WO2011010570A1 - Apparatus for measuring amount of aggregates, and method for measuring amount of aggregates - Google Patents

Abstract

Disclosed are an apparatus and a method for calculating the amount of aggregates within a short period and at low cost. In the apparatus for measuring the amount of aggregates, the amount (G) of aggregated blood cells can be calculated based on a velocity (V) of blood that passes through a microchip (2) and a velocity (V0) of blood cells that pass through any one gate (26) in the microchip (2).

Description

Aggregation amount measuring apparatus and aggregation amount measuring method

The present invention relates to an aggregation amount measuring apparatus and an aggregation amount measuring method.

In recent years, with increasing interest in health, blood fluidity has attracted attention as a health barometer. As a method for examining the blood fluidity, a method is known in which blood is passed through a microchip having a plurality of fine flow paths (gates) and the time required for the passage is measured (for example, Patent Document 1). , 2).

This fluidity of blood is related variously to the characteristics of blood cells in the blood. For example, in blood with low fluidity, agglomeration is likely to occur in which blood cells stay and bind in agglomerated form. Since the occurrence of this aggregation greatly affects the blood fluidity, it is desired to quantify the degree of the occurrence of aggregation as the amount of aggregation and to establish a highly accurate measurement method.

Therefore, a method of taking and analyzing a blood flow image of blood from which only white blood cells have been extracted and calculating the amount and area of accumulated white blood cells as an amount of aggregation (see, for example, Patent Document 3), There has been proposed a method (for example, see Patent Document 4) of taking and analyzing a blood flow image and calculating an aggregation rate (aggregation amount) of red blood cells.

Incidentally, blood cell aggregation is a phenomenon that occurs stochastically at a plurality of gates. Therefore, in order to calculate an accurate amount of aggregation by the methods disclosed in Patent Documents 3 and 4, it is necessary to perform photographing and image analysis for all the gates.

Japanese Patent No. 2685544 JP 2005-265634 A JP 2001-264318 A JP 2006-71475 A

However, if photographing and image analysis are generally performed on all thousands of gates, however, the measurement time required to calculate the amount of aggregation is increased, and the analysis cost is also increased.

The present invention has been made in view of the above circumstances, and an object of the present invention is to provide an aggregation amount measuring apparatus and an aggregation amount measuring method capable of calculating the amount of aggregation in a shorter time and at a lower cost than in the past.

In order to solve the above problems, the invention according to claim 1 is an aggregation amount measuring device,
A microchip having a plurality of gates through which a fluid (eg, blood) containing a plurality of particles (eg, blood cells) flows;
Fluid velocity calculating means for calculating the velocity of the fluid passing through the microchip;
Particle velocity calculating means for calculating the velocity of the particles flowing through any one of the plurality of gates;
Agglomeration amount calculating means for calculating the amount of aggregation of the particles based on the velocity of the fluid and the velocity of the particles;
It is characterized by providing.

The invention according to claim 2 is the aggregation amount measuring apparatus according to claim 1,
The aggregation amount calculation means calculates the aggregation amount of the particles based on a ratio or difference between the velocity of the fluid and the velocity of the particles.

The invention according to claim 3 is the aggregation amount measuring device according to claim 2,
The aggregation amount calculation means calculates the aggregation amount G of the particles satisfying the following formula (1) or formula (2) using the fluid velocity V and the particle velocity V _0. .

G = V ₀ / V (1)
G = (V ₀ −V) / V ₀ (2)
The invention according to claim 4 is the aggregation amount measuring apparatus according to any one of claims 1 to 3,
A speed recalculating means for recalculating the speed of the particles;
The speed recalculation means includes
Based on the positional relationship between the gate for which the velocity of the particle has been calculated by the particle velocity calculation means and a gate other than the gate, and the calculated velocity of the particle, the other particles at the other gate As well as guessing the speed,
An average value of the calculated velocity of the particles and the estimated velocity of the other gate particles is calculated as the velocity of the particles,
The aggregation amount calculating means calculates the aggregation amount of the particles based on the velocity of the particles calculated by the velocity recalculation means and the velocity of the fluid.

The invention according to claim 5 is the aggregation amount measuring apparatus according to any one of claims 1 to 4,
The fluid velocity calculation means and the particle velocity calculation means calculate the fluid velocity and the particle velocity at a plurality of time timings,
The aggregation amount calculating means calculates the aggregation amount of the particles at each time timing at which the velocity of the fluid and the velocity of the particles are calculated, and calculates a time integral value of the aggregation amount of the particles as a new aggregation amount of the particles. It is calculated as follows.

The invention according to claim 6 is the agglomeration amount measuring apparatus according to any one of claims 1 to 5,
A photographing means for photographing the flow of the fluid;
The particle velocity calculating unit analyzes the image captured by the imaging unit and calculates the particle velocity.

The invention described in claim 7 is the aggregation amount measuring apparatus according to any one of claims 1 to 6,
The fluid is blood, and the particles are blood cells.

The invention according to claim 8 is an aggregation amount measuring method,
Using a microchip having a plurality of gates through which a fluid containing a plurality of particles flows,
A fluid velocity calculating step for calculating a velocity of the fluid passing through the microchip;
A particle velocity calculating step for calculating a velocity of the particles flowing through any one of the plurality of gates;
An agglomeration amount calculating step for calculating the aggregation amount of the particles based on the velocity of the fluid and the velocity of the particles;
It is characterized by providing.

The invention according to claim 9 is the aggregation amount measuring method according to claim 8,
In the aggregation amount calculating step, the amount of aggregation of the particles is calculated based on a ratio or difference between the velocity of the fluid and the velocity of the particles.

The invention according to claim 10 is the aggregation amount measuring method according to claim 9,
In the aggregation amount calculating step, the particle aggregation amount G satisfying the following equation (1) or equation (2) is calculated using the fluid velocity V and the particle velocity V _0. .

G = V ₀ / V (1)
G = (V ₀ −V) / V ₀ (2)
The invention according to claim 11 is the aggregation amount measuring method according to any one of claims 8 to 10,
A speed recalculation step for recalculating the speed of the particles,
In the speed recalculation step,
Based on the positional relationship between the gate for which the velocity of the particle is calculated in the particle velocity calculation step and a gate other than the gate, and the calculated velocity of the particle, other particles in the other gate As well as guessing the speed,
An average value of the calculated velocity of the particles and the estimated velocity of the other gate particles is calculated as the velocity of the particles,
In the aggregation amount calculation step, the aggregation amount of the particles is calculated based on the velocity of the particles calculated in the velocity recalculation step and the velocity of the fluid.

The invention according to claim 12 is the aggregation amount measuring method according to any one of claims 8 to 11,
In the fluid velocity calculation step and the particle velocity calculation step, the velocity of the fluid and the velocity of the particles at a plurality of time timings are calculated,
In the aggregation amount calculating step, the amount of aggregation of the particles at each time timing at which the velocity of the fluid and the velocity of the particles are calculated is calculated, and a time integral value of the aggregation amount of the particles is calculated as a new aggregation amount of the particles. It is calculated as follows.

The invention described in claim 13 is the aggregation amount measuring method according to any one of claims 8 to 12,
A photographing step of photographing the flow of the fluid;
In the particle velocity calculating step, the velocity of the particles is calculated by analyzing the image captured in the imaging step.

The invention described in claim 14 is the aggregation amount measuring method according to any one of claims 8 to 13,
The fluid is blood, and the particles are blood cells.

According to the present invention, the amount of blood cell aggregation is calculated based on the velocity of fluid (blood) passing through the microchip and the velocity of particles (blood cells) flowing through any one of the plurality of gates. Therefore, it is not necessary to perform photographing and image analysis for all gates, and the degree of occurrence of aggregation can be quantitatively expressed by the speed of blood cells not including the influence of aggregation and the speed of blood including the influence of aggregation. . Therefore, the amount of aggregation can be calculated in a shorter time and at a lower cost than in the past.

In addition, based on the positional relationship between the gate where the blood cell velocity is calculated and another gate other than the gate, and the calculated blood cell velocity, the velocity of other blood cells at the other gate is estimated and calculated. When the average value of the blood cell velocity and the estimated blood cell velocity is calculated as the blood cell velocity, and the amount of blood cell aggregation is calculated using this blood cell velocity, the blood cell velocity depending on the gate position is calculated. It is possible to calculate the amount of aggregation taking into account the influence. Therefore, a more reliable aggregation amount can be calculated.

In addition, when the amount of blood cell aggregation at multiple time timings is calculated and the time integral value of this blood cell aggregation amount is calculated as the new blood cell aggregation amount, the influence on the occurrence of aggregation over time is taken into account. The amount of aggregation can be calculated. Therefore, a more reliable aggregation amount can be calculated.

It is a block diagram which shows the whole structure of the aggregation amount measuring apparatus. (A) It is a top view of a microchip, (b) It is a side view. It is the elements on larger scale of a microchip. (A) (b) It is a figure for demonstrating the gate of a microchip. It is a flowchart of the aggregation amount measuring method in an embodiment. It is the speed ratio table which displayed the relative velocity of the blood cell which flows through a gate. It is a flowchart of the aggregation amount measuring method in the 1st modification of embodiment. It is a flowchart of the aggregation amount measuring method in the 2nd modification of embodiment. It is a graph which shows the time change of the speed of the blood which passes a microchip.

Hereinafter, embodiments of the present invention will be described with reference to the drawings.

FIG. 1 is a block diagram showing the overall configuration of an aggregation amount measuring apparatus 1 according to the present invention.

As shown in this figure, the agglutination amount measuring apparatus 1 guides blood from a supply tank 10 through a microchip 2 to a discharge tank 11, and obtains an aggregation amount of blood cells in the blood from information acquired in the process. It is. In the present embodiment, the amount of aggregation refers to a quantitative value that will be described later indicating the degree of occurrence of aggregation, and aggregation refers to the accumulation of blood cells and binding in agglomerated form.

Specifically, the agglutination amount measuring apparatus 1 includes a microchip 2, a TV camera 3 that captures a blood flow in the microchip 2, a strobe 4 that irradiates light on the imaging range of the TV camera 3, and a microchip. 2, a flow sensor 5 that measures the amount of blood that has passed through 2, a personal computer (PC) 7 that calculates the amount of blood cells in the blood, a display 8 that displays a blood flow image, and the blood flow in the microchip 2. And a differential pressure control unit 9 for controlling the pressure.

The aggregating amount measuring apparatus 1 includes a plurality of solution bottles 13 connected to a blood flow path via a mixer 12 so that a liquid such as physiological saline or a physiologically active substance can be mixed with blood and guided to the microchip 2. Is further provided. The blood mixed with a liquid such as physiological saline or a physiologically active substance (hereinafter simply referred to as “blood”) is adjusted by the differential pressure control unit 9 to adjust the differential pressure before and after the microchip 2. A desired amount flows through the inside. In addition to the differential pressure control unit 9 and the mixer 12, the valve 10 a of the supply tank 10 is integrated and controlled by the sequence control unit 17.

FIG. 2A is a plan view of the microchip 2, and FIG. 2B is a side view.

As shown in this figure, the microchip 2 is formed by overlapping a rectangular glass flat plate 20 and a base plate 21.

The glass flat plate 20 is formed in a flat plate shape and covers the inner side surface of the base plate 21 (the upper surface in FIG. 2B).

The base plate 21 has depressions 210 and 211 at both ends, and a plurality of grooves 212 and so on between the depressions 210 and 211.

Among these, the hollow part 210 has a through-hole 210 a that communicates with the supply tank 10 and forms the blood inlet 27 on the bottom surface, and the upstream storage part 22 that stores blood is disposed between the glass plate 20 and the upstream storage part 22. Is formed.

Similarly, the recess 211 has a through-hole 211 a that communicates with the discharge tank 11 and forms the blood outlet 28 on the bottom surface, and the downstream reservoir 23 that stores blood is disposed between the glass flat plate 20. Is formed.

Further, the plurality of groove portions 212,... Are arranged so as to extend in parallel to the direction (X direction in the drawing) connecting the recess portion 210 and the recess portion 211 and extend in the X direction. Thus, it is partitioned in a direction (Y direction in the figure) perpendicular to the X direction. The plurality of grooves 212,... Alternately communicate with the depression 210 or the depression 211, whereby the upstream blood circuit 24 that allows blood to flow from the upstream reservoir 22 and the downstream reservoir 23. A downstream blood circuit 25 that allows blood to flow into the glass plate 20 is formed.

FIG. 3 is a partially enlarged view of the microchip 2, and FIGS. 4A and 4B are diagrams for explaining a gate 26 to be described later. 4A and 4B, the upper diagram is a plan view of the terrace portion 213, and the lower diagram is a side sectional view thereof. As shown in these drawings, a plurality of hexagonal bank portions 214 are arranged in the X direction at the upper end portion of the terrace portion 213 and are in contact with the glass flat plate 20 at the top surface.

The plurality of bank portions 214,... Form a canyon portion 215 between them. Between the canyon portions 215, a gate 26 is formed between the lower surface of the glass flat plate 20 as a fine flow path for flowing blood in a direction parallel to the Y direction (Z direction in the figure). This gate 26 has a rectangular cross-sectional shape, and its width is slightly narrower than the blood cell diameter of red blood cells (about 8 μm).

Thus, the width of the gate 26 is preferably narrower than the blood cell diameter of the blood cell type to be measured for measuring the amount of aggregation so that one blood cell can pass through while deforming. Thereby, it is possible to observe the deformation state of the blood cells passing through the gate 26. The width of the gate 26 is the same as or larger than the blood cell diameter of the blood cell type to be measured for measuring the amount of aggregation, and two blood cells are arranged in parallel. It can also be a width that cannot pass simultaneously.

In the microchip 2 having the above configuration, the blood introduced from the supply tank 10 is stored in the upstream storage unit 22, passes through the gate 26 and the downstream blood circuit 25 from the upstream blood circuit 24, and then downstream. It is stored in the side storage part 23 and discharged from the discharge tank 11. In this process, blood cells such as red blood cells in the blood flowing through the gate 26 pass through the gate 26 while being deformed.

Further, a pressure sensor E1 and a pressure sensor E2 for measuring blood pressure in the vicinity of the inlet and outlet of the microchip 2 are provided upstream and downstream of the microchip 2 (see FIG. 1). The pressure sensor E1 and the pressure sensor E2 output the measured tip upstream pressure P1 and tip downstream pressure P2 to the differential pressure control unit 9.

As shown in FIG. 1, the TV camera 3 is installed facing the glass flat plate 20 of the microchip 2 and photographs the flow of blood passing through the gate 26 through the glass flat plate 20. The TV camera 3 is a digital CCD camera, for example, and is a high-speed camera for photographing a blood flow or a camera capable of photographing a moving image. A blood flow image photographed by the TV camera 3 is output to the personal computer 7 and displayed on the display 8.

The strobe 4 irradiates light to the photographing range of the TV camera 3. In this embodiment, the strobe 4 irradiates light into the microchip 2 through the polarizing plate 41 through the glass plate 20.

The flow sensor 5 measures the flow rate of blood passing through the microchip 2 and outputs the measured blood flow rate to the personal computer 7.

The personal computer 7 controls the shooting timing of the TV camera 3 and the light emission timing of the strobe 4, and measures the time after blood starts flowing through the microchip 2. The personal computer 7 includes an arithmetic processing unit 70 that performs various calculations. The arithmetic processing unit 70 calculates the amount of blood cell aggregation based on the blood flow image input from the TV camera 3 and the blood flow rate input from the flow sensor 5.

The display 8 displays a blood flow image output from the TV camera 3, a calculation result calculated by the personal computer 7, and the like.

The differential pressure control unit 9 controls the differential pressure before and after the microchip 2 in accordance with a control command from the sequence control unit 17. Specifically, the differential pressure control unit 9 sets the pressure pump 15 upstream of the microchip 2 and the pressure reduction pump 16 downstream of the microchip 2 so that the chip upstream pressure P1 and the chip downstream pressure P2 become predetermined pressures, respectively. To control each. Note that the differential pressure control unit 9 and the sequence control unit 17 may be configured integrally with the personal computer 7.

Subsequently, an aggregation amount measuring method by the aggregation amount measuring apparatus 1 will be described.

FIG. 5 is a flowchart of the aggregation amount measuring method by the aggregation amount measuring apparatus 1.

As shown in this figure, first, blood to be measured is flowed to the microchip 2 (step S1). Specifically, blood to be measured is poured into the supply tank 10 and physiological saline or the like is added to the solution bottle 13 as necessary. Then, a predetermined differential pressure is applied to the microchip 2 by the differential pressure control unit 9, and blood flows to the microchip 2.

Next, the flow of blood passing through the gate 26 is photographed by the TV camera 3 (step S2). At this time, the shooting range of the TV camera 3 only needs to include any one of the plurality of gates 26.

Next, the velocity V ₀ of the blood cells flowing through the gate 26 is calculated (step S3). This step is performed by the arithmetic processing unit 70 of the personal computer 7 analyzing the blood flow image photographed in step S2. For calculating the blood cell velocity V ₀ , conventionally known methods described in, for example, JP-A-2006-223761 and JP-A-2002-148270 can be used. Here, the blood cell velocity V ₀ may be calculated for at least one gate 26. However, if aggregation has occurred at all the gates 26 in the blood flow image, that is, if the gates 26 in which blood cells are not flowing have been photographed, a different gate 26 is photographed again.

Next, the blood velocity V passing through the microchip 2 is calculated (step S4). In this step, the arithmetic processing unit 70 divides the blood flow rate output from the flow sensor 5 by the time from when the blood has started to flow and the cross-sectional area of the inlet 27 or outlet 28 to obtain the blood flow. The speed V is calculated. The calculation of the velocity V of the blood in the step S4 may be performed after the calculation of the velocity V ₀ which blood cells in step S3, may be performed in parallel.

Next, the blood cell aggregation amount G is calculated (step S5). In this step, the arithmetic processing unit 70, based on a ratio or difference between the speed V ₀ which blood cells which is calculated at a speed V and the step S3 of the blood that was calculated in step S4, and calculates the amount of agglutination G of blood cells. In the present embodiment, the arithmetic processing unit 70 uses the blood velocity V and the blood cell velocity V ₀ to calculate a blood cell aggregation amount G that satisfies the following equation (1) or equation (2).

G = V ₀ / V (1)
G = (V ₀ −V) / V ₀ (2)
The degree of occurrence of aggregation can be quantitatively represented by the blood cell aggregation amount G thus calculated. This is because the velocity V ₀ of the blood cell is the velocity of the blood cell flowing without staying in the gate 26 and does not include the influence of aggregation, whereas the velocity V of the blood is a plurality of gates 26, It is based on the fact that it includes the influence of blood cell aggregation at ... Therefore, when the blood cell aggregation amount G is calculated by the equation (1), it means that aggregation is not generated when the value is 1, and that the larger the value is, the larger the aggregation is. means. Further, when calculated by the equation (2), it means that aggregation is not generated when 0, and it means that aggregation is larger as the value approaches 1.

Note that the sum of the cross-sectional areas of the gates 26 in parallel facing the upstream blood circuit 24 connected to the inlet 27 or the downstream blood flow circuit 25 connected to the outlet 28 is the inlet 27 or The above relationship is obtained because the cross-sectional area of the outflow port 28 is substantially equal. However, when the relationship is not equal, when calculated by the equation (1), the closer to a predetermined value rather than 1, the larger the aggregation. Means that the aggregation is larger as the value exceeds a predetermined value. Similarly, in Formula (2), the closer to the first predetermined value, the less the aggregation occurs, and the larger the second predetermined value that is larger than the first predetermined value, the larger the aggregation occurs. Will mean.

According to the aggregation amount measuring apparatus 1 described above, it is not necessary to perform imaging and image analysis for all the gates 26, and from the blood cell velocity V ₀ not including the influence of aggregation and the blood velocity V including the influence of aggregation. The degree of occurrence of aggregation can be quantitatively represented by the calculated amount of blood cell aggregation G. Therefore, the amount of aggregation can be calculated in a shorter time and at a lower cost than in the past.

[Modification 1]
Then, the 1st modification of the said embodiment is demonstrated.

As shown in FIG. 1, the aggregation amount measuring apparatus 1A according to the first modification includes an arithmetic processing unit 70A instead of the arithmetic processing unit 70 in the above embodiment.

The arithmetic processing unit 70A has a speed ratio table T shown in FIG. 6 in addition to the configuration of the arithmetic processing unit 70 in the above embodiment. This speed ratio table T is a relative display of the speed of blood cells in the gate 26 in the preliminary measurement performed in advance. Each column of the speed ratio table T corresponds to the position of the gate 26 in the top view of the microchip 2, and the horizontal direction in the figure corresponds to the X direction and the vertical direction corresponds to the Y direction. . However, each column in the figure is illustrated with a part omitted, and the actual gate 26 and each column do not correspond one-to-one.

As shown in the speed ratio table T, the speed of blood cells at the plurality of gates 26 differs depending on the position of the gate 26. More specifically, the blood cell speeds at the plurality of gates 26 in a positional relationship arranged side by side in the X direction are fast at both ends and slow at the center in any Y direction position. In addition, the blood cell speed at the plurality of gates 26,.

It is noted that the blood cell speeds at the plurality of gates 26 are different from each other because the flow resistances around the gates 26 are different depending on the distances from the inlet 27 and the outlet 28. it is conceivable that. That is, the closer the gate 26 to the inflow port 27 or the outflow port 28 is, the lower the flow path resistance of the route through which the blood flows, the faster the blood cell speed.

Subsequently, an aggregation amount measuring method by the aggregation amount measuring apparatus 1A will be described.

FIG. 7 is a flowchart of the aggregation amount measuring method by the aggregation amount measuring apparatus 1A.

As shown in the figure, first, blood to be measured is flowed to the microchip 2 (step T1), and then the blood flow passing through the gate 26 is photographed by the TV camera 3 (step T2). Next, the velocity V ₀ of blood cells flowing through the gate 26 is calculated (step T3). Steps T1 to T3 are performed in the same manner as steps S1 to S3 in the above embodiment.

Next, the blood cell velocity is recalculated (step T4). In this step, the arithmetic processing unit 70A calculates a new blood cell velocity V ₁ using the blood cell velocity V ₀ calculated in step T3 and the velocity ratio table T.

Specifically, the arithmetic processing unit 70A first determines the position of the gate 26 (hereinafter also referred to as “calculation gate 26”) from which the blood cell velocity V ₀ was calculated in step T3 and the other gates 26 other than the gate 26. Based on the relationship and the calculated blood cell velocity V ₀ , other blood cell velocities Ve in all other gates 26 are estimated. More specifically, the arithmetic processing unit 70A specifies the corresponding speed ratio table T column from the position of the gate 26 where the blood cell velocity V ₀ is calculated, and from the relationship of the speed ratio between this column and other columns. The other blood cell velocities Ve at all other gates 26 are estimated. Then, the arithmetic processing unit 70A calculates an average value of the calculated blood cell velocity V ₀ in the calculation gate 26 and all the estimated blood cell velocities Ve, and calculates this as a new blood cell velocity V ₁ . To do.

In addition, regarding the above-described positional relationship of the “calculation gate 26” with respect to other gates, gates marked at predetermined intervals are provided, and the positional relationship of the calculation gate 26 is determined from the positional relationship with the gate. If the system has the same gate shape, it is possible to automatically or manually scroll the shooting range of the TV camera 3 to the end of the row of gates 26 before or after the measurement. You may make it grasp | ascertain the positional relationship of the calculation gate 26 from a relative position with the gate 26. FIG.

When the velocity Ve of another blood cell is estimated using the velocity ratio table T, the velocity ratio table T may be updated with the actually measured blood cell velocity V ₀ . The speed ratio table T is updated by the following procedure.

First, referring to the speed ratio table T, the blood cell speed Vmax at the gate 26 having the largest speed ratio and the blood cell speed Vmin at the gate 26 having the smallest speed ratio are measured in advance through steps T2 and T3. Keep it. Then, from these blood cell velocity Vmax and blood cell velocity Vmin and the velocity ratio R _{0 in} each column of the velocity ratio table T, a new velocity ratio R ₁ is obtained using the following equations (3) to (5). Is calculated.
Dmax = Vmax / Vmin (3)
D = (Dmax−Rmin) · (R ₀ −Rmin) / (Rmax−Rmin) + Rmin (4)
R ₁ = D × R ₀ (5)
Here, D is a correction ratio in each column of the speed ratio table T, and Dmax is a maximum correction ratio. Rmin is a value in the column of the speed ratio table T corresponding to the blood cell velocity Vmin, and Rmax is a value in the column of the speed ratio table T corresponding to the blood cell velocity Vmax. In equation (5), the correction ratio D in the corresponding column of the speed ratio table T is multiplied by the old speed ratio _R0 .

By updating the speed ratio table T in this way, it is possible to improve the estimation accuracy of the speed Ve of other blood cells.

Next, the velocity V of blood passing through the microchip 2 is calculated (step T5). This step is performed in the same manner as step S4 in the above embodiment.

Then, to calculate the aggregate amount _{G 1} of blood (step T6). In this step, the arithmetic processing unit 70A, the velocity V of the blood that was calculated in step T5, and by using the velocity V ₁ of the calculated blood cells in step T4, satisfies the following formula (6) or Formula (7) calculating the amount of agglutination G ₁ of blood.
G ₁ = V ₁ / V (6)
G ₁ = (V ₁ −V) / V ₁ (7)
According to the agglutination amount measuring apparatus 1A described above, the velocity Ve of the other blood cells in the other gate 26 is estimated from the actually measured blood cell velocity V ₀ and the velocity ratio table T, and a new blood cell that is an average value of these blood cells is estimated. because calculating the amount of agglutination G ₁ of blood cells using the velocity V _1, it is possible to calculate the aggregate amount G ₁ of blood cells in consideration of the influence of the velocity V ₀ which blood cells by the position of the gate 26. Therefore, a more reliable aggregation amount can be calculated.

In addition, when the speed ratio table T is updated using the actually measured blood cell velocity Vmax and blood cell velocity Vmin, the accuracy of estimating the velocity Ve of other blood cells can be improved. As a result, a more reliable amount of aggregation can be calculated.

[Modification 2]
Then, the 2nd modification of the said embodiment is demonstrated.

FIG. 8 is a flowchart of the aggregation amount measuring method in the second modification.

As shown in this figure, in the aggregation amount measuring method in the second modified example, first, blood to be measured is flowed to the microchip 2 as in step S1 in the above embodiment (step U1).

Next, the flow of blood passing through the gate 26 is photographed by the TV camera 3 (step U2). In this step, the TV camera 3 captures a blood flow at a plurality of preset time timings.

Next, at a plurality of time timings, the velocity V _{0 of} blood cells flowing through the gate 26 and the velocity V of blood passing through the microchip 2 are calculated (step U3). In this step, the arithmetic processing unit 70 analyzes the blood flow image at each time timing imaged in step U2, and calculates the blood cell velocity V ₀ at a plurality of time timings. At the same time, the arithmetic processing unit 70 calculates the blood velocity V at the plurality of time timings. The calculation method is the same as step S4 in the above embodiment.

Then, to calculate the amount of agglutination _{G 2} of the blood cell (Step U4). In this step, the arithmetic processing unit 70 first calculates the blood cell agglutination amount G at each time timing at which the blood velocity V and the blood cell velocity V ₀ are calculated, using Equation (1) or Equation (2) in the above embodiment. Calculated by Then, the arithmetic processing unit 70, the time integration value of the aggregate amount G of the blood cells, that is, the value obtained by integrating the amount of agglutination G of blood cells at each time the timing is calculated as aggregate amount G ₂ of the new blood cells. In the calculation of the blood cell aggregation amount G, the calculation method of the blood cell aggregation amount G ₁ in the first modification may be used instead of using the formula (1) or the formula (2).

Thus aggregated amount G ₂ new blood cells are calculated, the what is taken into account the effect on the occurrence of coagulation over time. Since the aggregation of blood cells expands the range with the passage of time, the blood velocity V gradually decreases with the passage of time, as shown in FIG. Therefore, the blood cell aggregation amount G is calculated by using the blood velocity V at a plurality of time timings, and by adding up each of the blood cell aggregation amounts G, the blood cell aggregation amount in consideration of the influence on the state of occurrence of aggregation over time is added. it can be calculated G _2.

According to the aggregation amount measuring method in the second modification described above, the blood cell aggregation amount G at a plurality of time timings is calculated, and the time integral value of the blood cell aggregation amount G is calculated as a new blood cell aggregation amount G. since calculated as _2, it is possible to calculate the amount of agglutination G ₂ of blood cells in consideration of the influence on the occurrence of coagulation over time. Therefore, a more reliable aggregation amount can be calculated.

It should be noted that the present invention should not be construed as being limited to the above-described embodiment and the first and second modifications thereof, and of course can be changed or improved as appropriate.

For example, in the above embodiment and the first and second modifications thereof, the aggregation amount measuring devices 1 and 1A calculate the aggregation amount of blood cells contained in blood. The amount of aggregation of a plurality of particles contained in can be calculated.

In the first modification of the above embodiment, the velocity Ve of other blood cells at all the other gates 26 for which the blood cell velocity V ₀ is not actually measured is estimated from the velocity ratio table T. It is also possible to estimate the velocity Ve of other blood cells, not just all the other gates 26, but only some gates 26.

1,1A Aggregation amount measuring device 2 Microchip 3 Camera (photographing means)
26 Gate 70, 70A arithmetic processing unit (fluid velocity calculation means, particle velocity calculation means, aggregation amount calculation means, velocity recalculation means)
G, G ₁ , G ₂ Aggregation amount V Blood velocity (fluid velocity)
V ₀ and V ₁ blood cell velocity (particle velocity)
Ve velocity of other blood cells (velocity of other particles)

Claims (14)

A microchip having a plurality of gates through which a fluid containing a plurality of particles flows;
Fluid velocity calculating means for calculating the velocity of the fluid passing through the microchip;
Particle velocity calculating means for calculating the velocity of the particles flowing through any one of the plurality of gates;
Agglomeration amount calculating means for calculating the amount of aggregation of the particles based on the velocity of the fluid and the velocity of the particles;
A coagulation amount measuring apparatus comprising: 2. The aggregation amount measuring apparatus according to claim 1, wherein the aggregation amount calculating means calculates the aggregation amount of the particles based on a ratio or difference between a velocity of the fluid and a velocity of the particles. The aggregation amount calculation means calculates the aggregation amount G of the particles satisfying the following formula (1) or formula (2) using the fluid velocity V and the particle velocity V _0. The aggregation amount measuring apparatus according to claim 2.
G = V ₀ / V (1)
G = (V ₀ −V) / V ₀ (2) A speed recalculating means for recalculating the speed of the particles;
The speed recalculation means includes
Based on the positional relationship between the gate for which the velocity of the particle has been calculated by the particle velocity calculation means and a gate other than the gate, and the calculated velocity of the particle, the other particles at the other gate As well as guessing the speed,
An average value of the calculated velocity of the particles and the estimated velocity of the other gate particles is calculated as the velocity of the particles,
The agglomeration amount calculation unit calculates the aggregation amount of the particles based on the velocity of the particles calculated by the velocity recalculation unit and the velocity of the fluid. The aggregation amount measuring apparatus according to any one of the above. The fluid velocity calculation means and the particle velocity calculation means calculate the fluid velocity and the particle velocity at a plurality of time timings,
The aggregation amount calculating means calculates the aggregation amount of the particles at each time timing at which the velocity of the fluid and the velocity of the particles are calculated, and calculates a time integral value of the aggregation amount of the particles as a new aggregation amount of the particles. The aggregation amount measuring device according to any one of claims 1 to 4, wherein A photographing means for photographing the flow of the fluid;
6. The aggregation amount measuring apparatus according to claim 1, wherein the particle velocity calculating unit calculates the velocity of the particles by analyzing an image captured by the imaging unit. The aggregation amount measuring apparatus according to any one of claims 1 to 6, wherein the fluid is blood and the particles are blood cells. Using a microchip having a plurality of gates through which a fluid containing a plurality of particles flows,
A fluid velocity calculating step for calculating a velocity of the fluid passing through the microchip;
A particle velocity calculating step for calculating a velocity of the particles flowing through any one of the plurality of gates;
An agglomeration amount calculating step for calculating the aggregation amount of the particles based on the velocity of the fluid and the velocity of the particles;
A method for measuring the amount of aggregation comprising the steps of: The aggregation amount measuring method according to claim 8, wherein, in the aggregation amount calculation step, the aggregation amount of the particles is calculated based on a ratio or difference between a velocity of the fluid and a velocity of the particles. In the aggregation amount calculating step, the particle aggregation amount G satisfying the following equation (1) or equation (2) is calculated using the fluid velocity V and the particle velocity V _0. The aggregation amount measuring method according to claim 9.
G = V ₀ / V (1)
G = (V ₀ −V) / V ₀ (2) A speed recalculation step for recalculating the speed of the particles,
In the speed recalculation step,
Based on the positional relationship between the gate for which the velocity of the particle is calculated in the particle velocity calculation step and a gate other than the gate, and the calculated velocity of the particle, other particles in the other gate As well as guessing the speed,
An average value of the calculated velocity of the particles and the estimated velocity of the other gate particles is calculated as the velocity of the particles,
The aggregation amount calculation step includes calculating the aggregation amount of the particles based on the velocity of the particles calculated in the velocity recalculation step and the velocity of the fluid. The aggregation amount measuring method according to any one of the above. In the fluid velocity calculation step and the particle velocity calculation step, the velocity of the fluid and the velocity of the particles at a plurality of time timings are calculated,
In the aggregation amount calculating step, the amount of aggregation of the particles at each time timing at which the velocity of the fluid and the velocity of the particles are calculated is calculated, and a time integral value of the aggregation amount of the particles is calculated as a new aggregation amount of the particles. The aggregation amount measuring method according to any one of claims 8 to 11, which is calculated as: A photographing step of photographing the flow of the fluid;
The aggregation amount measuring method according to any one of claims 8 to 12, wherein in the particle velocity calculating step, the velocity of the particles is calculated by analyzing the image photographed in the photographing step. 14. The aggregation amount measuring method according to claim 8, wherein the fluid is blood and the particles are blood cells.

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