JPH04120442A - Coagulation image discerning method - Google Patents

Abstract

PURPOSE:To automatically discern the coagulation from the non-coagulation of particles by detecting the border between the central part and peripheral part of a reaction pattern and obtaining the area of the border. CONSTITUTION:Many wells 11a having a conical concaved bottom surface are formed in matrix on a microplate 11. A solution which includes particles to be inspected is accommodated in each well 11a. A reaction pattern is formed by leaving the solution at rest. White the microplate 11 is illuminated by a fluorescent lamp 13, an image of the bottom surface of each well 11a is sequentially taken by a video camera 15. The image data is sent to an image processor 16. The processor 16 detects the boundary between the central part and peripheral part of the reaction pattern, and obtains the area X of the boundary. A data processor 19 compares the area X with a reference value thereby to discern the coagulation and non-coagulation of the particles. The result is displayed at 21. Accordingly, even the coagulation pattern with a small force of coagulation can be automatically discerned correctly without being erroneously judged as a non-coagulation pattern.

Description

Detailed Description of the Invention [Industrial Application Field] The present invention is used in clinical tests, etc. to optically measure the reaction pattern of test particles formed on the bottom of a reaction container to determine if they are agglomerated, non-agglomerated, or The present invention relates to an agglomerated image determination method for automatically determining other attributes.

[Conventional technology]

As a conventional agglomerated image determination method, for example, Japanese Patent Application Laid-open No. 58-10
As disclosed in Publication No. 5065, the ratio of brightness between the center and the periphery is determined from the measurement data of the reaction pattern of the particles to be tested, and aggregation or non-aggregation is determined based on that ratio. Also, as disclosed in Japanese Patent Application Laid-open Nos. 61-215948, 62-105031, 63-58237, and 63-256839, the optical state of the microplate is captured with a TV camera. The image data is processed to determine the area of particles that are agglomerated or that have fallen to low areas such as the center of the reaction vessel using the static method, and based on that area, it is determined whether the particles are agglomerated or not. There is something like this.

[Problem to be solved by the invention]

However, in the conventional aggregation image determination method described above, the agglomeration force is very weak, and when a reaction pattern that is almost the same as a non-aggregation pattern is formed, it is very difficult to automatically and accurately determine this. becomes difficult. As described above, in the conventional agglutinated image determination method described above, the reliability of the determination result is low, and therefore, in the automatic reaction pattern determination device that implements such agglutinated image determination method, the determination result is It is necessary for the operator or the like to check and correct each correction visually or by other means, which increases the burden on the operator and the like.

This invention was made by focusing on these conventional problems, and it provides an agglutination image that can automatically and accurately judge even agglutination patterns with very weak cohesive force, and therefore provides highly reliable judgment results. The purpose is to provide a determination method.

[Means for Solving the Problems] In order to achieve the above object, the present invention optically measures a reaction pattern of test particles formed on the bottom surface of a reaction vessel, and measures particles that pass approximately through the center of the reaction pattern. Obtain measurement data on a straight line or curve, detect the boundary between the center and the periphery of the reaction pattern based on the measurement data, calculate the number of data at the boundary, and calculate the number of data based on the number of data. Automatically determine agglutination, non-aggregation, or other attributes of sample particles.

Further, in the present invention, image data of the reaction pattern of the test particles formed on the bottom surface of the reaction container is obtained, and based on the image data, the area of the boundary between the center part and the peripheral part of the reaction pattern is determined, Aggregation, non-aggregation, or other attributes of the test particles are automatically determined based on the area.

[For production]

As shown in FIGS. 1A and B, a test solution containing test particles is placed in a reaction container 1 having a conically concave bottom such as a microplate, and a reaction pattern is formed by, for example, a standing method. Non-aggregation pattern shown in Figure 1A and Figure 1B
The agglomeration pattern shown in Fig. 1 may be almost the same as the aggregation pattern with very weak cohesive force. However, in the agglomeration pattern with a weak cohesive force shown in Fig. 1, the boundaries of the test particles settled in the center of the reaction vessel 1 are blurred and not clear compared to the non-aggregation pattern shown in Fig. 1A. There is a characteristic that it is not.

In this invention, such characteristics are utilized to distinguish between non-agglomerated patterns and agglomerated patterns with weak cohesive force.

That is, in FIGS. 1A and 1B, for example, if the amount of transmitted light on a straight line passing through the center of the reaction pattern is measured, the results will be as shown in FIGS. 2A and B, respectively. Here, the second
Focusing on the boundary between the center and the periphery of the reaction pattern indicated by the arrows in Figures A and B, the length of the straight line part in Figure 2B is longer than that in the non-aggregation pattern in Figure 2A. It can be seen that the aggregation pattern with weaker cohesive force is longer. Therefore, by extracting the measurement data of the boundary between the center and the periphery of the reaction pattern from the measurement data of the amount of transmitted light, and checking the number of measurement data at the boundary, that is, the length of the boundary, the cohesive force can be determined. It becomes possible to accurately distinguish between weakly agglomerated patterns and non-agglomerated patterns.

Similarly, if you capture image data of a reaction pattern with a video camera, etc., and check the area of the boundary between the center and periphery of the reaction pattern based on that image data, you can distinguish between agglomerated patterns with weak cohesive force and non-agglomerated patterns. It becomes possible to accurately distinguish between agglomeration patterns and agglomeration patterns.

[Embodiment] FIG. 3 is a block diagram showing the configuration of an example of an automatic aggregation image determination apparatus that implements the present invention. In this embodiment, a microplate 11 is used as a reaction vessel, and this microplate 11 is illuminated from the bottom side by a fluorescent lamp 13 connected to a fluorescent lamp power source 12.

As shown in FIG. 4, the microplate 11 is constructed by forming a large number of wells Ila each having a conically depressed bottom surface in a matrix, and each of the wells Ila contains a test solution containing test particles. A reaction pattern is formed on the bottom surface by a standing method.

An image of the bottom surface of each well lla of the microplate 11 illuminated by the fluorescent lamp 13 is sequentially captured by a video camera 15 and the image data is supplied to the image processing circuit 16.
Here, the boundary between the center and the periphery of the reaction pattern formed on the bottom of the well lla is detected based on the input image data, and its area is determined.

The image data of the reaction pattern of each well lla is
The microplate 11 and the video camera 15 are relatively moved in a two-dimensional direction within a horizontal plane to sequentially capture images.

Data processing in this image processing circuit 16 will be explained below.

The image processing circuit 16 first converts the image data of the reaction pattern of the well 11a from the video camera 15 into digital data. Note that the conversion of this image data into digital data is performed such that the value of bright data is large and the value of dark data is small. Next, from the image data converted into digital data, a certain amount of dark data is extracted from the preset well center area 17 shown in FIG. 5, and the average value C thereof is determined. 18 data are taken out and the average value P is determined. Thereafter, C is obtained by adding a predetermined positive value to the average value C, and p is obtained by subtracting a predetermined positive value from the average value P. Determine the range of data at the boundary between the center and the periphery of the reaction pattern.

Next, from the data of the well center area 17 in FIG.
Boundary data that satisfies the relationship p>x>c is extracted and the number of data, that is, the area X is determined.

The area X of the boundary obtained by the image processing circuit 16 as described above is supplied to the data processing circuit 19, where it is compared with a predetermined reference value to determine whether it is agglomerated or not. It is displayed on the display section 21 in response to instructions from the input section 20 such as a keyboard.

In this way, based on the image data of the reaction pattern formed in well lla, the area of the boundary between the center and the periphery of the reaction pattern is determined, and based on that area, the aggregation or non-aggregation of the reaction pattern is determined. By doing so, even an agglomerated pattern with a weak cohesive force can be automatically and accurately determined without being confused with a non-agglomerated pattern, and a highly reliable determination result can be obtained. Therefore, there is no need for the operator or the like to check and correct the determination result visually or by other means, unlike in the past, and the burden on the operator or the like can be significantly reduced.

In this embodiment, the microplate 11 and the video camera 15 are moved relatively in a two-dimensional direction within a horizontal plane to sequentially capture image data of the reaction pattern formed in each well 11a of the microplate H1. However, the image data of the entire microplate 11 may be taken in, and the image data of the reaction pattern of each well lla may be extracted from the image data and processed in the same manner.

FIG. 6 is a block diagram showing the configuration of another example of an automatic agglomerated image determination apparatus embodying the present invention. In this embodiment, the microplate 11 is spot-illuminated from the bottom side through a lens group 33 by a light source 32 connected to a power source 31.
The transmitted light is received by a light receiver 34. The output of the light receiver 34 is converted into a digital signal by a light reception data processing section 35 and supplied to a data processing section 36 . Further, the microplate 11 is moved linearly in a horizontal plane via the microplate transfer mechanism 37 under the control of the data processing section 36, thereby moving the well 11a in the diametrical direction as shown in FIG. 7A. 7B to obtain measurement data of the amount of transmitted light as shown in FIG. 7B. Note that the analog-to-digital conversion in the received light data processing section 35 is performed such that the value of bright data is large and the value of dark data is small.

In this example, the measurement data of the amount of transmitted light as shown in FIG. 7B is obtained for each well Ila as described above,
Based on the data, the data processing unit 36 detects the boundary between the center and the periphery of the reaction pattern formed on the bottom surface of the well lla, and calculates the number of data, that is, the length at the boundary.

Data processing in the data processing section 36 will be explained below.

The data processing unit 36 first calculates the average value C of the data at the center of the well set in advance for the measurement data of each well lla, and also calculates the average value P of the data at the periphery of the well. Thereafter, the average value C is multiplied by a predetermined positive value larger than l to obtain C, and the average value P is multiplied by a predetermined positive value smaller than 1 to obtain P.
The range of data at the boundary between the center and the periphery of the reaction pattern indicated by arrows in Figures A and B is determined.

However, the values by which the average values C and P are multiplied are p
> A value that satisfies the condition c. Next, the well l
From the measurement data of la, data satisfying p>x>c is extracted and the number of data, that is, the length X of the boundary part is determined. Thereafter, the length X is compared with a predetermined reference value to determine aggregation/non-aggregation.

After determining the reaction pattern in the data processing section 36 as described above, the determination result is transferred to the input section 3 such as a keyboard.
8 is displayed on the display section 39 in response to an instruction from 8.

In this way, the length of the boundary between the center and the periphery of the reaction pattern is determined from the measurement data on a straight line passing approximately through the center of the reaction pattern formed in well lla, and the reaction pattern is determined based on that length. If the agglomeration/non-aggregation is determined, as in the above-mentioned embodiment, even an aggregation pattern with a weak cohesive force can be automatically and accurately determined without being mistaken as a non-aggregation pattern. It is possible to obtain highly accurate judgment results. Therefore, there is no need for the operator or the like to check and correct the determination result visually or by other means, unlike in the past, and the burden on the operator or the like can be significantly reduced.

〔Effect of the invention〕

As described above, according to the present invention, the boundary between the center and the periphery of the reaction pattern is blurred and unclear in the agglomerated pattern, which has a weaker cohesive force than the non-agglomerated pattern. Based on the measurement data of the reaction pattern, the boundary part is detected and its area or length determined, and based on the area or length, it is automatically determined whether the target particle is agglomerated, non-agglomerated, or other attributes. Since the determination is made based on the following, it is possible to obtain highly reliable and accurate determination results. Therefore, according to the device implementing the present invention, there is no need for the operator, etc. to check and correct the determination result visually or by other means, as in the past, and the burden on the operator, etc. can be significantly reduced. can.

[Brief explanation of drawings]

1A and 2B and 2A and 2B are diagrams for explaining the present invention in detail; FIG. 3 is a block diagram showing the configuration of an example of an automatic agglutination image determination apparatus that implements the present invention; This figure shows the structure of the microplate shown in FIG. 3, FIG. 5 is a diagram for explaining the operation of FIG. 3, and FIG. A block diagram showing the configuration and FIGS. 7A and 7B are diagrams for explaining its operation. 1... Reaction container 11... Microplate 11a... Well 12... Fluorescent light power source 13... Fluorescent light 15... Video camera 16... Image processing circuit 17... Well center 18...Well peripheral area 19...Data processing circuit 20...Input section 21...Display section 31
... Power source 32... Light source 33... Lens group 34... Light receiver 35... Light reception data processing section 36... Data processing section 37... Microplate transfer mechanism 38... Input section 39...Display section Fig. 1A Fig. 2 Fig. 3 Fig. 4 U Fig. 5 1a Fig. 6

Claims (1)

[Claims] 1. Optically measuring a reaction pattern of test particles formed on the bottom surface of a reaction container to obtain measurement data on a straight line or curve passing approximately through the center of the reaction pattern; Based on the data, detect the boundary between the center and the periphery of the reaction pattern, calculate the number of data at the boundary, and determine whether the test particles are agglomerated, non-agglomerated, or other attributes based on the number of data. A method for determining an agglomerated image, characterized in that it automatically determines. 2. Obtaining image data of the reaction pattern of the test particles formed on the bottom surface of the reaction container, and determining the area of the boundary between the center and the periphery of the reaction pattern based on the image data,
A method for determining an agglomerated image, comprising automatically determining whether the test particles are agglomerated, non-agglomerated, or other attributes based on the area thereof.