JP2003323600A - Imaging apparatus and focusing method in imaging apparatus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an imaging apparatus and a focusing method in the imaging apparatus acquiring precise image data of an object subjected to an image pickup by a simple composition and accurately counting specimens in the image data. <P>SOLUTION: The imaging apparatus is provided with a base, an imaging means fixed to the base for picking up an image of the object subjected to the image pickup, a carriage moving two-dimensionally along an ordinate direction and an abscissa direction in an imaging area of the object, a stage provided on the carriage for mounting the object, and a focus changing means for changing focus with respect to the object during an image pickup of the object by the imaging means. The focus changing means is arranged in a predetermined position between the imaging means and the stage, and one of a plurality of glass bodies changing a focal length is positioned in the predetermined position. <P>COPYRIGHT: (C)2004,JPO

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an image pickup apparatus for picking up an image of a specimen such as bacteria or bacteria as an image pickup object and a focusing method in the image pickup apparatus, and more particularly, to moving a mounting table on which the image pickup object is placed. Thus, the present invention relates to an image pickup apparatus of an optical magnifying system that divides an image pickup surface into a plurality of areas and picks up an image, and a focusing method thereof.

[0002]

2. Description of the Related Art Conventionally, in the field of food hygiene, an apparatus for detecting or measuring microorganisms such as bacteria present in foods has been widely used. In the method for detecting microorganisms in these devices, a culturing method in which microbial cells are cultured on a flat medium solidified with agar or gelatin and the resulting colonies (a population of bacteria) are counted as viable cells,
AT for determining the presence or absence of microorganisms by detecting the fluorescence generated from the reaction of the firefly-derived photoprotein and its substrate
P (adenosine triphosphate) detection method, or a fluorescent staining method that uses fluorescent reagents with different wavelengths to detect different properties of bacteria in a single treatment using fluorescent staining, etc., to detect microorganisms such as bacteria Various methods are used depending on the environment.

As an example of such a device, Japanese Patent Laid-Open No. 2001-2001
JP-A-286296 discloses that a solution containing a microorganism is attached to a plate surface, a detection reagent is dropped, and then an excitation light source is turned on from a lower portion of the plate to cause the microorganism on the plate to fluoresce to be received by a fluorescence receiving section. Counting the number of living cells by counting the number of dead cells and the number of dead cells by counting the number of living cells by illuminating the individual microorganisms that emit fluorescence with a spectral film that transmits only a specific wavelength, and counting the number of single cells that emit light. A microorganism counting device for confirming the numbers of living cells and dead cells in a sample is disclosed.

In this type of device, in order to convert the amount of received light into an electric signal to obtain image data, the electric charge generated by the photodiode is stored in a capacitor and transmitted to the outside according to a predetermined clock. Charge-coupled device (hereinafter referred to as "C
Image pickup means (hereinafter referred to as "CCD" as appropriate) using a "CD"
"Camera") is used. Here, the CCD has
There is also a MOS type, but it is common in the basic principle of converting light into electric charges.

By the way, the CCD camera is widely used as an image pickup unit in various fields in addition to the above-mentioned microorganism measuring apparatus. For example, Japanese Patent Laid-Open No. 8-
Japanese Patent No. 241127 discloses an image processing apparatus that uses an image processing apparatus to generate an image signal obtained by illuminating a magnetic head as an object to be inspected (object to be imaged) with illumination light and capturing the transmitted light or reflected light with the image capturing apparatus. Disclosed is a surface inspection device which is processed, detects foreign matter or a defect of an inspection object, and further includes a positioning mechanism for the inspection object. Also in this conventional apparatus, a CCD camera is used as an image pickup unit, and in order to inspect (image) a plurality of positions of the magnetic head to be inspected (imaged), a stage on which the magnetic head is mounted is placed on a plane. In the vertical direction and the horizontal direction, the inspection (imaging) is performed at a plurality of predetermined positions. At that time, in order to correct the focus shift with respect to the inspection target area (imaging area) of the magnetic head due to the movement of the stage, an automatic focusing mechanism that functions so as to focus on the surface of each inspection area is provided.

[0006]

However, in the above-mentioned conventional apparatus, in order to eliminate the focal shift due to the movement of the image pickup object, the image pickup object is fixedly arranged without moving and a wide image pickup area is once provided. However, it is necessary to use an expensive CCD corresponding to high pixels under the condition that the resolution is fixed and a predetermined image quality is secured, in order to suppress the manufacturing cost of the imaging device. Has no choice but to move the object to be imaged. Further, in the above-described conventional apparatus, when focusing on the surface of the imaging region by the autofocus mechanism, the most common method is to use an objective lens configured as a part of the imaging unit as a magnetic field for imaging. Focusing is performed by moving the head in a direction away from and in contact with the head. In this case, assuming that the positional relationship between the two is in the vertical direction, the vertical movement of the objective lens described above is performed with perfect verticality by the objective lens installation mechanism or the installation method of the entire imaging unit. Is difficult In other words, by the vertical movement of the objective lens described above,
The relative positional relationship before and after the movement causes some blurring and displacement with respect to the verticality. Therefore, the microorganism counting device as described in the above-mentioned Japanese Patent Laid-Open No. 2001-286296 has a problem that it is not possible to accurately count the microorganisms in a predetermined imaging area.

The present invention has been made in view of the above problems of the prior art, and is a focus of a simple structure for acquiring precise image data of an object to be imaged moving in an image pickup area divided into a plurality of frames. An object of the present invention is to realize a cost reduction of an image pickup device obtained by a matching mechanism and to realize an image pickup device that accurately counts samples in the image data and a focusing method in the image pickup device.

[0008]

Therefore, according to the present invention, there is provided a base, an image pickup means fixed to the base for picking up an image of an object to be imaged, and a vertical axis direction in an image pickup area of the object to be imaged. A carriage that moves in a plane along the direction of the horizontal axis, a stage that is provided on the carriage and that mounts the object to be imaged, and a focus on the object to be imaged when the object is imaged by the imaging unit. A focus changing unit for changing the focus, wherein the focus changing unit is provided at a predetermined position between the image pickup unit and the stage.

In this image pickup apparatus, since the focus changing means is arranged at a predetermined position between the image pickup means and the stage as described above, it is possible to obtain precise image data which is in focus with a simple structure. This makes it possible to obtain high sample counting accuracy based on the image data.

As a first form of the focus changing means in the present image pickup apparatus, the focus changing means includes a belt-shaped member having a plurality of glass bodies having different thicknesses and movably mounting the plurality of glass bodies. , One of the plurality of glass bodies is positioned at the predetermined position by the movement of the belt-shaped member. As a result, in this device, focusing is realized by selecting a glass body having an optimum thickness in focusing from a plurality of glass bodies having different thicknesses, and an expensive automatic focus control mechanism is not required. Of.

Further, as a second form of the focus changing means in the present image pickup apparatus, the focus changing means comprises a movable support on which the plurality of glass bodies are placed and which performs a reciprocating or rotating operation in the horizontal direction. Even if focusing is achieved by configuring one of the plurality of glass bodies to be positioned at the predetermined position by reciprocating or rotating movement of the movable support, and selecting a glass body having an optimum thickness. good.

By the way, the image pickup means in the present apparatus takes a plurality of images in the predetermined image pickup area in correspondence with the plurality of glass bodies. Then, the present imaging device has a sample number measuring means for measuring the number of samples in the predetermined imaging area of the object to be imaged on the basis of the imaging data by the imaging means, and the imaging means performs the plurality of imaging operations. Among these, the glass body positioned at the predetermined position when the number of the specimens measured by the specimen number measuring means shows a maximum value is a focusing member that focuses on the imaging area. To judge.

This is because the contour of the specimen is clearly recognized in the imaged data in which the focus is matched.
This is because it can be considered that the glass body positioned at the predetermined position when the number of the specimens shows the maximum value is a focusing member that focuses on the imaging area.

As another method of selecting the focusing member in the present apparatus, among the plurality of times of imaging by the imaging means,
The glass body positioned at the predetermined position when the size of the sample in the predetermined size range preset by the sample size measuring unit replacing the sample number measuring unit shows the minimum value is focused on the imaging area. You may make it judge that it is a focusing member which matches.

This is because the glass body positioned at the predetermined position when the size of the sample shows the minimum value due to the fact that the outline of the sample does not become blurred in the focused imaging data is concerned. This is because it can be regarded as a focusing member that focuses on the imaging area.

Incidentally, the sample number measuring means or the sample size measuring means may be provided on the side of an external data processing device such as a personal computer to which the present imaging device is connected.

The present invention is also a focusing method for an image pickup device of an image pickup device for picking up an image pickup object in a predetermined image pickup area by means of an image pickup means fixedly mounted on a base. Upon image capturing, the plurality of transparent members are sequentially transferred to a predetermined position between the image capturing unit and the image-captured object for changing the focus of the image-captured body, and the images are captured. Focusing on the object to be imaged is performed by selecting one of the image pickup data based on the number of samples or the size of the sample that the object has in the predetermined imaging area. The present invention provides a focusing method in an image pickup apparatus.

When the image pickup area is changed, the image pickup object is picked up after the plurality of transparent members are transferred in the reverse order to the predetermined position. As a result, the number of times the plurality of transparent members are transferred to the predetermined position is reduced, and focus control can be performed in a shorter time.

Here, when the one imaged data is selected based on the number of the specimens, the number of specimens imaged and measured through the permeable member is transmitted through the immediately different permeable member. When it is confirmed that the number of images is reduced from the number of imaged specimens, the image data at the time of imaging through the different transmissive member immediately before is selected and determined to be the focus on the object to be imaged, Subsequent imaging in the imaging area is stopped. Further, when selecting the one imaged data based on the size of the sample, the number of samples in a preset predetermined size range measured by imaging through the permeable member is different from the previous one. When it is confirmed that the size is larger than the size of the specimen imaged through the member, it is determined that the image data at the time of imaging through the different transparent member immediately before is the focus on the object to be imaged. While being selected, subsequent imaging in the imaging area is stopped.

[0020]

DESCRIPTION OF THE PREFERRED EMBODIMENTS Before describing the details of the image pickup apparatus of the present invention, its technical background will be described.

The image pickup device according to the present invention collects a part of foods, cosmetics, etc., makes bacteria and fungi contained therein emit light by adding a special compound, and uses a CCD with a magnifying optical system. An image of the bacterium or fungus that has emitted light is captured by an imaging device, and the amount is counted.

The amount of light emitted from bacteria such as bacteria and fungi is very small, and although the amount of emitted light from bacteria is amplified by some image processing, long-time exposure of the image pickup device is essential. Since the time for long-time exposure of an image pickup device using a CCD is generally limited to about several seconds, it is necessary to keep the imaged object stationary during the time of about several seconds.

When a CCD is exposed for a long time longer than a predetermined time, the charge generated by heat is accumulated as noise in the CCD as the exposure time of the CCD becomes longer, which makes it indistinguishable from the charge accumulation due to the light emission of bacteria. I will end up. However, noise due to this cause can be prevented to some extent by cooling the image pickup unit, and when the image pickup unit is used, it is necessary to maintain the still state above the above.

The size of the bacteria to be imaged by the image pickup device applied to the present invention is about 1 micrometer in the case of a small one. Therefore, for example, when using a CCD in which one pixel is about 1.6 micrometers, one bacterium serves as information for one pixel of the CCD. There is no problem if the stationary state of the imaged object is completely stationary, but if there is some movement, the amount of emitted light will be dispersed to multiple pixels, resulting in a count error of bacteria. Alternatively, the charge may not be counted due to insufficient charge accumulation due to light emission. In such a case, the movement amount (blur amount) is required to be 1.6 μm or less. In other words, the maximum allowable range for blurring is
It is 1.6 micrometers.

On the other hand, it is conceivable to change the CCD used to one having a higher number of pixels.
If you reduce the number of pixels and increase the area of one pixel,
There is no problem if blurring occurs within the range of 1 pixel, but since multiple bacterial individuals will be included in 1 pixel,
There is a problem that the number of solids cannot be counted.

Further, when the number of pixels is increased and the area of one pixel is reduced, the light emission amount of one bacterium is received by a plurality of pixels, so that the light amount received by one pixel of the CCD is reduced and the exposure is performed. It becomes necessary to lengthen the time and increase the aperture diameter of the imaging optical system. Moreover, in this case, the apparatus becomes expensive and becomes large in size. Moreover, in this method, the amount of emitted light is dispersed to a plurality of pixels in accordance with the amount of blur, so that the amount of light per pixel is reduced, and it is not always possible to obtain high-quality image data.

Next, regarding the depth of focus of the image pickup optical system, it is necessary to enlarge the solid body of bacteria to about one pixel of the CCD as described above. In the case of a magnifying optical system, the depth of focus is generally shallow. The depth of focus in this embodiment is required to be about 60 micrometers.

Further, in the image pickup apparatus according to the present invention, when the image pickup unit having the magnifying optical system picks up an image of an individual such as a bacterium, the image pickup area is divided into a plurality of images and the image pickup is performed plural times. This is because by making the measurement area over a predetermined range, the measurement accuracy error factor due to the non-uniformity of the bacterial concentration distribution is reduced.

For this reason, the present image pickup apparatus employs a system in which the image pickup section is fixed and the carriage carrying the image pickup object can be moved to a plurality of locations.

On the other hand, under the condition that the depth of focus is 60 μm, it is necessary to always keep the depth of focus at this depth when moving the carriage to change the image pickup area of the object. This is extremely difficult in combination with the positioning accuracy of the image pickup, and in order to achieve high-accuracy imaging processing, and thus accurate counting processing of bacteria, focus on the object to be imaged in each imaging area. It is desired to match. That is, it is necessary to set the focal depth within the range of 60 micrometers for all the imaging areas.

For focusing in this case, when the objective lens configured as a part of the image pickup unit as in the above-mentioned conventional technique is moved in the vertical direction with respect to the object to be imaged, the objective lens installation mechanism is used. Since it is difficult to achieve perfect verticality by the installation method of the entire imaging unit,
As a result, the relative positions of the objective lens before and after the movement are displaced. That is, the imaging area (about 1 mm x 1
mm) shifts with each movement of the objective lens, resulting in double counting or missing of bacteria.

Further, since the lens itself may be shaken due to the movement depending on the mounting condition of the above-mentioned objective lens, it can be foreseen that the above-mentioned problems similarly occur.

It should be noted that the above-mentioned respective dimensions are merely cited as an example in order to easily understand the degree of precision of the required accuracy, and are not limited to the above-mentioned dimensions.

An embodiment of an image pickup device according to the present invention based on the above basic technique will be described below in detail with reference to the drawings.

FIG. 1 is a side view showing the entire image pickup device H, and FIG. 2 is a perspective view of a carriage 2 of the image pickup device. FIG. 3 shows a plan view of the stage 3 as seen from above in FIG. The main components of the present imaging apparatus are a stage 3 on which an object to be imaged is placed, a front cover 22 which is opened and closed for operations such as placing the object to be imaged on the stage 3, and an object to be imaged. The light source section 24 for irradiating, and the light source section 24
2 from the subject that was illuminated by
A CCD camera 20 that receives a three-dimensional image light and converts it into image data by an electric signal, a filter moving unit 23 that supports a plurality of filters and moves to switch the filters according to the switching of light emission, and further to the stage 3. It is composed of a first carriage moving mechanism section 13 and a second carriage moving mechanism section 12 for moving the stage 3 to the operation position when the object to be imaged is placed. And stage 3
Is formed between the upper surface of the second carriage 7 and the kit holder 27, and a kit such as an emblem kit or a film kit to which a sample is attached is attached thereto.

Further, the focus changing means having a plurality of glass bodies having different thicknesses is arranged between the CCD camera 20 and the stage, and a strip-shaped member for movably mounting the plurality of glass bodies is provided. (See FIG. 4).

The shape of the mounting table (stage) 3 on which the object to be imaged is placed is not limited to the shape shown in the attached drawings, and the object to be imaged is not limited to the shape shown in the accompanying drawings. As a configuration of mounting the sample on the stage 3, the mounting of the kit to which the sample is attached is illustrated in the present embodiment, but the present invention is not limited to this.

At the lower part of the first carriage 6, there is provided a first guide member 9 (guide rail 9a, which guides the first carriage 6 in the first direction (the direction of the vertical axis, which is the arrow Y shown in FIG. 1)).
9b) there is provided a first sleeve and a second sleeve (not shown) slidably fitted. Second carriage 7
At the lower part of the second guide member 10 for guiding it in the second direction (the direction of the horizontal axis indicated by the arrow X in FIG. 4).
A first sleeve 33 and a second sleeve 34, which are slidably fitted to the (guide rails 10a, 10b), are provided. As a result, the first carriage 6 and the second carriage 7 constitute a first carriage moving mechanism unit 13 (driving in the Y direction) and a second carriage moving mechanism unit 12 (driving in the X direction). The belt 36 pulls and moves.

When the apparatus is powered on by the power switch, first the initial operation starts. In the initial operation, the stage 3 moves to the reference position. This reference position is an image capturing position for capturing an image of the sample that is the imaged object,
It is set right under the CCD camera 20.

The stage 3 is arranged at a reference position in the X direction along the respective shaft directions of the first guide member 9 (guide rails 9a and 9b) and the second guide member 10 (guide rails 10a and 10b). The operated X-HP sensor and the Y-HP sensor arranged at the reference position in the Y direction move to a position where both are turned on.

When the initial operation is completed, the device enters the standby state. When the front cover 22 is opened, a signal indicating that the cover has been opened by the operator is emitted from a cover sensor (not shown). According to this signal (cover open signal), the stage 3 moves to the front position where the front cover 22 is open. Front cover 22
The open state is maintained by the coil spring.

When the object to be imaged (sample) is mounted on the stage 3 and the front cover 22 is closed by the operator, the signal from the cover sensor is sent to the data processing device connected to the interface, and the preparation is completed. Show that on the display. The operator instructs the data processing device to start imaging. The instructed data processing device sends an imaging start command to the imaging device. The control unit of the present imaging device controls the first carriage moving mechanism unit 13 to move the stage 3 to the home position for imaging, that is, the initial position of the imaging position.

The image pickup is repeated while the position of the stage 3 is changed little by little. There are a plurality of mounting kits for mounting the sample to be imaged, and there are differences between the case of measuring the emblem kit and the case of measuring the film kit. Here, the total area for imaging is 12 mm * 9 mm in the case of the emblem kit, and the area that can be acquired by one imaging is 1.2 mm * 0.9 mm, so 100 imaging frames are required. become.
In the imaging process, imaging is performed while moving the position 10 times in the Y direction, moving in the X direction, and repeating this.

Each time the image pickup of one frame is completed, the image data is transmitted to the data processing device. When the acquisition of the image data is completed, the data processing device requests the imaging device to acquire the next frame, and thus the imaging device performs the position movement and the re-imaging. In this way, imaging of required frames is repeated. After the imaging of all the necessary frames is completed, the stage 3 returns to the home position described above.

The imaged object PG is fixed on the stage 3 by a kit holder 27, and the stage 3 is provided on the carriage 2.

In this apparatus, the CCD camera 20 is not moved, but the stage 3 side on which the image pickup object PG is mounted is moved. Due to the movement of the stage 3, at the time of image pickup, the image formation point from the image pickup object PG is moved to the CCD camera
If it is not accurately focused on the CCD of 0, a precise image cannot be obtained, and as a result, the imaged object cannot be accurately recognized. Therefore, stage 3
Is always within the specified depth of focus range (for example, 6
Stage 3 to fit within 0 micrometer)
It is necessary to maintain the parallelism with respect to the image pickup means.

Therefore, in the present invention, in order to keep the relative distance between the CCD camera 20 side and the image pickup area A on the stage 3 constant, the stage attitude regulating means 4 for regulating the inclination attitude of the stage 3 and the fixed means. Fixing screw 5 as a means
And the connecting member 8 are arranged at a plurality of positions along the vertical axis direction (Y direction) and the horizontal axis direction (X direction) of the stage 3. As a result, the CCD camera 20 on the stage 3 and the CC
The parallelism in the plane direction with respect to the condenser lens arranged in front of the D camera 20 for obtaining the image of the imaged object is ensured.

In FIG. 2, the carriage 2 is composed of a first carriage 6 and a second carriage 7. The first carriage 6 is guided by a first guide member (guide rail) 9 fixed to the base 1 in a first direction (Y
Direction). The carriage 2 in the present device includes a second carriage 7 which is disposed on the first carriage 6 and is movable in a second direction (X direction) substantially orthogonal to the first direction (Y direction), and a second carriage 7. On the carriage 7, the stage 3 for mounting the imaged object, the connecting member 8 for connecting the stage 3 and the second carriage 7, and the stage 3
The first tilt adjusting means 4a and 4b which are the stage attitude regulating means 4 for adjusting the tilt of the stage 3 and the fixing screws 5a and 5b which are the fixing means for fixing the stage 3. Further, below the second carriage 7, second guide members (guide rails) 10a and 10b for guiding the second carriage 7 in a second direction (X direction), which is a moving direction, are provided. .

The first inclination adjusting means 4a and 4b constituting the stage attitude regulating means 4 are arranged in two pairs (a total of four) facing the side wall surface side of the stage 3, and the carriage 2 and the stage 3 are arranged. The relative position of is fixed by a fixing means (fixing screw) 5. As a result, the fixed positional relationship between the two becomes more reliable.

First carriage 6 and second carriage 7
Have separate drive motors and drive transmission mechanisms. This results in low maximum power consumption.

As shown in FIG. 1, the second carriage moving mechanism 12 moves the second carriage 7 mounted on the first carriage 6 in the horizontal axis direction (X direction). Drive source and a drive transmission unit. With such a configuration, it is possible to reduce the size of the carriage 2. In the present embodiment, both are not moved at the same time because of power consumption, but of course,
Both may be controlled to move at the same time.

In FIG. 3, the second carriage 7 faces the side wall surface of the stage 3 and faces the stage attitude regulating means 4.
2 pairs (total of 4), fixing means (fixing screws) 5 for fixing the relative positions of the carriage 2 and the stage 3, a connecting member 8 for connecting the stage 3 and the second carriage 7, And a second carriage moving mechanism portion 12 for moving the carriage 7. Further, the first carriage 6 can be moved in the first direction (Y direction) by being guided by the first guide member (guide rail) 9, and the second carriage 7 can be moved by the second guide member (guide rail). It is configured to be guided by a guide rail) 10 and movable in the second direction (X direction).

Between the stage 3 and the second carriage 7, springs 11, which are elastic members for urging the stage 3 upward with respect to the second carriage 7, are provided at the four corners of the stage 3, respectively. It is disposed so as to be wound around the axis at the position. Further, the connecting member 8 that connects the stage 3 to the upper portion of the second carriage 7 resists the upward biasing force of the spring 11 by the fixing screw 5 so that the stage 3 maintains a predetermined (horizontal) posture. And the connecting member 8 is fixed.

The object PG to be imaged is placed on the stage 3, and on the stage 3, the image pickup area A, and the height adjustment reference position p0 for adjusting the inclination of the stage and the distance from the CCD camera 20, And height adjustment positions p1, p2, p3,
p4 is shown. The attitude of the table is adjusted by using the height adjustment reference position, the height adjustment position, and an adjustment jig (not shown). This facilitates the work for fixing the relative position that realizes the flatness of the imaging surface with respect to the imaging device.

Further, since the depth of focus is extremely shallow (60 micrometers) in the image pickup apparatus H, the sample PG is actually only within the range of the depth of field of the objective lens 21 (FIG. 10) of the CCD camera 20. Imaging is not possible, and the imaging range is a very narrow range of several tens of micrometers depending on the objective lens used. Furthermore, since the stage 3 is moved with respect to the fixed CCD camera 20 and the image pickup area A is divided into a plurality of areas for image pickup, the parallelism of the stage 3 itself with respect to the CCD camera 20 is increased. It is very difficult to always keep it constant because of variations in accuracy of mounting parts and the presence of backlash, and as a result, the focus at the time of imaging the sample PG is deviated, and a precise image cannot be obtained. It is difficult to accurately count the number of bacteria based on image data with such a blurred focus.

The image pickup apparatus according to the present invention is provided with the focus changing means 60 for focusing on the object PG when moving the stage 3 to pick up a plurality of image pickup areas (shown in FIG. 11). The focus changing means 60 is arranged between the CCD camera 20 and the stage 3.

FIG. 4 is a front view showing the entire image pickup device H. In FIG. 4, the focus changing means 60 arranged at a predetermined position between the CCD camera 20 (imaging means) and the stage 3 arranged on the base 1 is a two-dimensional image light from the object PG. A plurality of glass bodies (transmissive members) 61 having different thicknesses for changing the optical path of the plurality of glass bodies, and a plurality of glass bodies 61 are placed and one of the plurality of glass bodies 61 is sequentially placed in the CCD camera 20
A chain 62, which is a belt-shaped member that is moved to a predetermined position between the stage 3 and the stage 3, and a sprocket wheel that is arranged on both sides of the CCD camera 20 and that meshes with the chain 62 to regulate the transfer path of the chain 62. 63 and a reel 64 which is provided above the two sprocket wheels 63 and which can reel out and wind the chain 62.
And so on. In the embodiment shown in FIG. 4, a chain 62 for transferring any one of the plurality of glass bodies 61 to a predetermined position between the CCD camera 20 and the stage 3 is extended or wound in the arrow Z direction. However, the present invention is not limited to this, and a chain formed by connecting the chains 62 in an annular shape can be used, and the sprocket wheel 63 described above can be used in place of the reel 64. In this case, the transport direction of the chain 62 may be any one direction, or may be both directions indicated by an arrow Z (reciprocating motion of the chain 62) as illustrated.

Here, the chain 6 used in this embodiment
2 will be briefly described. FIG. 5 shows a perspective view of the chain 62 used in this embodiment. As shown in FIG. 5, the chain 62 is made up of a steel plate eyebrow-shaped link plate 7
1, 73 are connected with the pin 74, and the bush 72
And a roller 75, the pin 74 is a pin link plate 71, and the bush 72 is a roller link plate 7.
The roller 75 may be a roller chain that can be freely rotated although it is fixed to the roller 3. By inserting the teeth of the sprocket wheel 63 between the rollers 75, the two are in mesh with each other. In the present embodiment, a roller chain with a substantially L-shaped attachment 68 integrally attached to the above-mentioned roller link plate 73 is used, and the plurality of glass bodies 61 in the present invention are substantially the same as the attachment 68. It will be attached to the horizontal part.

6 and 7 show a state in which a plurality of glass bodies 61 are attached to the chain 62.
FIG. 7 is a plan view showing a state where a plurality of glass bodies 61 are attached to a chain 62, and FIG. 7 is a side view showing a state where a plurality of glass bodies 61 are attached to the chain 62.
Note that the symbol A shown by the alternate long and short dash line in FIG. 6 represents a plurality of imaging areas (see FIG. 11) as one area, and indicates the entire imaging area of the object PG. As shown in FIG. 6, each of the glass bodies 61 is formed slightly larger so as to cover the imaging area A.

In the present invention, a plurality of glass bodies (transmissive members) 61 having different thicknesses, which are main parts of the focus changing means 60, are vertically held by a resin member 65 in a sandwich state, These resin members 65 are attached to a substantially horizontal portion of a substantially L-shaped attachment 68 integrally attached to a part of the chain 62 with a fixing screw 66 and a nut 67.

In this embodiment, the chain 62
Is arranged in two rows, and each of the above-mentioned attachment members is provided on each chain 62, and the glass body 61 is held in a pair at two opposite positions (double-sided construction). However, the chain 62 may be configured in one row, and only one position on each side of each glass body 61 may be held by the above-mentioned mounting member (cantilevered configuration).

In this embodiment, the glass body 61 is attached to the chain 62 by holding one side portion (one side) of the glass body 61 at one place, but the present invention is not limited to this, and two attachments are used. It may be configured to hold two locations.

As in this embodiment, the chain 6
When 2 is configured in two rows, two sprocket wheels 63 and reels 64 shown in FIG. 4 are arranged in the depth direction of the image pickup device H, two each.

Further, in this embodiment, the chain 62
Although the configuration in which a plurality of glass bodies 61 are transferred is shown by using, when the imaging device H allows the arrangement space, a plurality of glass bodies 61 are replaced with the chain 62 as shown in FIG. Table 6 as a movable support for
9a, or as shown in FIG. 9, it is also possible to configure the rotary table 69b as a movable support for the plurality of glass bodies 61.

In the horizontal table 69a shown in FIG. 8, a plurality of glass bodies 61 having different thicknesses are placed in a line in the horizontal direction and reciprocally moved in the direction of arrow W, whereby the glass bodies 61 are moved.
One of them is placed at a predetermined position.

The rotary table 69b shown in FIG.
By placing a plurality of glass bodies 61 having different thicknesses in a circle in the horizontal direction and rotating the glass bodies 61 in the direction of arrow S, one of the glass bodies 61 is positioned at a predetermined position.
This rotational movement may be in both directions as shown, or may be any unidirectional rotation.

The broken lines shown in FIGS. 8 and 9 indicate image pickup positions, and one of the plurality of glass bodies 61 is positioned at a predetermined position for image pickup.

The above-mentioned three embodiments (chain 6)
In the 2 / horizontal table 69a / rotary table 69b), a description of a specific configuration for transferring the plurality of glass bodies 61 is omitted, but even in the case of automatic conveyance by a conventionally known drive motor and drive transmission mechanism. Alternatively, manual transport may be performed manually, or a transport mechanism including both may be configured.

Next, the relationship between the thickness of the plurality of glass bodies 61 and the focal length will be described with reference to FIG. Figure 10
[Fig. 3] is a sectional view of a focus changing means for explaining the relationship between the thickness of a plurality of glass bodies and the focal length.

As described above, the structure in which the plurality of glass bodies 61 are sequentially transferred between the stage 3 and the CCD camera 20 and positioned at a predetermined position for image pickup has been described. Focusing on that the focal length extends about 1/3 of the thickness of the glass body when the glass body is positioned between the CCD camera 20 and the CCD camera 20 applied to the present invention. A glass body 61 having a different thickness is sequentially positioned between the objective lens 21 provided at the tip of the and the image-captured body PG so that the focal lengths are different.

In FIG. 10, 0.01 mm (10
7 glass bodies 6 from 0.10 mm to 0.16 mm with different thickness for each micrometer)
1a, 61b, 61c, 61d, 61e, 61f, 61
Although g is shown, according to the above-mentioned principle, the focal length is increased by 3.3 micrometers. Here, a glass body 61d having a thickness of 0.13 mm
When set as standard, the value is ± 1
The focal length can be changed in the range of 0 μm, and when the image of the object PG is picked up, the plurality of glass bodies 61a to 61g are sequentially replaced to change the focal length in seven steps, that is, seven steps. The focus position can be adjusted. In the present embodiment, the seven glass bodies 61a to 61g are described, but the glass body 6 is used.
There is no limitation on the number of glass bodies 61, and if the number of glass bodies 61 having different thicknesses is increased, the allowable execution range of the focus position adjustment described above is expanded.

FIG. 11 is a schematic plan view showing a state in which the image pickup area A of the object PG is divided into a plurality of areas. In FIG. 11, these plural areas are designated as a1 to a3 and b.
It is configured as 9 sections of 1 to b3 and c1 to c3. In this device, the two-dimensional image light from the imaged object PG emitted from the light source unit 24 (24a, 24d in FIG. 4) that irradiates the target imaged object PG and emits the excitation light by the reagent. The image pickup apparatus includes a CCD camera 20 that receives light and converts it into image data by an electric signal. The light source unit 24 described above includes a light emitting diode (LED) that emits light rays in different wavelength regions on the substrate 50 (in FIG.
0a, 50d), respectively, to receive the above-mentioned two-dimensional image light of a specific wavelength corresponding to a light beam (excitation light) from each light emitting diode, convert it into image data, and count the single imaged image. Then, for example, the object PG
It is possible to confirm the number of live and dead cells of the microorganism (bacteria) or the number of only dead cells. In this case, when confirming the living cells, the calculation process (the process of subtracting the number of dead cells from the number of dead cells) is performed, but as described above,
In order to confirm live cells, it is necessary to perform imaging processing at least twice in the same imaging area in order to perform imaging by irradiating excitation light in different wavelength regions and receiving two-dimensional image light corresponding thereto.

In the present invention, the stage 3 shown in FIG. 11 is moved in the directions of the arrows X and Y by the above-mentioned moving mechanism to set one of the divided image pickup areas to the position P (the position of the area b2). The image pickup is performed by positioning the image pickup device. Further, in order to adjust the focus position at the time of image pickup described above, image pickup is further performed a plurality of times in the same image pickup area corresponding to a plurality of glass bodies 61 having different thicknesses. It is a thing.

A chain 62, a horizontal table 69a or the like is used as a structure for transferring a plurality of glass bodies 61 arranged in a row when the image pickup area is changed and different image pickup areas are positioned at predetermined positions for image pickup. When reciprocating with a plurality of glass bodies 6, the transfer direction is reversed.
If the positioning order of 1 is reversed, the processing time can be shortened.

Next, based on the image data of the imaged object PG obtained by applying a plurality of glass bodies 61 and imaging a plurality of times, the system configuration of the image pickup processing relating to the focus determination processing for the image pickup area will be described. Description will be made based on the following block diagram.

FIG. 12 is a block diagram relating to the system configuration including the functions of the host device connected to the above-mentioned image pickup device.

First, the control section 110 of the image pickup apparatus H is for controlling the operation of each section of the apparatus.
U is used and is connected to the host device. The control unit 110 described above uses, for example, the imaging device 140, or the first moving member (carriage) 6 and the second moving member (carriage) 7 based on an operation instruction signal from the host device.
The exposure control means 111 controls the CCD 96 to operate the driving means 13 and 12 for moving the
In response to this, a signal relating to the start or reset of the clock relating to the image pickup (exposure) time control is transmitted, and the drive control means 115 makes the first in the Y-axis direction between the image pickup object attachment / detachment area and the image pickup area. ON / OFF to the drive motor as the first drive means 13 in order to execute the movement of the first moving member (carriage) 6 which is the moving means of
In order to send a signal and execute the movement of the second moving member 7 which is the second moving means in the image pickup area in the X-axis direction, the drive motor as the second drive means 12 is driven ON / OFF. Send an OFF signal. It should be noted that by selectively controlling the drive of the two drive motors described above, the power consumption of the device can be suppressed and the configuration relating to the power supply can be miniaturized. Good. In this case, the stage 3 is quickly moved to the predetermined position, and the processing time can be shortened.

In the present apparatus, the above-mentioned control section controls the drive control means 115 and the exposure control means 111 at individual timings, so that the carriage 2 provided with the stage 3 is moved to a predetermined position which is an image pickup execution position. The operation is controlled so that the photoelectric conversion is performed in a stationary state after the movement. As a result, the timing of the desired imaging process is operated correctly, the interface with the external device is facilitated, and the CC
As D96, the function of a sensor (plane sensor or area sensor) in which a plurality of photoelectric conversion elements are arranged in a plane can be sufficiently achieved.

A general personal computer having a desired application can be appropriately used as a host device for performing various data processing. Various operations of the entire image processing system are performed by the C as the central control unit 136.
Various operation command signals that are executed by the PU and are sent to the imaging device side, status signals that indicate an operation state including image data or operation errors from the imaging device side, and the like are communicated via the interface unit 130.

When an operation instruction relating to exposure is given to the control section 110 in the image pickup apparatus H through the above-mentioned communication path, the exposure control means 111 gives a command to the CCD 96. Around this, the control unit 110 in the image pickup apparatus H is instructed to move the stage 3 in a predetermined image pickup area. By repeating the operations of the image pickup means and the drive means based on both of these instructions a predetermined number of times, image data in a plurality of divisions (image pickup areas) shown in FIG. 11 can be obtained. The obtained image data is transferred from the photoelectric conversion unit 96 to the A / D converter 112 of the control unit 110, and the analog video signal is converted into digital image data. In this apparatus, the image data imaged in one imaging execution area and digitally converted is temporarily stored in the image data storage unit 113 such as an SD-RAM, and then transferred to the upper apparatus side.

The image data transferred from the reading device H side is sent to the image data storage unit 131 via the interface unit 130 and stored therein. The image data processing unit 132 reads the image data from the image data storage unit 131 and performs a predetermined process.

In the present invention, the image data processing unit 13
Reference numeral 2 determines whether or not the focus is on the object PG to be imaged based on the image data read from the image data storage unit. This determination process may be performed based on the number of specimens (bacteria) measured from the captured image data, or may be performed based on the size of the specimen measured from the captured image data. Therefore, the image data processing unit 132 also has a function as a sample number measuring unit that measures the number of samples or a sample size measuring unit that measures the size of the sample. The sample number measuring means or the sample size measuring means may be provided in the image pickup apparatus, but as shown in FIG. 12, it is provided in an external data processing apparatus such as a personal computer. It may be configured.

Then, the image data conversion unit 133 in the external device converts the image data processed by the image data processing unit 132 into a format for monitor display (including, for example, content for converting again into analog data).
Then, it is displayed on the monitor 134, or RGB (light) data is converted into CMY (color) data for printing by the printer 135 and printed by the printer 135.

The central control unit 136 described above gives various commands to the image pickup apparatus side through the interface unit 130, and, as described above, instructs the image data processing unit 132 from the image data storage unit. Based on the read-out image data, a command is given to determine the focus match with respect to the imaged object PG that is the imaging target, and the above-mentioned various data conversion is executed to the above-mentioned image data conversion unit 133. It gives a command.

In the present embodiment, the image pickup device H
Although the area sensor is used as the photoelectric conversion means 96, the present invention can be applied to a line sensor, and the type of the image pickup object as the image pickup object does not matter.

Here, the determination of focus matching with the object PG to be imaged based on the image data in the present invention will be described.

First, a brief description will be given of the case where the measurement is performed based on the number of specimens measured from the imaged image data. Bacteria as a sample in the imaged object PG have a size of about 1 micrometer in a small one, and if a plurality of bacteria are present in one imaging area described above,
CCD camera 20 (objective lens 2 for the object PG)
When the focal length of 1) does not match, the number of bacteria will be smaller than the actual number because adjacent bacteria overlap each other. Therefore, a plurality of image data whose focal lengths have been changed by sequentially interposing a plurality of glass bodies 61 are acquired, the number of bacteria is measured from each of those image data, and the image data when the maximum value is measured. Is determined and adopted as appropriate data, and the glass body 61 used at this time is applied as a focusing member in the imaging area. At that time, as the glass body 61 was sequentially changed, the measured value of the number of bacteria based on the acquired image data gradually increased, and when it was confirmed that it decreased at a certain point, the measurement immediately before the transition to the decreased state was performed. The value is determined to be the maximum value, and it is determined that the image capturing by the glass body 61 at the time of acquiring this image data is in focus. In this case, it may be determined that the image pickup using the other glass body 61 in the image pickup area is unnecessary, and the subsequent image pickup in the image pickup area may be stopped and the process may be shifted to the next process.

Next, a case will be described in which it is performed based on the size of the sample (bacteria) measured from the imaged image data. As described above, the bacterium as the imaged object PG has a size of about 1 micrometer when it is small. In the case where a plurality of image pickup areas exist in one image pickup area described above, in the state where the focal length of the CCD camera 20 (objective lens 21) does not match the image pickup object PG, adjacent bacteria actually overlap each other. Will be larger than the size of.

Therefore, a plurality of image data whose focal lengths are changed by sequentially interposing a plurality of glass bodies 61 are acquired, and the size of bacteria is measured from each of the image data, and the minimum size is obtained within a predetermined range. The image data at the time of measuring the value is determined and adopted as appropriate data, and the glass body 61 used at this time is applied as a focusing member in the imaging area. In this case, since it is necessary to limit specific bacteria as the target for measuring the size, for example, when the size of the target bacteria is 1.0 to 1.3 micrometers, for example, The measurement range to be set is set to 0.8 to 1.5 micrometers in advance,
With the minimum value of bacteria measured within this range, the imaging data is judged to be proper data and adopted.

In the present embodiment, the number of specimens measuring means or the specimen size measuring means described above, and the image data processing section 132 as the judging means for judging the focus match with the object PG to be imaged are included. The processing unit that performs various data processing has been described by taking a general personal computer having a desired application, which is a higher-level device of the image pickup apparatus H, as an example. However, these configurations are applied to the image pickup apparatus H according to the present invention. It may be configured to be functionalized and provided in the control unit.

Further, although the glass body 61 has been described as a constituent element of the focus changing means 60 in the present invention, the composition is not limited to glass, and a plastic body having a similar transparency is also used. May be.

As described above, the image pickup apparatus according to the present invention includes the base, the image pickup means fixed to the base for picking up an image of the object to be imaged, the vertical axis direction and the horizontal axis in the image pickup area of the object to be imaged. A carriage that moves in a plane along the direction, a stage that is provided on the carriage for mounting an imaged object, and a focus changing unit that changes the focus of the imaged object when the imaged object is imaged by the imaging unit. Equipped with. The focus changing means is arranged at a predetermined position between the image pickup means and the stage, and one of the plurality of glass bodies for changing the focal length is positioned at the predetermined position. As a result, the present invention realizes the cost reduction of the image pickup apparatus by obtaining the fine image data of the image pickup object moving in the image pickup area divided into a plurality of frames by the focusing mechanism having a simple structure. Thus, it is possible to accurately count the samples in the image data thus obtained.

[Brief description of drawings]

FIG. 1 shows a side view of an image pickup apparatus according to the present invention.

FIG. 2 is a perspective view of a carriage in the image pickup apparatus according to the present invention.

FIG. 3 is a plan view of the carriage shown in FIG. 2 seen from above.

FIG. 4 shows a front view of an imaging device according to the present invention.

FIG. 5 shows an example of a belt-shaped member on which a plurality of glass bodies are movably mounted.

FIG. 6 shows a plan view of a belt-shaped member on which a plurality of glass bodies are movably mounted.

FIG. 7 shows a side view of the belt-shaped member shown in FIG.

FIG. 8 shows a plan view of a horizontal table as a movable support for a plurality of glass bodies.

FIG. 9 shows a plan view of a rotary table as a movable support for a plurality of glass bodies.

FIG. 10 is a sectional view of a focus changing means for explaining the relationship between the thickness of a plurality of glass bodies and the focal length.

FIG. 11 is a schematic plan view showing a state where the imaging area A is divided into a plurality of areas.

FIG. 12 shows a block diagram relating to a system configuration of an imaging device.

[Explanation of symbols]

1 base 2 carriage 3 stages 6 First carriage (first moving member) 7 Second carriage (second moving member) 9 First guide member (guide rail) 10 Second guide member (guide rail) 12 Second carriage moving mechanism section (driving means) 13 First Carriage Moving Mechanism Section (Drive Unit) 20 CCD camera 21 Objective lens 24 Light source 60 Focus changing means 61 glass body 62 chains 63 sprocket wheel 64 reel 69a horizontal table 69b rotating table H imaging device

   ─────────────────────────────────────────────────── ─── Continued front page    (72) Inventor Masahiro Kono             1 430 Kobayashi, Masuho-cho, Minamikoma-gun, Yamanashi Prefecture             Within ska corporation F-term (reference) 2H052 AF02 AF14                 5C022 AA08 AB21 AC26 AC78 CA01                 5C054 CC06 CD03 CF01 CF05 EA01                       HA05 HA12

Claims (13)

[Claims] 1. A base, an image pickup means fixed to the base for picking up an image of an object to be imaged, and a carriage moving in a plane along a vertical axis direction and a horizontal axis direction in an image pickup area of the object to be imaged. A stage for mounting the object to be imaged, which is provided on the carriage; and a focus changing unit for changing a focus on the object to be imaged when the imager captures the object to be imaged, The imaging device, wherein the focus changing means is arranged at a predetermined position between the imaging means and the stage. 2. The image pickup apparatus according to claim 1, wherein the focus changing unit has a plurality of glass bodies having different thicknesses. 3. The focus changing means comprises a belt-shaped member on which the plurality of glass bodies are movably mounted, and one of the plurality of glass bodies is positioned at the predetermined position by the movement of the belt-shaped member. The image pickup apparatus according to claim 2, which is characterized in that. 4. The focus changing means includes a movable support body on which the plurality of glass bodies are mounted and which performs a reciprocating operation or a rotating operation in a horizontal direction, and the plurality of glass bodies are moved by the reciprocating operation or the rotating operation of the movable support body. The image pickup apparatus according to claim 2, wherein one of the glass bodies is positioned at the predetermined position. 5. The image pickup apparatus according to claim 3, wherein the image pickup unit picks up an image a plurality of times corresponding to the plurality of glass bodies in a predetermined image pickup area. 6. A sample number measuring unit for measuring the number of samples in a predetermined imaging area of the object to be imaged based on imaged data by the imaging unit, Determining that the glass body positioned at the predetermined position when the number of the specimens measured by the specimen number measuring means shows the maximum value is a focusing member that focuses on the imaging area. The image pickup apparatus according to claim 5, wherein 7. The sample size measuring means for measuring the size of the sample in the predetermined imaging area of the object to be imaged based on the imaged data by the imaging means, , The glass body positioned at the predetermined position when the size of the sample within a preset predetermined size range measured by the sample size measuring means exhibits a minimum value is focused on the imaging area. The image pickup apparatus according to claim 5, wherein the image pickup apparatus is determined to be a matching focusing member. 8. The imaging device is connected to an external data processing device having a sample number measuring device for measuring the number of samples in the predetermined imaging area of the object to be imaged based on the imaged data of the imaging device. Of the plurality of times of imaging by, the glass body positioned at the predetermined position when the number of the samples measured by the sample number measuring means shows the maximum value is focused on the imaging area. The imaging device according to claim 5, wherein the imaging device is determined to be a focusing member. 9. The image pickup device is connected to an external data processing device having a sample size measuring unit for measuring the size of a sample in a predetermined image pickup area based on image pickup data obtained by the image pickup unit. Among the plurality of times of imaging by the, the glass body positioned at the predetermined position when the size of the sample within the preset predetermined size range measured by the sample size measuring unit shows the minimum value is The imaging device according to claim 5, wherein the imaging device is determined to be a focusing member having a focus in the imaging area. 10. A focusing method for an image pickup device, wherein the image pickup device picks up an image pickup target in a predetermined image pickup area by an image pickup unit fixed to a base, wherein the image pickup unit picks up the image pickup target. The plurality of transmissive members capable of changing the focus on the imaged object are sequentially transferred to a predetermined position between the image pickup means and the imaged object and imaged,
Focusing on the object to be imaged is performed by selecting one of the imaging data for each of the plurality of transparent members based on the number of specimens or the size of the specimen in the predetermined imaging area. A focusing method in an image pickup apparatus, which is characterized by being executed. 11. When changing the imaging area,
The focusing method in the imaging apparatus according to claim 10, wherein the imaging target is imaged after the plurality of transmissive members are transferred to the predetermined position in reverse order. 12. When selecting the one image data based on the number of the specimens, the number of specimens imaged and measured through the permeable member is imaged through a different permeable member immediately before. By confirming that the number of specimens is smaller than the number of specimens, the image data at the time of imaging via the immediately preceding different transparent member is determined to be the focus on the object to be imaged and selected. The focusing method in the imaging apparatus according to claim 10, wherein subsequent imaging in the imaging area is stopped. 13. The number of specimens within a preset predetermined size range measured by imaging through the permeable member is immediately before when the one piece of imaging data is selected based on the size of the specimen. By confirming that the size of the specimen imaged through the different transparent members is different, the image data at the time of imaging through the immediately previous different transparent member has a focus on the object. 2. The image pickup device is determined to be selected, and the subsequent image pickup in the image pickup area is stopped.
0. The focusing method in the imaging device according to 0.