JPH0697203B2 - Particle aggregation determination device - Google Patents

Description

TECHNICAL FIELD The present invention relates to a particle agglutination determining apparatus for clinical examination, and more particularly to an agglomeration pattern determining apparatus for a microtiter method.

[Conventional technology]

At present, in the microtiter method in immunological measurement, it is widely performed to detect the presence or absence of aggregation on a microplate and to measure a trace amount of an immune component. The presence or absence of aggregation is visually determined by the naked eye. In this visual determination, the presence or absence of aggregation is regarded as the area of a portion of the distribution of particles in the well which is equal to or less than the brightness. Further, the judgment is made by comprehensively combining judgments such as comparison with the standard agglutination pattern and the standard non-aggregation pattern, and consideration of the relation with the state of the adjacent well. Therefore, the visual judgment requires a high degree of skill, and since it is a sensuous test method, there are individual differences among the judges, and there is a drawback that the same judge lacks reproducibility.

Automating this visual judgment by a device not only saves labor, but also makes the judgment result objective and can be expected to greatly improve the measurement accuracy. Furthermore, the automatic printing of the inspection results makes it possible to completely eliminate errors in the transcription of the measurement results. The particle agglutination reaction has many test items that can be applied, is easy to operate, has high detection sensitivity, and is suitable for large-volume sample processing, but the only drawback is that the final judgment is not automated. there were. Therefore, it is extremely important for clinical examinations to solve this drawback and to develop an automatic determination method of an agglutination reaction with high accuracy, and it will greatly contribute to the development of medicine.

[Problems to be solved by the invention]

However, mechanization of the same judgment as the visual judgment based on the combination of complicated factors which is currently performed makes the device extremely complicated, economically expensive and impractical. Therefore, it is a technical point to limit the number of judgment factors in the automatic judgment by the device and how to match the result with the visual judgment. All of the automatic judgment devices that have been attempted so far use a photometer and try to correspond the absorbance at the center of each well with the agglutination, or the light quantity ratio between the center of the well and the peripheral edge of the well corresponds to the agglutination. However, all of them have only the ability as an auxiliary means for visual judgment, and none of them have the ability to replace visual judgment.

Also, because the conventional device measures wells one by one,
It took a long time to perform the measurement, and a moving device for the plate was necessary.

Further, in the conventional automatic determination device, the transmitted light is used, but this method can achieve only extremely insufficient measurement, as described below. That is, the sample of the particle agglutination reaction contains various proteins such as serum as well as serum, and these components are often precipitated separately from the particle agglutination reaction, so that the entire solution becomes cloudy.

As a result of careful observation of an extremely large number of test results, the present inventors have found that this phenomenon is caused by an extremely complicated combination of the sample serum and the reagent solution. For the first time, I found that a high concentration occurs in an unignorable number.

However, in the conventional device, no countermeasure was taken against this white turbidity, and only insufficient measurement was possible. That is, in the measurement of transmitted light, the amount of light is remarkably reduced due to diffused reflection due to precipitates.Therefore, the amount of transmitted light and the particle agglutination reaction do not correspond, and the agglutination automatic determination device that converts the particle concentration into brightness can accurately determine the agglomeration There wasn't.

In order to solve the above-mentioned problems, the inventors of the present invention have proposed a device for irradiating light from above the microplate and utilizing the reflected light and the transmitted light to determine the aggregation of particles (Japanese Patent Application No. 60 -57880). However, although the influence of white turbidity was eliminated in this device, the amount of light in the wells of the microplate could not be increased only by illuminating from above so that the contrast was sufficient. That is, if the amount of light in the well is to be increased as much as the contrast is sufficient, the amount of light from the light source must be extremely increased, heat is often generated, and it is necessary to provide a cooling device such as a fan. It was expensive. In addition, the power consumption was also high. Furthermore, since the dynamic range of the TV camera is limited, the amount of light can be increased only until the upper surface of the microplate reaches a predetermined brightness, and the well cannot be sufficiently brightened. Furthermore, an image of the light source appears on the liquid surface in the well, and this may overlap with the agglutination image, making measurement difficult.

SUMMARY OF THE INVENTION It is an object of the present invention to solve the above problems and provide a particle aggregation determination device that is simpler than the conventional device and has an accuracy of visual determination or higher.

[Means for solving problems]

By the way, an image processing apparatus for processing an image signal from a TV camera is currently used in a wide field. For medical use, for example, an X-ray tomographic image is analyzed and an image of white blood cells in blood obtained by a microscope is used. For analysis and classification and number measurement, and for industrial use, there are inspection of substrates used for electronic circuits, non-destructive inspection of X-rays, and the like.

The image processing method used in this image processing device extracts only the image of the object to be measured from the captured image, and the area, peripheral length, number, angle Used for the purpose of measuring etc. In order to extract the image to be measured, if there is a brightness difference between the image to be measured and other images, the brightness difference is used to set the brightness of images below a certain brightness level to 0, and the images above that brightness level. Is the highest brightness in the brightness range (binarization process), and the method of extracting the target image, or taking the difference in brightness with respect to the image to be measured, and differentiating the difference There is a method of obtaining the gradient, obtaining the point having the largest gradient, and connecting the points having the largest gradient obtained to extract the contour of the target image. However, in order to perform these processes, there is a clear difference in brightness between the image to be measured and other parts in the original image to be processed, and the object to be measured was irradiated. It is required that the brightness of light be uniform.

However, in order to make the same judgment as the visual judgment by the combination of complicated factors that is currently performed and to limit the number of judgment factors in order to make the device economically inexpensive, there is a clear difference in brightness. Achieving a clear image formation with uniform illumination was not practical because the device was expensive.

Further, in the method of obtaining the contour of the image obtained by differentiating the brightness difference and extracting the image, it is difficult to extract only the contour of the aggregation pattern in the determination of the particle aggregation of the microplate. That is, in the well of the microplate, the brightness of the portion where the aggregation pattern is formed is
The transmitted light from the bottom is blocked, and the degree of reflection of the light from above is less than the reflection of the microplate itself, so there is a difference in brightness between the aggregation pattern and the surrounding donut-shaped portion. . On the other hand, outside the well, the top surface of the microplate is flat, so the light from the bottom enters the TV camera directly above, and the light from the top also reflects.
Incident on a TV camera. On the other hand, since the shape of the edge of the well is slanted downward, part of the light from the bottom is reflected and does not enter the TV camera, and the light from above is also reflected to the TV camera. Incidents only a little. Therefore, there is a brightness difference between the edge of the well and the flat part of the other microplate. In this way, in the determination of particle aggregation of the microplate, the aggregation pattern and the edge of the well are extracted as an image. In addition, if there is a scratch inside the well, that part will also be extracted with a brightness difference, and in order to distinguish it from other plates on the top surface of the microplate, the item to be measured, the date to be measured, the number of plates, etc. If you enter, it will also be extracted.

As described above, it is difficult to accurately determine the area of the aggregation pattern in the well by the generally known image processing method.

As a result of earnest research, the inventors of the present invention focused on the fact that the aggregation pattern is circular, and accurately determined the contour of the aggregation pattern by determining the brightness difference with a radius in a concentric circle from the center position of the well. It was made possible. In more detail, the agglomeration to be measured, the contour of the image formed by the pattern is not always clear, but this image is always circular, and the center of the circle is always dark and gradually brightens at the periphery, and the contour part The rate of change to brighten becomes maximum. Further, since the image is always formed in the well on the microplate, the interval between the aggregation pattern and the sedimentation pattern is constant. The present invention takes full advantage of these peculiarities. In the present invention, it is important to determine the center position of the particle aggregation pattern. If the center position can be determined, a derivative operator is operated from this center position toward the periphery to detect the point with the maximum differential coefficient that emphasizes the circular component that changes from dark to light, and the contour of the aggregation pattern is selectively extracted. Because you can do it. However, it is not always easy to accurately obtain the center position of the particle aggregation pattern in advance for the reason described below.
In other words, in order to keep the positional relationship between the TV camera and the microplate mechanically constant, the TV camera and the microplate must be configured with highly accurate parts for positioning, which makes the device extremely large, heavy, small, and lightweight. It is far from the measuring device. Moreover, if a mechanical deviation occurs due to aging or the like, the measurement accuracy becomes extremely inaccurate immediately.

Therefore, we devised a method in which a positioning hole is provided on the table on which the microplate is placed, the coordinates of this hole are measured, and then the center position of each well of the microplate, that is, the center position of the aggregation pattern is determined from this coordinate. . This way
Even if the relative positions of the TV camera and the microplate are slightly displaced, the center of the aggregation pattern can be accurately calculated. This method is extremely important for reducing the size and weight of the device, and is effective for maintaining the performance for a long period of time. The contour of the aggregation pattern obtained using the center position accurately obtained by the above method is "dark".
To "Ming".

Since it is obtained by the maximum value of the differential coefficient emphasizing the circular component, it achieves a wide low range of noise in the vicinity and does not necessarily require the uniformity of the illumination of the device, so that the economical effect of the device is great.

That is, the particle agglutination determination apparatus of the present invention is a plate mounting table that is light-transmissive on which a microplate is mounted and is provided with a positioning hole for determining the center position of each well of the microplate, and the plate mounting table. An image capturing device having light sources arranged above and below and a TV camera provided so that light emitted from the upper side of the microplate mounted on the plate mounting table is incident, and an image input from the TV camera An image processing device that processes a signal to determine the aggregation state, the image processing device determines the center position of each well by the transmitted light of the positioning hole, and then images the wells from the center position. The value is the sum of the differences in the brightness from the pixels in the outward direction at each divided part, and is the differential value of each pixel. The differential value of each pixel is sequentially compared in one direction and the higher order The feature is that the aggregation state is determined by counting the number of pixels between two differential values.

The plate mount of the particle aggregation determination device of the present invention is for positioning the microplate at a predetermined position.

In order to place the microplate in a predetermined position, even if a recess is formed on the plate table so that the entire lower part of the microplate fits, a well recess is formed so that each well of the microplate slides down and stops at a predetermined position. You may form. The well recess may have a conical shape, a spherical shape, or a through hole as long as the well is depressed and fixed.

The plate holder is formed to be light-transmissive so that light from a light source provided below can enter the microplate. Therefore, at least the portion on which the microplate is placed is formed of a transparent body or a semitransparent body, and the portion corresponding to the well may be cut to form a through hole.

Further, the plate mounting table is provided with a positioning hole for determining the center position of each well of the microplate. This positioning hole provides a reference line segment when the center position of the well is determined by the image processing apparatus, and at least two positioning holes are required.

This plate holder may be provided so as to be movable inside and outside the image capturing device. By movably providing the plate holder, the microplate can be automatically fed and discharged efficiently.

The microplate mounted on the plate mounting base is not particularly limited in kind, size, number of wells, etc. of rigid, permanent, etc., but it is required that it is formed to be light transmissive, that is, transparent or translucent.

A main light source is provided above the plate table.
This main light source irradiates the microplate with light and causes the reflected light to be taken into the TV camera as optical state information. The main light source is appropriately selected and used, but a fluorescent lamp with a high color temperature is preferable because it can be used for measuring various colors of particles, and a transistor inverter is used as the power supply unit because it has little flicker and good image reproducibility. It is preferred to use a high frequency fluorescent lamp with.

On the other hand, an auxiliary light source is provided below the plate stand. This auxiliary light source emits light from the back side of the microplate and allows the transmitted light to be captured by the TV camera as information on the optical state. It is for correcting the brightness in the well. The quality of light of the auxiliary light source may be the same as that of the main light source, but the amount of light is preferably 1/2 or less of the main light source on the microplate, and most preferably about 1/3 of the main light source. . Therefore, it is preferable that the auxiliary light source is capable of adjusting the amount of light arbitrarily, and most preferably, the amount of light can be adjusted without changing the color temperature of light and without causing flicker.

Further, a TV camera is provided so that light emitted from the upper side of the plate holder, that is, the upper side of the microplate is incident. This TV camera is for taking in the optical state of each well and outputting an image signal to an image processing device. As long as the TV camera is as far as possible from the microplate so that the optical states of all the wells can be captured at one time, the light may be incident on the TV camera after being bent through a mirror or the like. By providing the mirror, the TV camera can be provided at any position, and the device can be downsized.

This TV camera is preferably a solid-state camera from the viewpoint of accuracy and stability of operation. The lens of the TV camera is a lens with less geometric distortion, and a filter can be attached. It is preferable to use a color having a complementary color relationship with the color of the particles, because the agglutination image can be efficiently distinguished from other colors.

An image processing device is connected to the TV camera. This image processing device is a device for processing an image signal input from a TV camera to determine an aggregation state. The image processing apparatus includes a video input unit that receives the output from the TV camera, a digital image memory unit that records the data of each pixel, a video output unit that sends the contents to a computer as necessary, and this output. It is composed of a microcomputer for calculation processing, and further has a monitor TV for video input and a printer for recording calculation results.

A light quantity adjusting plate may be provided between the main light source and the auxiliary light source and the plate holder. The light amount adjusting plate is for adjusting the amount of light emitted from the light source to the microplate and for irradiating the entire microplate with a uniform amount of light, and is formed of a translucent body such as a milky plate. As described above, the light quantity adjusting plate for the auxiliary light source can achieve the same effect by forming the plate mounting base with a semitransparent body.

The TV camera is preferably configured so that only the light emitted from the microplate is captured. That is, a hood is provided in the passage of light emitted from the microplate,
The entire device may be provided inside the box, which makes it possible to obtain more accurate and better contrast without introducing unnecessary light into the TV camera for measurement.

It is sufficient that the image of the well is radially divided into a plurality of divisions by the image processing device as long as it is at least four divisions or more, and the division number can be increased as long as the differential value described below can be appropriately determined. The differential value specific to each pixel determined for each pixel is the value obtained by subtracting the brightness of the pixel for which the differential value is determined from the brightness of the pixel outside the pixel for which the differential value is determined for each pixel outside. It is the total value. The number and arrangement of the pixels located outside which are the targets of the brightness calculation for determining the differential value are appropriately changed depending on the number of divisions and the positions of the divided portions.

The differential value of each pixel is sequentially compared in one direction as long as all of the pixels to be compared are arranged in the same direction. It may be.

[Work]

In the particle aggregation determination device of the present invention, the main light source irradiates each well of the microplate with light of uniform intensity, and the irradiated light is incident on the diluting liquid in the well. The incident light maintains the same range of the particle concentration region where the turbidity can be determined even when the diluting liquid has white turbidity as in the case where the cloudiness does not occur.

On the other hand, the auxiliary light source irradiates the lower surface of the microplate with light of uniform intensity, and the light from below makes the inside of the well bright, so that it is possible to make the inside of the well brighter with a smaller amount of light than when illuminating from above. Further, when the light is irradiated from above, the flat upper surface of the microplate, which is close to the light source, is extremely bright compared to the inside of the well, but the light irradiated from below makes the inside of the well brighter than the upper surface of the microplate. . Therefore, the light from the lower side of the auxiliary light source has a higher contrast than the light from the upper side.

Then, the light emitted from each well is incident on the TV camera, and the optical state of all the wells is captured by the TV camera. The captured optical state is sent as an image signal to the image processing apparatus, and this image processing apparatus differentiates the brightness of each pixel to find the edge of the agglomerated portion, counts the pixels between each edge, and determines the reference value. The particle agglomeration is determined in comparison with.

〔Example〕

An embodiment of the particle aggregation determination device according to the present invention will be described with reference to FIGS. 1, 2, and 3.

FIG. 1 is a diagram showing an outline of a particle agglutination determining apparatus, FIG. 2 is an enlarged cross-sectional view of a plate placing table portion of the above, and FIG. 3 is a plan view of the plate placing table of the same.

In FIG. 1, reference numeral 1 is an image capturing device, and reference numeral 2 is an image processing device.

The image capturing device 1 is formed in a light-tight manner by a sealed box body 3, and a movable plate stand 5 is provided on a non-reflective plate 4 made of a black plate inside thereof. The plate holder 5 is made of transparent plastic, and on its upper surface, as shown in FIG. 2, substantially conical well recesses 6 ... 6 are formed at the same intervals as the wells 8 of the microplate 7. . The well recesses 6 ... 6 are provided in the microplate 7
This is for allowing the well 8 to slide down into the well recess 6 and to be in a predetermined position when the is mounted.

Further, on the outside of the well recessed portion 6, positioning holes 9 ...
The light on the lower side is transmitted to the upper side of the plate table 5.

A rack (not shown) is formed at one end of the plate stand 5 so that the rack moves forward and backward by engaging with a pinion (not shown) provided on the non-reflection plate 4.

A main light source 10 is provided above the plate stand 5, and an auxiliary light source 11 smaller than the main light source 10 is provided below the plate stand 5. A light amount adjusting plate 12 made of a milky white plate is provided between the main light source 10 and the plate stand 5, and the light amount adjusting plate 12 and the non-reflecting plate 4 are connected to reach the floor of the box body 3.
Is provided.

A portion of the light quantity adjusting plate 12 corresponding to the micro plate 7 is provided with a hood 14 having a substantially rectangular tube shape whose inner surface is painted black. The hood 14 is bent in a substantially "U" shape, and the TV camera 15 is connected to the end of the hood 14. Mirrors 16 and 16 are provided at the bent portions of the hood 14 so that light from the microplate 7 and the positioning hole 9 can enter the TV camera 15. Further, the hood 14 is provided with an optical filter 17. The TV camera 15 is a solid-state TV camera equipped with a 24 mm camera lens.

The image processing device 2 includes a video input unit 18 for receiving the output of the TV camera 15, a digital image memory unit 19 for recording the data of each pixel, and a video output unit 20 for sending the contents to a computer as necessary. A microcomputer 21 for calculating and processing this output, a monitor TV 22, and a printer 23 are further provided.

Next, a method for automatically determining agglomeration by using the above-described particle agglomeration determination device will be described.

(1) A microplate for determining agglutination is placed on the microplate table.

(2) Select the TV camera filter and adjust the squeeze to capture the image.

(3) The captured image is subjected to image processing as it is, or it is once recorded on a floppy disk and loaded into the computer 21 as necessary to perform image processing and calculation processing.

Next, a method of image processing will be described.

1. Determine the center position of each well.

(1) Measure the coordinates of the positioning hole.

Now 9 ₁ (x ₁ , y ₁ ), 9 ₂ (x ₂ , y ₂ ), 9 ₃ (x ₃ , y ₂ ), 9 ₄ (x ₄ ,
When the position of the positioning holes in the coordinate of y ₄₎ is measured, the linear equation _{Y 1 = a 1 x + b} 1 is obtained connecting the positioning holes 9 _1, 9 ₂ from the positioning holes 9 _1, 9 ₂ coordinates. Similarly, the positioning holes 9 ₃ , 9 ₄ , the positioning holes 9 ₁ , 9 ₃ and the positioning holes 9 ₂ , 9 ₄
Two linear equations Y ₂ , Y ₃ and Y ₄ are obtained, and a total of four linear equations are obtained.

(2) Next, based on the obtained linear equation and the position of the positioning hole 9 on the straight line, for example, the positions of the positioning holes 9 ₁ and 9 ₂ on the straight line of Y ₁ = a ₁ x + b ₁ , a predetermined ratio Then, the straight line is divided into 11 pieces at equal intervals to obtain 12 division points k ₁ ... k ₁₂ .
Similarly, dividing points h ₁ ... h ₁₂ are obtained on a straight line of Y ₂ = a ₂ x + b ₂ and these dividing points are connected from above to obtain 12 straight lines y ₁ , y ₂ ... y.
_Ask for _twelve .

(3) Further, in the remaining linear equations Y ₃ and Y ₄ , similarly, dividing points m ₁ ... M ₈ and n ₁ ... N ₈ are obtained, and eight straight lines x ₁ ... X ₈ connecting these are obtained.

(4) (2) obtained 12 linear y ₁ ... y ₁₂ and obtains the eight intersections of the straight line x ₁ ... x ₈ obtained in (3), to the intersection with the center of the well.

2. Next, as shown in FIG. 3, the center of the well is radially divided into eight fan-shaped divided portions S ₁ , S ₂ ... S _8, and each of these divided portions S ₁ ... S ₈ is divided. Determine the derivative value of the pixel. This differential value is a value obtained by summing the differences from the brightness of three pixels adjacent to the outside, and in the divided portion S ₁ , the left side, the oblique upper left side, and the oblique lower left side of the pixel whose differential value is to be determined. The pixel adjacent to is the target for determining the differential value. Also,
In the divided portion S ₂ , pixels adjacent to the upper side, the left side, and the upper left side of the pixel for which the differential value is to be determined are targeted, and in the divided portion S ₃ , the upper side of the pixel for which the differential value is to be determined, the diagonal Pixels adjacent to the upper left side and the diagonally upper right side are targeted.

Now, the differential value of each pixel of the divided portion S ₂ will be calculated based on the partially enlarged view of the divided portion S ₂ displaying the luminance of each pixel in FIG. Note that reference numerals A to F and numbers I to VI in FIG. 4 indicate the rows and columns of the pixels in FIG. 4 for convenience, and the numbers attached to the upper left of the pixels indicate the luminance. . For example, if the differential value of pixel D-III is to be obtained, pixel D
-III has a brightness of "32" and has pixels D-II, C-II and C-III.
Since the brightness of each pixel is "44", "47", and "42", the difference between the pixel and the pixel D-II is 44-32.
= 12, the difference from the pixel C-II is 47-32 = 15, and the difference from the pixel C-III is 42-32 = 10. Therefore, the sum of these differences becomes the differential value of the pixel D-III, and the value is 12 + 15 + 10.
= 37. In this way, the differential value of each pixel is determined and given to each pixel.

3. When the differential value of each pixel is determined, the differential values of the pixels arranged in a line horizontally are compared to detect the top two and the corresponding pixel is selected as the edge of the particle agglomeration portion.

If the differential values shown in FIG. 6 are given, for example, f
In the row, the differential values “34” and “33” are the top two, so
Pixels corresponding to this correspond to edges. Similarly, when the edges are selected for all rows, the edges shown by the thick line in the figure are obtained. The chain line in the figure shows the actual edge of the particle aggregation portion. Then, the number of pixels between each edge is counted, and the area of the agglutination portion obtained by this is compared with a reference value to determine whether the agglutination reaction is positive or negative.

7 and 8 are schematic views showing another example in which the configuration of the hood is changed.

FIG. 7 shows an example in which the hood 14 is formed in a linear shape so that the light emitted from the microplate 7 is directly incident on the TV camera 15. FIG. 8 shows a hood 14 which is formed by bending the hood 14 once in a substantially “L” shape so that the light emitted from the microplate 7 is reflected by the mirror 16 once and then enters the TV camera 15. It is an example.

Next, the measurement results comparing the effects of white turbidity when measuring transmitted light and when measuring reflected light will be described.

FIG. 9 shows the measurement result of the relationship between the particle concentration and the brightness when the transmitted light was measured. In this figure, the relationship between the well brightness and the particle concentration is represented by ABC in the sample that does not cause white turbidity. OB is a region where particle concentration and brightness correspond,
BC is a portion where the particle concentration is high and light does not pass, and where the particle concentration and brightness do not correspond. K represents the amount of transmission reduced by diffuse reflection due to turbidity generated in the well. Therefore, the relationship between the brightness and particle concentration in a well with turbidity is represented by DEC. As a result, the change in particle concentration changes the brightness in the well only in the OE region, and the brightness does not change between EB even if the particle concentration changes. When the transmitted light is used as described above, the region in which the change in the particle concentration cannot be measured spreads in the sample accompanied by the precipitate, which leads to an error in the judgment of aggregation.

On the other hand, FIG. 10 shows the measurement results of the relationship between the particle concentration and the brightness when the reflected light was measured. In this measurement of reflected light, the same as in Fig. 9 for the sample without white turbidity.
It is represented by ABC, but this is DEC 'for samples with cloudiness. As can be seen from this result, in the measurement of the reflected light, the area in which the brightness does not change does not increase with respect to the particle concentration change due to white turbidity.

Since the measurement by the apparatus of FIG. 1 consists of the measurement by the reflected light of the main light source 10 and the measurement by the transmitted light of the auxiliary light source 11, the addition of FIGS. 9 and 10 is the apparatus of FIG. Will be measured. Therefore, the effect of white turbidity is largely offset and virtually eliminated. For this reason, it is a factor that enables more stable determination of turbid samples than when only transmitted light is used.

Next, the measurement results comparing the contrasts in the case of only the reflected light from the main light source 10 and the cases of both the reflected light and the transmitted light by the main light source 10 and the auxiliary light source 11 will be described. FIG. 11 is a diagram showing the measurement results. In the figure, a represents the intensity of the observed light in the case of reflected light. In this case, for example, the contrast at the point where the transmittance is 0.6 is the magnitude shown by A in the figure. B in the figure represents the intensity of the light observed when both the reflected light and the transmitted light are used, and the contrast at the point where the transmittance is 0.6 is the magnitude shown in B in the figure. Therefore, it is understood that the contrast B in the case of both the reflected light and the transmitted light is larger than the contrast A in the case of only the reflected light, and the contrast is stronger. That is, this increase in contrast is due to the increase in the amount of light due to the transmitted light, and the increase in contrast according to the increase amount.

〔The invention's effect〕

Since the present invention is configured as described above and the TV camera captures the optical state of the wells, data of all the wells of one microplate can be captured in an extremely short time, for example, 1/60 second. Therefore, a large amount of samples can be processed quickly. Further, since the optical states of all the wells are taken in at one time, the plate moving device is not required and the operation can be simplified. Furthermore, not only the information about the particle concentration in the well as a photometer but also the information about the two-dimensional spread of particle aggregation can be obtained at the same time. Compared with the case of using a conventional photometer, various kinds of data can be obtained. Obtained at high speed.

Further, in the present invention, light sources are provided above and below the plate stand, and both the reflected light from the upper light source and the transmitted light from the lower light source are captured in the TV camera to determine aggregation, so that the conventional lower The effect of white turbidity generated when using the transmitted light from the light source can be eliminated, and the brightness in the well, which was limited when using the reflected light from the upper light source, is sufficient with the lower light source. Brightness can be increased and contrast can be increased.

Further, since the brightness of the pixel is differentiated and the edge of the agglomerated portion is determined based on the magnitude of the value, the edge can be accurately determined.
Therefore, the aggregation of particles can be determined extremely accurately.

[Brief description of drawings]

FIG. 1 is a schematic view of an embodiment of the particle aggregation determination device of the present invention, FIG. 2 is a partial cross-sectional view of the plate holder of the above, and FIG. 3 is a plan view of the plate holder of the same. FIG. 4 is a schematic diagram showing a state in which the well is divided, FIG. 5 is a schematic diagram of a part of the well in which the brightness and the differential value are displayed in each pixel, and FIG. 6 is an outline of the well in which the differential value is displayed in each pixel. It is a schematic diagram of half. 7 and 8 are partial schematic diagrams showing another example of the image capturing device, FIG. 9 is a diagram showing the relationship between particle concentration and brightness when the transmitted light is measured, and FIG. 10 is the reflected light. FIG. 11 is a diagram showing the relationship between particle concentration and brightness when measured, and FIG. 11 is a diagram showing the results of contrast measurement. 1 ... Image capturing device, 2 ... Image processing device, 5 ... Plate stand, 7 ... Microplate, 8 ... Well,
9 ... Positioning hole, 10 ... Main light source, 11 ... Auxiliary light source, 12
...... Light intensity control plate, 14 hood, 15 ...... TV camera, 18 ...
… Video input, 21… Microcomputer.

 ─────────────────────────────────────────────────── --Continued from the front page (56) Reference JP 62-105031 (JP, A) JP 60-168339 (JP, A) JP 58-32142 (JP, A) JP 58- 19540 (JP, A) Actual development Sho 59-82841 (JP, U)

Claims (2)

[Claims] 1. A plate mounting base on which a microplate is mounted and which is light-transmissive and has a positioning hole for determining the center position of each well of the microplate, and arranged above and below the plate mounting base. Image capturing device having a light source that is provided and a TV camera provided so that light emitted from the upper side of the microplate mounted on the plate mounting table is incident, and an image signal input from the TV camera is processed. And an image processing device for determining a state of aggregation. 2. The image processing device determines the center position of each well by the light transmitted through the positioning hole, and then divides the image of each well radially from this center position into a plurality of parts, and at each divided part The value obtained by summing the difference in brightness from the pixels in the direction is set as the differential value of each pixel, and the differential value of each pixel is sequentially compared in one direction to count the number of pixels between the top two differential values to determine the aggregation state. The particle aggregation determination device according to claim 1, wherein the determination is made.