KR20220153908A - Blood Coagulation Measuring Device - Google Patents

Abstract

A blood coagulation measuring device according to one embodiment of the present invention comprises: a substrate; a microchannel disposed on the substrate; a plurality of electrodes disposed to be spaced apart from each other on the substrate and having a part thereof disposed between the substrate and the microchannel; a graphene sheet disposed between the substrate and the microchannel, having a first region disposed on the plurality of electrodes, and having a second region extending from the first region and disposed on the substrate; a first through hole passing through the microchannel to be in contact with the second region; a second through hole disposed apart from the first through hole in a first direction and in contact with the substrate while penetrating the microchannel; and a connection pipe disposed between the substrate and the microchannel and connecting the first through hole and the second through hole. The present invention may take a short time of less than 15 minutes without a reagent using a blood coagulation measuring device.

Description

Blood coagulation measuring device {Blood Coagulation Measuring Device}

The present invention relates to a blood coagulation measuring device, and more particularly, to a blood coagulation measuring device capable of analyzing and evaluating electrical characteristics of blood using a transistor element capable of using graphene as an active layer.

Fibrin formation is generally determined by changes in the mechanical and optical properties of a plasma sample. In addition to processing large amounts of sample prior to testing to obtain plasma only, conventional equipment available in clinical laboratories takes a long time to obtain results related to clot formation.

In an emergency where the patient's medical background is unknown and prompt action is required to stop bleeding or proceed with surgery, not only the diagnosis and response time is delayed, but also a large amount of blood is required for each test.

Registered Patent Publication No. 10-1221296 (Title of Invention: Blood Coagulation Measuring Device and Manufacturing Method Thereof)

The present invention has been made to solve the above problems, and an object of the present invention is to provide a blood coagulation measuring device capable of providing detailed information on a coagulation process by measuring a blood sample without prior treatment and a large amount of blood.

The technical objects to be achieved in the present invention are not limited to the above-mentioned matters, and other technical problems not mentioned are those of ordinary skill in the art to which the present invention belongs from the embodiments of the present invention to be described below. can be considered by

Hereinafter, a blood coagulation measuring device will be described as embodiments of the present invention.

An apparatus for measuring blood coagulation according to an embodiment of the present invention includes a substrate, a microchannel disposed on the substrate, a plurality of electrodes disposed on the substrate and spaced apart from each other, some of which are disposed between the substrate and the microchannel; a graphene sheet disposed between the substrate and the microchannel, having a first region disposed on the plurality of electrodes, and having a second region extending from the first region and disposed on the substrate; a first through-hole passing through and in contact with the second region, a second through-hole disposed spaced apart from the first through-hole in a first direction, and passing through the microchannel and in contact with the substrate, and the substrate and the microchannel and a connection pipe disposed therebetween and connecting the first through hole and the second through hole.

In addition, the plurality of electrodes include a first electrode disposed elongated in a second direction crossing the first direction, a second electrode disposed elongated in the first direction and bent at least once, and disposed in the first direction. It may include providing a third electrode that is disposed long and spaced apart from the second electrode in the second direction and is bent at least once.

In addition, the first electrode may be disposed between the first through hole and the second through hole and intersect the connection pipe.

In addition, the first through hole may include being disposed between the first electrode and the second electrode or between the first electrode and the third electrode.

The first region of the graphene sheet is disposed on the second electrode and the third electrode, and the second region of the graphene sheet is exposed to the outside through the first through hole. can do.

In addition, the blood sample injected into the microchannel may include an injection of 10 μl or more and less than 20 μl.

The above-described aspects of the present invention are only some of the preferred embodiments of the present invention, and various embodiments in which the technical features of the present invention are reflected are the details of the present invention to be detailed below by those skilled in the art. It can be derived and understood based on the description.

According to embodiments of the present invention, the following effects can be obtained.

The present invention provides detailed information on the coagulation process by measuring a blood sample without prior treatment and a large amount of blood, so that rapid and accurate results can be obtained.

The present invention can be extended to the entire application range of the coagulation process based on the interaction of blood factors (coagulation factors and anticoagulants) using a blood coagulation measuring device.

According to the present invention, the state of coagulation factors and anticoagulants according to the doping effect of the graphene sheet can be quickly and accurately measured using the blood coagulation measuring device.

Since the present invention can use a blood sample in a small amount (less than 20 μl) using the blood coagulation measuring device, it can be used more conveniently.

The present invention can be used more quickly because there is no prior treatment before the test using the blood coagulation measuring device and the whole blood sample can be collected with a fingerstick.

The present invention can take a short time of less than 15 minutes without reagents using a blood coagulation measuring device.

In the present invention, PT and aPPT tests can be performed when a specific reagent is mixed with a whole blood sample on a graphene surface using a blood coagulation measuring device.

The present invention can evaluate the degree of coagulation factors and anticoagulants using a blood coagulation measuring device.

Effects obtainable in the embodiments of the present invention are not limited to the above-mentioned effects, and other effects not mentioned are common in the art to which the present invention pertains from the description of the following embodiments of the present invention. can be clearly derived and understood by those with knowledge of That is, unintended effects according to the practice of the present invention may also be derived by those skilled in the art from the embodiments of the present invention.

Included as part of the detailed description to aid understanding of the present invention, the accompanying drawings provide various embodiments of the present invention. In addition, the accompanying drawings, together with the detailed description, are used to describe the embodiments of the present invention.
1 is a perspective view for explaining a blood coagulation measuring device according to an embodiment of the present invention.
2 is a plan view illustrating an apparatus for measuring blood coagulation according to an embodiment of the present invention.
3 is a cross-sectional view for explaining a blood coagulation measuring device according to an embodiment of the present invention.
4 to 5 are diagrams for explaining evaluation of electrical characteristics of a blood sample during a coagulation process using a blood coagulation measuring device according to an embodiment of the present invention.
6 is a diagram for explaining the operation of a transfer characteristic curve of a blood sample during a coagulation process using the blood coagulation measuring device according to an embodiment of the present invention.
7 and 8 are views for explaining graphs obtained by measuring blood samples using the blood coagulation measuring device according to an embodiment of the present invention.

Hereinafter, a blood coagulation measuring device will be described as embodiments of the present invention.

The following embodiments combine the elements and features of the present invention in certain forms. Each component or feature may be considered optional unless explicitly stated otherwise. Each component or feature may be implemented in a form not combined with other components or features. In addition, an embodiment of the present invention may be configured by combining some components and/or features. The order of operations described in the embodiments of the present invention may be changed. Some components or features of one embodiment may be included in another embodiment, or may be replaced with corresponding components or features of another embodiment.

In the description of the drawings, parts, devices, and/or components that may obscure the subject matter of the present invention are not described, and parts, devices, and/or components that can be understood by those skilled in the art are not described. In addition, parts referred to using the same reference numerals in the drawings mean the same components or steps in the apparatus configuration or method.

Throughout the specification, when a part is said to "comprising" or "including" a certain element, it means that it may further include other elements, not excluding other elements, unless otherwise stated. do. In addition, terms such as "?? unit" or "?? group" described in the specification mean a unit that processes at least one function or operation. Also, "a or an", "one", "the" and similar related words in the context of describing the invention (particularly in the context of the claims below) Unless indicated or otherwise clearly contradicted by context, both the singular and the plural can be used.

In addition, specific terms and / or symbols used in the embodiments of the present invention are provided to help the understanding of the present invention, and the use of these specific terms may be used in other forms without departing from the technical spirit of the present invention. can be changed to

1 is a perspective view for explaining a blood coagulation measuring device according to an embodiment of the present invention, FIG. 2 is a plan view for explaining a blood coagulation measuring device according to an embodiment of the present invention, and FIG. It is a cross-sectional view for explaining a blood coagulation measuring device according to an embodiment.

1 to 3, the blood coagulation measuring device 100 according to an embodiment of the present invention includes a substrate 110, a microchannel 150, a plurality of electrodes 130, a graphene sheet 170, and the like. can include

The substrate 110 may have a certain thickness and a certain area. The substrate 110 may be formed as a flat plate with both upper and lower surfaces flat. For example, the substrate 110 may be formed of a biocompatible material such as glass or polyamide. The substrate 110 may deposit a plurality of electrodes 130 or graphene sheets 170 thereon.

The microchannel 150 may be disposed on the substrate 110 . The microchannel 150 may be disposed or fabricated on top of one electrode among the graphene sheet 170 and the plurality of electrodes 130 . The microchannel 150 can be used to deliver a biological sample (whole blood sample) that interacts with the non-functionalized graphene surface. For example, the microchannel 150 may be referred to as polydimethylsiloxane, PDMS, or silicone oil. When viewed optically, the microchannel 150 formed of PDMS has a transparent color and may be an inert material. An inert material may be a material that has a very stable chemical state and maintains its state without reaction or change even when in contact with other materials. Accordingly, the microchannel 150 can well transfer the biological sample in a stable state. For example, the biological sample may be blood.

In addition, the microchannel 150 may be formed in a rectangular parallelepiped shape. For example, the microchannel 150 may be formed in a rectangular parallelepiped shape having a first direction, a second direction crossing the first direction horizontally, and a third direction perpendicularly crossing the first and second directions. . For example, the first direction may be referred to as a front-back direction, the second direction may be referred to as a left-right direction, and the third direction may be referred to as a vertical direction.

As described above, the microchannel 150 of the blood coagulation measuring device 100 according to an embodiment of the present invention is between the graphene sheet 170 (active layer) and the first electrode 130a, which will be described later. By being fabricated or placed on top of a region commonly referred to as the gate terminal, it is possible to transfer a biological sample (whole blood sample) that interacts with the non-functionalized graphene surface.

The plurality of electrodes 130 are disposed spaced apart from each other on the substrate 110, and some may be disposed between the substrate 110 and the microchannel 150. The plurality of electrodes 130 may include a conductive material such as gold, silver, or metal. The plurality of electrodes 130 may include a first electrode 130a, a second electrode 130b, and a third electrode 130c.

The first electrode 130a may be disposed elongately in a second direction crossing the first direction. That is, the first electrode 130a is disposed elongated in the left-right direction. A portion of the first electrode 130a may overlap the microchannel 150 . The first electrode 130a may be spaced apart from the second electrode 130b and the third electrode 130c by a predetermined distance. For example, the first electrode 130a may be spaced apart from the second electrode 130b and the third electrode 130c in a first direction or in a front-back direction by a predetermined distance.

Also, the first electrode 130a may be spaced apart from the graphene sheet 170 by a predetermined distance. The first electrode 130a may be referred to as a gate electrode or a gate terminal.

The second electrode 130b may be disposed elongated in the first direction and bent at least once. That is, the second electrode 130b may be disposed long in the front-back direction. The second electrode 130b may be formed to extend in the first direction and then refract and progress in the second direction. A part of the second electrode 130b may overlap the microchannel 150 . The second electrode 130b may be disposed apart from the first electrode 130a in a first direction or in a front-back direction, and may be disposed apart from the third electrode 130c in a second direction or in a left-right direction. The second electrode 130b may be referred to as a source electrode or a source terminal.

The third electrode 130c may be disposed elongated in the first direction and bent at least once. That is, the third electrode 130c may be disposed long in the front-back direction. The third electrode 130c may be formed to extend in the first direction and then refract and progress in the second direction. That is, the third electrode 130c may be disposed long in the first direction, spaced apart from the second electrode 130b in the second direction, and bent at least once. The third electrode 130c may be refracted in a direction opposite to that of the second electrode 130b. The third electrode 130c may be formed symmetrically with the second electrode 130b. The third electrode 130c may be referred to as a drain electrode or a drain terminal. It is not limited thereto, and when the second electrode 130b is a drain electrode, the third electrode 130c may be a source electrode.

As described above, the blood coagulation measuring device 100 according to an embodiment of the present invention may have three electrodes/terminals that may be made of a conductive and biocompatible material such as gold or silver. That is, the first electrode 130a to the third electrode 130c may have a structure substantially similar to that of a field effect transistor (FET) device.

The graphene sheet 170 may be disposed between the substrate 110 and the microchannel 150 . The graphene sheet 170 may be formed by being laminated on the substrate 110 , and the microchannel 150 may be formed on the graphene sheet 170 .

The graphene sheet 170 may be referred to as graphene, a graphene active layer, an active layer, or a graphene layer. The graphene sheet 170 may include a first region and a second region. The first region of the graphene sheet 170 may be disposed on the plurality of electrodes 130 . That is, the first region of the graphene sheet 170 may be stacked on the substrate 110, and a portion of the first region overlaps a portion of the second electrode 130b and a portion of the third electrode 130c. have. That is, the graphene sheet 170 may operate as an active layer by being transferred to a first region, which is a region between two electrodes identified as a drain electrode or a source electrode.

The second region of the graphene sheet 170 may extend from the first region and be disposed on the substrate 110 . The second region of the graphene sheet 170 may be disposed between the substrate 110 and the microchannel 150 .

As described above, the graphene sheet 170 may contact the substrate 110, the second electrode 130b, the third electrode 130c, and the microchannel 150. That is, the first region of the graphene sheet 170 is disposed on the second electrode 130b and the third electrode 130c, and the second region of the graphene sheet 170 has the first through hole 151 can be exposed to the outside.

The first through hole 151 may be disposed through the microchannel 150 . The first through hole 151 may be formed to contact the second region of the graphene sheet 170 by penetrating the microchannel 150 in a third direction or a vertical direction. For example, the first through hole 151 may have a cylindrical shape having a constant diameter or a cone shape whose diameter gradually decreases as the diameter approaches the substrate 110 .

The first through hole 151 may be disposed between the first electrode 130a and the second electrode 130b or between the first electrode 130a and the third electrode 130c.

The second through hole 152 may be disposed through the microchannel 150 . The second through hole 152 may be spaced apart from the first through hole 151 in the first direction. The second through hole 152 may be formed to contact the substrate 110 by penetrating the microchannel 150 in a third direction or a vertical direction. For example, the second through hole 152 may have a cylindrical shape having a constant diameter or a cone shape whose diameter gradually decreases as the diameter approaches the substrate 110 .

The diameter of the second through hole 152 measured from the top or upper surface of the microchannel 150 may be smaller than that of the first through hole 151 .

The connection pipe 153 may be disposed between the substrate 110 and the microchannel 150 . The connection pipe 153 is disposed on the substrate 110 and may pass through the microchannel 150 in a first direction or in a front-back direction. Accordingly, the connection pipe 153 may be formed to connect the first through hole 151 and the second through hole 152 . The connection pipe 153 may transfer the biological sample (whole blood sample) introduced through the first through hole 151 to the second through hole 152 .

Also, the aforementioned first electrode 130a may be disposed between the first through hole 151 and the second through hole 152 . The first electrode 130a may be disposed crossing the connection pipe 153 by being formed long in the second direction or in the left and right directions. Accordingly, the first electrode 130a may detect or measure a biological sample (whole blood sample) flowing through the connection pipe 153 .

The blood coagulation measurement device 100 according to one embodiment of the present invention shown in FIGS. 1 to 3 is not limited thereto, and the structure of the electrode and the microchannel 150 may be changed.

4 to 5 are diagrams for explaining evaluation of electrical characteristics of a blood sample during a coagulation process using a blood coagulation measuring device according to an embodiment of the present invention.

Referring to FIGS. 4 and 5 , the blood coagulation measurement device according to an embodiment of the present invention may be electrically connected to an analysis device 300 capable of measuring electrical characteristics of a blood sample.

The analysis device 300 may be electrically connected to the first to third electrodes of the blood coagulation measuring device. The analyzer 300 may be electrically connected to the first to third electrodes, supply a predetermined voltage to at least one of the electrodes, and evaluate or measure a Dirac point and drain current from these.

The analysis device 300 may measure the drain current (ID) while maintaining the drain voltage (VD) constant and applying different bias voltages (VG) to the common gate terminal of the blood coagulation measuring device of the present invention. The drain voltage (VD) may be 03 to 0.5V.

As shown in FIGS. 4 and 5 , a whole blood sample may be injected into the blood coagulation measuring device according to an embodiment of the present invention. For example, the whole blood sample may be injected into the microchannel of the blood coagulation measuring device using the syringe 500 . At this time, approximately 10 to 20 μL of the whole blood sample may be injected into the microchannel.

The whole blood sample injected into the microchannel may come into contact with the graphene sheet and the gate electrode while being moved to the first through-hole connecting pipe and the second through-hole.

6 is a diagram for explaining the operation of a transfer characteristic curve of a blood sample during a coagulation process using the blood coagulation measuring device according to an embodiment of the present invention.

Referring to FIG. 6 , a drain current (ID)-gate voltage (VG) characteristic curve according to an embodiment of the present invention is shown.

Figure 6 (a) shows immediately before the biological sample is tested through the blood coagulation measuring device, and Figure 6 (b) shows a biological sample tested after a predetermined time has elapsed through the blood coagulation measuring device. will be.

In the present invention, in the case of an experiment using a biological sample to obtain more accurate results, the transfer curve was repeatedly measured several times for 15 minutes at 30 second intervals.

As shown in FIG. 6, the present invention can evaluate the Dirac point and drain current for each device in the course of the experiment. That is, the present invention measured an electrical signal through a blood coagulation measuring device, and the measured electrical signal indicates that a doping effect occurred in a biological process, and due to an increase or decrease in charge density on the surface of the active layer, which is a graphene sheet, Dirac ) can measure the movement of a point to the left or right.

For the above-described experiment, in order to evaluate the electrical properties of the blood sample during the coagulation process, the transfer characteristics of the graphene sheet or graphene were measured after the blood sample was injected (less than 20 μl). At this time, the drain current of the blood coagulation measuring device of the present invention was measured while applying different bias voltages to the common gate electrode and maintaining the drain voltage constant. For example, the drain current (ID) was measured while holding the drain voltage (VD) constant at 0.4 V while applying different bias voltages (VG) to the common gate terminal.

The shift of the curve during measurement of the transfer characteristics shown in FIG. 6 may represent the doping effect of graphene. Changes in the Dirac point can be proportional to the effects induced by the levels of coagulation factors and anticoagulants in the blood sample. In addition, the present invention can evaluate changes in the concentration of factors that inhibit or accelerate the blood coagulation process (cascade process) using the blood coagulation measuring device.

7 and 8 are views for explaining graphs obtained by measuring blood samples using the blood coagulation measuring device according to an embodiment of the present invention.

Referring to FIG. 7 , the value of the Dirac point was measured immediately after blood samples were taken and injected into each microchannel containing an activator and an inhibitor.

Dirac point voltage (0 min) was measured using raw and treated blood samples with activators (VitK, TR and CaCl2) and inhibitors (LMWHE and LMWHP).

Here, HWB: clean blood, VitK: vitamin K, TR: thromboplastin reagent, CaCl2: calcium chloride, LMWHE: enoxaparin sodium, LMWHP: parnaparin sodium.

Dirac voltage values for liquefaction of blood samples mixed with different activators and inhibitors and measured every 15 minutes. HWB: clean blood, VitK: vitamin K, TR: thromboplastin reagent, CaCl2: calcium chloride, LMWHE: enoxaparin sodium; LMWHP: parnaparin sodium.

In addition, referring to FIG. 8, when using untreated and treated samples in the present invention, the concentration of charge carriers that change for each case is evaluated based on factors that inhibit or accelerate the cascade process. and expressed graphically.

Here, samples treated with inhibitors (enoxaparin sodium and parnaparin sodium) exhibited higher Dirac point voltages because clot formation was delayed due to the presence of low molecular weight heparin.

And for samples treated with active agents (vitamin K, thromboplastin reagent, calcium chloride), the change in Dirac point was higher compared to clean blood for all three treated samples.

For example, each activator has a unique role in the coagulation process, resulting in different values of change. Therefore, it was found that the acceleration changed when the concentration of a specific coagulation factor increased. For example, clot formation appeared faster than other samples because the thromboplastin reagent contained tissue factor with a large blood coagulation effect.

The above-described embodiments of the present invention may be embodied in other specific forms without departing from the essential characteristics of the present invention. Accordingly, the foregoing detailed description should not be construed as limiting in all respects and should be considered illustrative. The scope of the present invention should be determined by reasonable interpretation of the appended claims, and all changes within the equivalent scope of the present invention are included in the scope of the present invention. In addition, claims that do not have an explicit citation relationship in the claims may be combined to form an embodiment or may be included as new claims by amendment after filing.

100: blood coagulation measuring device
110: substrate
130: multiple electrodes
150: micro channel
170: graphene sheet

Claims (7)

Board;
a microchannel disposed on the substrate;
a plurality of electrodes disposed spaced apart from each other on the substrate, some of which are disposed between the substrate and the microchannel;
a graphene sheet disposed between the substrate and the microchannel, having a first region disposed on the plurality of electrodes, and having a second region extending from the first region and disposed on the substrate;
a first through hole passing through the microchannel and contacting the second region;
a second through hole disposed apart from the first through hole in a first direction and contacting the substrate while penetrating the microchannel; and
a connection pipe disposed between the substrate and the microchannel and connecting the first through hole and the second through hole;
Blood coagulation measuring device comprising a. According to claim 1,
A plurality of electrodes,
A first electrode extending in a second direction crossing the first direction;
A second electrode disposed elongated in the first direction and bent at least once; and
Blood coagulation measuring device having a third electrode disposed in the first direction and spaced apart from the second electrode in the second direction and bent at least once. According to claim 2,
The first electrode is
disposed between the first through hole and the second through hole;
A blood coagulation measuring device disposed to cross the connection pipe. According to claim 2,
The first through hole,
Blood coagulation measuring device disposed between the first electrode and the second electrode or between the first electrode and the third electrode. According to claim 3,
The first region of the graphene sheet,
disposed on the second electrode and the third electrode;
The second region of the graphene sheet,
Blood coagulation measuring device exposed to the outside by the first through hole. According to claim 1,
The blood sample injected into the microchannel,
A blood coagulation measuring device that is injected with more than 10 μl and less than 20 μl. According to claim 6,
The blood sample is
A device for measuring blood coagulation in which an activator and an inhibitor are mixed and injected into the microchannel.

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