WO2010047191A1 - 血球変形能計測装置 - Google Patents

血球変形能計測装置 Download PDF

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Publication number
WO2010047191A1
WO2010047191A1 PCT/JP2009/065813 JP2009065813W WO2010047191A1 WO 2010047191 A1 WO2010047191 A1 WO 2010047191A1 JP 2009065813 W JP2009065813 W JP 2009065813W WO 2010047191 A1 WO2010047191 A1 WO 2010047191A1
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Prior art keywords
blood
gate
deformability
blood cell
volume
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PCT/JP2009/065813
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English (en)
French (fr)
Japanese (ja)
Inventor
修司 一谷
Original Assignee
コニカミノルタオプト株式会社
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
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Application filed by コニカミノルタオプト株式会社 filed Critical コニカミノルタオプト株式会社
Priority to CN200980141499.5A priority Critical patent/CN102187217B/zh
Priority to JP2010534755A priority patent/JP5093357B2/ja
Publication of WO2010047191A1 publication Critical patent/WO2010047191A1/ja

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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
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1429—Signal processing
    • G01N15/1433—Signal processing using image recognition
    • 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
    • G01N15/01—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials specially adapted for biological cells, e.g. blood cells
    • G01N2015/012—Red blood cells
    • 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
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N2015/1493—Particle size
    • 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
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N2015/1493—Particle size
    • G01N2015/1495—Deformation of particles

Definitions

  • the present invention relates to a blood cell deformability measuring apparatus.
  • the deformation index is defined using the major and minor diameters of elliptical red blood cells from the diffraction image.
  • a method for calculating the deformation index as a quantitative value of the deformability of red blood cells has been proposed (see, for example, Patent Documents 1 and 2).
  • Patent Documents 1 and 2 are methods that can be applied only to red blood cells that retain an elliptical shape, and cannot be applied to amoeba-like white blood cells that can be deformed into various shapes.
  • moving blood cells can be displayed as an image, it is difficult to visually grasp how the blood cells move while deforming because the blood cells flow at a uniform speed on the image.
  • the present invention has been made in view of the above circumstances, and can measure each deformability when red blood cells and white blood cells pass through capillaries and can easily grasp the state of blood cells moving while deforming. It is an object to provide a measuring device.
  • a blood cell deformability measuring device for flowing blood from an upstream terrace having a width wider than the blood cell diameter toward a gate having a width narrower than the blood cell diameter, and measuring the deformability of the blood cells in the blood, Imaging means for imaging the flow of blood passing through the upstream terrace and the gate; From the blood flow image obtained by the imaging means, speed calculation means for calculating the speed of blood cells passing through the upstream terrace or the gate; From the blood flow image obtained by the imaging means, volume calculating means for calculating the volume of blood cells passing through the gate; Deformability calculating means for calculating the deformability of the blood cell from the velocity of the blood cell and the volume of the blood cell; It is characterized by providing.
  • the invention according to claim 2 is the blood cell deformability measuring device according to claim 1,
  • the deformability calculating means calculates D 1 or D 2 satisfying the following formula (1) or formula (2) as the deformability of the blood cell.
  • the invention according to claim 4 is the blood cell deformability measuring apparatus according to any one of claims 1 to 3, It has blood flow image display means for displaying a blood flow image obtained by the photographing means on a display screen.
  • the invention according to claim 5 is the blood cell deformability measuring apparatus according to any one of claims 1 to 4, A plurality of the gates; Occlusion state calculation means for calculating the occlusion ratio of the gate occluded by the blood cells, wherein the blood cell speed obtained by the speed calculation means is zero,
  • the deformability calculating means calculates the deformability of the blood cell from the occlusion ratio of the gate and the volume of the blood cell closed by the volume calculating means.
  • the invention according to claim 6 is the blood cell deformability measuring device according to claim 5,
  • the occlusion state calculating means calculates the occlusion ratio of the gate that is occluded by blood cells of the volume according to the volume of the blood cells,
  • the deformability calculation means calculates a D 1 satisfies the following equation (3).
  • the invention according to claim 7 is the blood cell deformability measuring device according to claim 5 or 6,
  • the plurality of gates are arranged in the blood flow direction and are formed in a plurality of different widths so as to gradually narrow from the upstream side to the downstream side in the blood flow direction, or orthogonal to the blood flow direction.
  • the occlusion state calculating means detects, as a maximum occlusion width, the largest one of the widths of the gates occluded by the blood cells, from the blood flow image obtained by the imaging means,
  • the deformability calculation means calculates the deformability of the blood cell from the maximum occlusion width of the gate and the volume of the blood cell that occludes the gate corresponding to the maximum occlusion width calculated by the volume calculation means.
  • the invention according to claim 8 is the blood cell deformability measuring apparatus according to claim 5 or 6,
  • the plurality of gates form a plurality of sets for each width
  • the blockage state calculating means obtains the blockage rate of the gate blocked by the blood cells according to the width of the gate, and detects the width of the gate that maximizes the blockage rate as the maximum blockage width
  • the deformability calculation means calculates the deformability of the blood cell from the maximum occlusion width of the gate and the volume of the blood cell that occludes the gate corresponding to the maximum occlusion width calculated by the volume calculation means.
  • the invention according to claim 9 is the blood cell deformability measuring device according to claim 5 or 6,
  • the occlusion state calculating means detects, as a maximum occlusion width, the width of the gate at the occlusion position of the blood cell in the gate occluded with the blood cell from the blood flow image obtained by the imaging means,
  • the deformability calculating means calculates the deformability of the blood cell from the maximum occlusion width of the gate and the volume of the blood cell closed by the volume means.
  • the invention according to claim 10 is the blood cell deformability measuring apparatus according to any one of claims 7 to 9,
  • the deformability calculating means calculates D 2 or D 3 satisfying the following formula (4) or formula (5) as the deformability of the blood cell.
  • the invention according to claim 11
  • a blood cell deformability measuring apparatus for flowing blood to a plurality of gates formed with a narrower width than the blood cell diameter and measuring the deformability of the blood cells in the blood, Photographing means for photographing the flow of blood passing through the gate; From the blood flow image obtained by the imaging means, an occlusion state calculation means for calculating the occlusion ratio of the gate occluded by the blood cells, From the blood flow image obtained by the imaging means, a volume calculation means for calculating the volume of the blood cell
  • the invention according to claim 12 is the blood cell deformability measuring device according to claim 11,
  • the occlusion state calculating means calculates the occlusion ratio of the gate that is occluded by blood cells of the volume according to the volume of the blood cells,
  • the deformability calculation means calculates a D 1 satisfies the following equation (3).
  • the invention according to claim 13 is the blood cell deformability measuring device according to claim 11 or 12, It has blood flow image display means for displaying a blood flow image obtained by the photographing means on a display screen.
  • the invention according to claim 14 is the blood cell deformability measuring device according to any one of claims 11 to 13,
  • the plurality of gates are arranged in the blood flow direction and are formed in a plurality of different widths so as to gradually narrow from the upstream side to the downstream side in the blood flow direction, or orthogonal to the blood flow direction.
  • the occlusion state calculating means detects, as a maximum occlusion width, the largest one of the widths of the gates occluded by the blood cells, from the blood flow image obtained by the imaging means,
  • the deformability calculation means calculates the deformability of the blood cell from the maximum occlusion width of the gate and the volume of the blood cell that occludes the gate corresponding to the maximum occlusion width calculated by the volume calculation means.
  • the invention according to claim 15 is the blood cell deformability measuring apparatus according to any one of claims 11 to 13,
  • the plurality of gates form a plurality of sets for each width
  • the blockage state calculating means obtains the blockage rate of the gate blocked by the blood cells according to the width of the gate, and detects the width of the gate that maximizes the blockage rate as the maximum blockage width
  • the deformability calculation means calculates the deformability of the blood cell from the maximum occlusion width of the gate and the volume of the blood cell that occludes the gate corresponding to the maximum occlusion width calculated by the volume calculation means.
  • the invention according to claim 16 is the blood cell deformability measuring apparatus according to any one of claims 11 to 13,
  • the occlusion state calculating means detects, as a maximum occlusion width, the width of the gate at the occlusion position of the blood cell in the gate occluded with the blood cell from the blood flow image obtained by the imaging means,
  • the deformability calculating means calculates the deformability of the blood cell from the maximum occlusion width of the gate and the volume of the blood cell closed by the volume means.
  • the invention according to claim 17 is the blood cell deformability measuring apparatus according to any one of claims 14 to 16,
  • the deformability calculating means calculates D 2 or D 3 satisfying the following formula (4) or formula (5) as the deformability of the blood cell.
  • the deformability of the blood cell is calculated from the velocity of the blood cell and the volume of the blood cell. Therefore, the deformability of the blood cell is not limited to the red blood cell having an elliptical shape, and even if it is an amoeba-like white blood cell. Can be measured. Further, since the deformability of blood cells passing through a gate having a width narrower than the blood cell diameter is calculated, the deformability when blood cells pass through the capillaries can be measured by simulating capillaries by the gate.
  • the invention since the deformability of the blood cell is calculated from the occlusion ratio or the maximum occlusion width of the gate and the volume of the blood cell, the invention is not limited to the red blood cell having an elliptical shape, but an amoeba-like shape. Even leukocytes can be measured for deformability. Further, since the deformability of blood cells passing through a gate having a width narrower than the blood cell diameter is calculated, the deformability when blood cells pass through the capillaries can be measured by simulating capillaries by the gate.
  • the gate having a plurality of sets for each width, the maximum obtained as the gate width at which the blocking ratio for each width of the gate is maximized. Since the deformability of the blood cell is calculated from the occlusion width, the deformability is calculated using more gates, and a more stable calculation result is obtained.
  • FIG. 3A is a plan view of the flow path portion
  • FIG. 3B is a side sectional view.
  • It is a flowchart of deformability measurement in an embodiment. It is a figure which shows an example of the velocity map of a blood cell. It is a graph of a deformability. It is a flowchart of the deformability measurement in the modification of embodiment. It is a figure which shows a mode that white blood cells pass a gate. It is a graph which shows the time change of the density
  • FIG. 11 (a) is a graph showing the distribution range of measurement results of soft blood cells, with the vertical and horizontal axes representing changes in the vertical and horizontal widths of blood cells
  • FIG. 11 (b) shows the distribution ranges of measurement results of hard blood cells. It is the shown graph.
  • FIG. 12A is a graph showing measurement results of soft blood cells
  • FIG. 12B is a graph showing measurement results of hard blood cells, with the vertical width and the horizontal axis representing the change width and area of the vertical width of the blood cells.
  • FIG. 15 (a) is a processing image diagram in edge extraction processing
  • FIG. 15 (b) is a processing image diagram in monochrome / binarization processing
  • FIG. 15 (c) is a processing image diagram in noise processing
  • FIG. d) It is a process image figure in a morphological process
  • FIG.15 (e) is a process image figure in a blood cell residence part determination process. It is a figure which shows a mode that some gates are obstruct
  • FIG. 20 (a) is a figure which shows the state in which the blood containing many soft blood cells was poured
  • FIG. 20 (a) is a figure which shows the state in which the blood containing many soft blood cells was poured
  • FIG.20 (b) contains many hard blood cells.
  • FIG. 20 (b) contains many hard blood cells.
  • FIG. 20 (b) contains many hard blood cells.
  • FIG. 20 (b) contains many hard blood cells.
  • FIG. 1 is a block diagram showing an overall configuration of a blood cell deformability measuring apparatus 1 according to the present invention.
  • the blood cell deformability measuring apparatus 1 guides blood from a supply tank 10 through a filter 2 to a discharge tank 11, and measures the deformability of blood cells in the blood from information acquired in the process.
  • blood cells indicate red blood cells and / or white blood cells.
  • the blood cell deformability measuring apparatus 1 mainly includes a filter 2, a TV camera 3 for photographing a blood flow in the filter 2, and a deformability based on a blood flow image photographed by the TV camera 3.
  • a personal computer (PC) 7 that measures the blood flow
  • a display 8 that displays a blood flow image
  • a differential pressure control unit 9 that controls the blood flow in the filter 2.
  • a plurality of liquids such as physiological saline and physiologically active substances that are connected to the flow path via the mixer 12 so as to be mixed with blood and guided to the filter 2.
  • a solution bottle 13 or the like is further provided.
  • the differential pressure control unit 9 controls the pressurization pump 15 and the decompression pump 16 to adjust the differential pressure across the filter 2. By doing so, the filter 2 flows by a desired amount.
  • the valve 10 a of the supply tank 10 and the like 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 in which the silicon single crystal substrates 22 and 22 are juxtaposed (the X direction in the drawing).
  • a plurality of hexagonal bank portions 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.
  • FIGS. 3A and 3B are diagrams for explaining the flow path section 25.
  • 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.
  • 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 width t of the gate 25a is narrower than the blood cell diameter (about 8 ⁇ m) of the red blood cells R in the present embodiment.
  • the height h of the gate 25a is formed to be narrower than the blood cell diameter of the red blood cells R (about 8 ⁇ m).
  • the width t and height h may be formed narrower than the blood cell diameter of white blood cells W (about 10 to 20 ⁇ m).
  • the lengths la, lb, and lc in the width direction (Y direction in the drawing) of the raised portion 22a in the upstream terrace 25b, the gate 25a, and the downstream terrace 25c are all formed to be about 30 ⁇ m.
  • the upstream terrace 25b should just be formed in the width
  • the blood introduced from the supply tank 10 through the introduction hole 21a passes through the flow path portion 25 and is then discharged to the discharge tank 11 through the discharge hole 21b. More specifically, blood cells in blood flowing through the flow path section 25, for example, red blood cells R, first pass through the upstream terrace 25b, pass through the gate 25a, and finally pass through the downstream terrace 25c. It becomes.
  • 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 sufficient for photographing a blood flow.
  • the TV camera 3 is installed opposite to the glass flat plate 24 in the filter 2 and photographs the blood flow passing through the flow path portion 25 over the glass flat plate 24.
  • the photographing range may be a range including at least the plurality of gates 25a and the upstream terrace 25b.
  • 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 personal computer 7 is connected to the TV camera 3 and includes an arithmetic processing unit 70 capable of calculating a plurality of types of blood characteristics from image information output from the TV camera 3.
  • the blood characteristics are various characteristic values indicating the properties of blood and the like, and include those relating to fluidity such as blood cell velocity and volume as well as blood cell deformability.
  • the arithmetic processing unit 70 can detect the gate 25a clogged with blood cells. As such an arithmetic processing part 70, a conventionally well-known thing can be used.
  • the display 8 is connected to the personal computer 7 and displays a blood flow image output from the TV camera 3 and blood characteristics calculated by the personal computer 7.
  • the differential pressure control unit 9 is connected to the sequence control unit 17, the pressure pump 15, and the pressure reduction 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 deformability measurement by the blood cell deformability measuring apparatus 1.
  • the blood flow passing through the flow path portion 25 is imaged (step S1). Specifically, first, 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 filter 2 by the differential pressure control unit 9 so that blood flows through the filter 2, and at the same time, the blood flow passing through the flow path unit 25 is photographed with the TV camera 3. The photographed blood flow image is displayed on the display 8.
  • the blood flow image is processed by the arithmetic processing unit 70, and a blood cell velocity map is created (step S2).
  • a blood cell velocity map is created (step S2).
  • FIG. As shown, blood cells are detected at each intersection of the grid set on the blood flow image of the upstream terrace 25b, and a velocity map is obtained that calculates the velocity vector of the blood cells at the intersection where the blood cells are detected.
  • the created velocity map may not be obtained from the velocity vector of blood cells passing through the upstream terrace 25b, but may be obtained from the velocity vector of blood cells passing through the gate 25a.
  • the blood cell type of the blood cell detected on the speed map is determined by the arithmetic processing unit 70 (step S3).
  • the red blood cell R is determined as a white portion in the red hue range.
  • the white blood cells W may be discriminated using luminance, or may be discriminated as white portions with few holes or white portions with a small number of edges per unit area using the fact that they are larger than other blood cell types.
  • the blood cell type is discriminated using a known method described in, for example, JP-A-10-48120, JP-A-10-90163, and JP-A-10-274652. Can do. If the blood to be measured includes only one of red blood cells R and white blood cells W, this blood cell type discrimination step is omitted.
  • the blood cell velocity S is calculated by the arithmetic processing unit 70 (step S4).
  • the average value of the velocity vector of the blood cell calculated in step S2 is calculated by dividing into the red blood cell R and the white blood cell W determined in step S3, so that the average value of the red blood cell R and the white blood cell W is obtained as the average value.
  • a speed S is calculated.
  • the blood cell volume V is calculated by the arithmetic processing unit 70 (step S5).
  • the area of a blood cell passing through the gate 25a is calculated using a known method described in Japanese Patent Laid-Open No. 5-79970, and the area V is multiplied by the height h of the gate 25a to obtain the volume V of the blood cell. Calculated.
  • step S6 the deformability of the blood cell is calculated by the arithmetic processing unit 70 (step S6).
  • the speed S and the volume V of the blood cell of the blood cell which is calculated in step S4 and step S5, D 1 satisfies the following equation (1) is calculated as a deformability of blood cells.
  • ⁇ ⁇ / (S ⁇ V) (1)
  • ⁇ is a predetermined coefficient set in a measurement test performed in advance. More specifically, ⁇ is calculated by the blood cell deformability measuring apparatus 1 with the deformability of red blood cells measured by another apparatus as positive. The coefficient is set so that the deformability is calculated from the velocity S of the red blood cells and the volume V of the blood cells. If ⁇ is not constant with respect to the product S ⁇ V of blood cell velocity S and volume V in a measurement test performed in advance, ⁇ may be used as a lookup table for the product S ⁇ V. .
  • D 2 satisfying the following equation (2) may be calculated as the deformability of the blood cell.
  • D 2 1 / (S ⁇ V) / (1 / (S 0 ⁇ V 0 )) (2) here, S 0 : velocity of blood cells in the blood when a predetermined reference blood is passed to the upstream terrace 25b or the gate 25a, V 0 : The volume of blood cells in the blood when the predetermined reference blood is passed to the gate 25a, and the predetermined reference blood is blood of a standard health level.
  • D 1 calculated as deformability shows that blood cells stiff greater the value.
  • D 2 indicates that the blood cell has a standard hardness when the value is 1, and indicates that the blood cell is harder as it is larger than 1 and softer as it is smaller.
  • D 1 or D 2 calculated as deformability is arranged as a graph relating to the blood cell volume V as shown in FIG. 6, and the graph for each blood cell type of red blood cells R and white blood cells W is displayed on the display 8. Is done.
  • FIG. 6 is an example in which measurement results for two types of blood A and B are displayed together.
  • the blood cell deformability measuring device 1 is not limited to the red blood cell R having an elliptical shape, and is an amoeba-like white blood cell. Even if it is W, its deformability can be measured. Further, since the deformability of the blood cells passing through the gate 25a having a width smaller than the blood cell diameter is calculated, the deformability when the blood cells pass through the capillaries can be measured by simulating the capillaries by the gate 25a.
  • the blood flow passing through the gate 25a having a width narrower than the blood cell diameter is displayed on the display 8, the blood cell having the velocity S decreased can be confirmed by the gate 25a, and the blood cell has a uniform velocity on the image. Compared to the conventional flow that has flowed in step 1, it is possible to visually grasp how the blood cells move while deforming.
  • the blood cell deformability measuring apparatus 1A includes a personal computer 7A instead of the personal computer 7 in the above embodiment, and the personal computer 7A includes an arithmetic processing unit 70A instead of the arithmetic processing unit 70. Yes.
  • the arithmetic processing unit 70A is configured to be able to measure a passage time T required for blood cells to pass through the gate 25a instead of the velocity S of the blood cells in the blood.
  • the blood cell deformability measuring apparatus 1A includes a filter 2A instead of the filter 2 in the above embodiment, and the filter 2A is replaced with a flow path portion 25 as shown in FIG. 25A is provided.
  • the shape of the flow path portion 25A is equivalent to that of the flow path portion 25.
  • FIG. 7 is a flowchart of deformability measurement by the blood cell deformability measuring apparatus 1A.
  • step T1 the blood flow passing through the flow path portion 25 is photographed. This step is performed in the same manner as step S1 in the above embodiment.
  • the blood flow image is processed by the arithmetic processing unit 70A, and blood cells are detected (step T2).
  • a blood cell flowing in the upstream terrace 25b is detected and tracked by using a known method described in JP-A-2001-264318.
  • step T3 the blood cell type of the blood cell detected in step T2 is determined by the arithmetic processing unit 70A (step T3). This step is performed in the same manner as step S3 in the above embodiment.
  • step T4 the blood cell volume V is calculated by the arithmetic processing unit 70A (step T4). This step is performed in the same manner as step S5 in the above embodiment.
  • the passage time T required for the blood cells to pass through the gate 25a is measured by the arithmetic processing unit 70A (step T5).
  • the passage time T is measured by detecting the density change of the image color in the gate 25a.
  • FIGS. 8A to 8D are views showing how the white blood cells W pass through the gate 25a
  • FIG. 9 is a graph showing the change over time of the color density of the blood flow image at that time.
  • the deformability of the blood cell is calculated by the arithmetic processing unit 70A (step T6).
  • D 3 satisfying the following equation (30) is calculated as the deformability of the blood cell from the blood cell volume V and the passage time T calculated in step T4 and step T5.
  • ⁇ ⁇ ⁇ T / V (30)
  • ⁇ is a predetermined coefficient set in a measurement test performed in advance, and is a coefficient set in the same manner as ⁇ in the above equation (1).
  • D 4 satisfying the following formula (40) may be calculated as the deformability of the blood cell.
  • T 0 is a transit time required for blood cells in a predetermined reference blood to pass through the gate 25a.
  • D 3 and D 4 calculated from the equations (30) and (40) are the reciprocals of the passage time T instead of the blood cell velocity S with respect to the equations (1) and (2) in the above embodiment. It is none other than D 1 and D 2 when. Therefore, D 3 and D 4 can be used for the evaluation of deformability in the same manner as D 1 and D 2 .
  • D 3 or D 4 calculated as deformability is arranged as a graph relating to blood cell volume V as shown in FIG. 6, and the graph for each blood cell type of red blood cells R and white blood cells W is displayed on the display 8. Is done. If the blood to be measured includes only one of red blood cells R and white blood cells W, the calculated values in steps T4 to T6 may be the average value of all blood cells detected in step T1. .
  • the deformability is calculated from the volume V of the blood cells passing through the gate 25a, etc.
  • the deformability can also be obtained from a blood flow image when passing through the upstream terrace 25b or the downstream terrace 25c.
  • the blood cells passing through the upstream terrace 25b or the downstream terrace 25c change their shapes in various ways depending on their deformability as shown in FIGS. 10 (a) to 10 (c), for example. Therefore, the changes in the vertical and horizontal widths of the blood cells are calculated from the blood flow image, and as shown in FIGS. 11A and 11B, the coordinates of the vertical and horizontal axes corresponding to the data of each of the plurality of blood cells are obtained. Create a plotted graph.
  • the vertical width and horizontal width of a blood cell be the length of the Y direction of a flow-path part 25, and a X direction, for example.
  • the vertical width of the blood cell passing through the upstream terrace 25b or the downstream terrace 25c and the area of the blood cell are expressed as the vertical axis and the horizontal axis.
  • the vertical width of the blood cell on the vertical axis may be the horizontal width of the blood cell.
  • the blood cell deformability measuring apparatus 1E includes a personal computer 7E instead of the personal computer 7 in the above embodiment, and the personal computer 7E includes an arithmetic processing unit 70E instead of the arithmetic processing unit 70. Yes.
  • the arithmetic processing unit 70E is configured to determine that the gate 25a where the blood cell velocity S is zero is a closed gate 25a, and to calculate the closed ratio.
  • FIG. 13 is a flowchart of deformability measurement by the blood cell deformability measuring apparatus 1E
  • FIG. 14 is a flowchart when the closed gate 25a is detected.
  • blood to be measured is flowed to the filter 2E (step S10). 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 filter 2E by the differential pressure control unit 9, and blood flows through the filter 2E.
  • a blood flow passing through the flow path portion 25E is photographed, and a blood flow image is collected (step S20).
  • the TV camera 3 takes a moving image of the blood flow that passes through the flow path portion 25E.
  • the photographed blood flow image is displayed on the display 8.
  • FIGS. 15A to 15E are diagrams showing an example of the processed image in each step of FIG. However, FIGS. 15A to 15E are for facilitating the understanding of the processing in this step, and are not processed blood flow images in the gate 25a.
  • a Sobel filter is applied to the collected blood flow image in both the vertical and horizontal directions to extract the edge of the staying blood cell portion (hereinafter referred to as a blood cell staying portion).
  • the image is grayscaled and binarized with a predetermined threshold value, and the blood cell retention portion is displayed in white (step S32).
  • noise and blood flow shadows that are misrecognized as edges of the blood cell retention portion are removed from the white portion (step S33).
  • a white portion having an area smaller than a preset threshold is set as noise, and a white portion whose length ratio between the X direction and the Y direction is outside a predetermined range is blackened as a blood flow shadow.
  • step S34 The image from which noise or the like has been removed is expanded and contracted by morphological processing, and the gaps in the white portion are filled (step S34). And the white part which remained so far is determined as a blood cell retention part (step S35).
  • the blood cell retention part is detected, and the gate 25a having the blood cell retention part is detected as the closed gate 25a.
  • the volume V of the blood cell retention part is calculated by the arithmetic processing part 70E (step S40).
  • the area of the blood cell retention portion in the gate 25a is calculated using a known method described in JP-A-5-79970, and this area is multiplied by the height h of the gate 25a, whereby the gate 25a is The volume V of the blood cell retention part being obstructed is calculated.
  • the blood cell type of the blood cell retention part is determined by the arithmetic processing unit 70E (step S50).
  • the red blood cell R is determined as a white portion in the red hue range.
  • the white blood cells W may be discriminated using luminance, or may be discriminated as white portions with few holes or white portions with a small number of edges per unit area using the fact that they are larger than other blood cell types.
  • the blood to be measured contains only one of red blood cells R and white blood cells W, this blood cell type discrimination step is omitted.
  • the blockage rate C of the gate 25a is calculated by the arithmetic processing unit 70E (step S60).
  • the ratio of the gate 25a blocked by the blood cell retention part of the volume V is calculated as the blockage ratio C for each volume V of the blood cell retention part.
  • the occlusion ratio C is preferably a ratio with respect to the total number of gates 25a. However, when all the gates 25a are not included in the blood flow image, the ratio is as a ratio with respect to the number of gates 25a included in the blood flow image. Also good.
  • the deformability of the blood cell is calculated by the arithmetic processing unit 70E (step S70).
  • a graph as shown in FIG. 17 is drawn with the horizontal axis and the vertical axis respectively from the volume V and the blockage rate C of the blood cell retention portion calculated in step S40 and step S60, and the blockage rate C of the graph is shown.
  • the deformability of the blood cell is calculated by multiplying the reciprocal of the integrated value by a predetermined coefficient. That, D 1 satisfies the following equation (3) is calculated as a deformability of blood cells.
  • ⁇ 2 is a predetermined coefficient set in a measurement test performed in advance. More specifically, ⁇ 2 is calculated by the blood cell deformability measuring apparatus 1E with the red blood cell deformability measured by another apparatus as positive. The coefficient is set so that the deformability is calculated from the volume V and the occlusion ratio C of the erythrocytes. In the measurement test performed in advance, if ⁇ 2 is not constant with respect to the integral value of the blocking ratio C with respect to the volume V, a lookup table for the integral value may be used.
  • step S60 when the blockage ratio C averaged about all the blood cell retention parts was calculated by step S60, you may calculate D1A which satisfy
  • ⁇ 2 A is a predetermined coefficient set in a measurement test performed in advance, and is a coefficient set in the same manner as ⁇ 2 in the above equation (3).
  • D 1 or D 1A calculated as the deformability indicates that the larger the value, the softer the blood cell and the higher the deformability.
  • the deformability of the blood cells is calculated from the blockage ratio C of the gate 25a and the volume V of the blood cell retention portion. Therefore, the blood cell deformability measuring device 1E is not limited to the red blood cells R having an elliptical shape. Even if it is an amoeba-like leukocyte W, its deformability can be measured. Further, since the deformability of the blood cells passing through the gate 25a having a width smaller than the blood cell diameter is calculated, the deformability when the blood cells pass through the capillaries can be measured by simulating the capillaries by the gate 25a.
  • the flow of blood passing through the gate 25a having a narrower width than the blood cell diameter is displayed on the display 8, the blood cells whose speed has been reduced by the gate 25a can be confirmed, and the blood cells are displayed at a uniform speed on the image. Compared to the conventional flow, it is possible to visually grasp how the blood cells move while deforming.
  • the blood cell deformability measuring apparatus 1F includes a filter 2F instead of the filter 2E in the above embodiment, and the filter 2F is replaced with a flow path portion 25E as shown in FIG. A flow path portion 25F is provided.
  • the flow path portion 25F has a plurality of gates 25aF formed between a plurality of bank portions 22bF,.
  • the widths of the plurality of gates 25aF are narrower than the blood cell diameter and are different from each other.
  • the width is gradually changed in the arrangement direction of the bank portions 22bF.
  • FIG. 19 is a flowchart of deformability measurement by the blood cell deformability measuring apparatus 1F.
  • step T1 blood to be measured is flowed to the filter 2F (step T1), and a blood flow image of the flow path portion 25E is collected (step T2).
  • steps S10 and S20 blood to be measured is flowed to the filter 2F (step T1), and a blood flow image of the flow path portion 25E is collected (step T2).
  • the blood flow image is processed by the arithmetic processing unit 70E, and the blood flow image of the gate 25aF is extracted for each different width (step T3).
  • the blood flow image is divided and extracted by each gate 25aF.
  • step T4 After the gate 25aF closed by the blood cell retention part is detected by the arithmetic processing unit 70E (step T4), the volume V of the blood cell retention part is calculated (step T5), and then the blood cell retention part Are determined (step T6). These steps are performed in the same manner as steps S30 to S50 in the above embodiment.
  • the arithmetic processing unit 70E detects the maximum width of the gate 25aF blocked by the blood cell retention part as the maximum blocking width L (step T7).
  • the deformability of the blood cell is calculated by the arithmetic processing unit 70E (step T8).
  • D satisfying the following equation (4) from the maximum occlusion width L of the blood cell retention portion calculated in step T7 and the volume V of the blood retention portion that occludes the gate 25aF corresponding to the maximum occlusion width L: 2 is calculated as the deformability of blood cells.
  • ⁇ is a predetermined coefficient set in a measurement test performed in advance, and is a coefficient set in the same manner as ⁇ 2 in the above equation (3).
  • the D 3 that satisfies the following equation (5) may be calculated as a deformability of blood cells.
  • L 0 is the volume of the blood cell in the maximum occlusion width L for each volume of the blood cell in the gate 25aF blocked by the blood cell in the blood when a predetermined reference blood is passed to the gate 25aF. It is a value when it is V in Formula (4), and the predetermined reference blood is blood of a standard health level.
  • D 2 calculated as deformability shows that blood cells stiff greater the value.
  • D 3 indicates that the blood cell has a standard hardness when the value is 1, and indicates that the blood cell is harder as it is larger than 1 and softer as it is smaller. If the D 3 is calculated, since deformability is shown as a relative value with respect to blood cells of the standard health, it is possible to understand the degree of uniquely deformability regardless of the volume V of the blood cell trapping portion.
  • the gate 25aF is described as having a width that gradually changes in the arrangement direction of the bank portion 22bF.
  • FIGS. A plurality of stages of gates 25a 1 ,... Having respective widths gradually narrowing in the blood flow direction (Y direction in the figure) may be provided.
  • the gates 25a 1 to 25a 4 formed between the four bank portions 22b 1 to 22b 4 having different arrangement intervals at each stage are provided in the direction of blood flow.
  • Each step has a width that gradually decreases.
  • the blood cell deformability measuring apparatus 1B includes a filter 2B instead of the filter 2E in the above embodiment, and the filter 2B is replaced with a flow path portion 25E as shown in FIG. A flow path portion 25B is provided.
  • the flow path portion 25B has a plurality of gates 25aB formed between a plurality of bank portions 22bB,.
  • the plurality of gates 25aB are formed to have a width narrower than the blood cell diameter and form a plurality of sets for each width.
  • three gates 25aB constitute a set of the bank portion 22bB. It is gradually changing in the arrangement direction.
  • the blood cell deformability measuring apparatus 1B having the above configuration can calculate the deformability of blood cells by the same operation as the blood cell deformability measuring apparatus 1F in the third modified example.
  • the maximum blocking width L is detected (step T7), as shown in FIG. 22, the blocking ratio C of the gate 25aB is obtained for each width of the gate 25aB, and the width of the gate 25aB at which the blocking ratio C is maximized. Is preferably detected as the maximum closing width L.
  • the number of gates 25aB closed is determined according to the width of the gate 25aB, and the gate 25aB width obtained by averaging the blockage numbers or the width of the gate 25aB having the maximum blockage number is maximum blocked.
  • the width L may be detected.
  • the size of blood cells contained in the blood flowing through the gate 25aB is not uniform but has a distribution variation, so that a distribution peak occurs at an intermediate gate width.
  • the blood cell deformability measuring apparatus 1B it has the gate 25aB that forms a plurality of sets for each width as well as the same effect as that of the above-described embodiment. Since the deformability of blood cells can be calculated from the maximum occlusion width L obtained as the width of the gate 25aB where the occlusion ratio C by width of 25aB is the maximum, the deformability is calculated using more gates 25aB, and is more stable. A calculation result is obtained.
  • the blood cell deformability measuring apparatus 1C includes a filter 2C instead of the filter 2E in the above embodiment, and the filter 2C is replaced with a flow path portion 25E as shown in FIG. A flow path portion 25C is provided.
  • the flow path portion 25C has a plurality of gates 25aC formed between a plurality of bank portions 22bC,... Which are tapered in the blood flow direction (Y direction in the drawing). .
  • the width of the plurality of gates 25aC is gradually narrowed in the blood flow direction and narrower than the blood cell diameter.
  • the blood cell deformability measuring apparatus 1C having the above configuration can calculate the deformability of blood cells by the same operation as the blood cell deformability measuring apparatus 1F in the third modified example.
  • the width of the gate 25aC at the position where the blood cell retention portion is retained is the maximum occlusion width L. Detect as.
  • the blood flow image in the gate 25aC formed with a width narrowing in the blood flow direction can be obtained as well as the same effect as the above embodiment. Since it is displayed on the display 8, it is easy to visually grasp the width of the gate 25aC blocked by the blood cell retention portion.
  • red blood cells R and / or white blood cells W is calculated, but among leukocytes W, granulocytes, lymphocytes and monocytes are further discriminated, Each of these deformability may be calculated. In this way, more detailed blood diagnosis can be performed.
  • a known method described in, for example, JP-A-2001-174456 may be used.
  • Blood cell deformability measuring device 3 TV camera (photographing means) 7, 7A, 7E PC (blood flow image display means) 8 Display (display screen) 25, 25A, 25B, 25C, 25E, 25F Channel portion 25a Gate 25b Upstream terrace 70, 70A Arithmetic processing section (speed calculation means, volume calculation means, deformability calculation means) 70E arithmetic processing unit (speed calculation means, volume calculation means, deformability calculation means, blockage state calculation means, blockage ratio calculation means, blockage width detection means) S Blood cell velocity T Passing time V Blood cell volume C Blockage ratio L Maximum blockage width

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PCT/JP2009/065813 2008-10-24 2009-09-10 血球変形能計測装置 WO2010047191A1 (ja)

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WO2010137470A1 (ja) * 2009-05-29 2010-12-02 コニカミノルタオプト株式会社 変形能計測装置及び変形能計測方法
US20190114790A1 (en) * 2017-10-13 2019-04-18 University Of Rochester Rapid assessment and visual reporting of local particle velocity

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CN108918898A (zh) * 2018-05-16 2018-11-30 南方医科大学 体外检测脂多糖对红细胞变形能力影响的方法及其应用

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JP2685544B2 (ja) * 1988-11-11 1997-12-03 株式会社日立製作所 血液フィルタおよび血液検査方法並びに血液検査装置
JP2720161B2 (ja) * 1988-02-01 1998-02-25 株式会社アドバンス 細胞変形能測定装置
JP2001507122A (ja) * 1996-11-26 2001-05-29 コールター インターナショナル コーポレイション 個々の赤血球の形状を決定する装置及び方法

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JP2720161B2 (ja) * 1988-02-01 1998-02-25 株式会社アドバンス 細胞変形能測定装置
JP2685544B2 (ja) * 1988-11-11 1997-12-03 株式会社日立製作所 血液フィルタおよび血液検査方法並びに血液検査装置
JP2001507122A (ja) * 1996-11-26 2001-05-29 コールター インターナショナル コーポレイション 個々の赤血球の形状を決定する装置及び方法

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Publication number Priority date Publication date Assignee Title
WO2010137470A1 (ja) * 2009-05-29 2010-12-02 コニカミノルタオプト株式会社 変形能計測装置及び変形能計測方法
US20190114790A1 (en) * 2017-10-13 2019-04-18 University Of Rochester Rapid assessment and visual reporting of local particle velocity
US10803601B2 (en) * 2017-10-13 2020-10-13 University Of Rochester Rapid assessment and visual reporting of local particle velocity

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