WO2010137470A1 - Dispositif de mesure de la déformabilité et méthode de mesure de la déformabilité - Google Patents

Dispositif de mesure de la déformabilité et méthode de mesure de la déformabilité Download PDF

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Publication number
WO2010137470A1
WO2010137470A1 PCT/JP2010/058180 JP2010058180W WO2010137470A1 WO 2010137470 A1 WO2010137470 A1 WO 2010137470A1 JP 2010058180 W JP2010058180 W JP 2010058180W WO 2010137470 A1 WO2010137470 A1 WO 2010137470A1
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Prior art keywords
red blood
deformability
blood cells
hematocrit
velocity
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Ceased
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PCT/JP2010/058180
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English (en)
Japanese (ja)
Inventor
修司 一谷
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Konica Minolta Opto Inc
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Konica Minolta Opto Inc
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Priority to JP2011515974A priority Critical patent/JP5541280B2/ja
Priority to CN201080022844.6A priority patent/CN102449480B/zh
Publication of WO2010137470A1 publication Critical patent/WO2010137470A1/fr
Anticipated expiration legal-status Critical
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials

Definitions

  • the present invention relates to a deformability measuring apparatus and a deformability measuring method.
  • the deformability of the blood cells can be quantified using the velocity of the blood cells.
  • the moving distance of each blood cell is obtained from a plurality of blood flow images obtained by continuously photographing blood vessels, and the velocity of each blood cell is determined from the moving distance and the frame rate value of the camera.
  • a calculation method has been proposed (see, for example, Patent Document 1).
  • a method for measuring the velocity of fine particles other than blood cells can be applied to blood cells to determine the velocity.
  • red blood cells in blood have a characteristic that the speed changes depending on the hematocrit value of blood.
  • the present invention has been made in view of the above circumstances, and an object of the present invention is to provide a deformability measuring apparatus and a deformability measuring method capable of measuring the deformability of erythrocytes with high reliability independent of the hematocrit value.
  • the invention according to claim 1 is a deformability measuring apparatus, Speed calculating means for calculating the speed of red blood cells in the blood flowing through a flow path having a narrower width than the blood cell diameter of red blood cells; As the hematocrit value of the blood, hematocrit measuring means for obtaining a volume ratio of red blood cells occupying the blood, Deformability calculating means for calculating the deformability of the red blood cells by correcting the speed of the red blood cells calculated by the speed calculating means based on the hematocrit value obtained by the hematocrit measuring means; It is characterized by providing.
  • the deformability measuring apparatus of Claim 1 is the deformability measuring apparatus of Claim 1, Comprising:
  • the deformability calculating means calculates the red blood cell deformability D that satisfies the following formula (1) or formula (2).
  • Invention of Claim 4 is a deformability measuring apparatus of Claim 3, Comprising: An imaging means for imaging blood flowing through the flow path, The speed calculation means uses the blood flow image photographed by the photographing means, tracks the red blood cells in the blood flow image, calculates the speed of the red blood cells, The hematocrit measurement means uses the blood flow image used to calculate the velocity of the red blood cells, extracts a region containing red blood cells from the blood flow image based on a difference in concentration, and obtains the hematocrit value. .
  • the invention according to claim 5 is a deformability measuring method, A speed calculating step for calculating the speed of red blood cells in the blood flowing through a flow path having a narrower width than the blood cell diameter of red blood cells; As the hematocrit value of the blood, a hematocrit measurement step for determining the volume ratio of red blood cells in the blood, Based on the hematocrit value obtained in the hematocrit measurement step, a deformability calculation step for calculating the deformability of the red blood cells by correcting the speed of the red blood cells calculated in the speed calculation step; It is characterized by providing.
  • Invention of Claim 6 is a deformability measuring method of Claim 5, Comprising: In the deformability calculation step, the deformability D of the red blood cells satisfying the following formula (1) or formula (2) is calculated.
  • Invention of Claim 8 is a deformability measuring method of Claim 7, Comprising: Comprising a photographing step of photographing blood flowing through the flow path; In the velocity calculating step, using the blood flow image taken in the imaging step, tracking the red blood cells in the blood flow image to calculate the velocity of the red blood cells, In the hematocrit measurement step, the blood flow image used for calculating the velocity of the red blood cells is used, and a region containing red blood cells is extracted from the blood flow image based on a difference in concentration to obtain the hematocrit value. .
  • the speed of the red blood cells is corrected based on the hematocrit value of the blood containing the red blood cells, so the deformability of red blood cells is calculated while taking into account the influence of the hematocrit value. can do. Therefore, it is possible to measure the deformability of erythrocytes with high reliability independent of the hematocrit value.
  • the blood extracted for the measurement operation for calculating the velocity and the hematocrit value are thereby obtained. It is possible to prevent the hematocrit values from being different from each other for blood extracted for measurement work. Since red blood cells are not uniformly contained in blood, hematocrit values may be different from each other even if they are extracted from the same blood. According to the invention, it is possible to prevent such a case from occurring and reliably correct the speed of the red blood cells based on the corresponding hematocrit value, so that it is possible to measure the deformability of the red blood cells with higher reliability.
  • FIG. 1 is a block diagram showing an overall configuration of a deformability measuring apparatus 1 according to the first embodiment of the present invention.
  • the deformability measuring device 1 guides blood from the supply tank 10 through the filter 2 to the discharge tank 11, calculates the blood cell velocity in the blood from the information acquired in the process, Blood is supplied from the supply tank 10 to the hematocrit measuring device 4 to measure the hematocrit value, and the deformability (ease of deformation) of red blood cells is obtained from the velocity of the blood cells and the hematocrit value.
  • the deformability measuring apparatus 1 mainly includes a filter 2, a TV camera 3, a hematocrit measuring instrument 4, a personal computer (PC) 7, a display 8, and a differential pressure control unit 9.
  • the deformability measuring apparatus 1 according to the first embodiment is connected to a 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 filter 2.
  • a plurality of solution bottles 13 and the like are further provided.
  • the blood mixed with a liquid such as physiological saline or a physiologically active substance (hereinafter referred to as blood) is operated by the differential pressure control unit 9 with the pressure pump 15 and the pressure reduction pump 16 with the first valve 10a opened.
  • this blood is sent to the hematocrit measuring instrument 4 by a desired amount when the differential pressure control unit 9 drives the pressurizing pump 15 with the second valve 10b opened.
  • the differential pressure control unit 9, the mixer 12, the first valve 10 a, and the second valve 10 b are integrated and controlled by the sequence control unit 17.
  • FIG. 2 is a sectional view of the filter 2.
  • the filter 2 includes a base plate 21, silicon single crystal substrates 22 and 22, an outer plate 23, and a glass flat plate 24.
  • the base plate 21 is formed in a flat plate shape, and has an introduction hole 21a that communicates the upper surface near the center and the outer surface, and a discharge hole 21b that communicates the upper surface near one side end and the outer surface. .
  • the introduction hole 21a and the discharge hole 21b are connected to the supply tank 10 and the discharge tank 11 from the outer surface of the base plate 21 via a blood tube (not shown).
  • the two silicon single crystal substrates 22 and 22 are both formed in a substantially flat plate shape, and are arranged in parallel on the upper surface of the base plate 21 with a predetermined gap therebetween.
  • An introduction hole 21 a of the base plate 21 is opened in the gap between the two silicon single crystal substrates 22 and 22.
  • a protruding portion 22a extends in the direction of arrow X (hereinafter referred to as X direction) at the upper end portions of the silicon single crystal substrates 22 and 22, and the upper end portion of the protruding portion 22a has a hexagonal shape.
  • a plurality of banks 22b are arranged in the X direction with the top surface in contact with the glass flat plate 24 (see FIG. 3).
  • the outer plate 23 is fixed to the upper surface end of the base plate 21 so as to surround the silicon single crystal substrates 22 and 22.
  • a predetermined gap is provided between the outer plate 23 and the silicon single crystal substrates 22, 22, and a discharge hole 21 b of the base plate 21 is opened in this gap.
  • the glass flat plate 24 is formed in a flat plate shape and is fixed to the upper surface of the outer plate 23. Further, between the lower surface of the glass flat plate 24 and the upper surface of the raised portion 22a, a channel portion 25 of a fine channel group is formed.
  • FIG. 3A is a view (plan view) of the flow path portion 25 as viewed from above, and FIG. 3B is a side sectional view thereof.
  • the flow path portion 25 includes a plurality of gates 25a formed between a plurality of bank portions 22b at the upper end of the raised portion 22a, and the gate 25a.
  • the upper terrace 25b is a space on the center side of the filter 2 (upper side in the drawing) and the downstream terrace 25c is a space outside the filter 2 (lower side in the drawing) with respect to the gate 25a.
  • the gate 25a is formed in a width t narrower than the blood cell diameter (about 8 ⁇ m) of the red blood cell R in the first embodiment.
  • the lengths la, lb, and lc in the direction of the arrow Y (hereinafter referred to as the Y direction) on the upstream terrace 25b, the gate 25a, and the downstream terrace 25c are all formed to be about 30 ⁇ m. .
  • the blood introduced from the supply tank 10 through the introduction hole 21a passes through the flow path portion 25 from the center side to the outside of the filter 2 and then is discharged through the discharge hole 21b. It will be discharged into the tank 11. More specifically, blood cells, for example, red blood cells R, in the blood flowing through the flow path section 25 first pass through the upstream terrace 25b, pass through the gate 25a while being deformed, and finally pass through the downstream terrace 25c. .
  • pressure sensors E1 and E2 are provided upstream and downstream of the filter 2, and the pressure sensors E1 and E2 are configured to provide a difference between the measured filter upstream pressure P1 and filter downstream pressure P2.
  • the pressure is output to the pressure control unit 9.
  • the TV camera 3 is a digital CCD camera, for example, and is a high-speed camera having a resolution and a shutter speed sufficient for photographing a blood flow.
  • the TV camera 3 is installed to face the glass flat plate 24 in the filter 2 and photographs the blood flowing through the flow path portion 25 over the glass flat plate 24.
  • the blood flow image obtained by the TV camera 3 is output to the personal computer 7 and displayed on the display 8.
  • the TV camera 3 is not particularly limited, but is a camera capable of shooting a moving image.
  • the hematocrit measuring device 4 measures the volume ratio of the red blood cells R in the blood as the hematocrit value of the blood.
  • the hematocrit measuring instrument 4 includes a high-speed centrifuge (not shown), and measures a hematocrit value by a so-called micro hematocrit method. Specifically, the blood supplied from the supply tank 10 through the second valve 10b is sealed in a glass capillary, centrifuged with a high-speed centrifuge, and then the value when packed to a certain volume by this centrifugation is not shown. The hematocrit value is measured by reading with a reader.
  • the hematocrit measuring device 4 may be any device as long as it can measure the hematocrit value, and may use a known method described in, for example, Japanese Patent Application Laid-Open No. 11-118794.
  • the personal computer 7 includes an arithmetic processing unit 70.
  • the arithmetic processing unit 70 analyzes the blood flow image input from the TV camera 3 to calculate the velocity of the red blood cell R, and corrects the velocity based on the hematocrit value measured by the hematocrit measuring device 4.
  • the deformability of red blood cells R is calculated.
  • an arithmetic processing unit 70 for example, a CPU (Central Processing ⁇ ⁇ Unit) capable of calculating the deformability of the red blood cell R with a required accuracy can be used.
  • a CPU Central Processing ⁇ ⁇ Unit
  • the display 8 displays a blood flow image input from the TV camera 3, an analysis image analyzed by the personal computer 7, and the like.
  • the differential pressure control unit 9 is connected to the sequence control unit 17, the pressurization pump 15 and the decompression pump 16, and controls the differential pressure before and after the filter 2 in accordance with a control command from the sequence control unit 17. Yes. More specifically, the differential pressure control unit 9 controls the pressure pump 15 upstream of the filter 2 and the pressure reduction pump 16 downstream of the filter 2 so that the filter upstream pressure P1 and the filter downstream pressure P2 become predetermined pressures. To do. Note that the differential pressure control unit 9 and the sequence control unit 17 may be configured integrally with the personal computer 7.
  • FIG. 4 is a flowchart of the deformability measuring method by the deformability measuring apparatus 1.
  • step S1 blood to be measured is passed through the filter 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, the first valve 10 a is opened by the sequence control unit 17 and a predetermined differential pressure is applied to the filter 2 by the differential pressure control unit 9, so that blood flows through the filter 2.
  • the blood flowing through the flow path portion 25 is photographed by the TV camera 3 (step S2).
  • the TV camera 3 takes a moving image of the blood flow so that the same red blood cell R flowing through the gate 25a can be captured in at least two frames.
  • step S3 the speed of red blood cells R in the blood flowing through the gate 25a is calculated (step S3).
  • This step is performed by the arithmetic processing unit 70 of the personal computer 7 analyzing the blood flow image obtained in step S2. Specifically, the arithmetic processing unit 70 uses the blood flow images of a plurality of frames to track the red blood cells R in the blood flow images, thereby obtaining the moving distance of the same red blood cells R flowing through the gate 25a.
  • the speed of the red blood cell R is calculated by dividing by the shutter speed.
  • imaging of blood flow and calculation of velocity in steps S2 and S3 are not limited to the above methods, and the methods described in Patent Documents 1 and 2 and Non-Patent Document 1 described above may be used.
  • the speed is measured by measuring the passage time when a predetermined amount of blood is flowed to the gate 25a without taking the blood flow at S2, and dividing the flow amount of blood by the cross-sectional area and the passage time of the gate 25a. You may use the method of calculating
  • step S4 blood to be measured is set in the hematocrit measuring instrument 4 (step S4).
  • the first valve 10a is closed and the second valve 10b is opened by the sequence control unit 17, and the pressurizing pump 15 is further driven, so that the same blood flowed to the filter 2 in step S1 is hematocritized. It is sent to the measuring device 4 and set.
  • the hematocrit value of the blood set in the hematocrit measuring device 4 is measured (step S5).
  • the measured hematocrit value is output to the personal computer 7.
  • the process related to the measurement of the hematocrit value in steps S4 and S5 may be performed before the process related to the calculation of the speed in steps S2 and S3, or may be performed in parallel.
  • step S6 the deformability of the red blood cell R is calculated.
  • the arithmetic processing unit 70 calculates the deformability of the red blood cell R by correcting the speed of the red blood cell R calculated in step S3 based on the hematocrit value measured in step S5.
  • the arithmetic processing unit 70 calculates the deformability D of the red blood cell R as a value satisfying the following formula (1) or formula (2).
  • V V / V 0 (1)
  • V V ⁇ V 0 (2)
  • V is the velocity of the red blood cell R calculated in step S3
  • V 0 is the red blood cell R in the reference blood when the reference blood having the hematocrit value H measured in step S5 is flowed to the gate 25a.
  • Speed the reference blood refers to the blood of a healthy person
  • the hematocrit value H and the velocity V 0 of the red blood cell R in the reference blood have a relationship as shown in FIG. 5, for example. Therefore, by storing a conversion formula or conversion table corresponding to the relationship between the hematocrit value H of FIG.
  • the hematocrit value H measured in step S5 is stored. From this, the velocity V 0 of the red blood cell R in the reference blood can be obtained.
  • the blood to be measured can be identified as being closer to the reference blood as it is closer.
  • the velocity V 0 of the red blood cell R in the reference blood generally has a constant velocity range ⁇ V 0 with respect to the same hematocrit value H as shown in FIG.
  • the average value may be the speed V 0 .
  • the deformability range ⁇ D corresponding to the speed range ⁇ V 0 the blood to be measured is determined as being within the reference blood range and being good if within the deformability range ⁇ D. You may make it do.
  • the deformability D shown on the vertical axis in FIG. 6 is a unit and a scale when calculated using the above formula (1).
  • the deformability measuring apparatus 1 in the first embodiment described above when calculating the deformability D of the red blood cell R, the velocity V of the red blood cell R is corrected based on the hematocrit value H of the blood containing the red blood cell R. Therefore, the deformability D of the red blood cell R can be calculated while considering the influence of the hematocrit value H. Therefore, it is possible to measure the deformability D of the red blood cell R with high reliability that does not depend on the hematocrit value H.
  • FIG. 7 is a block diagram showing the overall configuration of the deformability measuring apparatus 1A.
  • the deformability measuring apparatus 1A is different from the deformability measuring apparatus 1 in the first embodiment in that it does not include the hematocrit measuring instrument 4 and the second valve 10b, and instead of the personal computer 7. And a personal computer 7A.
  • the personal computer 7A includes an arithmetic processing unit 70A in place of the arithmetic processing unit 70 in the first embodiment.
  • the arithmetic processing unit 70A analyzes the blood flow image input from the TV camera 3 to calculate the velocity V of the red blood cell R and the hematocrit value H of the red blood cell R, and also calculates the red blood cell R from the speed V and the hematocrit value H of the red blood cell R.
  • the deformability D is calculated.
  • FIG. 8 is a flowchart of the deformability measuring method by the deformability measuring apparatus 1A.
  • step T1 blood to be measured is passed through the filter 2 (step T1), the blood flowing through the flow path section 25 is photographed (step T2), and then the speed of the red blood cells R in the blood flowing through the gate 25a is obtained. V is calculated (step T3).
  • steps T1 to T3 are performed in the same manner as steps S1 to S3 in the first embodiment.
  • the processing unit 70A performs the same processing as the processing unit 70 in the first embodiment, whereby the velocity V of the red blood cell R is calculated.
  • the hematocrit value H of the blood to be measured is calculated (step T4).
  • the arithmetic processing unit 70A uses the blood flow image used for calculating the velocity V of the red blood cell R, and calculates the red blood cell R from the blood flow image based on the difference in density between the red blood cell R portion and the non-red blood cell portion.
  • the hematocrit value H which is the volume ratio of blood cells in the blood, is obtained by extracting the included region.
  • the arithmetic processing unit 70A calculates a ratio of the region in the entire blood flow image by setting a region where the density of the image color is equal to or greater than a predetermined threshold as a region including the red blood cells R. Such a region is preferably in the region of the upstream terrace 25b upstream of the gate 25a.
  • the velocity V and the hematocrit value H of the red blood cells R are calculated using the blood flow image captured in step T2, that is, the blood flow imaging in step T2.
  • the velocity V and the hematocrit value H of the red blood cell R are calculated through the same measurement work common to each other.
  • step T5 the deformability D of the red blood cell R is calculated (step T5).
  • the arithmetic processing unit 70A uses the velocity V and hematocrit value H of the red blood cells R calculated in steps T3 and T4, and is the same as the arithmetic processing unit 70 in step S6 in the first embodiment. By performing the processing, the deformability D of the red blood cell R is calculated.
  • the velocity V of the red blood cell R can be obtained through the same measurement work common to each other as well as the same effect as that of the first embodiment. Since the hematocrit value H is calculated, it is possible to prevent the hematocrit value H from being different between the blood extracted for the measurement operation for calculating the velocity V and the blood extracted for the measurement operation for obtaining the hematocrit value H. Since the red blood cells R are not uniformly contained in the blood, the hematocrit values H may be different from each other even if the same blood is extracted separately.
  • the deformability measuring apparatus 1A According to the deformability measuring apparatus 1A, such a case can be prevented and the velocity V of the red blood cell R can be reliably corrected based on the corresponding hematocrit value H. Therefore, the deformability of the red blood cell R is more reliable. D can be measured.
  • the velocity V of the red blood cell R is corrected based on the hematocrit value H.
  • the velocity V of the red blood cell R or the amount of hemoglobin may be corrected, or a combination thereof may be used. It may be corrected.
  • the conversion equation or conversion table for converting the age or hemoglobin amount in the healthy person into the velocity of the red blood cell R is stored in the arithmetic processing units 70 and 70A, so that the red blood cell R as in the case of the hematocrit value H is stored. Can be corrected.
  • 1,1A Deformability measuring device 3 TV camera (photographing means) 4 Hematocrit measuring instrument (hematocrit measuring means) 25a Gate (flow path) 70 arithmetic processing unit (speed calculation means, deformability calculation means) 70A arithmetic processing unit (speed calculation means, hematocrit measurement means, deformability calculation means) D Deformability of red blood cells H Hematocrit value R Red blood cells V Red blood cell velocity V 0 Red blood cell velocity in reference blood

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Abstract

La déformabilité des érythrocytes peut être mesurée avec une haute fiabilité indépendamment des valeurs d'hématocrite. Pour cela, une unité de calcul (70) calcule la vitesse (V) des érythrocytes (R) dans le sang s'écoulant à travers une ouverture (25a), dont la largeur (t) est plus petite que le diamètre cellulaire des érythrocytes (R), et détermine le rapport en volume des érythrocytes (R) dans le sang en tant que valeur d'hématocrite (H) du sang. En outre, l'unité de calcul (70) corrige la vitesse (V) des érythrocytes (R) sur la base de la valeur d'hématocrite (H) et détermine ainsi la déformabilité (D) des érythrocytes (R).
PCT/JP2010/058180 2009-05-29 2010-05-14 Dispositif de mesure de la déformabilité et méthode de mesure de la déformabilité Ceased WO2010137470A1 (fr)

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JP2011515974A JP5541280B2 (ja) 2009-05-29 2010-05-14 変形能計測装置及び変形能計測方法
CN201080022844.6A CN102449480B (zh) 2009-05-29 2010-05-14 变形能测量装置及变形能测量方法

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JPWO2019240061A1 (ja) * 2018-06-15 2021-05-13 株式会社 レオロジー機能食品研究所 赤血球老化度の評価方法
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US12025548B2 (en) 2021-09-29 2024-07-02 Orange Biomed Ltd., Co. Apparatus for measuring properties of particles in a solution and related methods
US12517109B2 (en) 2022-12-09 2026-01-06 Orange Biomed Ltd., Co. Sample-testing system for measuring properties of red blood cells

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JP2005106778A (ja) * 2003-10-02 2005-04-21 Fuji Photo Film Co Ltd フィルターチップ及びそれを用いた細胞及び粒子の流れ特性観察方法
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