possible to obtain the dark output. there is a possibility.

  Therefore, in order to frequently acquire dark output, it is conceivable to provide a period for turning off the light source and acquiring the dark output of the color sensor separately from the period for acquiring the spectral data of the subject. However, the light source provided in the conventional virtual slide device is generally a halogen lamp or the like, and it takes time until the light quantity becomes stable after the light source is turned on. There is a problem that it takes time to acquire the data and the dark output of the color sensor. When correcting dark output, as described above, it is necessary to turn off the light source and acquire dark output separately from the acquisition period of the subject spectral data, which is necessary for shooting one slide. There is a problem that the shooting time becomes long.

  The present invention has been made to solve the above problems, and an object thereof is to provide a virtual slide device that can acquire more accurate spectrum data in a shorter time.

  The present invention includes a stage for holding a sample, a light source for irradiating the sample with light, an imaging device for capturing an image according to light incident from the sample, and spectral data of light incident from the sample. A color sensor to be acquired, an optical system that guides light from the sample to the imaging device and the color sensor, a light source controller that turns on the light source during a normal sampling period, and turns off the light source during a dark sampling period; A spectrum for correcting the color of the image captured by the image sensor based on the spectral data acquired by the color sensor during the normal sampling period and the spectral data acquired by the color sensor during the dark sampling period. A data correction unit and the stage and the optical system during the dark sampling period; A stage control unit for moving the relative position, a virtual slide apparatus comprising a.

  In the virtual slide device of the present invention, the length of the dark sampling period is (1 / N) times the length of the normal sampling period, and the spectrum data correction unit Using the value of the spectral data acquired by the color sensor and the value obtained by multiplying the value of the spectral data acquired by the color sensor during the dark sampling period by N times, the color of the image of the image sensor is corrected, N is a real number greater than one.

  Further, the present invention is the virtual slide device characterized in that the dark sampling period is provided every time the stage control unit moves the relative position between the stage and the optical system a predetermined number of times.

  Further, the present invention is the virtual slide device characterized in that the dark sampling period is provided every time the stage control unit moves the relative position between the stage and the optical system.

  In addition, the present invention provides the dark sampling period each time the stage control unit moves the relative position between the stage and the optical system, and the spectral data correction unit performs the color sampling in the dark sampling period. The spectrum data acquired by the sensor is averaged every predetermined number of acquisition times, and the imaging device captures an image based on the averaged spectrum data and the spectrum data acquired by the color sensor during the normal sampling period. The virtual slide device corrects the color of the image.

  The present invention also includes a second image sensor that captures an entire image corresponding to light incident from the sample, and a second optical system that guides light from the entire sample to the entire image sensor. The virtual slide apparatus is characterized in that the dark sampling period is provided based on the whole image captured by the second image sensor.

  In the virtual slide device of the present invention, the second image sensor shares at least a part with the image sensor.

  In the virtual slide device of the present invention, the second optical system shares at least a part with the optical system.

  According to the present invention, the stage holds a sample. The light source irradiates the sample with light. The imaging element captures an image corresponding to light incident from the sample. The color sensor acquires spectral data of light incident from the sample. The optical system guides light from the sample to the image sensor and the color sensor. The light source controller turns on the light source during the normal sampling period and turns off the light source during the dark sampling period. The spectral data correction unit corrects the color of the image captured by the imaging device based on the spectral data acquired by the color sensor during the normal sampling period and the spectral data acquired by the color sensor during the dark sampling period. Further, the stage control unit moves the relative position between the stage and the optical system during the dark sampling period.

  Thereby, since it is possible to acquire the spectrum data when the light source is not lit using the time during which the relative position of the stage and the optical system is moved, more accurate spectrum data is acquired in a shorter time. be able to.

It is the schematic which showed the structure of the virtual slide apparatus in the 1st Embodiment of this invention. It is the schematic which showed the operation | movement timing at the time of the virtual slide apparatus in the 1st Embodiment of this invention imaging a some tiling image. It is the schematic which showed the operation | movement timing at the time of the virtual slide apparatus in the 2nd Embodiment of this invention imaging a some tiling image. It is the schematic which showed the structure of the virtual slide apparatus in the 3rd Embodiment of this invention. In the 3rd Embodiment of this invention, it is the schematic which showed the example of the tiling image area | region with little light quantity from a sample, and the tiling image area | region with much light quantity from a sample. It is the flowchart which showed the operation | movement procedure of the virtual slide apparatus in the 3rd Embodiment of this invention. It is the schematic which showed the structure of the conventionally known virtual slide apparatus. It is the schematic which showed the range of the visual field range of the conventionally known color sensor, and the range of the image area of an RGB image sensor.

(First embodiment)
Hereinafter, a first embodiment of the present invention will be described with reference to the drawings. FIG. 1 is a schematic diagram illustrating a configuration of a virtual slide device 1 according to the present embodiment. In the illustrated example, the virtual slide apparatus 1 includes an overall control unit 100, a stage 102, a stage control unit 103, a light source 104, a light source control unit 105, an objective lens 106, a half mirror 107, and an imaging unit 108. (Imaging device), a pinhole 109, a color sensor 110, a correction unit 111 (spectrum data correction unit), and an entire image generation unit 112.

  The overall control unit 100 controls each unit included in the virtual slide device 1. Further, the overall control unit 100 sets a normal sampling period that is a period for acquiring the spectrum data of the sample 101 (sample) and a dark sampling period that is a period for acquiring the dark output of the color sensor 110.

  The stage 102 is a table on which the sample 101 is placed. The stage control unit 103 drives the stage 102 in a horizontal direction and a vertical direction (three-dimensional direction) with respect to the imaging surface of the imaging unit 108 based on the control of the overall control unit 100. The light source 104 is, for example, a white LED (Light Emitting Diode) or the like, and generates light that irradiates the sample 101. In addition, the light source 104 can quickly switch between turning on and off. The light source control unit 105 controls turning on and off of the light source 104. Specifically, the light source control unit 105 turns on the light source 104 during the normal sampling period and turns off the light source 104 during the dark sampling period.

  The objective lens 106 condenses the light beam from a predetermined region of the sample 101 and irradiates the condensed light to the half mirror 107. The half mirror 107 reflects a part of the light from the objective lens 106 in the direction of the pinhole 109 and transmits a part thereof.

  The imaging unit 108 includes a light receiving surface and is disposed at a position to receive light transmitted through the half mirror 107. In addition, the imaging unit 108 photoelectrically converts the received light into an electrical signal corresponding to the intensity of the received light. With this configuration, the imaging unit 108 can capture a tiling image (an image of a predetermined region, a partial image) of the sample 101. The objective lens 106 and the half mirror 107 that guide light from the sample 101 to the imaging unit 108 are optical systems.

  The pinhole 109 is disposed between the half mirror 107 and the color sensor 110, and blocks light in a part of the light from the half mirror 107 and transmits light in a part of the area. The light transmitted through the pinhole 109 enters the color sensor 110. The color sensor 110 acquires spectral data of light transmitted through the pinhole 109 out of light from the sample 101 during a normal sampling period. In addition, the color sensor 110 acquires a dark-time output that is spectrum data in a state where no light is incident on the color sensor 110 (dark state) during the dark sampling period.

  The correction unit 111 corrects dark output using the spectral data acquired during the dark sampling period in order to improve the accuracy of the spectral data acquired by the color sensor 110 during the normal sampling period. Then, the correction unit 111 performs color correction of the tiling image of the sample 101 captured by the imaging unit 108 using the spectrum data that has been corrected for dark output. The overall image generation unit 112 recognizes adjacent tiling images for each of a plurality of tiling images captured by the imaging unit 108 and color-corrected by the correction unit 111, and further, a margin between adjacent tiling images. And recognize all the tiling images based on the recognized result to generate one whole image.

  Next, the operation timing when the virtual slide device 1 captures a plurality of tiling images will be described. FIG. 2 is a schematic diagram illustrating operation timing when the virtual slide device 1 according to the present embodiment captures a plurality of tiling images. In the illustrated example, the operation timing when the tiling images of the first tile to the third tile are captured is shown. Note that the operation timing when the tiling images after the fourth tile are taken is the same as the operation timing when the tiling images of the first to third tiles are taken.

  The imaging period of each tiling image includes a dark sampling period 201 and a normal sampling period 202. The dark sampling period 201 is a period for acquiring the dark output of the color sensor 110. The normal sampling period 202 is a period for acquiring the spectrum data of the sample 101. Note that the length of the normal sampling period 202 is the same as the length at which the imaging unit 108 captures the tiling image. The dark sampling period 201 and the normal sampling period 202 have the same length. Therefore, in the present embodiment, the overall control unit 100 sets the length of the normal sampling period 202 to the same length as the tiling image capturing period, and sets the length of the dark sampling period 201 to the normal sampling. The length is set to the same length as the period 202.

  In the dark sampling period 201, the light source controller 105 turns off the light source 104, and the color sensor 110 acquires the dark output. In order to capture a tiling image of a desired area of the sample 101 within the dark sampling period 201, the stage control unit 103 drives the stage 102 to move the position of the sample 101. In addition, the stage control unit 103 drives the stage 102 to move the position of the sample 101 to a desired position, and then stops the stage 102. In the dark sampling period 201, the imaging unit 108 does not perform any particular processing.

  In the normal sampling period 202, the light source control unit 105 turns on the light source 104, and the color sensor 110 acquires the spectrum data of the sample 101. In the normal sampling period 202, the imaging unit 108 captures a tiling image of the sample 101. Note that in the normal sampling period 202, the stage control unit 103 does not perform any particular processing.

  As described above, since the virtual slide device 1 drives the stage 102 while acquiring the dark output, the time required only for acquiring the dark output can be further shortened.

  Note that in order to improve the accuracy of spectral data acquired by the color sensor 110 during the normal sampling period, the correction unit 111 corrects the dark output using the spectral data acquired during the dark sampling period, and the dark time. The timing at which the correction unit 111 performs color correction of the tiling image of the sample 101 captured by the imaging unit 108 using the spectrum data subjected to output correction may be performed at any timing. For example, after the tiling image has been captured, the color correction of the captured tiling image may be performed while another tiling image is captured, or all tiling images have been captured. After that, the color correction of each tiling image may be performed.

  In the above-described example, the dark sampling period is provided every time the stage is moved. However, the present invention is not limited to this, and the dark sampling period may be provided every time the stage is moved a predetermined number of times. If a dark sampling period is not provided when tiling images are captured and the dark output is not acquired, the dark output is corrected using the dark output acquired in the immediately preceding dark sampling period. You may make it perform.

  Moreover, you may make it improve the precision of each spectrum data acquired in a normal sampling period using the average value of several dark time output. For example, the average value of all dark output acquired at the time of capturing each tiling image is calculated, and the accuracy of spectrum data acquired at the time of capturing each tiling image is improved using the calculated dark output value. May be. This makes it possible to make the joints of the tiling images inconspicuous when generating an entire image from the tiling images that have been color-corrected using the spectrum data. Further, an average value of dark output may be calculated for each of a plurality of images, and the accuracy of spectrum data acquired at the time of capturing each tiling image may be improved using the calculated dark output value. Further, an average value of dark output may be calculated for each row or column of the tiling image, and the accuracy of spectrum data acquired at the time of capturing each tiling image may be improved using the calculated dark output value. .

  In addition, the timing at which the entire image generation unit 112 generates a single entire image by combining a plurality of tiling images may be any timing. For example, the entire image generation unit 112 may generate the entire image immediately after the correction unit 111 performs color correction on all tiling images. Further, the entire image generation unit 112 may generate the entire image immediately before the display unit (not shown) displays the entire image. In addition, the entire image generation unit 112 may generate one entire image before performing color correction on each tiling image, and then the correction unit 11 may perform color correction on the entire image. In this case, for example, the correction unit 11 performs color correction of the entire image using the average value of the spectrum data of each tiling image.

  As described above, the virtual slide device 1 needs to move the stage 102 on which the sample 101 is placed for alignment of the sample 101 before capturing a tiling image. At least during the acquisition of dark output (dark sampling period), the stage control unit 103 drives the stage 102 to move the position of the sample 101. In other words, the virtual slide device 1 acquires the dark output of the color sensor 110 without wasting the driving time of the stage 102. Therefore, the virtual slide device 1 can acquire more accurate spectrum data in a shorter time.

(Second Embodiment)
Next, a second embodiment of the present invention will be described with reference to the drawings. The configuration of the virtual slide device 1 in the present embodiment is the same as the configuration of the virtual slide device 1 in the first embodiment. The difference between this embodiment and the first embodiment is that, in this embodiment, the dark-time output is acquired within the time for moving the stage 102 on which the sample 101 is placed for alignment of the sample 101. is there.

  Next, the operation timing when the virtual slide device 1 captures a plurality of tiling images will be described. FIG. 3 is a schematic diagram illustrating operation timing when the virtual slide device 1 according to the present embodiment captures a plurality of tiling images. In the illustrated example, the operation timing when the tiling images of the first tile to the third tile are captured is shown. Note that the operation timing when the tiling images after the fourth tile are taken is the same as the operation timing when the tiling images of the first to third tiles are taken.

  The imaging period of each tiling image includes a dark sampling period 301 and a normal sampling period 302. In addition, the processing performed by each unit of the virtual slide device 1 in each sampling period is the same as that in the first embodiment. The difference between this embodiment and the first embodiment is that in this embodiment, the stage control unit 103 acquires the dark output during a period in which the stage 102 is driven to move the position of the sample 101. In other words, the length of the dark sampling period 301 is the same as the length of the period in which the stage controller 103 drives the stage 102 to move the position of the sample 101. In general, the length of the period during which the stage 102 is moved for alignment of the sample 101 is shorter than the length of the period during which the imaging unit 108 captures the tiling image. Therefore, the length of the dark sampling period 301 is shorter than the length of the normal sampling period 302. In the illustrated example, when the length of the normal sampling period 302 is “1”, the length of the dark sampling period 301 is “1 / N” (N is a real number greater than 1).

  Therefore, the integration time of the dark output acquired in the dark sampling period 301 is “1 / N”, the integration time of the spectrum data acquired in the normal sampling period 302 is “1”, and each integration time is different. However, when correcting dark output, the conditions of the dark output integration time and the spectral data integration time must match. Therefore, in this embodiment, the dark output value used for dark correction is a value obtained by multiplying the dark output value acquired by the color sensor 110 by N times.

  In order to improve the accuracy of the spectrum data acquired in the normal sampling period, the correction unit 111 corrects the dark output using the spectrum data acquired in the dark sampling period, and the correction of the dark output. The timing at which the correction unit 111 performs color correction of the tiling image of the sample 101 imaged by the imaging unit 108 using the performed spectrum data may be any timing as in the first embodiment. Also, the timing at which the overall image generation unit 112 generates a single overall image by combining a plurality of tiling images may be any timing as in the first embodiment. Similarly to the first embodiment, the average value of a plurality of dark outputs may be used to improve the accuracy of each spectrum data acquired during the normal sampling period.

  As described above, the virtual slide device 1 needs to move the stage 102 on which the sample 101 is placed for alignment before capturing a tiling image. Therefore, the dark output is acquired during the period in which the stage 102 on which the sample 101 is placed is moved. When correcting the dark output, the dark output value obtained by the color sensor 110 is set to N times in order to unify the conditions of the dark output integration time and the spectral data integration time. Use the value. In other words, the virtual slide device 1 acquires the dark output of the color sensor 110 without wasting the driving time of the stage 102. Therefore, the virtual slide device 1 can acquire more accurate spectrum data in a shorter time.

(Third embodiment)
Next, a third embodiment of the present invention will be described with reference to the drawings. FIG. 4 is a schematic diagram showing the configuration of the virtual slide device 2 in the present embodiment. In the illustrated example, the virtual slide device 2 includes an overall control unit 100, a stage 102, a stage control unit 103, a light source 104, a light source control unit 105, a first objective lens 206, and a second objective lens. 207, a half mirror 107, a first imaging unit 208 (imaging device) that captures a high-magnification image, a second imaging unit 209 (second imaging device) that captures a low-magnification image, and a pin A hole 109, a color sensor 110, a correction unit 111, an entire image generation unit 112, and an area determination unit 211 are provided.

  Overall control unit 100, stage 102, stage control unit 103, light source 104, light source control unit 105, half mirror 107, pinhole 109, color sensor 110, correction unit 111, and overall image generation unit Reference numeral 112 denotes the same parts as those in the first embodiment.

  The first objective lens 206 is an objective lens used for enlarging and capturing the tiling image of the sample 101, and is an objective lens having a higher magnification than the second objective lens 207. The second objective lens 207 is an objective lens that is used to capture the entire image of the sample 101 and is an objective lens that has a lower magnification than the first objective lens 206. In addition, at a temporary point, either the first objective lens 206 or the second objective lens 207 can be disposed so as to face the sample 101. When the first objective lens 206 is disposed so as to face the sample 101, the first objective lens 206 condenses the light flux from a predetermined region of the sample 101 and irradiates the condensed light to the half mirror 107. When the second objective lens 207 is disposed so as to face the sample 101, the second objective lens 207 condenses the light flux from the entire area of the sample 101 and irradiates the condensed light to the half mirror 107. The half mirror 107 reflects a part of the light from the first objective lens 206 or the second objective lens 207 in the direction of the pinhole 109 and transmits a part thereof.

  The first imaging unit 208 has a larger number of pixels than the second imaging unit 209, and captures a high-resolution image by performing photoelectric conversion into an electric signal corresponding to the intensity of received light. The second imaging unit 209 has a smaller number of pixels than the first imaging unit 208, and captures a low-resolution image by performing photoelectric conversion into an electrical signal corresponding to the intensity of received light. In addition, at a temporary point, either the first imaging unit 208 or the second imaging unit 209 can be arranged at a position where the light transmitted by the half mirror 107 is received. In the present embodiment, when the first objective lens 206 is disposed so as to face the sample 101, the first imaging unit 208 is disposed at a position for receiving the light transmitted by the half mirror 107. Further, when the second objective lens 207 is disposed so as to face the sample 101, the second imaging unit 209 is disposed at a position for receiving the light transmitted by the half mirror 107. Accordingly, the first imaging unit 208 can capture the tiling image of the sample 101 with high resolution, and the second imaging unit 209 can capture the entire image of the sample 101 with low resolution.

  The optical system that guides the light in the tiling image area of the sample 101 to the first imaging unit 208 is the first optical system, and the optical system that guides the light in the entire image area of the sample 101 to the second imaging unit 209. The second optical system is assumed. In the illustrated example, the first optical system and the second optical system share the half mirror 107.

  The area determination unit 211 determines an area with a large amount of light and an area with a small amount of light among the tiling image areas. In the present embodiment, the area determination unit 211 calculates an average value of pixel values for each tiling image area using the pixel values of one whole image captured by the second imaging unit 209, and calculates the calculated average value. Based on the above, a tiling image region with a small amount of light from the sample 101 and a tiling image region with a large amount of light from the sample 101 are determined. For example, when the calculated average value of the pixel values is equal to or greater than a predetermined threshold value, it is determined that the tiling image region has a large amount of light from the sample 101. Further, when the calculated average value of the pixel values is less than a predetermined threshold value, it is determined that the tiling image region has a small amount of light from the sample 101.

  In the illustrated example, the virtual slide device 2 includes the first imaging unit 208 and the second imaging unit 209 separately. However, the present invention is not limited to this, and the second imaging unit 209 includes the first imaging unit 209. A part or all of the image capturing unit 208 may be shared.

  In the illustrated example, the virtual slide device 2 includes the first imaging unit 208 and the second imaging unit 209, but is not limited thereto, and includes only the first imaging unit 208. By changing the arrangement of the objective lens 206 and the second objective lens 207, the tiling image of the sample 101 and the entire image may be captured.

  Next, in the present embodiment, a tiling image area for acquiring dark output will be described. The output value when the color sensor 110 acquires the spectrum data of the tiling image region with a small amount of light from the sample 101, and the output value when the spectrum data of the tiling image region with a large amount of light from the sample 101 is acquired. When the spectrum data of the tiling image area with a small amount of light from the sample 101 is acquired, the output value is smaller. For this reason, when spectral data of a tiling image region with a small amount of light from the sample 101 is acquired, the dark output of the color sensor 110 is highly likely to greatly affect the accuracy of the spectral data.

  Therefore, in this embodiment, in the tiling image region where the dark output of the color sensor 110 is highly likely to have a great influence on the accuracy of the spectral data, the dark output of the color sensor 110 is acquired and dark correction is performed. . Further, in the tiling image region where the dark output of the color sensor 110 is unlikely to have a great influence on the accuracy of the spectrum data, the dark output of the color sensor 110 is not acquired and the dark correction is not performed. Thereby, the virtual slide apparatus 2 can acquire more accurate spectrum data in a shorter time.

  FIG. 5 is a schematic diagram illustrating an example of a tiling image region with a small amount of light from the sample 101 and a tiling image region with a large amount of light from the sample 101 in the present embodiment. In the illustrated example, tiling image areas 501 to 515 imaged by the first imaging unit 208 and an entire image area 601 imaged by the second imaging unit 209 are illustrated. Further, tiling image regions 501, 503, 505, 508, 509, and 512 are shown as tiling image regions with a small amount of light from the sample 101. Further, tiling image regions 502, 504, 506, 507, 510, 511, 513 to 515 are shown as tiling image regions having a large amount of light from the sample 101. In the present embodiment, since the color sensor 110 acquires the dark output of the tiling image area with a small amount of light from the sample 101, only the tiling image areas 501, 503, 505, 508, 509, and 512 are output in the dark. To get.

Next, the imaging procedure of the sample 101 by the virtual slide device 2 will be described. FIG. 6 is a flowchart showing an operation procedure of the virtual slide device 2 in the present embodiment.
(Step S101) The overall control unit 100 arranges the second objective lens 207 at a position facing the sample 101, arranges the second imaging unit 209 at a position to receive the light transmitted by the half mirror 107, The entire image of the sample 101 is captured using the second objective lens 207 and the second imaging unit 209. Thereafter, the process proceeds to step S102.
(Step S <b> 102) The region determination unit 211 uses the entire image of the sample 101 captured by the second imaging unit 209 in the process of step S <b> 101, and a tiling image region with a small amount of light from the sample 101 and the sample 101 A tiling image area with a large amount of light is determined. Thereafter, the process proceeds to step S103.

(Step S103) The overall control unit 100 arranges the first objective lens 206 at a position facing the sample 101, and arranges the first imaging unit 208 at a position for receiving the light transmitted by the half mirror 107. Thereafter, the process proceeds to step S104.
(Step S <b> 104) The overall control unit 100 determines whether or not the first imaging unit 208 has captured tiling images in all tiling image regions of the sample 101. If the overall control unit 100 determines that the first imaging unit 208 has captured the tiling images of all the tiling image areas of the sample 101, the process proceeds to step S107. Otherwise, the process proceeds to step S105. Proceed to processing.

  (Step S105) The overall control unit 100 controls the stage 102 (sample 101) by controlling the stage control unit 103 so that the first imaging unit 208 can capture a tiling image of a tiling image area that has not yet been captured. ) In the horizontal direction. Further, when the tiling image region with a small amount of light from the sample 101 passes on the optical axis of the first objective lens 206 during the movement of the stage 102, the overall control unit 100 sets the dark sampling period 301, The light source 104 is turned off, and the color sensor 110 is made to acquire a dark output. The color sensor 110 outputs the acquired dark output to the correction unit 111. Note that the areas where the color sensor 110 acquires the dark output at this time are, for example, tiling image areas 501, 503, 505, 508, 509, and 512 shown in FIG. Thereafter, the process proceeds to step S106.

  (Step S106) The overall control unit 100 sets the normal sampling period 302, causes the first imaging unit 208 to capture a tiling image, and causes the color sensor 110 to acquire spectral information. The first imaging unit 208 captures a tiling image and outputs the captured tiling image to the correction unit 111. In addition, the color sensor 110 acquires spectral information and outputs the acquired spectral information to the correction unit 111. When the color sensor 110 acquires the dark output in the process of step S105, the correction unit 111 corrects the dark output of the spectral data, and the spectral data acquired by the color sensor 110 during the normal sampling period. Improve accuracy. When the color sensor 110 has not acquired the dark output in the process of step S105, the correction unit 111 does not correct the dark output of the spectrum data. Then, the correction unit 111 performs color correction of the tiling image of the sample 101 captured by the imaging unit 108 using the spectrum data. Thereafter, the process returns to step S104.

  (Step S107) The overall image generation unit 112 generates a single overall image by pasting the tiling images that have been subjected to color correction by the correction unit 111 in the process of step S106. Thereafter, the process ends.

  As described above, when the color sensor 110 acquires the spectrum data of the tiling image region with a small amount of light from the sample 101 and the spectrum data of the tiling image region with a large amount of light from the sample 101. When the spectrum data of the tiling image region with a small amount of light from the sample 101 is acquired, the output value is smaller. For this reason, when spectral data of a tiling image region with a small amount of light from the sample 101 is acquired, the dark output of the color sensor 110 is highly likely to greatly affect the accuracy of the spectral data.

  Therefore, in this embodiment, in the tiling image region where the dark output of the color sensor 110 is highly likely to have a great influence on the accuracy of the spectral data, the dark output of the color sensor 110 is acquired and dark correction is performed. . Further, in the tiling image region where the dark output of the color sensor 110 is unlikely to have a great influence on the accuracy of the spectrum data, the dark output of the color sensor 110 is not acquired and the dark correction is not performed. Thereby, the virtual slide apparatus 2 can acquire more accurate spectrum data in a shorter time.

  Although the first to third embodiments of the present invention have been described in detail with reference to the drawings, the specific configuration is not limited to this embodiment and does not depart from the gist of the present invention. Range design etc. are also included.

  For example, in the above-described embodiment, the relative position between the stage and the optical system is moved by moving the stage. However, the present invention is not limited to this, and the relative position between the stage and the optical system is moved by moving the optical system. You may make it move, You may make it move the relative position of a stage and an optical system by moving both a stage and an optical system.

  DESCRIPTION OF SYMBOLS 1, 2 ... Virtual slide apparatus, 100 ... Overall control part, 101 ... Sample, 102 ... Stage, 103 ... Stage control part, 104 ... Light source, 105 ... Light source control 106, objective lens, 107 ... half mirror, 108 ... imaging unit, 109 ... pinhole, 110 ... color sensor, 111 ... correction unit, 112 ... overall image Generating unit, 206 ... first objective lens, 207 ... second objective lens, 208 ... first imaging unit, 209 ... second imaging unit, 211 ... area determination unit

Claims (8)

A stage to hold the sample;
A light source for irradiating the sample with light;
An image sensor that captures an image according to light incident from the sample;
A color sensor for acquiring spectral data of light incident from the sample;
An optical system for guiding light from the sample to the image sensor and the color sensor;
A light source controller that turns on the light source during a normal sampling period and turns off the light source during a dark sampling period;
A spectrum for correcting the color of the image captured by the image sensor based on the spectral data acquired by the color sensor during the normal sampling period and the spectral data acquired by the color sensor during the dark sampling period. A data correction unit;
A stage control unit that moves a relative position between the stage and the optical system during the dark sampling period;
A virtual slide device comprising: The length of the dark sampling period is (1 / N) times the length of the normal sampling period,
The spectrum data correction unit uses a value of the spectrum data acquired by the color sensor during the normal sampling period and a value obtained by multiplying the value of the spectrum data acquired by the color sensor during the dark sampling period by N times. To correct the image color of the image sensor,
The virtual slide device according to claim 1, wherein the N is a real number greater than one. 2. The virtual slide apparatus according to claim 1, wherein the dark sampling period is provided each time the stage control unit moves the relative position between the stage and the optical system a predetermined number of times. 2. The virtual slide device according to claim 1, wherein the dark sampling period is provided each time the stage control unit moves a relative position between the stage and the optical system. Each time the stage control unit moves the relative position between the stage and the optical system, the dark sampling period is provided,
The spectrum data correction unit averages the spectrum data acquired by the color sensor during the dark sampling period every predetermined number of acquisition times, and the color sensor acquires the averaged spectrum data and the normal sampling period. The virtual slide device according to claim 1, wherein the color of the image captured by the image sensor is corrected based on the spectrum data. A second imaging element that captures an entire image according to light incident from the sample;
A second optical system for guiding light from the entire sample to the entire image sensor;
With
The virtual slide device according to claim 1, wherein the dark sampling period is provided based on the entire image captured by the second image sensor. The virtual slide device according to claim 6, wherein the second image sensor shares at least a part with the image sensor. The virtual slide device according to claim 6, wherein the second optical system shares at least a part with the optical system.