TW201331582A - Device for detecting blood coagulation and manufacturing method thereof - Google Patents

Abstract

A device for detecting blood coagulation includes a substrate layer, a middle layer, an upper layer and two electrodes. The middle layer is disposed on the substrate layer. The upper layer is disposed on the middle layer. A fluidic channel is defined by the substrate layer, the middle layer and the upper layer. The electrodes are disposed on the upper layer and the substrate layer respectively. The present invented provides a manufacturing method of the device for detecting blood coagulation as well. The present invention is advantageous for obtaining the blood clotting time more objectively and accurately.

Description

Blood coagulation detecting device and manufacturing method thereof

The present invention relates to a detecting device and a method of manufacturing the same, and more particularly to a detecting device for blood coagulation and a method of manufacturing the same.

Because blood coagulation test is of high importance in clinical application, in addition to the screening and diagnosis of bleeding diseases, it is also often used for the examination of the coagulation status of patients before surgery, as well as the medication guidance and prognosis estimation of various anticoagulant patients.

However, the traditional blood coagulation test must be done in the laboratory, and the blood test sample needs to be the plasma obtained by centrifugation. Not only the required sample size is large, but also the pretreatment of the sample is quite time consuming, so it is necessary to know the test at the first time. The result is quite inconvenient for the medical staff. In addition, the instruments used are bulky and must be operated by professionals, and it is almost impossible for the general public to self-test. Since then, several small instruments for blood coagulation testing have been introduced, but their use is still limited to patients taking anticoagulant drugs based on cost.

In recent years, the technology of biomedical testing devices has developed rapidly, especially in the design of wafers. These emerging tools replace the bulky old-fashioned instrumentation, providing better applicability and operability. Of course, these technologies are also increasingly relied on in the field of blood-related testing.

One development focus of biomedical test wafers is the driving and mixing of fluids, in which the operation of high viscosity fluids, especially blood, is more difficult. In the past, external forces were used to drive fluid movement in the wafer flow path. From early human-powered syringes to mechanical micro-pulse drives, they have been quite mature and common. However, no matter how to improve the pump volume, the additional components still occupy a certain space, which will make the wafer device lose its small volume advantage, and also increase the production cost and complexity.

In addition, there are still no objective criteria for the determination of blood coagulation tests, and most of them can only be carried out using a more subjective method, such as visual judgment or microscopic observation, without being quantified into data comparison, which cannot provide objective and Accurate analysis.

Therefore, how to provide a device that is simple in operation, quick in detection, and capable of objectively judging the detection result, and thus achieves the immediacy and accuracy of blood coagulation detection has become one of the important topics.

In view of the above problems, an object of the present invention is to provide a blood coagulation detecting device which is simple in operation, quick and instantaneous in detecting, and capable of objectively judging the detecting result, and a manufacturing method thereof, thereby improving the immediacy and accuracy of the detecting.

To achieve the above object, a blood coagulation detecting device according to the present invention comprises a substrate layer, an intermediate layer, an upper cap layer and two electrodes. The intermediate layer is disposed on the substrate layer. The upper cover layer is disposed on the intermediate layer. The substrate layer, the intermediate layer and the upper cover layer together define a first-class track. The two electrodes are respectively disposed on the upper cover layer and the substrate layer, respectively.

In one embodiment, the blood coagulation detecting device is a capacitive or electrical impedance blood coagulation detecting wafer.

In one embodiment, the material of the substrate layer and the upper cap layer comprises glass, indium tin oxide, antimony oxide, or twin crystal.

In an embodiment, the material of the intermediate layer comprises an optical glue or a photoresist material.

In an embodiment, the upper cover layer has a first opening, a second opening, and an outlet connected to the flow channel.

In one embodiment, the flow prop has a mixing portion and a communication portion, and the second opening and the mixing portion are in communication with each other.

In one embodiment, the flow prop has a capillary portion and the capillary portion is wound around the communication portion.

In an embodiment, the flow passage has a larger inner diameter near one end of the first opening.

In an embodiment, the flow passage has at least one turning portion, and the turning portion has an arc shape, a hairpin shape, or a horseshoe shape.

In one embodiment, one of the two electrodes is disposed over the upper cap layer and the other of the two electrodes is disposed below the substrate layer.

In an embodiment, the flow channel is sandwiched between the two electrodes.

A method for manufacturing a blood coagulation detecting device according to the present invention includes the steps of: forming a first opening, a second opening, and an outlet on an upper cap layer; forming an electrode on the upper cap layer and a substrate layer; An optical glue or a photoresist material is formed on the substrate layer to form an intermediate layer; the intermediate layer is processed; and the upper cap layer, the intermediate layer and the substrate layer are combined to define a first-class track.

In one embodiment, the formation of the electrode includes providing a heat resistant material or another photoresist material on the upper cap layer and the substrate layer, defining a pattern of the electrodes, and a process of plating the film.

In one embodiment, the definition of the pattern includes a laser processing or lithography process.

In one embodiment, the processing of the intermediate layer forms part of the flow path and includes procedures for laser processing, lithography, or machining.

In one embodiment, the combination of the upper cap layer, the intermediate layer, and the substrate layer includes a procedure for low temperature bonding.

The term "blood coagulation" or "coagulation" as used in the present invention refers to a process in which blood changes from a liquid to a solid, and may also be referred to as "blood coagulation" or "coagulation". .

In summary, the blood coagulation detecting device according to the present invention transmits the first-class track jointly defined by the substrate layer, the intermediate layer and the upper cap layer for blood input detection, and uses the correspondingly disposed electrodes while guiding the flow. It detects the change of physical properties during blood coagulation, thereby providing the functions of simple operation and rapid detection, and has the results of data analysis for objective judgment.

Compared with the prior art, the detecting device and the manufacturing method thereof are suitable for using a hydrophilic substance as a material of the substrate layer and the upper cap layer, so as to achieve the effect of self-driving the blood to be tested without external force, and not only the pump is eliminated. Demand, effectively reducing the size of the device and manufacturing costs, more ready-to-use, no need for cleaning and special pre-processing advantages.

In addition, the device and the manufacturing method thereof can specifically utilize the characteristics of capacitance or electrical impedance change in the flow channel when the blood is coagulated, and objectively analyze the data blood coagulation time, which is significantly improved compared with the conventional image analysis method. Sexuality and applicability.

Hereinafter, a blood coagulation detecting device and a method of manufacturing the same according to a preferred embodiment of the present invention will be described with reference to the accompanying drawings, wherein the same elements will be described with the same reference numerals.

1 is an exploded perspective view of a blood coagulation detecting device according to a preferred embodiment of the present invention, FIG. 2A is a combined appearance view of the blood coagulation detecting device shown in FIG. 1, and FIG. 2B is a perspective view of the blood coagulation detecting device shown in FIG. 2A. schematic diagram. As shown in FIG. 1 to FIG. 2B, in the present embodiment, the blood coagulation detecting device 1 includes an upper cap layer 11, an intermediate layer 12, a substrate layer 13, and two electrodes 14 from top to bottom, wherein the substrate layer 13. The intermediate layer 12 and the upper cover layer 11 together define a first-class track CH. However, in other embodiments, the structures of the blood coagulation detecting device 1 may also change the order relationship of the settings, or may be Other structures are included between or outside the structures, and the present invention is not limited thereto.

The material of the upper cap layer 11 and the substrate layer 13 includes glass, indium tin oxide, antimony oxide, or twin crystal, which are all hydrophilic materials. Preferably, the upper cap layer 11 and the substrate layer 13 are made of hydrophilic glass. By the hydrophilic property of the above material, it is possible to enhance the capillary action force on the blood input into the blood coagulation detecting device 1 and to cause self-driving effect. In addition, in other embodiments, the upper cap layer 11 and the substrate layer 13 may also be made of indium tin oxide (ITO), yttrium oxide, twin crystal, or a combination thereof. Among them, the use of indium tin oxide as a material can enhance the detection effect of the electrode 14 function and the blood coagulation detecting device 1.

The upper cover layer 11 is provided with a first opening 111, a second opening 112 and an outlet 113 for communicating the flow path CH to respectively inject and discharge blood or other liquid. The first opening 111, the second opening 112, and the outlet 113 may have different sizes and shapes according to requirements. The present invention is not limited thereto, but preferably all of them are circular, and the inner diameter of the first opening 111 is For about 11 mm, the inner diameter of the second opening 112 is about 2 mm, and the inner diameter of the outlet 113 is about 2 mm. The structures of the upper cover layer 11 can be formed by water-assisted laser processing.

In this embodiment, the material of the intermediate layer 12 comprises an optical adhesive or a photoresist material, preferably an optically transparent film double-sided adhesive material or a JSR photoresist material, which is not limited thereto. The intermediate layer 12 is selected from the materials to further increase the capillary force of the flow channel CH, which helps to improve the liquid transport efficiency, and can also be thermally bonded by the photoresist material at a lower temperature, and the cover layer of the fixed glass material is fixed. 11 and the substrate layer 13, thereby simplifying the process and improving efficiency.

A portion of the intermediate layer 12 is vertically penetrated to form a continuous reciprocating and bent extending channel structure 121. After being combined with the substrate layer 13 and the upper cap layer 11, the channel structure 121 can be formed by being closed up and down only to be added at a specific position. Or discharge the liquid flow path CH.

Referring to FIG. 1 , the channel structure 121 has an injection hole 122 at a position corresponding to the first opening 111 . When combined with the substrate layer 13 , a cavity can be formed after the first opening 111 is injected into the cavity. The sample size, and adjust the gap between the amount of blood injected and the amount of flow into the flow, improve the ease of operation. The channel structure 121 further includes a mixing hole 123 which can also be combined with the substrate layer 13 to form a mixing portion for mixing with blood to be mixed with the additionally added calcium chloride solution. The diameter of the mixing hole 123 is about 4 mm in this embodiment, which is larger than the second opening 112, and the two are connected by a communicating portion 124. The communication portion 124 is located directly under the second opening 112. In addition, in one aspect of the embodiment, the channel structure 121 is further sandwiched between the substrate layer 13 and the upper cover layer 11 and connected. The annular capillary portion is disposed around the communicating portion 124, and the capillary portion can provide a capillary force to the liquid preparation to be mixed which is added from the second opening 112 to be sucked into the capillary portion and mixed with the blood. Specifically, the liquid preparation may be a calcium chloride solution.

As for the processing of the above structure of the intermediate layer 12, an optical glue or a photoresist material may be first disposed and adhered to the top surface of the substrate layer 13, and the injection hole 122 and the mixing hole 123 may be processed by water-assisted laser processing. And the communication portion 124 and the like, but this is not limitative. The material of the intermediate layer 12 is preferably a hydrophilic optical transparent film double-sided adhesive material or a JSR photoresist material. In addition to the laser, the method of processing includes a lithography process or a mechanical process, and the present invention is not limited thereto.

The two electrodes 14 are respectively disposed on the upper cover layer 11 and the substrate layer 13 respectively. In detail, as shown in FIG. 1 , one of the electrodes 14 may be disposed on the upper cover layer 11 , and the other one of the electrodes 14 may be disposed under the substrate layer 13 , and the flow channel CH may be disposed on the two electrodes 14 . between. Therefore, the relationship between the two electrodes 14 is upper and lower corresponding but not in contact, and is not in direct contact with the blood in the flow channel CH, thereby reducing the risk of qualitative change of the blood sample during the detection process. In addition, the electrodes 14 each have a contact portion 141 and a detecting portion 142, wherein the contact portion 141 is connected to an external power source to supply power required for operation; and the detecting portion 142 substantially enables the flow channel CH to be in a uniform electric field. In order to sense the physical changes in blood coagulation. It should be noted that, in this embodiment, the detecting portion 142 of the electrode 14 has the same shape as the portion of the channel structure 121 behind the second opening 112, that is, the portion of the flow channel CH behind the second opening 112. The same shape, and the parallel flow paths CH are respectively disposed so that the flow path CH is in a uniform electric field environment, which is not limitative.

The invention is not limited to the method for fabricating the electrode 14. The heat-resistant material or a photoresist material may be disposed on the upper cap layer 11 and the substrate layer 13 and defined on the upper cap layer 11 and the substrate layer 13 by laser processing or a yellow photolithography process. After the electrode patterns are formed, they are separately or collectively coated. The photoresist material may be the same as or different from the photoresist material used in the intermediate layer 12.

Referring to FIG. 2B, the upper cap layer 11, the intermediate layer 12, the substrate layer 13, and the electrode 14 are stacked and fixed to form a blood coagulation detecting device 1. Since the intermediate layer 12 can be an optically transparent film double-sided tape or a photoresist material, it is inherently viscous and thus can be thermally bonded in low temperature fabrication. At this time, the substrate layer 13, the upper cap layer 11 and the intermediate layer 12 together define a continuous bending and extending flow channel CH, respectively connecting the first opening 111 and the outlet 113 at opposite ends of the flow channel CH, and adjacent to the first opening. The second opening 112 of 111.

In this embodiment, after the external power supply of the electrode 14 is performed, the flow path CH can be placed under a uniform electric field because of the corresponding relationship between the shape and the size. At this time, if the blood in the flow path CH starts to solidify, the measured physical change changes, and in particular, the capacitance or the electrical impedance value rapidly increases, so that the blood coagulation detecting device 1 can judge the blood coagulation time.

3A is a schematic enlarged plan view of the intermediate layer shown in FIG. 1, and FIG. 3B is a schematic view showing a variation of the intermediate layer of FIG. 3A. Referring first to FIG. 3A, the channel structure 121 has a larger inner diameter near one end of the first opening 111. Specifically, the inner diameter of the channel structure 121 between the injection hole 122 and the mixing hole 123 is gradually enlarged due to the proximity of the injection hole 122, so that it is roughly flared or tapered (the area CP is shown in the figure), and there is a speaker at the end. Extended shape or cone (marked area EP in the figure). In this way, the effect of extending the volume of the blood sample injection can be achieved, and the capillary force can be enhanced to drive the blood to flow by itself. In addition, this flared or tapered design also reduces flow resistance and facilitates blood flow. Further, the turning portion 126 of the passage structure 121 has an arc shape, a hairpin shape, a horseshoe shape or the like to further reduce the flow resistance.

Of course, in addition to the above-described form of the channel structure, the channel structure of the present invention can have various variations, as exemplified below. Referring to FIG. 3B, the difference between the intermediate layer 12' and the intermediate layer 12 of the foregoing embodiment is that the channel structure 121' has substantially the same inner diameter between the injection hole 122' and the mixing hole 123'. Straight strip (labeled area SP in the figure) instead of the flared shape shown in Figure 3A. Moreover, the turning portion 126' of the channel structure 121' is designed at a right angle. In contrast, the embodiment of FIG. 3B has no flare or cone and its end is extended, and the turning portion 126' cannot make a blood sample. The flow is smooth, but the production is simple and the cost is cheap, so there is still application value.

4 is a schematic view showing the appearance of different aspects of an electrode of a blood coagulation detecting device according to another embodiment of the present invention. Referring to FIG. 4, in the present embodiment, the blood coagulation detecting device 4 and the foregoing embodiment have substantially the same components, connection manners, and effects as those of the foregoing embodiments, except that the electrode 44 is shaped like an upper cap layer. 41. The intermediate layer 42 and the substrate layer 43 have the same rectangular plate shape, and are respectively disposed in parallel with the upper cover layer 41 and the substrate layer 43. Therefore, although the electrode 44 of the present embodiment is different from the configuration of the flow channel, it also produces a uniform electric field, and the same effect can be achieved. Of course, in other embodiments, the electrodes may be of any shape or size according to the principle, and the invention is not limited thereto.

According to a preferred embodiment of the present invention, the shape and size of the electrodes are correspondingly designed so that the flow path is under a uniform electric field, which is advantageous for the detection operation. However, in other embodiments, an uneven electric field caused by asymmetric or incomplete symmetry of the electrode shape may be applied, and the present invention is not limited thereto.

Hereinafter, the operation method and the detection flow of the blood coagulation detecting device 1 will be described with reference to Figs. 1 to 2B. However, the parameters or numerical values used below are all references and are not intended to limit the present invention.

4.5 mL of whole blood just taken out from the human body and a sodium citrate anticoagulant having a concentration of 0.129 M are mixed according to a ratio of 9:1, so that the blood becomes citrated blood, and the coagulation is lost; Blood is injected from the first opening 111. When the blood flows through the mixing portion below the second opening 112 and the mixing hole 123, the calcium chloride solution is dropped from the second opening 112, and the citrated blood and the calcium chloride solution are uniformly mixed through the communicating portion 124 and the capillary portion. . When the blood is mixed with the calcium chloride solution, it is called blood recalcification. The blood is subjected to the capillary force during the recalcification process, and the self-driving effect is generated, and the blood moves in the flow channel CH. In addition, since the blood coagulation reaction starts from the blood until it is completely solidified, since the electrode 14 of the blood coagulation detecting device 1 can be connected to the impedance analyzer HP 4194A Impedance/Gain-phase Analyzer (Hewlett-Packard Co. USA), the impedance analyzer can be The change in capacitance in the flow path CH of the blood coagulation detecting device 1 is taken, and the amount of change in the capacitance is used as a criterion for judging the blood coagulation reaction time.

5A and 5B are diagrams showing experimental results of blood coagulation detection using the blood coagulation apparatus shown in Fig. 1. 5A and 5B, in detail, in the above operation, the test frequency used for the impedance analyzer for detecting the change of the capacitance value may be 51 kHz, and the specific calculation method of the blood coagulation time may include the following several programs. . First, blood is injected or dripped from the first opening 111, and the capacitance value rapidly increases as shown in the area a of Fig. 5A. Next, the calcium chloride solution is added from the second opening 112, and the capacitance value rapidly drops as shown in the area b of FIG. Referring again to region c of Figure 5A, after the blood is completely mixed with the calcium chloride solution, the capacitance value tends to be stable, which can be defined as the start time of blood coagulation.

Thereafter, please refer to the area d of FIG. 5A and FIG. 5B, wherein FIG. 5B is an enlarged schematic view of the area d of FIG. 5A. Since the area d, the capacitance value increases steadily, but the amplitude is quite small, and the capacitance value does not increase until 15 minutes and 25 seconds. This time point defines that the blood does not solidify completely. The solidification starting point was subtracted from the solidification time point to measure the solidification time, which was 9 minutes and 08 seconds in total. This time is consistent with the normal clotting time interval.

Figure 6 is a flow chart showing a method of manufacturing a blood coagulation detecting device in accordance with a preferred embodiment of the present invention. Referring to FIG. 6 , in the embodiment, the method for manufacturing a blood coagulation detecting device comprises the steps of: forming a first opening, a second opening and an outlet on an upper cover layer (S61); a substrate layer respectively forming an electrode (S63); an optical glue or a photoresist material is disposed on the substrate layer to form an intermediate layer (S65); processing the intermediate layer (S67); and bonding the upper cap layer, the intermediate layer and the substrate layer To jointly define the first-class road (S69). Wherein, the formation of the electrode includes providing a heat resistant material or another photoresist material on the upper cap layer and the substrate layer, defining a pattern of the electrode, and a process of coating the film. Preferably, the definition of the pattern includes a laser processing or a lithography process. . In addition, the processing of the intermediate layer forms part of the flow path and includes procedures for laser processing, lithography, or machining. Further, the combination of the upper cap layer, the intermediate layer, and the substrate layer includes a procedure for low temperature bonding. However, the respective steps of the components and the manufacturing method of the blood coagulation detecting device have been described in detail in the foregoing embodiments, and will not be described herein.

As described above, the blood coagulation detecting device according to the present invention transmits the first-class track jointly defined by the substrate layer, the intermediate layer and the upper cap layer for blood input detection, and simultaneously guides the flow, and uses the correspondingly disposed electrodes. It detects the change of physical properties during blood coagulation, thereby providing the functions of simple operation and rapid detection, and has the results of data analysis for objective judgment.

Compared with the prior art, the detecting device and the manufacturing method thereof are suitable for using a hydrophilic substance as a material of the substrate layer and the upper cap layer, so as to achieve the effect of self-driving the blood to be tested without external force, and not only the pump is eliminated. Demand, effectively reducing the size of the device and manufacturing costs, more ready-to-use, no need for cleaning and special pre-processing advantages.

In addition, the device and the manufacturing method thereof can specifically utilize the characteristics of the capacitance change in the flow channel when the blood is coagulated, and objectively analyze the data blood coagulation time, which significantly improves the accuracy and application compared with the conventional image analysis method. Sex.

1, 4. . . Blood coagulation detecting device

11, 41. . . Upper cover

111. . . First opening

112. . . Second opening

113. . . Export

12, 12', 42. . . middle layer

121, 121’. . . Channel structure

122, 122’. . . Injection hole

123, 123’. . . Mixing hole

124, 124’. . . Connecting part

126, 126’. . . Steering section

13,43. . . Substrate layer

14, 44. . . electrode

141. . . Contact

142. . . Detection department

a, b, c, d, CP, EP, SP. . . region

CH. . . Runner

S61～S69. . . step

1 is an exploded perspective view of a blood coagulation detecting device according to a preferred embodiment of the present invention;

2A is a schematic view showing the combined appearance of a blood coagulation detecting device according to a preferred embodiment of the present invention;

2B is a schematic perspective view of the blood coagulation detecting device shown in FIG. 2A;

3A and 3B are top views of different aspects of an intermediate layer of a blood coagulation detecting device according to a preferred embodiment of the present invention;

4 is a schematic view showing the appearance of different aspects of an electrode of a blood coagulation detecting device according to a preferred embodiment of the present invention;

5A and 5B are diagrams showing experimental results of blood coagulation detection of a blood coagulation device according to a preferred embodiment of the present invention;

Figure 6 is a flow chart showing a method of manufacturing a blood coagulation detecting device according to a preferred embodiment of the present invention.

1. . . Blood coagulation detecting device

11. . . Upper cover

111. . . First opening

112. . . Second opening

113. . . Export

12. . . middle layer

121. . . Channel structure

122. . . Injection hole

123. . . Mixing hole

124. . . Connecting part

13. . . Substrate layer

14. . . electrode

141. . . Contact

142. . . Detection department

Claims (16)

A blood coagulation detecting device comprises: a substrate layer; an intermediate layer disposed on the substrate layer; an upper cap layer disposed on the intermediate layer, wherein the substrate layer, the intermediate layer, and the upper cap layer jointly define a first-class track And two electrodes are respectively disposed on the upper cap layer and the substrate layer. The blood coagulation detecting device according to claim 1 is a capacitive or electrical impedance blood coagulation detecting wafer. The blood coagulation detecting device according to claim 1, wherein the material of the substrate layer and the upper cap layer comprises glass, indium tin oxide, antimony oxide, or twin crystal. The blood coagulation detecting device according to claim 1, wherein the material of the intermediate layer comprises an optical glue or a photoresist material. The blood coagulation detecting device according to claim 1, wherein the upper cap layer has a first opening, a second opening and an outlet connected to the flow channel. The blood coagulation detecting device according to claim 1, wherein the flow prop has a mixing portion and a communicating portion, and the second opening and the mixing portion communicate with each other through the communicating portion. The blood coagulation detecting device according to claim 6, wherein the flow prop has a capillary portion, and the capillary portion surrounds the communication portion. The blood coagulation detecting device according to claim 1, wherein the flow path has a larger inner diameter near one end of the first opening. The blood coagulation detecting device according to claim 1, wherein the flow path has at least one turning portion, and the turning portion has an arc shape, a hairpin shape, or a horseshoe shape. The blood coagulation detecting device according to claim 1, wherein one of the electrodes is disposed on the upper cap layer, and the other one is disposed under the substrate layer. The blood coagulation detecting device according to claim 1, wherein the flow channel is interposed between the electrodes. A method for manufacturing a blood coagulation detecting device, comprising the steps of: forming a first opening, a second opening and an outlet on an upper cap layer; forming an electrode on the upper cap layer and a substrate layer; and providing an optical glue or a photoresist material on the substrate layer to form an intermediate layer; processing the intermediate layer; and bonding the upper cap layer, the intermediate layer and the substrate layer to collectively define a first-class track. The manufacturing method of claim 12, wherein the forming of the electrode comprises providing a heat resistant material or another photoresist material on the upper cap layer and the substrate layer, defining a pattern of the electrodes, and a process of coating the film. The manufacturing method of claim 13, wherein the definition of the patterns includes a laser processing or a lithography process. The manufacturing method of claim 12, wherein the processing of the intermediate layer forms part of the flow path and includes a process of laser processing, lithography, or machining. The manufacturing method of claim 12, wherein the bonding of the upper cap layer, the intermediate layer, and the substrate layer comprises a process of low temperature bonding.