KR20200059671A - Apparatus and method for multiple measurement of blood biophysical property using microfluidic device - Google Patents

Abstract

The present invention relates to an apparatus for measurement of multiple blood biophysical properties using a microfluidic device, and a method therefor. The apparatus for measurement of multiple blood biophysical properties using a microfluidic device comprises a microfluidic device in which a channel through which a fluid to be measured passes is formed. The microfluidic device comprises: an injection hole through which the fluid to be measured is injected; a main channel communicating with the injection hole and allowing the fluid to be measured to pass therethrough; an expansion channel communicating with the main channel and having a diameter wider than that of the main channel; a branch channel communicating with the expansion channel and allowing the fluid to be measured to pass therethrough; a first discharge hole through which the fluid to be measured is discharged; a first connection channel separated from the branch channel and communicating with the first discharge hole through which the fluid to be measured is discharged; an expansion chamber communicating with the expansion channel through the branch channel and including micro channels formed therein and separated from each other by a micro gap; a second discharge hole through which the fluid to be measured is discharged; and a second connection channel communicating with the expansion channel and the second discharge hole. A method for measurement of multiple blood biophysical properties using a microfluidic device is configured to measure multiple blood biophysical properties using the apparatus for measurement of multiple blood biophysical properties using a microfluidic device.

Description

{Apparatus and method for multiple measurement of blood biophysical property using microfluidic device}

The present invention relates to a multi-biological value measuring device and a measuring method of blood using a microfluidic device, and more specifically, based on the microfluidic device, measuring three blood biologic values-red blood cell deformability, red blood cell aggregation, and hematocrit at the same time. It relates to a multi-biological value measuring device and a measuring method for blood using a microfluidic device capable of.

Recently, the number of cardiovascular disease patients such as hypertension, diabetes, and heart disease is steadily increasing. In particular, cardiovascular diseases such as arteriosclerosis, myocardial infarction, and brain hemorrhage occur suddenly without prior symptoms, causing death or serious complications. have. However, there is a problem in that early diagnosis of such diseases is difficult only with the conventional biochemical detection method.

Therefore, biophysical diagnostic technology has been developed to effectively cope with such diseases. Recently, many studies have been conducted on blood bioactivity-viscosity, viscoelasticity, hematocrit, deformability, cohesiveness, etc., and blood bioactivity is relatively related to symptoms such as hypertension, diabetes, and smoking, which are known as risk factors for cardiovascular diseases. It turned out to be high.

Therefore, measuring blood bioactivity plays an important role in the early diagnosis of cardiovascular disease patients who develop without prior symptoms, and accordingly, biosensors capable of measuring blood bioactivity have been developed. However, the conventional biosensor has the following problems.

Conventionally, biosensors capable of measuring red blood cell deformability, red blood cell aggregation, and hematocrit among blood biologic values have been developed. However, such a conventional biosensor could not measure erythrocyte deformability, erythrocyte cohesiveness, and hematocrit at the same time, and there is a problem in that each blood bioactivity must be measured separately.

The present invention was created to solve the above-mentioned problems, and more specifically, based on the microfluidic device, uses three microbial devices that can simultaneously measure three blood biologic values-red blood cell deformability, red blood cell aggregation, and hematocrit. It relates to a multi-biological value measuring device and a measuring method for blood.

The apparatus for measuring multiple biological properties of blood using a microfluidic device for solving the above-described problem includes a microfluidic device in which a channel through which the measurement target fluid passes is formed, wherein the microfluidic device is the target fluid. Inlet can be injected; A main channel in communication with the inlet and through which the fluid to be measured passes; An expansion channel communicating with the main channel and formed with a diameter larger than the diameter of the main channel; A branch channel in communication with the expansion channel and through which the fluid to be measured passes; A first outlet through which the fluid to be measured can be discharged; A first connection channel that is separated from the branch channel and communicates with a first discharge port through which the fluid to be measured is discharged; An expansion chamber in communication with the expansion channel through the branch channel, and having a micro channel disposed at a fine interval therein; A second outlet through which the fluid to be measured can be discharged; It is characterized in that it comprises a second connection channel in communication with the second outlet, while communicating with the expansion chamber.

The apparatus for measuring multiple biological properties of blood using a microfluidic device for solving the above-described problem may be connected to the injection port and include a syringe pump capable of injecting the measurement target fluid into the injection port.

The syringe pump of the multi-biological value measuring device for blood using a microfluidic device for solving the above-described problems may be periodically operated and stopped to inject the fluid to be measured into the inlet, and the syringe pump The measurement target fluid and a reference fluid may be injected into the injection port.

The second outlet of the multi-biological value measuring device for blood using a microfluidic device for solving the above-described problem is connected to a tube filled with a reference fluid, and the tube may be blocked on one side through a pinch valve.

The apparatus for measuring multi-biological values of blood using a microfluidic device for solving the above-described problem may include an image acquisition device capable of measuring the image intensity of a specific region of interest, and the red blood cell deformability index of the branch channel It can be derived through the flow rate (<U _B >) and the cross-sectional area (A _C ) of the branch channel.

A first specific region of interest is selected in the expansion chamber of the multi-biological value measuring device for blood using a microfluidic device for solving the above-described problem, and the first of the first specific region of interest is obtained through the image acquisition device. The image intensity (<I _MP >) is measured, and the red blood cell cohesiveness index can be derived through the first image intensity (<I _MP >), and a second specific region of interest is selected inside the expansion channel, and the The second image intensity (<I _BC >) of the second specific region of interest is measured by the image acquisition device, and the hematocrit index can be derived through the second image intensity (<I _BC >).

In order to solve the above-mentioned problem, a method for measuring multi-biological values of blood using a microfluidic device is a device for measuring multi-biological values of blood using a microfluidic device that simultaneously measures red blood cell deformability, red blood cell cohesiveness, and hematocrit of a fluid to be measured. As a method for measuring the multi-biological value of blood through, the apparatus for measuring multi-biological value of blood using the microfluidic device includes a microfluidic device in which a channel through which the measurement target fluid passes is formed, and the microfluidic device The device may include an injection port through which the fluid to be measured is injected; A main channel in communication with the inlet and through which the fluid to be measured passes; An expansion channel communicating with the main channel and formed with a diameter larger than the diameter of the main channel; A branch channel in communication with the expansion channel and through which the fluid to be measured passes; A first outlet through which the fluid to be measured can be discharged; A first connection channel that is separated from the branch channel and communicates with a first discharge port through which the fluid to be measured is discharged; An expansion chamber in communication with the expansion channel through the branch channel, and having a micro channel disposed at a fine interval therein; A second outlet through which the fluid to be measured can be discharged; An injection step of injecting the measurement target fluid into the microfluidic device through the injection port, the second connection channel communicating with the expansion chamber while communicating with the expansion chamber; Filling the microfluidic device with the fluid to be measured; A measurement step of measuring a flow velocity or image intensity at a designated point of the microfluidic device; It characterized in that it comprises a; deriving step of deriving the erythrocyte deformability index, erythrocyte cohesiveness index, hematocrit index through the flow rate or image intensity measured through the measurement step.

The apparatus for measuring multiple biological properties of blood using the microfluidic device of the method for measuring multiple biological properties of blood using a microfluidic device for solving the above-described problem is connected to the inlet, and the target fluid is measured at the inlet It may include a syringe pump capable of injecting.

In the injection step of the method for measuring a multi-biological value of blood using a microfluidic device for solving the above-described problem, the measurement target fluid may be injected into the injection port while periodically operating and stopping the syringe pump, In the injection step, the measurement target fluid and the reference fluid are injected into the injection port through the syringe pump, and in the filling step, the measurement target fluid and the reference fluid may be filled in the microfluidic device.

The second outlet of the method for measuring multiple biological properties of blood using a microfluidic device for solving the above-described problem is connected to a tube filled with a reference fluid, and the tube may be blocked on one side through a pinch valve.

The apparatus for measuring multiple biological properties of blood using the microfluidic device of the method for measuring multiple biological properties of blood using the microfluidic device for solving the above-described problem acquires an image capable of measuring the image intensity of a specific region of interest It may include a device, the measuring step, the flow rate of the branch channel (<U _B >) is measured, the red blood cell deformability index in the derivation step, the flow rate of the branch channel (<U _B >) and the It can be derived through the cross-sectional area (A _C ) of the branch channel.

In the measuring step of a method for measuring multiple biological properties of blood using a microfluidic device for solving the above-described problem, a first specific region of interest is selected inside the expansion chamber, and the first specific is obtained through the image acquisition device. The first image intensity (<I _MP >) of the region of interest is measured, and in the derivation step, the erythrocyte cohesiveness index can be derived through the first image intensity (<I _MP >), and the measurement step comprises: A second specific region of interest is selected in the extended channel, and the second image intensity (<I _BC >) of the second specific region of interest is measured through the image acquisition device, and the hematocrit index in the deriving step is: It may be derived through the second image intensity (<I _BC >).

The present invention relates to a device for measuring and measuring multiple biological properties of blood using a microfluidic device capable of simultaneously measuring three blood biologic values-erythrocyte deformability, erythrocyte cohesion, and hematocrit based on a microfluidic device. It is possible to measure both deformability and erythrocyte cohesion at the same time, and also has the advantage of simultaneously measuring hematocrit affecting red blood cell deformability and erythrocyte cohesion.

In the present invention, three blood biologic values-red blood cell deformability, erythrocyte cohesion, and hematocrit are simultaneously measured, and thus, red blood cell deformability, protein contribution in plasma, and the effect of red blood cell number can be simultaneously examined, through which blood bioactivity values for each circulatory disease There is an advantage that can effectively evaluate the impact of the change.

1 is a view showing a device for measuring multiple biological properties of blood using a microfluidic device according to an embodiment of the present invention.
Figure 2 is the flow rate of the main channel of the apparatus for measuring multi-biological values of blood using the microfluidic device according to an embodiment of the present invention (<U _M >), the flow rate of the branch channel (<U _B >), the first specific interest This is a diagram showing an area and a second area of interest.
3 is a view showing the main dimensions of a multi-biological value measuring device for blood using a microfluidic device according to an embodiment of the present invention.
4 is a flow rate of a main channel (<U _M >) and a flow rate of a branch channel (<U _B >) when blood is injected into a device for measuring multi-biological values of blood using a microfluidic device according to an embodiment of the present invention. ), The first image intensity (<I _MP >), the second image intensity (<I _BC >) and the erythrocyte deformability index (DI), erythrocyte aggregation index (AI _N ), and hematocrit index () H _iBC ).
5 is a flow chart of a method for measuring multiple biological properties of blood using a microfluidic device according to an embodiment of the present invention.
6 and 7 are red blood cell deformability of the blood through multiple biological Toughness Measurement device and measurement of the blood using a microfluidic device according to an embodiment of the invention index (DI), red blood cell cohesion index (RBC aggregation index, AI _N ), Hematocrit index (H _iBC ) is an example of measuring at the same time.

Hereinafter, various embodiments of the present invention will be described in connection with the accompanying drawings. Various embodiments of the present invention may have various modifications and various embodiments, and specific embodiments are illustrated in the drawings and related detailed descriptions are described. However, this is not intended to limit the various embodiments of the present invention to specific embodiments, and should be understood to include all modifications and / or equivalents or substitutes included in the spirit and scope of the various embodiments of the present invention. In connection with the description of the drawings, similar reference numerals have been used for similar elements.

Expressions such as “comprises” or “can include” that may be used in various embodiments of the present invention indicate the existence of a corresponding function, operation, or component that has been invented, and additional one or more functions, operations or The components and the like are not limited. Further, in various embodiments of the present invention, terms such as “include” or “have” are intended to indicate that there are features, numbers, steps, operations, components, parts, or combinations thereof described in the specification, It should be understood that one or more other features or numbers, steps, operations, components, parts or combinations thereof are not excluded in advance.

When it is mentioned that a component is "connected" to another component, the other component may be directly connected to the other component, but another new component between the other component and the other component It should be understood that may exist. On the other hand, when it is mentioned that an element is "directly connected" or "directly connected" to another component, it will be understood that no other new component exists between the one component and the other components. You should be able to.

Terms used in various embodiments of the present invention are only used to describe specific specific embodiments, and are not intended to limit various embodiments of the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person skilled in the art to which various embodiments of the present invention pertain.

Terms, such as those defined in a commonly used dictionary, should be interpreted as having meanings consistent with meanings in the context of related technologies, and are ideally or excessively formal unless explicitly defined in various embodiments of the present invention. It is not interpreted as meaning.

The present invention relates to a multi-biological value measuring apparatus and a measuring method for blood using a microfluidic device, and based on the microfluidic device, three blood biologic values-red blood cell deformability, red blood cell cohesion, and hematocrit capable of simultaneously measuring hematocrit It relates to a multi-biological value measuring device and a measuring method for blood using a fluid element. Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

Referring to FIG. 1, a device for measuring multi-biological values of blood using a microfluidic device according to an embodiment of the present invention includes a microfluidic device 100, wherein the microfluidic device 100 is an inlet 110, Main channel 120, expansion channel 130, branch channel 140, first outlet 150, first connection channel 160, expansion chamber 170, second outlet 180, second connection channel (190).

The fluid to be measured according to an embodiment of the present invention may be blood, and the fluid to be measured is filled while being injected into the microfluidic device 100, and the flow velocity and image intensity of a designated point inside the microfluidic device 100 Red blood cell deformability, red blood cell cohesiveness, and hematocrit may be measured by measuring.

The injection port 110 is a point where the target fluid can be injected into the microfluidic device 100, and the target fluid is injected into the microfluidic device 100 through the injection port 110.

Referring to FIG. 1, the injection port 110 may be connected to a syringe pump 210. The syringe pump 210 is a place where the measurement target fluid can be stored, and the measurement target fluid stored in the syringe pump 210 is injected into the injection port 110.

Here, the reference fluid may be stored together with the measurement target fluid in the syringe pump 210, and the measurement target fluid and the reference fluid may be injected into the injection port through the syringe pump 210. Specifically, the fluid to be measured may be blood, the reference fluid may be saline, and blood and saline may be injected into the inlet 110 through the syringe pump 210. (However, the saline solution is injected at a constant flow rate.)

According to an embodiment of the present invention, prior to the experiment through the microfluidic device 100, the reference fluid may be injected into the inlet 110 through the syringe pump 210, and the microfluidic fluid When an experiment is performed through the device 100, the measurement target fluid may be injected into the injection port 110 through the syringe pump 210.

When the fluid to be measured (or the reference fluid) is injected through the syringe pump 210, the syringe pump periodically operates and stops to inject the fluid to be measured into the injection port. Can be. Referring to FIG. 1, the syringe pump 210 includes a period (T _on ) for injecting the object to be measured and a period (T _off ) for not injecting the object to be measured (T _off ). While forming, the measurement target fluid may be injected into the injection port 110.

In one embodiment of the present invention, the cycle (T) of the syringe pump 210 may be 240s, and the section (T _on ) injected by the fluid to be measured through the syringe pump 210 may be 210s. In addition, a section (T _off ) in which the fluid to be measured is not injected through the syringe pump 210 may be 30 s. However, the present invention is not limited thereto, and the cycle T of the syringe pump 210 may be changed as necessary.

2 and 3, the main channel 120 is in communication with the injection port 110, and is a channel through which the measurement fluid injected through the injection port 110 can pass. The measurement target fluid passing through the main channel 120 may have a flow velocity (<U _M >), and the flow velocity of the main channel 120 may be measured through micro-PIV (Particle Image Velocimetry). . (The flow rate of blood in the microfluidic device 100 may be measured by various flow rate measurement devices, and may be measured through the micro-PIV (Particle Image Velocimetry).)

2 and 3, the expansion channel 130 is in communication with the main channel 120 and may be formed to have a larger diameter than the diameter of the main channel 120. Specifically, the main channel 120 may be formed in a tubular shape, and the expansion channel 120 may be formed in a circular shape. Referring to FIG. 3, when the expansion channel 120 is formed in a circular shape, the diameter of the expansion channel 130 may be 1500 μm.

As will be described later, a specific region of interest is selected from the extension channel 130 in a circular shape, and an image intensity of the specific region of interest is measured to derive an index of hematocrit through the image intensity.

1 to 3, the branch channel 140 is in communication with the expansion channel 130, and the measurement target fluid passing through the expansion channel 130 can pass through. Specifically, the main channel 120 is a channel connecting the inlet 110 and the expansion channel 130, the branch channel 140 is the expansion channel 130 and the expansion chamber 170 to be described later Is to connect it. The measurement target fluid that has passed through the expansion channel 130 passes through the branch channel 140 and moves to the expansion chamber 170. Referring to FIG. 3, the diameter of the branch channel 140 may be 250 μm.

1 to 3, the first outlet 150 is a point at which the measurement object fluid can be discharged, and the first connection channel 160 is split from the branch channel 140 and the measurement object It is in communication with the first outlet 150 through which fluid can be discharged. The first connection channel 160 is branched from the branch channel 140 and is in communication with the first outlet 150, and the measurement target fluid passing through the branch channel 140 is the first connection channel. It may be discharged to the first discharge port 150 through (160).

The expansion chamber 170 is in communication with the expansion channel 130 through the branch channel 140, and is formed with a micro channel 171 disposed at minute intervals therein. The expansion chamber 170 is disposed at minute intervals, and the micro-channel 171 having a gap between channels is disposed. A specific region of interest is selected within the expansion chamber 170, and the region of the specific region of interest is selected. By measuring the image intensity, it is possible to derive the red blood cell cohesiveness index through the image intensity. (Here, according to an embodiment of the present invention, the dimension of the expansion chamber 170 may be made as shown in FIG. 3)

Referring to FIG. 1, the second outlet 180 is a point at which the fluid to be measured can be discharged, and the second connection channel 190 includes the expansion chamber 170 and the second outlet 180. In communication, the expansion chamber 170 and the second outlet 180 may be connected. The fluid to be measured passing through the expansion chamber 170 passes through the second connection channel 190 and is discharged to the second outlet 180.

The second outlet 180 may be connected to a tube 220 filled with a reference fluid to effectively measure red blood cell cohesiveness at the entrance of the expansion chamber 170 (second specific region of interest to be described later). The tube 220 is installed for back-and-forth blood flow in the expansion chamber 170, and for this purpose, the tube 220 is filled with a reference fluid, and then the tube (through the pinch valve 221) The one side of the tube 220 is blocked while clamping one side of 220). Here, the reference fluid may be saline, or other fluid, if necessary.

The apparatus for measuring multiple biological properties of blood using a microfluidic device according to an embodiment of the present invention may further include an image acquisition device capable of measuring the image intensity of a specific region of interest, and the microstructure according to an embodiment of the present invention The apparatus for measuring multi-biological values of blood using a fluid device measures image intensity measured through an image acquisition device and blood flow rate inside the microfluidic device 100 to simultaneously measure red blood cell deformability, red blood cell cohesiveness, and hematocrit. have. Here, according to an embodiment of the present invention, the blood flow rate inside the microfluidic device 100 may be measured through micro-PIV (Particle Image Velocimetry).

According to an embodiment of the present invention, the red blood cell deformability index (DI) may be derived through a flow rate (<U _B >) of the branch channel 140 and a cross-sectional area (A _C ) of the branch channel 140. Specifically, the red blood cell deformability index (DI) may be derived as shown in Equation 1 below.

Here, A _C denotes the cross-sectional area of the branch channel 140, T denotes the cycle of the syringe pump 210, and <U _B > denotes the flow rate of the branch channel 140.

According to an embodiment of the present invention, the red blood cell aggregation index (RBC aggregation index, AI _N ) may be derived as follows. A first specific region of interest 310 (ROI-585 × 60 pixels) is selected inside the expansion chamber 170, and the first image intensity of the first specific region of interest 310 through the image acquisition device (< I _MP >). Here, the first specific region of interest 310 is a point shown inside the expansion chamber 170 of FIG. 2 as an entrance point of the micro-channel 171 inside the expansion chamber 170.

The erythrocyte aggregation index (RBC aggregation index, AI _N ) may be derived through the first image intensity (<I _MP >). Specifically, the red blood cell cohesiveness index (AI _N ) may be derived as shown in Equation 2 below.

Here, <I _MP > is the first image intensity of the first specific region of interest 310, t _o is when the measurement target fluid is injected from the syringe pump 210, the <I _MP > It is a time point at which the value is increased, and t _s indicates the start time point of a section in which the target fluid is not injected in the syringe pump 210.

According to an embodiment of the present invention, the hematocrit index (H _iBC ) may be derived as follows. A second specific region of interest 320 (ROI-220 × 220 pixels) is selected inside the extension channel 130, and a second image intensity of the second specific region of interest 320 (<I _BC >). Here, the second specific region of interest 320 is a point inside the extension channel 130 as a point inside the extension channel 130 of FIG. 2.

The hematocrit index (H _iBC ) may be derived through the second image intensity (<I _BC >). Specifically, the hematocrit index (H _iBC ) can be derived as shown in Equation 3 below.

Here, <I _BC > is the second image intensity of the second specific region of interest 320, and T is the cycle of the syringe pump 210.

The apparatus for measuring multi-biological values of blood using a microfluidic device according to an embodiment of the present invention includes the red blood cell deformability index (DI), the red blood cell cohesiveness index (RBC aggregation index) through the microfluidic device 100 as described above. AI _N ), the hematocrit index (H _iBC ) can be measured at the same time.

4 is a flow rate (<U _M >) of the main channel when injecting the measurement target fluid while forming a cycle T through the syringe pump 210 according to an embodiment of the present invention, the branch The flow rate of channel 140 (<U _B >), the first image intensity (<I _MP >), the second image intensity (<I _BC >) and the red blood cell deformability index (DI) measured accordingly, the Red blood cell aggregation index (RBC aggregation index, AI _N ), it represents the hematocrit index (H _iBC ). Specifically, it is an experimental result when the fluid to be measured is blood (normal RBCs in dextran solution [20 mg / mL], Hct = 50%).

Referring to FIG. 5, a method for measuring multi-biological values of blood using a microfluidic device according to an embodiment of the present invention includes an injection step (S110), a filling step (S120), a measurement step (S130), and a derivation step (S140). It includes. The method for measuring the multi-biological value of blood using the microfluidic device according to the embodiment of the present invention is the multi-biological value of blood through the apparatus for measuring the multi-biological value of blood using the microfluidic device according to the embodiment of the present invention described above. As a method of measuring, a detailed description of the apparatus for measuring multiple biological properties of blood using a microfluidic device is omitted.

The injection step (S110) is a step of injecting the fluid to be measured into the microfluidic device 100 through the injection port 110. Here, the fluid to be measured may be blood. Referring to FIG. 1, the injection port 110 may be connected to a syringe pump 210. The syringe pump 210 is a place where the fluid to be measured can be stored, and in the injection step (S110), the fluid to be measured stored in the syringe pump 210 is injected into the injection port 110. do.

Here, the reference fluid may be stored together with the measurement target fluid in the syringe pump 210, and in the injection step (S110), the measurement target fluid and the reference fluid are through the syringe pump 210. It can be injected into the inlet. Specifically, the fluid to be measured may be blood, and the reference fluid may be saline, and blood and saline may be injected into the inlet 110 through the syringe pump 210. (However, the saline solution is injected at a constant flow rate.)

When the measurement target fluid and the reference fluid are injected into the injection port 110 through the syringe pump 210 in the injection step (S110), in the filling step (S120), the measurement target fluid and the reference fluid Is filled in the microfluidic device 100.

When the measurement target fluid (or the reference fluid) is injected into the injection port 110 through the syringe pump 210 in the injection step (S110), the measurement target is periodically operated and stopped. Fluid may be injected into the inlet 110. Referring to FIG. 1, the syringe pump 210 includes one cycle T including a section T _on injecting the target fluid and a section T _off not injecting the target fluid. While forming, the measurement target fluid may be injected into the injection port 110.

In one embodiment of the present invention, the cycle (T) of the syringe pump 210 may be 240s, and the section (T _on ) injected by the fluid to be measured through the syringe pump 210 may be 210s. In addition, a section (T _off ) in which the target fluid is not injected through the syringe pump 210 may be 30 s. However, the present invention is not limited thereto, and the cycle T of the syringe pump 210 may be changed as necessary.

The filling step (S120) is a step of filling the microfluidic device 100 with the measurement target fluid, and filling the microfluidic device 100 through the measurement target fluid injected through the injection step (S110).

The measuring step (S130) is a step of measuring the flow rate or image intensity of the designated point of the microfluidic device 100, and the deriving step (S140) is the flow rate or image intensity measured through the measuring step (S130). This is the step of deriving the red blood cell deformability index, red blood cell cohesiveness index, and hematocrit index.

The apparatus for measuring multiple biological properties of blood using a microfluidic device according to an embodiment of the present invention is an image acquisition device capable of measuring the image intensity of a specific region of interest and the blood flow rate inside the microfluidic device 100. It may further include a micro-PIV (Particle Image Velocimetry), and in the measurement step (S130), the designated point of the microfluidic device 100 through the image acquisition device and the Micro-PIV (Particle Image Velocimetry). Can measure the flow velocity or image intensity.

The deriving step (S140) in the red blood cell deformability index (DI) via a flow rate or image intensity measured by the measuring step (S130), the red blood cell cohesion index (RBC aggregation index, AI _N) in accordance with an embodiment of the present invention , The hematocrit index (H _iBC ) is derived.

The red blood cell deformability index (DI), said measuring step (S130) the flow rate of the branch channel in the (<U _B>) the measurement, the flow rate of the branch channel in the deriving step _{(S140) (<U B>} ) and It can be derived through the cross-sectional area (A _C ) of the branch channel. Specifically, in the derivation step (S140), the red blood cell deformability index (DI) may be derived as shown in Equation 4 below.

Here, A _C denotes the cross-sectional area of the branch channel 140, T denotes the cycle of the syringe pump 210, and <U _B > denotes the flow rate of the branch channel 140.

The red blood cell aggregation index (RBC aggregation index, AI _N ) selects a first specific region of interest (310) (ROI) inside the expansion chamber 170 in the measurement step (S130), and through the image acquisition device The first image intensity (<I _MP >) of the first specific region of interest 310 may be measured, and may be derived through the first image intensity (<I _MP >) in the deriving step S140. Here, the first specific region of interest 310 is a point shown inside the expansion chamber 170 of FIG. 2 as an entrance point of the micro-channel 171 inside the expansion chamber 170. Specifically, in the derivation step (S140), the red blood cell aggregation index (RBC aggregation index, AI _N ) may be derived as shown in Equation 5 below.

Here, <I _MP > is the first image intensity of the first specific region of interest 310, t _o is when the measurement target fluid is injected from the syringe pump 210, the <I _MP > It is a time point at which the value is increased, and t _s indicates the start time point of a section in which the target fluid is not injected in the syringe pump 210.

The hematocrit index (H _iBC ) selects a second specific region of interest 320 (ROI) inside the extended channel 140 in the measurement step (S130), and the second specific region of interest through the image acquisition device. By measuring the second image intensity (<I _BC >) of 320, it may be derived through the second image intensity (<I _BC >) in the derivation step (S140). Here, the second specific region of interest 320 is a point inside the extension channel 130 and is a point shown inside the extension channel 130 of FIG. 2. Specifically, the hematocrit index (H _iBC ) may be derived as in Equation 6 below.

Here, <I _BC > is the second image intensity of the second specific region of interest 320, and T is the cycle of the syringe pump 210.

The method for measuring multi-biological values of blood using a microfluidic device according to an embodiment of the present invention includes the erythrocyte modification index (DI), the erythrocyte aggregation index (AI _N ), and the hematocrit index through the above-described method. It is possible to measure (H _iBC ) simultaneously.

6 and Fig. 7 is an embodiment in accordance with an embodiment of the invention measures the blood erythrocyte deformability index (DI), red blood cell cohesion index _{(RBC aggregation index, AI N)} , the hematocrit index (H _iBC) at the same time.

6, normal RBCs are fixed with GA (glutaraldehyde) solution and injected into plasma (autologous plasma). Here, the hematocrit was set to 50%. 6 (a) shows a change in the red blood cell deformability index (DI) when the red blood cell deformability is reduced using a GA (glutaraldehyde) solution. As the concentration of _GA (C _GA ) increases, the deformability of RBC deteriorates, and as a result, <U _B > is significantly lowered. Accordingly, the red blood cell deformability index (DI) shows a tendency to decrease significantly with increasing GA concentration (C _GA ). The concentration of _GA (C _GA ) after 2 μL / mL shows a certain trend.

FIG. 6 (b) shows changes in the erythrocyte aggregation index (AI _N ) and hematocrit index (H _iBC ) according to the GA concentration (C _GA ). The red blood cell aggregation index (RBC aggregation index, AI _N ) shows a tendency to decrease gradually as the GA concentration (C _GA ) increases. Referring to FIG. 6 (b), the hematocrit index (H _iBC ) is affected by the red blood cell deformability index (DI). As described above, the apparatus for measuring and measuring multiple biological properties of blood using a microfluidic device according to an embodiment of the present invention has an advantage of simultaneously measuring three blood biological properties.

FIG. 7 shows a blood sample prepared by partially mixing red blood cells cured to GA concentration (C _GA ) = 2 μL / mL and normal red blood cells when the blood contains partially deformable red blood cells. Here, the mixing ratio (ψ) = V _H / (V _H + V _N ), V _H is the volume of the blood containing the cured red blood cells, V _N is the volume of the blood containing the normal red blood cells.

Referring to FIG. 7 (a), the red blood cell deformability index (DI) is significantly reduced as the mixing ratio increases. In particular, for a mixing ratio of 10% or more, the red blood cell deformability index (DI) is remarkably low and shows the same tendency. That is, it is possible to sufficiently detect red blood cell deformability through an apparatus for measuring and measuring multiple biological properties of blood using a microfluidic device according to an embodiment of the present invention.

Figure 7 (b) shows the change in the erythrocyte aggregation index (RBC aggregation index, AI _N ), hematocrit index (H _iBC ) according to the mixing ratio. Referring to FIG. 7 (b), it can be seen that the erythrocyte aggregation index (AI _N ) and the hematocrit index (H _iBC ) are affected according to the mixing ratio. As shown in the above results, the three blood biomass indexes (RBC deformability index (DI), RBC aggregation index (AI _N ), and hematocrit index (H _iBC )) are affected by the proportion of erythrocytes with poor deformability. You can see that

The apparatus for measuring and measuring multiple biological properties of blood using a microfluidic device according to an embodiment of the present invention has the following effects.

The apparatus for measuring and measuring multiple biological properties of blood using a microfluidic device according to an embodiment of the present invention can simultaneously measure red blood cell deformability and red blood cell cohesiveness based on the microfluidic device 100, together with red blood cell deformability and There is an advantage in that hematocrit affecting red blood cell cohesion can be measured simultaneously.

The apparatus for measuring and measuring multiple biological properties of blood using a microfluidic device according to an embodiment of the present invention evaluates three blood biological properties-red blood cell deformability, erythrocyte cohesion, and hematocrit, simultaneously measuring red blood cell deformability and protein contribution in plasma , The effect of the number of red blood cells can be reviewed at the same time, and through this, it is possible to effectively evaluate the effect of changes in blood biologic values for each circulatory disorder.

The present invention has been described in detail with reference to preferred embodiments, but the present invention is not limited to the above embodiments, and various modifications may be provided without departing from the scope of the present invention. Therefore, the true technical protection scope of the present invention should be determined by the technical spirit of the appended claims.

100 ... fine fluid element 110 ... inlet
120 ... main channel 130 ... extended channel
140 ... branch channel 150 ... first outlet
160 ... first connection channel 170 ... expansion chamber
171 ... micro channel 180 ... second outlet
190 ... second connection channel 210 ... syringe pump
220 ... tube 221 ... pinch valve
310 ... first specific area of interest 320 ... second specific area of interest
S110 ... Injection stage
S120 ... Filling step
S130 ... Measurement step
S140 ... During phase

Claims (18)

A device for measuring multi-biological values of blood using a microfluidic device that simultaneously measures red blood cell deformability, red blood cell cohesiveness, and hematocrit of a fluid to be measured,
And a microfluidic device in which a channel through which the measurement target fluid passes is formed,
The microfluidic device,
An injection hole into which the measurement target fluid is injected;
A main channel in communication with the inlet and through which the fluid to be measured passes;
An expansion channel communicating with the main channel and formed with a diameter larger than the diameter of the main channel;
A branch channel in communication with the expansion channel and through which the fluid to be measured passes;
A first outlet through which the fluid to be measured can be discharged;
A first connection channel that is separated from the branch channel and communicates with a first discharge port through which the fluid to be measured is discharged;
An expansion chamber in communication with the expansion channel through the branch channel, and having a micro channel disposed at a fine interval therein;
A second outlet through which the fluid to be measured can be discharged;
A device for measuring multiple biological properties of blood using a microfluidic device, characterized in that it comprises a second connection channel in communication with the second outlet while communicating with the expansion chamber. According to claim 1,
It is connected to the inlet, and the multi-biological value measuring device for blood using a microfluidic device, characterized in that it comprises a syringe pump capable of injecting the fluid to be measured into the inlet. According to claim 2,
The syringe pump is a device for measuring multi-biological values of blood using a microfluidic device, characterized in that the fluid to be measured is injected into the inlet while periodically operating and stopping. According to claim 2,
The syringe pump is a device for measuring multiple biological values of blood using a microfluidic device, characterized in that the measurement target fluid and a reference fluid are injected into the injection port. According to claim 1,
The second outlet is connected to a tube filled with a reference fluid, the tube is a multi-biological value measuring device for blood using a microfluidic device, characterized in that one side is blocked through a pinch valve. According to claim 1,
Multi-biological value measurement device for blood using a microfluidic device, characterized in that it comprises an image acquisition device capable of measuring the image intensity of a specific region of interest. According to claim 3,
The red blood cell deformability index,
Multi-biological value measuring device for blood using a microfluidic device, characterized in that derived from the flow rate (<U _B >) of the branch channel and the cross-sectional area (A _C ) of the branch channel. The method of claim 6,
A first specific region of interest is selected inside the expansion chamber, and the first image intensity (<I _MP >) of the first specific region of interest is measured through the image acquisition device,
The erythrocyte cohesiveness index,
A device for measuring multiple biological properties of blood using a microfluidic device, characterized in that it is derived through the first image intensity (<I _MP >). The method of claim 6,
A second specific region of interest is selected within the extended channel, and the second image intensity (<I _BC >) of the second specific region of interest is measured through the image acquisition device,
Hematocrit index,
Multi-biological value measurement device for blood using a microfluidic device, characterized in that derived through the second image intensity (<I _BC >). As a method for measuring multi-biological values of blood through a multi-biological value measuring device of blood using a microfluidic device that simultaneously measures red blood cell deformability, erythrocyte cohesiveness, and hematocrit of the fluid to be measured
The apparatus for measuring multiple biological properties of blood using the microfluidic device,
And a microfluidic device in which a channel through which the fluid to be measured passes is formed, wherein the microfluidic device comprises: an injection port through which the fluid to be measured is injected; A main channel in communication with the inlet and through which the fluid to be measured passes; An expansion channel communicating with the main channel and formed with a diameter larger than the diameter of the main channel; A branch channel in communication with the expansion channel and through which the fluid to be measured passes; A first outlet through which the fluid to be measured can be discharged; A first connection channel that is separated from the branch channel and communicates with a first discharge port through which the fluid to be measured is discharged; An expansion chamber in communication with the expansion channel through the branch channel, and having a micro channel disposed at a fine interval therein; A second outlet through which the fluid to be measured can be discharged; While communicating with the expansion chamber, and includes a second connection channel in communication with the second outlet,
An injection step of injecting the target fluid into the microfluidic device through the injection hole;
Filling the microfluidic device with the fluid to be measured;
A measurement step of measuring a flow velocity or image intensity at a designated point of the microfluidic device;
A derivation step of deriving a red blood cell deformability index, a red blood cell cohesiveness index, and a hematocrit index through a flow rate or image intensity measured through the measuring step; a method for measuring multi-biological value of blood using a microfluidic device. The method of claim 10,
The apparatus for measuring multiple biological properties of blood using the microfluidic device,
A method for measuring multiple biological properties of blood using a microfluidic device, characterized in that it comprises a syringe pump that is connected to the inlet and can inject the fluid to be measured into the inlet. The method of claim 11,
In the injection step,
A method for measuring a multi-biological value of blood using a microfluidic device, characterized by injecting the measurement target fluid into the injection port while periodically operating and stopping the syringe pump. The method of claim 11,
In the injection step,
The measurement target fluid and a reference fluid are injected into the injection port through the syringe pump,
In the filling step,
A method for measuring multiple biological properties of blood using a microfluidic device, characterized in that the microfluidic device is filled with the target fluid and the reference fluid. The method of claim 10,
The second outlet is connected to a tube filled with a reference fluid, the tube is a multi-biological value measurement method of blood using a microfluidic device, characterized in that one side is blocked through a pinch valve. The method of claim 10,
The apparatus for measuring multiple biological properties of blood using the microfluidic device,
A method for measuring multiple biological properties of blood using a microfluidic device, comprising an image acquisition device capable of measuring the image intensity of a specific region of interest. The method of claim 12,
The measuring step measures the flow rate (<U _B >) of the branch channel,
The red blood cell deformability index in the derivation step,
Method for measuring multi-biological value of blood using a microfluidic device, characterized in that it is derived through the flow rate (<U _B >) of the branch channel and the cross-sectional area (A _C ) of the branch channel. The method of claim 15,
In the measuring step, a first specific region of interest is selected in the expansion chamber, and a first image intensity (<I _MP >) of the first specific region of interest is measured through the image acquisition device,
The red blood cell cohesiveness index in the derivation step,
Method for measuring multiple biological properties of blood using a microfluidic device, characterized in that it is derived through the first image intensity (<I _MP >). The method of claim 15,
In the measuring step, a second specific region of interest is selected in the extended channel, and a second image intensity (<I _BC >) of the second specific region of interest is measured through the image acquisition device,
In the deriving step, the hematocrit index,
Method for measuring multiple biological properties of blood using a microfluidic device, characterized in that it is derived through the second image intensity (<I _BC >).

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