JP2013088570A - Microscope apparatus - Google Patents

Abstract

PROBLEM TO BE SOLVED: To enable acquisition of an image better focused on a sample more quickly than conventional techniques.SOLUTION: An imaging element 110 outputs an image signal on the basis of an image formed on a first surface. An AF element 111 outputs a contrast signal on the basis of light incident on a second surface. An optical system 109 forms an image of a second zone that is a part of a first zone, on the first surface, and causes light to enter the second surface, the light coming from a third zone that is set at a position at least shifted to a first direction from the center of the second zone and is in the second zone. A focal point detection part 112 calculates the focal point of the second zone. A control part 114 performs controls so that the imaging element outputs a plurality of image signals, while matching the second zone to the focal point on the basis of the focal point and changing a relative position between the optical system 109 and a stage 102 so that the second zone is scanned over the first zone in the first direction. An image signal generation part 115 generates an image on the basis of the plurality of image signals.

Description

  The present invention relates to a microscope apparatus.

  Conventionally, in an optical instrument such as a microscope, a light receiving element is irradiated with light from a sample such as a biological specimen, the contrast value of the sample is detected from a signal output from the light receiving element, and the sample is based on the detected contrast value. Contrast AF (Autofocus) control for determining the in-focus point is used. For example, as one of the contrast AF methods, the contrast value of the image signal acquired by the image sensor is detected by the contrast detector while moving the lens along the optical axis direction, and the position where the contrast value is maximized is adjusted. A hill-climbing servo system for determining a focal point is known (for example, see Patent Document 1).

  In addition, as a contrast AF method, light receiving elements of a front pin sensor and a rear pin sensor are arranged at two front and rear positions with respect to a predetermined focal point, and a difference in the contrast value of a sample between the front pin sensor and the rear pin sensor is detected and detected. There is known an optical path difference method in which a position where the contrast value difference is 0 is determined as a focal point (see, for example, Patent Document 2).

  Further, a microscope apparatus that captures a sample to be observed using the above-described contrast AF technique and acquires a digital image with a wide field of view and a high resolution is known (for example, see Patent Document 3). FIG. 7 is a schematic diagram showing the configuration of a conventionally known microscope apparatus. In the illustrated example, the microscope apparatus 700 includes a stage 702, a transmission light source 703, a condenser lens 704, an objective lens 705, an imaging lens 706, an image sensor 707, a focus detection unit 708, and a control unit 709. And.

  The stage 702 is a table on which the sample 701 is placed. Under the control of the control unit 709, the horizontal direction (the X axis direction and the Y axis direction orthogonal to each other in the figure) and the optical axis direction (the Z axis direction in the figure). ) And move to. The transmitted light source 703 emits light. The condenser lens 704 collects the light emitted from the transmissive light source 703 and irradiates the sample 701 with it. Thereby, the sample 701 installed on the stage 702 is irradiated with light from the transmission light source 703 through the condenser lens 704.

  The objective lens 705 is disposed so as to face the sample 701. The objective lens 705 condenses the light beam from the sample 701. The imaging lens 706 images the light from the sample 701 on the imaging surface of the imaging element 707. As a result, the transmitted light from the sample 701 is incident on the image sensor 707 through the objective lens 705 and the imaging lens 706. The imaging element 707 captures a partial image of a predetermined region of the sample 701 imaged on the imaging surface by the imaging lens 706. Note that the partial image captured by the image sensor 707 is input to the in-focus detection unit 708.

  The in-focus detection unit 708 detects the contrast value in the center area among the partial image areas captured by the image sensor 707, and samples the position of the sample 701 (position of the stage 702) when the contrast value is the largest. Calculated as the focal point of 701. Then, the in-focus detection unit 708 generates a drive signal for moving the stage 702 in the optical axis direction so that the position of the sample 701 is moved to the position where the in-focus is achieved based on the calculated in-focus. The in-focus detection unit 708 inputs the generated drive signal to the control unit 709. The control unit 709 moves the stage 702 in the optical axis direction based on the drive signal, and then causes the image sensor 707 to capture a partial image. After the partial image focused by the image sensor 707 is captured, the control unit 709 moves the stage 702 in the horizontal direction in order to capture the next partial image.

  The microscope apparatus 700 can capture a partial image of the sample 701 by using the contrast AF technique by repeatedly executing the above-described operation. Further, the microscope apparatus 700 can acquire a digital image obtained by capturing the entire sample 701 with high resolution by pasting the captured partial images.

  Next, an area of the sample 701 (hereinafter referred to as “imaging area”) captured by the microscope apparatus 700 and an area of the sample 701 (hereinafter referred to as “AF area”) from which the in-focus detection unit 708 detects a contrast value. explain. FIG. 8 is a schematic diagram illustrating an imaging area of a sample 701 captured by a conventionally known microscope apparatus 700 and an AF area of the sample 701 in which a focus detection unit 708 detects a contrast value. In the illustrated example, the AF area 803 is arranged in the center area of the imaging area 802 on the focal plane 801 of the sample 701.

  Next, a method in which the microscope apparatus 700 captures a plurality of partial images of the sample 701 and generates one whole image will be described. FIG. 9 is a schematic diagram showing a relationship between a partial image captured by a conventionally known microscope apparatus 700 and a sample 701. In the illustrated example, nine partial images X1Y1 to X3Y3 captured by the microscope apparatus 700 and a sample 701 are illustrated. Further, each AF area 901 in the area of nine partial images X1Y1 to X3Y3 is shown.

  First, the microscope apparatus 700 divides an area including the sample 701 to be observed into nine small areas of partial images X1Y1 to X3Y3 in accordance with the imaging area of the imaging element 707. Next, the microscope apparatus 700 sequentially captures partial images X1Y1 to X3Y3 of each small region while performing AF control for each small region. In the illustrated example, the microscope apparatus 700 captures a partial image X1Y1, a partial image X2Y1, a partial image X3Y1, a partial image X3Y2, a partial image X2Y2, a partial image X1Y2, a partial image X1Y3, a partial image X2Y3, and a partial image X3Y3. Here, an operation in which the imaging element captures the nth partial image is referred to as an “nth shot”. That is, an operation in which the imaging element captures the partial image X1Y1 is referred to as a first shot, and an operation in which the partial image X2Y1 is captured is referred to as a second shot.

  Subsequently, the microscope apparatus 700 captures nine partial images X1Y1 to X3Y3, and then generates one digital image in which the entire sample 701 is captured with high resolution by using an image pasting technique using software.

  In the example shown in FIG. 9, the sample 701 exists in the AF area 901 in the area of the partial image other than the partial images X3Y1 and X3Y3. Therefore, since the microscope apparatus 700 can detect the contrast value of the sample 701 when capturing a partial image other than the partial images X3Y1 and X3Y3, the in-focus point of the sample 701 can be calculated. However, the sample 701 does not exist in the AF area 901 in the areas of the partial images X3Y1 and X3Y3. Therefore, the microscope apparatus 700 cannot detect the contrast value of the sample 701 and cannot calculate the focal point of the sample 701 when capturing the partial images X3Y1 and X3Y3. Accordingly, when the microscope apparatus 700 captures the partial images X1Y1 to X3Y3 of the sample 701 as shown in FIG. 9 and generates one digital image, the focal point of the sample 701 is calculated in the partial image. The image of the entire sample 701 may not be generated.

  As a method for solving this problem, when capturing a partial image in which the focal point of the sample 701 cannot be calculated by contrast AF, a technique for capturing an image using the focal point of the previously captured partial image is known. (For example, refer to Patent Document 4). For example, in the example illustrated in FIG. 9, when the partial image X3Y1 for which the focal point of the sample 701 cannot be calculated is captured, the focal point obtained when the partial image X2Y1 is captured is used to capture the partial image X3Y3. The image is picked up using the focal point when the partial image X2Y3 is picked up. This is based on the premise that the height of the sample 701, that is, the focal point changes gradually, and the focal point in the adjacent partial images does not change abruptly.

  Also, when the first shot partial image for capturing a partial image is captured for the first time, the center of the sample 701 is set in advance as a reference position for calculating the focal point, and before the first shot partial image is captured. In addition, a method is also known in which the stage 702 is moved so that the contrast value at the center of the sample 701 can be calculated, and the focal point at the center of the sample 701 is calculated (see, for example, Patent Document 4).

JP 2004-170481 A JP-A-8-160284 JP 2011-95500 A Japanese Patent Laid-Open No. 11-264937

  However, the above-described method has the following two problems. The first is that the stage is moved to the center of the sample before imaging the first shot in order to calculate the focal point of the reference position, so that it takes time to acquire a partial image. is there. Second, since the height of the sample changes gradually, the focal point in the center of the sample matches the focal point of the first shot even though there is little difference in the focal point with the adjacent image. Is not limited. Therefore, when there is no sample in the AF area in the first shot, even if the focus information at the center of the sample is used, it is different from the actual focus in the first shot. There is a possibility of acquiring a captured image of the first shot.

  The present invention has been made to solve the above-described problems, and an object of the present invention is to provide a microscope apparatus that can acquire an image focused on a sample more quickly.

  The present invention relates to an imaging element that outputs an image signal based on an image formed on a first surface, an AF element that outputs a contrast signal based on light incident on a second surface, and an entire sample. An image of a second region that is a part of the first region is formed on the first surface, is in the second region, and is at least in the first direction from the center of the second region An optical system for injecting light from a third region set at a shifted position onto the second surface, a stage on which the sample is installed, and a focal point of the second region based on the contrast signal An in-focus detection unit for calculating the in-focus position, and the second region is adjusted to an in-focus position based on the in-focus point calculated by the in-focus detection unit, and the second region is positioned on the first region. The relative position of the optical system and the stage is changed so as to scan in one direction. And a control unit that controls the image pickup device to output a plurality of the image signals, and an image signal generation unit that creates an image of the sample based on the plurality of the image signals. It is a microscope apparatus.

  In the microscope apparatus of the present invention, the third region is set at a position further shifted in the second direction from the center of the second region, and the control unit is configured so that the second region is the first region. The relative position between the optical system and the stage is changed so that the region of 1 is further scanned in the second direction, and the first direction and the second direction are perpendicular to each other.

  In the microscope apparatus of the present invention, when the in-focus detection unit cannot calculate the in-focus, the control unit adjusts the second region based on the in-focus calculated by the in-focus detection unit last time. It is characterized by being adjusted to the focal position.

  In the microscope apparatus of the present invention, the second surface is the same as the first surface, and the third region is at least a part of the second region.

  According to the present invention, the image sensor outputs an image signal based on the image formed on the first surface. The AF element outputs a contrast signal based on the light incident on the second surface. In addition, the optical system forms an image of the second region, which is a part of the first region including the entire sample, on the first surface, and is in the second region from the center of the second region. Light from the third region set at a position shifted in at least the first direction is incident on the second surface. In addition, a sample is installed on the stage. The in-focus detection unit calculates the in-focus point of the second region based on the contrast signal. In addition, the control unit optically adjusts the second region to the in-focus position based on the focal point calculated by the focal point detection unit, and scans the second region in the first direction on the first region. The image sensor is controlled to output a plurality of image signals while changing the relative position between the system and the stage. The image signal generation unit creates a sample image based on the plurality of image signals.

  With this configuration, since the third region is set at a position shifted at least in the first direction from the center of the second region, the contrast value output by the AF element is highly likely to be the contrast value of the sample. Therefore, it is possible to acquire an image that is more focused on the sample more quickly.

It is the schematic which showed the structure of the microscope apparatus in the 1st Embodiment of this invention. It is the schematic which showed the relationship between the partial image which the microscope apparatus in the 1st Embodiment of this invention images, and a sample. It is the schematic which showed the imaging area of the sample which the microscope apparatus in the 1st Embodiment of this invention images, and the AF area of the sample from which an AF element detects a contrast value. It is the schematic which showed the imaging area of the sample which the microscope apparatus in the 2nd Embodiment of this invention images, and the AF area of the sample from which an AF element detects a contrast value. It is the schematic which showed the relationship between the partial image which the microscope apparatus in the 2nd Embodiment of this invention images, a sample, and AF area | region. In the 2nd Embodiment of this invention, the focus detection result of the sample at the time of imaging the partial image of a sample is shown. It is the schematic which showed the structure of the microscope apparatus known conventionally. It is the schematic which showed the imaging area | region of the sample imaged with the microscope apparatus conventionally known, and the AF area | region of the sample from which a focusing point detection part detects a contrast value. It is the schematic which showed the relationship between the partial image which a conventionally known microscope apparatus images, and a sample.

(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 microscope apparatus 100 according to the present embodiment. In the illustrated example, the microscope apparatus 100 includes a stage 102, a transmission light source 103, a condenser lens 104, an optical system 109, an imaging element 110, an AF element 111, a focal point detection unit 112, and a storage unit 113. The control unit 114 and the image signal generation unit 115 are provided. The optical system 109 includes an objective lens 105, an optical path splitting device 106, a first imaging lens 107, and a second imaging lens 108.

  The stage 102 is a stage on which the sample 101 is placed, and is controlled in the horizontal direction (the orthogonal X-axis direction (first direction) and Y-axis direction (second direction) in the figure) by the control of the control unit 113. ) And the optical axis direction (Z-axis direction in the figure). The transmitted light source 103 emits light. The condenser lens 104 collects the light emitted from the transmissive light source 103 and irradiates the sample 101. As a result, the sample 101 placed on the stage 102 is irradiated with light from the transmission light source 103 through the condenser lens 104.

  The objective lens 105 is disposed so as to face the sample 101. The objective lens 105 condenses the light beam from the sample 101 and irradiates the optical path dividing device 106. Thereby, the light transmitted through the sample 101 is irradiated to the optical path dividing device 106. The optical path splitting device 106 is a half mirror or the like, which reflects part of the transmitted light of the sample 101 in the direction of the second imaging lens 108 and part of it in the direction of the first imaging lens 107. To do.

  The first imaging lens 107 forms an image on the imaging surface (first surface) of the imaging element 110 with the light from the sample 101 transmitted through the optical path dividing device 106. As a result, part of the transmitted light from the sample 101 enters the image sensor 110 via the objective lens 105, the optical path splitting device 106, and the first imaging lens 107. The imaging element 110 captures a partial image of a predetermined region of the sample 101 imaged on the imaging surface by the first imaging lens 107. A partial image (image signal) captured by the image sensor 110 is input to the image signal generation unit 115.

  The second imaging lens 108 images the light from the sample 101 reflected by the optical path dividing device 106 on the light receiving surface (second surface) of the AF element 111. As a result, part of the transmitted light from the sample 101 is incident on the AF element 111 via the objective lens 105, the optical path splitting device 106, and the second imaging lens 108. The AF element 111 detects a contrast value (contrast signal) of a predetermined AF area of the sample 101 imaged on the light receiving surface by the second imaging lens 108. Note that the contrast value detected by the AF element 111 is input to the in-focus detection unit 112.

  The focal point detection unit 112 calculates the focal point of the sample 101 in the imaging region where the imaging element 110 captures a partial image by analyzing the contrast value input from the AF element 111 using a hill-climbing servo system or the like. Specifically, the in-focus detection unit 112 calculates the position of the sample 101 (position of the stage 102) when the contrast value input from the AF element 111 is the largest as the in-focus point. Then, the in-focus detection unit 112 generates a drive signal for moving the stage 102 in the optical axis direction so that the position of the sample 101 moves to a position where the in-focus point is based on the calculated in-focus point. The stage 102 drive signal generated by the in-focus detection unit 112 is input to the control unit 114.

  The control unit 114 moves the stage 102 in the optical axis direction based on the drive signal input from the in-focus detection unit 112. Note that the control unit 114 causes the storage unit 113 to store position information of the stage 102 after being moved in the optical axis direction. The control unit 114 moves the stage 102 in the horizontal direction (X-axis direction and Y-axis direction) based on a horizontal signal transmitted from a horizontal drive signal generation unit (not shown).

  The storage unit 113 stores the position information of the stage 102 input from the control unit 114 as the focal point information of the sample 101. The focal point information stored in the storage unit 113 is read by the focal point detection unit 112 when the focal point detection unit 112 cannot calculate the focal point of the sample 101. For example, if the sample 101 does not exist in the AF area where the AF element 111 detects the contrast value and the focus value detection unit 112 cannot calculate the focus point of the sample 101 because the contrast value is low, the focus detection unit 112 stores the focus value. The in-focus information stored in the unit 113 is read out. Then, the focus detection unit 112 generates a drive signal based on the read focus information. Thereby, even when the sample 101 does not exist in the AF area, the stage 102 can be moved to a position where the position of the sample 101 becomes a focal point.

  The imaging device 110 captures a partial image of the sample 101 imaged on the imaging surface by the first imaging lens 107 after the focusing operation is completed. The image sensor 110 inputs the captured partial image to the image signal generation unit 115. The image signal generation unit 115 generates a single digital image in which the entire sample 101 is captured at a high resolution by pasting together a plurality of partial images captured by the image sensor 110.

  Next, a method in which the microscope apparatus 100 captures a plurality of partial images of the sample 101 and generates one whole image will be described. FIG. 2 is a schematic diagram illustrating a relationship between a partial image captured by the microscope apparatus 100 and the sample 101 according to the present embodiment. In the example shown in the figure, the area of four partial images 201 to 204 captured by the microscope apparatus 100 and the sample 101 are shown.

  First, the microscope apparatus 100 divides an area (first area) including the sample 101 to be observed into four small areas (second areas) of the partial images 201 to 204 according to the imaging area of the imaging element 110. Area). Next, the microscope apparatus 100 sequentially captures the partial images 201 to 204 of each small region while performing AF control for each small region. In the example illustrated, the microscope apparatus 100 captures a partial image 201, a partial image 202, a partial image 203, and a partial image 204 in this order. Here, an operation in which the imaging element captures the nth partial image is referred to as an “nth shot”. That is, an operation in which the imaging element captures the partial image 201 is referred to as a first shot, and an operation in which the partial image 202 is captured is referred to as a second shot.

  Specifically, in order to capture the partial image 201 in the first shot, the control unit 114 moves the stage 102 in the X direction and the Y direction so that the area of the partial image 201 enters the imaging area of the image sensor 110. Let Subsequently, the AF element 111 detects the contrast value of the AF area (third area) of the partial image 201. Subsequently, the focus detection unit 112 calculates the focus of the sample 101 based on the contrast value detected by the AF element 111.

  Subsequently, when the in-focus detection unit 112 can calculate the in-focus point of the sample 101, the control unit 114 moves the stage 102 in the optical axis direction (Z direction) using the drive signal generated by the in-focus detection unit 112. Move to. In addition, the control unit 114 causes the storage unit 113 to store the position information of the stage 102 after being moved in the optical axis direction. Thereafter, the imaging device 110 captures the partial image 201 of the sample 101 imaged on the imaging surface by the first imaging lens 107 after the above-described focusing operation is completed.

  On the other hand, in the operation of the first shot, when the sample 101 does not exist in the AF area of the partial image 201 and the focus detection unit 112 cannot calculate the focus of the sample 101, the position of the sample 101 is the focus. A drive signal for moving the stage 102 in the optical axis direction so as to move to a certain position cannot be generated. If the second shot or later, since the storage unit 113 stores the position information of the stage 102 when the partial image was captured last time, the control unit 114 stores the position information stored in the storage unit 113. Can be used to generate a drive signal. In the present embodiment, by increasing the probability that the sample 101 exists in the AF area in the processing of the first shot, the captured images of the partial images 201 to 204 of the sample 101 can be clearly acquired in a focused state. Increase the possibility.

  Note that the direction in which the stage 102 moves when capturing the partial image 201 of the first shot is determined in advance for each microscope apparatus 100. Further, the order of imaging the partial images 201 to 204 of the sample 101 (the order in which the stage 102 moves) is also predetermined for each microscope apparatus 100. The microscope apparatus 100 according to the present embodiment captures a partial image 201, a partial image 202, a partial image 203, and a partial image 204 in this order. In general, the sample 101 exists in the central area of the entire image. From the above, in the first shot imaging area, the area where the sample 101 is most likely to exist is an area closer to the second shot imaging area than the center of the first shot imaging area. .

  That is, in the imaging area of the first shot, the region with the highest probability of the sample 101 being present is the scanning direction (X-axis direction) that is closer to the center of the sample 101 than the center of the imaging area of the first shot. This is an area located on the side (opposite to the traveling direction of the stage 102). Therefore, by providing the AF area in an area located on the scanning direction side of the center of the imaging area, the probability that the sample 101 exists in the AF area can be increased, and the focal point of the sample 101 can be calculated. Probability can be increased. Thus, in this embodiment, by providing the AF area in the area on the center side of the sample 101 in the area of the partial image 201 that is the imaging area of the first shot, the AF area is processed in the first shot processing. It is possible to increase the probability that the sample 101 exists.

  Further, when the partial image 201 of the first shot is captured, there is a high probability that the sample 101 exists in the AF area. Therefore, before performing the processing of the first shot as in the prior art, the central portion of the sample 101, etc. It is not necessary to measure the contrast value at a predetermined position and calculate the focal point. Therefore, the partial image 201 of the first shot can be captured quickly.

  In addition, when the partial image 201 of the first shot is captured using the central focal point of the sample 101 as in the related art, the distance between the first shot region and the central portion of the sample 101 is large. The in-focus point at the center of the sample 101 does not always coincide with the in-focus point of the first shot, and there is a possibility of capturing the partial image 201 of the first shot that is not in focus. However, in the present embodiment, since there is a high probability that the focal point of the sample 101 can be calculated in the imaging region of the first shot, the partial image 201 of the first shot in focus can be captured. Therefore, the microscope apparatus 100 according to the present embodiment can capture the partial image 201 focused on the sample 101 in the first shot.

  FIG. 3 is a schematic diagram illustrating an imaging area of the sample 101 captured by the microscope apparatus 100 according to the present embodiment and an AF area of the sample 101 where the AF element 111 detects a contrast value. In the illustrated example, the focal plane 301 of the microscope apparatus 100, the imaging area 302 in which the imaging element 110 captures the partial images 201 to 204, and the AF area 303 in which the AF element 111 detects a contrast value, It is shown. Further, in the focal plane 301 of the sample 101, the AF area 303 is not the central area of the imaging area 302, but the scanning direction (X-axis direction) side that is the central side of the sample 101 (the direction opposite to the traveling direction of the stage 102). Arranged in the area.

  After completing the imaging of the partial image 201 in the first shot, in order to capture the partial image 202 in the second shot, the control unit 114 causes the area of the partial image 202 to enter the imaging area of the image sensor 110. The stage 102 is moved in the direction opposite to the X direction. Subsequently, the AF element 111 detects the contrast value of the AF area of the partial image 202. Subsequently, the focus detection unit 112 calculates the focus of the sample 101 based on the contrast value detected by the AF element 111. In the second shot operation, the in-focus detection unit 112 does not include the sample 101 in the AF area of the partial image 202, and the in-focus detection unit 112 cannot calculate the in-focus of the sample 101. Since the storage unit 113 stores the position information of the stage 102 when the image 201 is captured as the focal point information, the drive signal is generated using the focal point information stored in the storage unit 113.

  Subsequently, the control unit 114 drives the stage 102 in the optical axis direction (Z-axis direction) using the drive signal generated by the in-focus detection unit 112. In addition, the control unit 114 causes the storage unit 113 to store the position information of the stage 102 after being moved in the optical axis direction. After the focusing operation described above is completed, the image sensor 110 captures the partial image 202 of the sample 101 imaged on the imaging surface by the first imaging lens 107. Note that the partial images 203 and 204 are captured in the same manner as the partial image 202 is captured.

  After the image sensor 110 captures the partial images 201 to 204, the image signal generation unit 115 combines a plurality of partial images captured by the image sensor 110 so as to capture one sample of the entire sample 101 with high resolution. Generate an image.

  As described above, according to the present embodiment, the AF area is provided in the area on the center side of the sample 101 among the areas of the partial image 201 that is the imaging area of the first shot, that is, the stage is set from the center of the imaging area. By providing the AF area 303 in an area located on the opposite side of the traveling direction of 102, the probability that the sample 101 exists in the AF area 303 is increased when the first shot partial image 201 is captured. Accordingly, since the probability that the contrast value of the sample 101 of the first shot can be calculated and the in-focus point of the sample 101 can be calculated, the partial image 201 in focus in the first shot is captured. be able to.

  Further, when the partial image 201 of the first shot is captured, since there is a high probability that the sample 101 exists in the AF area 303, the central portion of the sample 101 is processed before the processing of the first shot as in the prior art. It is not necessary to measure the contrast value at a predetermined position such as, and calculate the focal point. Therefore, the partial image 201 of the first shot can be captured quickly.

  In addition, when the partial image 201 of the first shot is captured using the central focal point of the sample 101 as in the related art, the distance between the first shot region and the central portion of the sample 101 is large. The in-focus point at the center of the sample 101 does not always coincide with the in-focus point of the first shot, and there is a possibility of capturing the partial image 201 of the first shot that is not in focus. However, in the present embodiment, since there is a high probability that the focal point of the sample 101 can be calculated in the imaging region of the first shot, the partial image 201 of the first shot in focus can be captured. Therefore, the microscope apparatus 100 according to the present embodiment can capture the partial image 201 focused on the sample 101 in the first shot.

  Thereby, the microscope apparatus 100 can acquire the partial images 201 to 204 in focus on the sample 101 more quickly, and the entire sample 101 is high-resolution by pasting the acquired partial images 201 to 204. It is possible to acquire a digital image captured in

  Even if the sample 101 does not exist in the AF area 303 after the second shot, and the in-focus detection unit 112 cannot calculate the in-focus based on the contrast value of the sample 101, the previous partial images 201 to 201 can be obtained. Since the storage unit 113 stores in-focus information that is position information of the stage 102 when the image 204 is captured, the drive signal can be generated using the in-focus information stored in the storage unit 113.

(Second Embodiment)
Next, a second embodiment of the present invention will be described. The configuration of the microscope apparatus 100 in the present embodiment is the same as the configuration of the microscope apparatus 100 in the first embodiment. The difference between the present embodiment and the first embodiment is that, in this embodiment, when capturing a partial image, the control unit 114 adds the stage 102 to the X-axis direction shown in the first embodiment. The driving is also performed in the axial direction, and the AF area is arranged at a position shifted in the X-axis direction and the Y-axis direction from the center of the imaging area in accordance with the horizontal driving of the stage 102.

  FIG. 4 is a schematic diagram illustrating an imaging area of the sample 101 captured by the microscope apparatus 100 according to the present embodiment and an AF area of the sample 101 where the AF element 111 detects a contrast value. In the illustrated example, a focal plane 401 of the microscope apparatus 100, an imaging region 402 where the imaging element 110 captures a partial image, and an AF region 403 where the AF element 111 detects a contrast value are shown. ing. In the focal plane 401 of the sample 101, the AF area 403 is not located in the center area of the imaging area 402 but at a position shifted in the X-axis direction as the first direction and the Y-axis direction as the second direction. ing.

  Next, the relationship between the partial image captured by the microscope apparatus 100 according to the present embodiment, the sample 101, and the AF area will be described. FIG. 5 is a schematic diagram illustrating the relationship between the partial image captured by the microscope apparatus 100 according to the present embodiment, the sample 101, and the AF area. In the illustrated example, a region of 35 partial images 501 to 535 captured by the microscope apparatus 100, a sample 101, and an AF region 403 are illustrated. Moreover, the broken line which showed the order in which the microscope apparatus 100 images the partial images 501-535 is shown. The first method is a method in which the microscope apparatus 100 captures the partial images 501 to 535 and a method of generating a single digital image in which the entire sample 101 is captured at a high resolution by pasting the partial images 501 to 535. This is the same as each method in the embodiment.

  In the example illustrated, the microscope apparatus 100 captures the partial image 501 in the first shot process. The partial image 501 is an image of the lower left area of the sample 101. Therefore, in the area of the partial image 501 that is the imaging area of the first shot, the area with the highest probability that the sample 101 exists is the upper right area that is the area on the center side of the sample 101. Therefore, in the present embodiment, the AF area 403 is provided in the upper right area of the imaging area 402.

  As described above, among the areas of the partial image 501 that is the imaging area of the first shot, the area on the center side of the sample 101, that is, the scanning direction in the X direction after obtaining the partial image 501 from the center of the imaging area and Y By providing the AF area 403 in the area on the scanning direction side, the probability that the sample 101 exists in the AF area in the first shot process can be increased. In addition, since the probability that the sample 101 exists in the AF area 403 can be increased in the processing of the first shot, the probability that the focal point of the sample 101 can be calculated can be increased.

  FIG. 6 shows a focus detection result of the sample 101 when the partial images 501 to 535 of the sample 101 shown in FIG. 5 are captured. In the example shown in the figure, a circle indicates that the in-focus detection unit 112 can calculate the in-focus of the sample 101. Further, Δ mark indicates that the in-focus detection unit 112 cannot calculate the in-focus point of the sample 101, but generates drive information using the in-focus information when the partial images 501 to 535 are captured last time. In the illustrated example, when the partial images 504, 507, 508, 521, 522, 529, 534, and 535 are imaged, the focal point detection unit 112 cannot calculate the focal point of the sample 101. However, when the partial image 501 that is the imaging area of the first shot is captured, the focal point detection unit 112 can calculate the focal point of the sample 101.

  As described above, according to the present embodiment, the area on the center side of the sample 101 among the areas of the partial image 501 that is the imaging area of the first shot, that is, the scanning direction in the X direction after obtaining the partial image 501 and By providing the AF area 403 in the Y-direction scanning area, it is possible to increase the probability that the sample 101 exists in the AF area 403 in the first shot process. In addition, since the probability that the sample 101 exists in the AF area 403 can be increased in the processing of the first shot, the probability that the focal point of the sample 101 can be calculated can be increased. Therefore, the microscope apparatus 100 can capture the partial image 501 in focus in the first shot.

  In addition, since the probability that the sample 101 exists in the AF area 403 is high when the partial image 501 of the first shot is captured, the central portion of the sample 101 is processed before the processing of the first shot as in the past. It is not necessary to measure the contrast value at a predetermined position such as, and calculate the focal point. Therefore, the microscope apparatus 100 according to the present embodiment can quickly capture the partial image 501 of the first shot.

  Further, when the partial image 501 of the first shot is captured using the central focal point of the sample 101 as in the prior art, the distance between the first shot region and the central portion of the sample 101 is large. The in-focus point at the center of the sample 101 and the in-focus point of the first shot do not always coincide with each other, and there is a possibility of capturing the partial image 501 of the first shot that is not in focus. However, in the present embodiment, since there is a high probability that the focal point of the sample 101 can be calculated in the imaging region of the first shot, the partial image 501 of the first shot that is in focus can be captured. Therefore, the microscope apparatus 100 according to the present embodiment can capture the partial image 501 in which the sample 101 is in focus in the first shot.

  In addition, when the microscope apparatus 100 captures the partial images 502 to 535 in the processes after the second shot, the sample 101 does not exist in the AF area 403, and the in-focus detection unit 112 calculates the in-focus point of the sample 101. Even if it cannot be performed, the stage 102 is moved in the optical axis direction using the in-focus information (position information of the stage 102) stored in the storage unit 113 when the partial images 501 to 535 are captured last time. Therefore, the partial images 502 to 535 in focus can be captured.

  As a result, the microscope apparatus 100 can acquire the partial images 501 to 535 focused on the sample 101 more quickly, and the high resolution of the entire sample 101 by pasting the acquired partial images 501 to 535. It is possible to acquire a digital image captured in

  Although the first embodiment and the second embodiment 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 first embodiment and the second embodiment, the microscope apparatus 100 includes the image sensor 110 and the AF element 111. However, the present invention is not limited thereto, and at least a part of the area of the image sensor 110 is an AF element. 111 may be used. In this case, since it is not necessary to divide the light from the sample in the direction of the imaging element 110 and the AF element 111, the optical path dividing device 106 is also unnecessary. In addition, the microscope apparatus 100 uses the transmissive light source 103 as a light source, but is not limited thereto, and an epi-illumination light source or a reflection light source may be used. The microscope apparatus 100 moves the stage 102 to focus the sample 101. However, the present invention is not limited to this, and the sample 101 may be focused by moving the objective lens 105. Further, the method for detecting the focal point may be a hill-climbing servo method or an optical path difference method.

  DESCRIPTION OF SYMBOLS 100,700 ... Microscope apparatus, 101,701 ... Sample, 102,702 ... Stage, 103,703 ... Transmission light source, 104,704 ... Condenser lens, 105,705 ... Objective Lens, 106... Optical path splitting device, 107... First image forming lens, 108... Second image forming lens, 109. ... AF element 112,708 ... focus detection unit 113 ... storage unit 114,709 ... control unit 115 ... image signal generation unit 201-204, 501-535 Partial image, 301, 401, 801 ... focal plane, 302, 402, 802 ... imaging area, 303, 403, 803 ... AF area, 706 ... imaging lens

Claims (4)

An image sensor that outputs an image signal based on an image formed on the first surface;
An AF element that outputs a contrast signal based on light incident on the second surface;
An image of a second region that is part of the first region including the entire sample is formed on the first surface, and is in the second region and is at least first from the center of the second region. An optical system for entering light from the third region set at a position shifted in the direction of
A stage on which the sample is placed;
An in-focus detection unit that calculates an in-focus point of the second region based on the contrast signal;
Based on the in-focus point calculated by the in-focus detection unit, the second area is aligned with the in-focus position, and the second area scans the first area in the first direction. A control unit that controls the imaging device to output a plurality of the image signals while changing a relative position between an optical system and the stage;
An image signal generation unit that creates an image of the sample based on a plurality of the image signals;
A microscope apparatus comprising: The third region is set at a position further shifted in the second direction from the center of the second region;
The control unit changes a relative position between the optical system and the stage so that the second region further scans the first region in the second direction,
The microscope apparatus according to claim 1, wherein the first direction and the second direction are perpendicular to each other. The control unit adjusts the second region to a focus position based on the focus calculated previously by the focus detection unit when the focus detection unit cannot calculate the focus. The microscope apparatus according to claim 1 or 2. The second surface is identical to the first surface;
The microscope apparatus according to any one of claims 1 to 3, wherein the third region is at least a part of the second region.

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