JPS5998708A - Method for judging particle agglomeration pattern - Google Patents

Abstract

PURPOSE:To always accurately judge a particle agglomeration pattern, by judging the pattern on the basis of the thickness ratio of particle layers of the patterns in a part including the lowermost point of the bottom surface of a reaction vessel and the circumferential part separated therefrom. CONSTITUTION:The image formed to the central part including the lowermost point of the bottom surface 1b of a reaction vessel 1a is received in a first light receiving region 6a by a light receiving element 6 and the image of the circumferential part separated from the central part of the bottom surface 1b is received in a second light receiving regions 6b. The outputs of these regions 6a, 6b are supplied to an operation apparatus 9 through amplifiers 7a, 7b and an A-D converter 8 to judge the particle agglomeration pattern formed to the bottom surface 1b and the result thereof is displayed by a display apparatus 10. In the next step, the thickness ratio of particle layers of the patterns in the aforementioned two parts is calculated and this ratio is compared to a reference value to judge the pattern. Therefore, the particle agglomeration pattern can be always accurately judged.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for determining a particle aggregation pattern using an immunological agglutination reaction.

As a method for determining such a particle aggregation pattern, for example, a reaction vessel whose bottom surface is formed in the shape of an inverted cone is used, and an image of the center portion of the bottom surface and an image of the peripheral portion are imaged on two separate light receiving elements. There is a method that determines the agglomeration pattern based on the difference between these outputs. In this way, when using a reaction vessel with an inverted conical bottom, when the reaction solution containing the contained particles undergoes an aggregation reaction, the settled particles coagulate and bond uniformly to the inverted conical bottom. 1. A uniform deposition pattern is formed, and if no agglomeration reaction occurs, the particles settle in a discrete manner, and when they reach the bottom, they roll down the slope and collect at the lowest point in the center. is formed, the output of the light-receiving element that receives the image at the center of the bottom surface is E+, and the output of the light-receiving element that receives the image at the periphery is E?
Then, their difference ΔE=lE2-E+l changes depending on the aggregation pattern. In other words, when the agglomeration pattern is a uniform deposition pattern as shown in FIGS. 1A and B and the particle amount distribution is shown in FIG. 1C, ΔE is small and the second Diagram A.

In the case of accumulation patterns such as those shown in B and C, 8E becomes large. Therefore, by comparing the two experimentally determined reference values V+, V2 (Vl > V2) and ΔF, when 8E<V2 (Fig. 1) is [) agglomeration (+) J, ΔE >V+ Togi (Figure 2) is “non-aggregation (
-)", but as shown in Figure 3 A, B, and C, when the degree of particle accumulation is intermediate, that is, when V2≦ΔF<, Vl, it is subject to re-examination.
? ” can be determined.

However, such conventional determination methods have the following drawbacks. In other words, if the amount of particles in the reaction solution dispensed into the reaction container is small and an accumulation pattern is formed as shown in FIG. 4, B and C, 1. ΔF is small due to the small amount of particles gathered in the center, and Δ
If F≦V1, it may be determined as "?" as in FIG. 3, or in an extreme case, ΔE<V2, and as in the case of FIG. 1, it may be determined as "+". In addition, if the amount of particles in the dispensed reaction solution is large and aggregation reaction is occurring, even if the agglutination reaction is equal to or greater than that shown in Figure 1, the reaction partner Particles with a higher power than the components do not combine, but roll down the inclined bottom surface and gather in the center, forming an agglomeration pattern as shown in FIG. 5A, B, and C.

In this case, since a normal agglutination reaction is occurring, it should be determined that r0J, but ΔF is not necessarily 8E<V2, and ΔE>V2 or ΔE>V
The result may be erroneously determined as "?" or "-".

SUMMARY OF THE INVENTION An object of the present invention is to solve the various problems mentioned above and to provide a method for determining a particle aggregation pattern that can always accurately determine an aggregation pattern.

The present invention provides a method for photoelectrically detecting and determining a particle aggregation pattern formed on the bottom surface by precipitation of particles in a reaction solution contained in a reaction container with a small bottom surface and a common part having an inclined surface. Based on measurements corresponding to a first portion including the lowest point of the bottom of the container and a peripheral second portion remote from this first portion, the thickness of the particle layer in these first and second portions; The method is characterized in that the aggregation pattern is determined based on this ratio.

The present invention will be described in detail below with reference to the drawings.

FIG. 6 is a diagram showing the configuration of an example of a particle aggregation pattern determination device implementing the present invention. In this example, the base is an inverted circle 1
A microplate 1 is used in which a plurality of reaction vessels having a shape of 1 [1] are formed in a matrix on a substrate made of transparent plastic or the like. The bottom surface of the microplate 7 is uniformly illuminated by light emitted from the light source 2 via the light beam 3 and the diffusion plate 4, and an image of the inverted conical bottom surface 1b of each reaction vessel 1a is formed. An image is formed on the light-receiving element 6 by the one-zoom lens 5. As shown in the plan view, the light-receiving element 6 has concentric first and second light-receiving areas 6a and 6b, and the first light-receiving area 6a covers the middle heating area including the lowest point of the bottom surface 1h. The second light-receiving area M61) receives the image at the periphery of the bottom surface 1h away from the center. These first and second
The outputs of the light-receiving regions 6a and 6b are respectively transmitted to an amplifier 7.
a and 7b, are supplied to the arithmetic unit 9 via the A-D converter 8, where the aggregation pattern formed on the bottom surface 1b is determined by performing the arithmetic operation described later, and the result is displayed on the display device 10.
and display it.

Here, the first. The intensity of the incident light to the central and peripheral parts of the inverted conical bottom surface 1b of the reaction vessel 1a corresponding to the second light-receiving areas 6a and 5b is J++, II2, and the average of the particle layer of the agglomeration pattern in the central and peripheral parts. The overall transmittance of the optical path substances other than the particle layer (sample liquid, reaction vessel material, etc.) is -I, -2, 1st. The average intensity of light incident on the second light receiving areas 6a and 6b is It, 1
2゜ Its light receiving area is S + , S 2 and its conversion efficiency is 1. 2. AI the amplification degree of amplifiers 7a and 7b.

If A2 and its output are El and E2, then Ei = K
i −At −3i ・Ii (i=+, z
) (i = ki−k −i ・f+ i, E! = Ki −Ai = 3i −kii(−
i −II i. Now, if the amplification degree Ai is adjusted so that the output Ei is 100 mV when the reaction solution containing no particles is stored in the reaction container 1a, that is, when ki = 1, the output when the reaction solution containing particles is stored is E
1 becomes F 1 = 100 · k i . Here, if we newly define ρ1=i−ki, we get
This expresses the degree to which light is scattered and absorbed by the particle layer in the center and periphery and does not pass through. and ρ-1=1(')0ρi=10
0-100k 1-100-Ei. In other words, ρ-1 is the value obtained by subtracting the output Ei when a reaction solution containing particles is contained from the output 100 mV when a reaction solution containing no particles is contained, and the decrease in output value due to particles is expressed as a percentage. It is determined by the thickness of the particle layer.

In this example, the arithmetic unit 9 calculates the above ρ-1 and r
−ρ−1/ρ′2 is calculated, and this ratio r and a predetermined reference value R
Compare blindness and R2 (R+ > R2). That is, as shown in FIGS. 1 and 5, when there is a particle bonding reaction, the ratio r of the thickness of the particle layer in the central part to that in the peripheral part is small, and as shown in FIGS. The sea urchin shown
When there is almost no particle binding reaction, the ratio r is large.

Further, as shown in FIG. 3, if there is a slight particle bonding reaction, the ratio will be an intermediate value. Therefore, by comparing this ratio r with two experimentally determined reference values R1°R2, we find that r>R1;"-" R1≧r≧R2:r? It is determined that J r <R2; r+J.

In this way, the various agglomeration patterns shown in FIGS. 1 to 5 can be accurately determined.

Note that the present invention is not limited to the above-mentioned example, and can be modified or modified in many ways.

For example, the reaction vessels are not formed into microplates, but can be individually separated, and their bottoms can have various shapes, such as a single-sided roof, a gable roof, and a pyramid. can. Also,
The bottom surface of the reaction vessel may be scanned to determine the thickness of a unique point or the average particle layer at a portion including the lowest point and at a peripheral portion away from this portion.

As mentioned above, in the present invention, the ratio of the thickness of the particle layer of the agglomerated pattern in the bottom of the reaction vessel is calculated based on the photoelectric output of the part including the lowest point and the peripheral part far away from the bottom of the reaction vessel. This ratio is compared with a reference value to determine the agglomeration pattern, so accurate determination can always be made.

[Brief explanation of the drawing]

Figures 1, B, C to Figure 5 A, B, and C are diagrams showing five aspects of agglomeration patterns, and Figure 6 is a pictorial diagram showing the configuration of an example of a particle aggregation pattern determination device implementing the present invention. It is. 1... Microplate 1a... Reaction container 1h.
... Bottom surface 2 ... Light source 3 ... Lens for illumination 4 ... Diffusion plate 5 ... Lens for imaging
6... Light receiving element 6a, 6h... Light receiving area
7a, 7b...Amplifier 8...A-D converter
9... Arithmetic device 10... Display device,

Claims (1)

[Claims] 1. In photoelectrically detecting and determining a particle aggregation pattern formed on the bottom surface by precipitation of particles in a reaction solution contained in a reaction container with a small bottom surface and a common part having an inclined surface, the bottom surface of the reaction container Based on the measurements corresponding to a first part including the lowest point of and a peripheral second part remote from this first part, the ratio of the thickness of the particle layer in these first and second parts A method for determining a particle aggregation pattern, characterized in that the aggregation pattern is determined based on this ratio.