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Title:
A MICROFLUIDIC CHIP AND A DIAGNOSTIC APPARATUS CONTAINING THE SAME
Document Type and Number:
WIPO Patent Application WO/2016/040620
Kind Code:
A1
Abstract:
The present invention provides a microfluidic chip comprising: a base layer wherein multiple electrodes are formed on both sides, and a first detection part and a second detection part, wherein a blood sample is injected to the first and the second detection parts; and the first and the second detection parts measuring different properties of the blood sample.

Inventors:
CHOI, Young Ho (15 Wedgewood Court, Belle Mead, New Jersey, 08502, US)
HAN, Byung Don (1 B-Dong, 401 Yangcheon-Ro,Gangseo-Gu, Seoul, 405-1406, KR)
RYU, Chi Un (1105-Ho, 156 Maebong-Ro,Dongjak-Gu, Seoul, KR)
HWANG, Yoon Ho (610-Ho, 10-20 Yangcheon-Ro 57-Gil,Gangseo-Gu, Seoul, KR)
KIM, Sae Ho (2 40, Sanghyeon-Ro42Beon-Gil, Suji-Gu,Yongin-Si, Gyeonggi-Do, 72-1201, KR)
YOON, Guk Hyun (9 Yongdap 9 Na-Gil, Seongdong-Gu, Seoul, KR)
Application Number:
US2015/049409
Publication Date:
March 17, 2016
Filing Date:
September 10, 2015
Export Citation:
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Assignee:
WELLS BIO, INC. (1 B-Dong, 401 Yangcheon-Ro,Gangseo-Gu, Seoul, 405-1406, KR)
CHOI, Young Ho (15 Wedgewood Court, Belle Mead, New Jersey, 08502, US)
International Classes:
G01N33/48; G01N33/487; G01N33/49
Foreign References:
US20130224781A12013-08-29
US20130118901A12013-05-16
US20120261067A12012-10-18
US20140183044A12014-07-03
US20070111353A12007-05-17
US7943089B22011-05-17
US20140083858A12014-03-27
Attorney, Agent or Firm:
CHOE, Yun H. (Lucas & Mercanti LLP, 30 Broad Street 21st Floo, New York New York, 10004, US)
Download PDF:
Claims:
CLAIMS

We Claim:

1. A microfluidic chip comprising:

a base layer wherein multiple electrodes are formed on both sides; and

a first detection part and a second detection part,

wherein a blood sample is injected to the first and the second detection parts; and the first and the second detection parts measuring different properties of the blood sample.

2. The microfluidic chip of claim 1, wherein the first detection part measures an enzyme activity level in a blood sample using photometric, electrochemical, or colorimetric techniques; and the second detection part measuring the amount of hemoglobin in the blood sample using photometric, electrochemical, or colorimetric techniques.

3. The microfluidic chip of claim 1, wherein the same blood sample is divided and injected into the first detection and to the second detention units.

4. The microfluidic chip of claim 1, further comprising:

an upper and a lower protective layers formed on both sides of the base layer; and an upper and a lower bonding layer adhering the upper and the lower protective layers on the base layer.

5. The microfluidic chip of claim 1, wherein the first and the second detection parts are defined by the opening formed on the upper and the lower bonding layers.

6. The microfluidic chip of claim 1, wherein the multiple electrodes comprises a contacting electrode that contacts each of the blood sample; a connecting electrode connected to a diagnostic apparatus; and a wiring electrode connecting the contacting electrode and the connecting electrode.

7. The microfluidic chip of claim 6, wherein the connecting electrode that is the part of the multiple electrode bends toward one side.

8. The microfluidic chip of claim 6, wherein the contacting electrodes and the connecting electrodes are formed with a bigger width compared to the wiring electrodes

9. The microfluidic chip of claim 1, wherein the protective layers and the upper and the lower bonding layers are form in a smaller area than the base layer so as for a part of the electrodes to be exposed

10. The microfluidic chip of claim 1, wherein an opening is formed on the top and the bottom protective layers; and the opening is made with a transparent substance.

11. The microfluidic chip of claim 4, wherein an auxiliary electrode is formed on the upper and the lower bonding layers in the position corresponding to that of the electrodes.

12. The microfluidic chip of claim 2, wherein the first detection and the second detection part comprising a multiple detection methods selected from photometric, an electrochemical, or colorimetric techniques, or the combination of them.

13. The microfluidic chip of claim 2, wherein the first and the second detection parts comprises electron transport medium and electrodes used for the electrochemical detection.

14. A diagnostic apparatus comprising:

a microfluidic chip measuring a property of a blood sample comprising:

a first and a second analysis units; and

a control unit which receives an analysis results from the first and the second analysis units and displays on a display unit, wherein the microfluidic chip measures different properties of a same blood sample.

15. The diagnostic apparatus of claim 14, wherein the microfluidic chip comprising: a base layer, wherein a multiple electrodes are formed on both sides; and a first and a second detection parts formed on both sides of the base layer,

wherein a blood sample is injected into the first and the second detection parts and the injected sample is dissolved so that the first and the second detection parts measures a different properties of the blood sample.

16. The diagnostic apparatus of claim 14, wherein the first detection part measures an enzyme activity level in the blood sample, and the second detection part measures an amount of hemoglobin in the blood sample. 17. The diagnostic apparatus of claim 16, wherein the control unit calculates and produces an activity level of the enzyme per gram of hemoglobin from the enzyme activity level and the amount of hemoglobin.

18. The diagnostic apparatus of claim 14, wherein the first and the second analysis units comprise an amplifier that amplifies a signal; and a converter which converts the measured value to digital signals.

19. The diagnostic apparatus of claim 14, further comprising a socket unit connecting to the microfluidic chip, wherein the socket unit is connected to a side of the microfluidic chip.

20. The diagnostic apparatus of claim 14, wherein the first and the second detection parts containing a surfactant that dissolves the blood sample.

21. The diagnostic apparatus of claim 20, wherein the surfactant is selected from the group consisting of a cationic surfactant, an anionic surfactant, an amphiphilic surfactant, a non-ionic surfactant, and a mixture thereof.

Description:
A MICROFLUIDIC CHIP AND A DIAGNOSTIC APPARATUS CONTAINING THE SAME

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of priority from Korean Patent Application KR 10- 2014-0119631, filed September 10, 2014, the contents of which are incorporated herein by reference in their entirety.

FIELD OF INVENTION

The present invention is related to a microfluidic chip and a diagnostic apparatus containing the same.

BACKGROUND OF THE INVENTION

Macroscopic, optical, or electrochemical detection techniques are generally utilized to detect the amount of a specific enzyme in blood. Among these techniques,

electrochemical detection processes can be influenced greatly by various artifacts in the blood sample such as oxidation-prone ascorbic acid, acetoaminophen and/or uric acids. In particular, hematocrit artifacts may cause significant measurement errors, which lead to a wrong interpretation.

Examples of known methods used to reduce the effect of hematocrit includes: compensating signal reduction using the electrical signals in erythrocytes detected from the substances determining hematocrit; using a sensing-membrane completed with reagents immobilized on electrodes based on screen printing technology, or using a blood separating unit; preventing the binding of erythrocytes and proteins from adhering on the electrode by usage of a thin-layer of the enzyme interacting with an analyte; correcting the results using mathematical manipulation of the results obtained by applying electric potential twice. Glucose-6-phosphate dehydrogenase (G6PD) plays a critical role in biochemical reactions. As a part of pentose phosphate cycle, it has been known to minimize the oxidative attack of activated oxygen species. Glucose-6-phosphate dehydrogenase is ubiquitous in human cells, especially exists at high concentration in erythrocytes which function as oxygen carriers, and are particularly prone to oxidative attacks. The glucose-6-phosphate dehydrogenase activity offers a highly efficient defense mechanism under the unwanted oxidation stress. In glucose-6-phosphate dehydrogenase deficiency, one can experience deleterious side-effects when strongly oxidizing agents like quinine as an antimalarial drug is administrated.

The conventional diagnostic methods for measurement of the glucose-6-phosphate dehydrogenase activity level is either a lateral flow kit utilizing the enzyme reaction, or a diagnostic kit that is based on fluorochrome analysis of a fluid system. These methods, however, often require expensive diagnostic devices, or have limitations in visual detection of a carrier.

An alternative methods for measurement of enzyme activities with improved accuracy and convenience are still desirable.

SUMMARY OF INVENTION

Disclosed is a microfluidic chip that can measure two properties of a blood sample simultaneously.

Disclosed is a diagnostic apparatus for measuring simultaneously or respectively the hemoglobin amount, and enzyme activity level, and enzyme activity per gram of hemoglobin in a blood sample.

DETAILED DESCRIPTION OF THE DISCLOSURE

Disclosed is a microfluidic chip comprising:

a base layer wherein multiple electrodes are formed on both sides; and

a first detection part and a second detection part,

wherein a blood sample is injected into the first and the second detecti parts;

wherein the first and the second detection parts measure different properties of the blood sample. The first detection part can measure an enzyme activity level in a blood sample using photometric, electrochemical, or colorimetric techniques; and the second detection part can measure the amount of hemoglobin in the blood sample using photometric, electrochemical, or colorimetric techniques.

The same blood sample can be divided and injected into the first detection and the second detention parts.

An upper and a lower protective layers formed on both sides of the base layer, and an upper and a lower bonding layer adhering the upper and the lower protective layers on the base layer are further disclosed.

Disclosed are first and the second detection parts defined by the opening area formed on the upper and the lower bonding layers. The multiple electrodes can comprise a contacting electrode that contacts each of the blood sample; a connecting electrode connected to a diagnostic kit; and a wiring electrode connecting the contacting electrode and the connecting electrode.

The connection of the electrodes among the multiple electrodes can be bent toward one side.

The contacting electrode and the connecting electrode can be wider than the wiring electrode.

The upper and the lower protective layers and the upper and the lower bonding layers can be formed in a smaller area than that of the base layer so that a part of electrodes to be exposed.

An opening can be formed on the upper and the lower protective layers; and the openings can be made of transparent substance.

An auxiliary electrode can be formed on the upper and the lower bonding layers in the corresponding areas of the electrodes. A diagnostic apparatus is disclosed comprising:

a microfluidic chip capable of detecting a property of a blood sample comprising: a first analysis unit and a second analysis units and

a control unit capable of receiving data from the first analysis unit and the second analysis unit and displays on the display unit,

wherein the microfluidic chip measures different properties of the same blood sample.

Disclosed is a microfluidic chip comprising:

a base layer, wherein multiple electrodes are formed on both sides; and a first and a second detection parts formed on both sides of the base layer,

wherein a blood sample is injected into the first and the second detection parts and the injected sample is dispersed such that the first and the second detection parts can respectively measure a different properties of the blood sampk

The first detection part can measure an enzyme activity level in the blood sample, and the second detection part can measure an amount of hemoglobin in the blood sample

The disclosed control unit calculates an activity level of the enzyme per gram using data for the enzyme activity level and the amount of hemoglobin.

The first and the second analysis units that respectively contain an amplifier that amplifies signals; and a converter that converts the measurements to digital signals.

A socket unit connecting the microfluidic chip is further disclosed, wherein the socket unit can be connected to one side of the microfluidic chip.

The disclosed microfluidic chip has electrodes placed on both sides of the base laye and the each side measures two properties of the blood sample to quickly determine any deficiency of glucose-6-phosphate dehydrogenase enzyme.

A disclosed diagnostic kit comprises: detection parts on both sides of the base layer, wherein each detection part is capable of measuring the amount of hemoglobin or the level of enzyme activity; and

the control unit sequentially calculates the results to produce the enzyme activity level per gram of hemoglobin, which allows a detection of glucose-6-phosphate

dehydrogenase deficiency.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 is a block diagram showing the diagnostic apparatus in Example 1.

Figure 2 is a schematic diagram illustrating the method to measure simultaneously the enzyme activity level and the amount of hemoglobin in a blood sample.

Figure 3 is the exploded oblique view of the microfluidic chip in Example 1.

Figure 4 is the drawing of the base layer of the microfluidic chip in Example 1.

Figure 5 is the top-view of the assembled microfluidic chip in Example 1.

Figure 6 is the cross-section of the microfluidic chip in Example 1, cut in A-A " direction.

Figure 7 is the diagram showing the method measuring the glucose-6-phosphate dehydrogenase activity in Example 1.

Figure 8 is the exploded oblique view of the microfluidic chip from the top, after the adhesive layer has been imprinted with the electrodes by the electrode on the base layer, as in Example 2.

Figure 9 is the exploded oblique view of the microfluidic chip from the bottom, after the adhesive layer has been imprinted with the electrodes by the electrode on the base layer, as in Example 2.

Figure 10 is the cross-section of the microfluidic chip in Example 1 or 2.

Figure 11 is the top view of the microfluidic chip in Example 3 layer by layer from top to bottom, from left to right.

Figure 12 is the top view of the assembled microfluidic chip in Example 3.

Figure 13 is the cross-section of the microfluidic chip in Example 3, cut in B-B' direction. EXAMPLES

Hereinafter, specific examples of the present invention will be described using Figures. But the disclosure is not limited to the examples.

The identical numbering is used for the explanations when indicating the

components.

EXAMPLE 1

Figure 1 is a block diagram showing the diagnostic apparatus according to Example 1.

Figure 1 disclosed a diagnostic apparatus (1) comprising a mounting unit (10), a socket unit (15), a first analysis unit (20), a second analysis unit (30), a control unit (40), and a display unit (50).

The mounting unit (10) can provide the way whereon the microfluidic chip (Figure 3, 100) can be mounted.

The socket unit can be electrically connected to the mounting unit (10).

The socket unit (15) can recognize the measured values detected by the microfluidic chip and transfer the data to the diagnostic apparatus (1)

The first analysis unit (20) and the second analysis unit (30) analyze the measured value transferred from the socket unit (15).

The first analysis unit (20) and the second analysis unit (30) can analyze the different measured values respectively.

The first analysis unit (20) can analyze the enzyme activity level detected by the microfluidic chip (100), and the second analysis unit (30) can analyze the amount of hemoglobin in the blood sample that is measured at the second analysis unit (30).

The first analysis unit (20) can obtain the enzyme activity measured value from the first detection part on the microfluidic chip (100), and the second analysis unit (30) can obtain from the second detection part (Figure 3, 151) the amount of hemoglobin in the blood sample. The control unit (40) can be electronically connected to, and controls the socket unit (15), the first analysis unit (20), the second analysis unit (30), and the display unit (50).

The control unit (40) can control the microfluidic chip (100). By the control of the control unit (40), the measured values data obtained from the microfluidic chip (100) is transferred to the first analysis unit (20) and the second analysis unit (30), and the measured values can be displayed on the display unit (50) by the first and second analysis units (20 and 30). The control unit (40) can calculate and produce the enzyme activity level per gram of hemoglobin from the values analyzed at the first analysis part (20) and the second analysis unit (30).

In other words, the control unit (40) can calculate and produce the enzyme activity level per a gram of hemoglobin from the enzyme activity value obtained from the first analysis unit (20) and the amount of hemoglobin value obtained from the second analysis unit (30).

The control unit (40) can display the enzyme activity level per a gram of hemoglobin on the display unit (50).

The display unit (50) converts the values analyzed at the the first and second analysis unit (20 and 30) to images.

The display unit (50) can display the analyzed values as images by the control of the control unit (40), and allows visual confirmation of the amount of hemoglobin and the enzyme activity in the blood sample.

Moreover, the display unit (50) can display the enzyme activity level per a gram of hemoglobin on the display (50) so that the apparatus to evidence the safety for

administering the agent, such as the administration of the antimalarial drug or apply for neonate medical examination program. Figure 2 is a schematic diagram showing the usage of the method in order to measure simultaneously the amount of hemoglobin and the enzyme activity in a blood sample.

Figure 2 discloses blood samples are injected into the microfluidic chip (1) that can be mounted on the mount unit (10) of the diagnostic apparatus (1) from the Example 1.

The blood sample placed into the microfluidic chip (100) is dispersed and the red blood cells lysed. The lysate moves to the first detection part (131) and the second detection part (151) on the microfluidic chip (100).

Surfactants can be placed into the vicinity of the first and second detection parts (131, 151).

A surfactant can dissolve the blood sample.

The surfactant can contain cationic surfactants, anionic surfactants, zwitterionic surfactants, or nonionic surfactants. The surfactant can be a combination of two or more of the surfactants listed in the previous sent.

The first detection part (131) can measure the first property of the blood sample lysate, and the second detection part (151) can measure the second property of the blood sample lysate.

The first and second properties can be different properties. The first property can be an enzyme activity, and the second property can be an amount of hemoglobin.

Alternatively, the first property can be an amount of hemoglobin and the second property can be an enzyme activity.

The first detection part (131) can transfer to the the first analysis data (20) an enzyme activity value measured in the blood sample lysate, and the second detection part (151) can transfer to the second analysis part (30) an amount of hemoglobin measured in the blood sample lysate.

The first analysis part (20) analyzes the enzyme activity level transferred from the first detection part (131) and transfers the results to the control unit (40). The first analysis unit (20) can comprise the first amplifier (21) and the first converter (23). The first amplifier (21) can amplify the measured values of the enzyme activity level received from the first detection part (131), and transfer the amplified values to the first converter (23). The first converter can covert the amplified values into digital signals and transfers the data to the control unit (40).

The second analysis unit (30) analyzes the measured values of the amount of hemoglobin transferred from the second detection part (151) and transfers the resulting data to the control unit (40).

The second analysis unit (30) comprises the second amplifier (31) and the second converter (33). The second amplifier (31) can amplify the measured values of the amount of hemoglobin received from the second detection part (151), and transfer the amplified values to the second converter (33). The second converter coverts the amplified values into digital signals and transfers to the control unit (40).

The control unit (40) can calculate and deduce the enzyme activity level per a gram of hemoglobin from the values analyzed at the second analysis part (30).

In other words, the control unit (40) can calculate and deduce the enzyme activity level per gram of hemoglobin from the enzyme activity value obtained from the first analysis unit (20) and the amount of hemoglobin value obtained from the second analysis unit (30).

The control unit (40) can control the display unit (50), which can display the amount of hemoglobin in the blood sample, the enzyme activity level, and the enzyme activity level per a gram of hemoglobin.

Figure 3 is the exploded oblique view showing the microfluidic chip in the Example 1. Figure 4discloses a drawing of the base layer of the microfluidic chip in the Example 1, Figure 5 is the top view of the microfluidic chip in the Example 1, and Figure 6 is the cross- section of the Figure 5 cross-secting in the A-A " direction. Referring to Figures 3 through 6, the microfluidic chip (100) can comprise as a layered structure of a base layer (100), an upper bonding layer (130), and an upper protective layer (140), a lower bonding layer (150), and a lower protective layer (160).

A plurality of electrodes can be formed on the base layer (101). A plurality of electrodes can be formed on each side of the base layer (101). On the upper face (103) of the base layer (101), a plurality of electrodes can be formed, on the lower face (105) of the base layer (101), a plurality of electrodes can be formed. The plurality of electrodes can comprise contacting electrodes, wiring electrodes, and connecting electrodes. The contacting, wiring, and connecting electrodes can comprise a single unit.

The length of the microfluidic chip is 30-80 mm, preferably 30-60 mm, or more preferably 40-60 mm, and the width of the microfluidic chip is 4 mm to 8 mm, preferably 4 mm to 7 mm, more preferably 4 mm to 6 mm. The thickness of the microfluidic chip is 0.5 mm to 1.5 mm, preferably 0.8 mm to 1.3 mm, more preferably 0.9 mm to 1.1 mm. The opening area (241) or (251) is 1 mm to 3 mm in width and 4 mm to 7 mm in length, preferably 5 mm to 6 mm. The thickness of the each layer in the microfluidic chip is 100 μιη to 400 μιη, preferably 100 μιη to 350 μιη. The thickness of the base layer (100), upper bonding layer (130) and lower bonding layer (150) is 100 μιη to 300 μιη and the thickness of the upper protective layer (140) and the lower protective layer (160) is 100 μιη to 150 μιη.

The first (111) and the second electrodes (121) can be formed on the upper layer (103) of the base layer (101).

The first electrodes (111) can comprise the first contacting electrodes (113), the first wiring electrodes (115), and the first connecting electrodes (117). The first wiring electrodes (115) are capable of electrically connecting the first contacting (113) and the first connecting electrodes (117). The first contacting electrodes (113), the first wring electrodes (115), and the first connecting electrodes (117) can be formed with the identical substances. The first contacting electrodes (113), the first wring electrodes (115), and the first connecting electrodes (117) can be formed with the identical substances. The first contacting electrodes (113), the first wiring electrodes (115), and the first connecting electrodes (117) can comprise a single unit.

The first wiring electrodes (115) can be narrower width than of the first contacting electrode (113). The first wiring electrodes (115) can also be formed with a narrower width than that of the first connecting electrode (117). The first contacting electrodes (113) can contact with the blood sample. The first connecting electrodes (117) can electrically connect with the socket unit (15) of the diagnostic apparatus (1). The contacting area of the first connecting electrodes (117) can wider than of the first wiring electrodes (115).

The second electrodes (121) can comprise the second contacting electrodes (123), the second wiring electrodes (125), and the second connecting electrodes (127). The second wiring electrodes (125) can electrically connect the second contacting electrodes (123) and the second connecting electrodes (127). The second contacting electrodes (123), the second wiring electrodes (125), and the second connecting electrodes (127) can be formed by identical substances. The second contacting electrodes (123), the second wiring electrodes (125), and the second connecting electrodes (127) can be formed as single unit.

The second wiring electrodes (125) can be narrower than the second contacting electrode (123) and the second connecting electrode (127). The second contacting electrodes (123) can contact the blood sample The second connecting electrodes (127) can electrically connect with the socket unit (15) of the diagnostic apparatus (l).The contacting area of the second connecting electrodes (127) can be wider width than the second wiring electrodes (125).

An upper bonding layer (130) can be formed on the upper face (103) of the base layer (101) comprising first electrodes (111) and the second electrodes (121). The upper bonding layer (130) can contain adhesive substances on each side.

The upper bonding layer (130) can comprise an opening. The opening can be at a position corresponding to the first contacting electrodes (113) and the second contacting electrodes (123). The opening can expose a part of the first contacting electrodes (113). The opening can expose a part of the the second contacting electrodes (123). The opening can correspond to the first detection part (131). The opening in the first bonding layer (130) forms a void that is the first detection part (131).

When a blood sample is injected, the first detection part (131) can measure enzyme activity in the blood sample lysate by electrochemical detection method.

The contents of the first detection part (131) can comprise electron transport medium. It is thought that Glucose-6-phosphate in the blood sample placed into the first detection part (131) undergoes a dehydrogenation reaction mediated by glucose-6- phosphate dehydrogenase, and forms 6-phosphogluconolacton in the first detection part (131) of the microfluidic chip (100). From the dehydrogenation reaction, it is thought that oxidized Nicotinamide Adenine Dinucleotide Phosphate (NADP+) and Nicotinamide Adenine Dinucleotide Phosphate H (NADPH) are formed in the detection part (131) of the microfluidic chip (100), and electrons produced from the process that NADPH converts into NADP+, move through electron transport medium in the first contacting (113) or the second contacting (123) units.

It is thought that depending on the activity level of the glucose-6-phosphate dehydrogenase, a different amount of electrons goes through the first contacting electrodes (113) or the second contacting electrodes (123) on the microfluidic chip (100). The amount of the glucose-6-phosphate dehydrogenase in a blood sample can be determined by measuring the amount of the electrons moving into the first contacting electrodes (113) and the second contacting electrodes (123) contained in the first detection part (121) on the microfluidic chip (100).

One of the electrodes from the first contacting electrodes (113) or the second contacting electrodes (123) can be an active electrode and the other can be a standard electrode. It is thought that, when a blood sample is introduced in the first detection part (131), a voltage difference between the first contacting electrode (113) and the second contacting electrode (123) forms, the voltage difference can be transferred to the first connecting electrodes (117) and the second connecting electrodes (127) electrodes through the first wiring electrodes (115) and the second wiring electrodes (125).

The first detection part (131) can determine the enzyme activity level in the blood sample using electrochemical, photometric, and colorimetric methods. When the first detection part (131) does not use an electrochemical method, the electrodes on the base layer (101) can be omitted.

The upper protective layer (140) can layered on the upper bonding layer (130). An upper opening (141) can be formed on the upper protective layer (140). The upper opening (141) can be formed over the upper detection part (131). A transparent substance can fill or partially fill the upper opening (141).

The upper opening (141) can be partially or totally filled with air. The first detection part (131) can be connected the exterior outside via an upper vent hole (143). That is, air inside the first detection part (131) can escape through the upper vent hole. When the blood sample is positioned near the first detection part (131) via the vent hole (143), it is thought that capillary effects will place the blood sample onto the first detection part (131).

The upper bonding layer (130) and the upper protective layer (140) can be narrower than of the base layer (101). In these embodiments, it is thought that the parts of the first connecting electrodes (117) and the second connecting (127) electrodes can be exposed. In embodiments in which the first connecting electrodes (117) and the second connecting electrodes (127) are exposed partly, the first connecting (117) and the second connecting electrodes (127) can be connected to the socket unit (15), when the microfluidic chip (100) is mounted on the mounting unit (10). In some embodiments, the exposed parts of the first connecting electrodes (117) and the second connecting electrodes (127) can be connected to the socket unit (15).

The second connecting electrodes (127) can bend in the direction of the first connecting electrodes (117). In those embodiments, the socket unit (15) can be positioned on a side of the microfluidic chip (100). In those embodiments, the socket resides on a side of the microfluidic chip (100) near the first connecting electrodes and is electrically connected to the first (117) and the second connecting electrodes (127). Because of the bend of the second (127) toward the first (117) electrodes, the length of the first (111) and the second electrodes (121) can be set identically.

As disclosed in the figures, the positions of the first (113) and the second contacting electrodes (123) allow the first wiring electrodes (115) have longer length than the second wiring electrodes (125). In these embodiments, this leads to form a bend in the second connecting electrodes (127). One of ordinary skill in the art would expect results for an unexpected results argument. The electrodes can be constructed so that the second connecting electrodes (127) is longer than the first connecting electrodes (117) so that both electrodes should have the identical length. In some embodiments in which the length of the first (111) and the second electrodes (121) are identical, the resistance difference in the first (111) and the second (121) is reduced and the distortion of the voltage transferred from the contacting electrodes to the connecting electrodes can be prevented.

A third electrode (171) and the fourth electrode (181) can be formed on the bottom of the base layer (101).

The third electrodes (171) can comprise the third contacting electrodes (173), the third wiring electrodes (175), and the third connecting electrodes (177). The third wiring electrodes (175) can electrically connect the third contacting (173) and the third connecting electrodes (177). The third contacting electrodes (173), the third wring electrodes (175), and the third connecting electrodes (177) can be formed by the identical substances. The third contacting electrodes (173), the third wiring electrodes (175), and the third connecting electrodes (177) can be formed as single unit.

The third wiring electrodes (175) can be narrower than the third contacting electrode (173) and the third connecting electrode (177). The third contacting electrodes (173) can contact the blood sample and the third connecting electrodes (177) can electrically connect with the socket unit (15) of the diagnostic apparatus (l).The contacting area of the third connecting electrodes (177) can be wider than the third wiring electrodes (175).

The fourth electrodes (181) can comprise the fourth contacting electrodes (183), the fourth wiring electrodes (185), and the fourth connecting electrodes (187). The fourth wiring electrodes (185) can electrically connect the fourth contacting (183) and the fourth connecting electrodes (187). The fourth contacting electrodes (183), the fourth wring electrodes (185), and the fourth connecting electrodes (187) can be formed by the identical substances. The fourth contacting electrodes (183), the fourth wiring electrodes (185), and the fourth connecting electrodes (187) can be formed as single unit.

The fourth wiring electrodes (185) can be narrower than the fourth contacting electrode (183) and the fourth connecting electrode (187). The fourth contacting electrodes (183) can contact with the blood sample, and the fourth connecting electrodes (187) can electrically connect with the socket unit (15) of the diagnostic apparatus (l).The contacting area of the fourth connecting electrodes (187) can be wider than the fourth wiring electrodes (185).

A lower bonding layer (150) can be layered on the lower face (103) of the base layer (101) that has third electrodes (171) and the fourth electrodes (181). The lower bonding layer (150) can comprise adhesive substances on each side so as to bond the lower protective layer (160) onto the base layer (101).

The lower bonding layer (150) can comprise an opening. The opening can be formed over the third contacting electrodes (173) and the fourth contacting electrodes (183). The opening can expose the part of the third contacting electrodes (173) and the fourth contacting electrodes (183). The opening can define the physical dimensions of a second detection part (151).

The opening of the second bonding layer (150) forms an empty space that is the second detection part (151).

In some embodiments, when a blood sample is injected, the second detection part (151) can measure the amount of hemoglobin in the blood sample using electrochemical, photometric, and colorimetric methods. In some embodiments, when the second detection part (151) does not use an electrochemical method, the electrodes on the base layer (101) can be omitted.

In embodiments in which the second detection part (151) measures the amount of hemoglobin using an electrochemical method, One of the electrodes from the third (173) or the fourth contacting (183) can be used as an active electrode and the other can be used as a standard electrode. It is believed that when a blood sample is injected in the second detection part (151), a voltage difference between the third contacting electrodes (173) and the fourth contacting electrodes (183) forms and that the voltage difference can be transferred to the third connecting electrodes (177) and the fourth connecting (187) electrodes through the third (175) wiring electrodes and the fourth wiring electrodes (185).

The lower protective layer (160) can be layered on the lower bonding layer (150). A lower opening (161) can be formed on the lower protective layer (160). The lower opening (161) can be formed over the lower detection part (151). A transparent substance can fill the lower opening (161)

The lower opening (161) can be partially or totally filled with air. The second detection part (151) can be connected the exterior outside via lower vent hole (163). That is, air inside the second detection part (151) can escape through the lower vent hole. When the blood sample is positioned near the second detection part (151) via the vent hole (163), it is thought that capillary effects will place the blood sample into the second detection part (151).

The lower bonding layer (150) and the lower protective layer (160) can be narrower width than the base layer (101). In some embodiments in which the lower bonding layer (150) and the lower protective layer (160) are narrower than the base layer (101), the parts of the third connecting (177) and the fourth connecting (187) electrodes can be exposed. In these embodiments, the third connecting electrodes (177) and the fourth connecting electrodes (187) are exposed partly, the third connecting (177) and the fourth connecting electrodes (187) can be easily connected to the socket unit (15), when the microfluidic chip (100) is mounted on the mounting unit (10). In these embodiments, the exposed parts of the third connecting electrodes (177) and the fourth connecting electrodes (187) can be connected to the socket unit (15). The fourth connecting electrodes (187) can bend in the direction of the third connecting electrodes (177). In those embodiments, the socket unit (15) can be positioned on a side of the microfluidic chip (100). In those embodiments, the socket resides on a side of the microfluidic chip (100) near the third connecting electrodes and is electrically connected to the third (177) and the fourth connecting electrodes (187).

Because of the bend of the fourth (187) toward the third (117) electrodes, the length of the third (111) and the fourth electrodes (181) can be set identically.

As disclosed in the figures, the positions of the third (173) and the fourth contacting electrodes (183) allow the third wiring electrodes (175) have longer length than the fourth wiring electrodes (185). In these embodiments, this leads to form a bend in the fourth connecting electrodes (187). The electrodes can be constructed so that the fourth connecting electrodes (187) longer than the third connecting electrodes (177) so that both electrodes should have the identical length. In some embodiments in which the length of the third (171) and the fourth electrodes (181) are identical, the resistance difference in the third (171) and the fourth (181) is reduced and the distortion of the voltage transferred from the contacting electrodes to the connecting electrodes can be prevented.

The third electrode (171) and the fourth electrode (181) can be formed

symmetrically to the base layer (101). In these embodiments, the fourth connecting electrodes (187) can bend in the same direction as the second connecting electrodes (127). As the third (171) and the fourth (181) are symmetrical tithe first (111) and the second (121) electrodes, respectively, the tip of the bend forms the socket unit (15) in a way that the socket is positioned on a side of the microfluidic chip.

In some embodiments, the first (131) and the second detection part (151) exist in symmetrical position. In these embodiments, when a blood sample that is injected into a side of the microfluidic chip near the first (131) and second detection part (151), the blood sample is divided and placed by capillary action into the first detection (131) and the second detection parts (151). In these embodiments, the first detection part (131) measures enzyme activity level, and the second detection part (151) measures the amount of hemoglobin from the identical blood sample. The first detection part (131) and the second detection part (151) can each comprise an electron transport medium. In some embodiments, the the electron transport medium can be used in electrochemical detection.

In some embodiments, there is a plurality of first detection parts (131) and the second detection parts (151) so that multiple detection methods among electrochemical, photometric, and colorimetric methods can be used.

EXAMPLE 2

Figure 8 and Figure 9 are exploded oblique views showing the microfluidic chip from the top or from the bottom, respectively, after the adhesive layer has been imprinted with the electrodes by the electrode on the base layer, as in Example 2.

Figure 10 is a cross-section according to Example 1 or 2.

Example 2 is identical to Example 1 except that electrodes are formed on the upper and lower bonding layers by imprinting with the electrodes on the base layer. Therefore, detailed descriptions about the identical components are omitted in Example 2.

In Figure 8 and Figure 9, a microfluidic chip (200) comprises a structure, a base layer (201), an upper bonding layer (230), an upper protective layer (240), a lower bonding layer (250), and a lower protective layer (260) being stacked.

The upper face of the base layer (201) comprise the first (211) and the second electrodes (221). The lower face of the base layer (2001) comprise the third (271) and the fourth electrodes (281).

The first electrode (211) comprises the first contacting electrodes (213), the first wiring electrodes (215), and the first connecting electrodes (217), and the second electrode (221) comprises the second contacting electrodes (223), the second wiring electrodes (225), and the second connecting electrodes (227).

The third electrode (271) comprises the third contacting electrodes (273), the third wiring electrodes (275), and the third connecting electrodes (277), and the fourth electrode (281) comprises the fourth contacting electrodes (283), the fourth wiring electrodes (285), and the fourth connecting electrodes (287). In some embodiments plurality of electrodes are formed on the upper (230) and the lower bonding (250) layers.

In some embodiments, a first auxiliary (232) and the second auxiliary electrodes (234) are formed on the upper bonding layer (230). A first auxiliary (232) and the second auxiliary electrodes (234) are formed on the lower bonding layer (230).

In some embodiments, the first auxiliary electrodes (232) are formed on the corresponding position of the first electrodes (211).

The second auxiliary electrodes (234) are formed on the corresponding position of the second electrodes (221).

In some embodiments, when the first bonding layer (230) attaches to the upper face of the base layer (101), the first auxiliary electrodes (232) can contact with the first electrodes (211), and the second auxiliary electrodes (234) can contact to the second electrodes (221). In these embodiments, the first auxiliary electrodes (232) contact with the first electrodes (211), and the second auxiliary electrodes (234) contact to the second electrodes (211), the area where electrons can pass through increases, and resistance decrease accordingly. The resistance decrease minimizes the voltage distortion transferred from the first (213) and the second contact (223) electrodes to the first (217) and the second contacting (227) electrodes, which allows one to measure the enzyme activity level more accurately.

In some embodiments, a third auxiliary (252) and the fourth auxiliary electrodes

(254) are formed on the lower bonding layer (250). A third auxiliary (252) and the fourth auxiliary electrodes (254) are formed on the lower bonding layer (250).

In some embodiments, the third auxiliary electrodes (252) are formed on the corresponding position of the third electrodes (271).

The fourth auxiliary electrodes (254) are formed on the corresponding position of the fourth electrodes (281).

In some embodiments, when the third bonding layer (250) attaches to the upper face of the base layer (101), the third auxiliary electrodes (252) can contact with the third electrodes (271), and the fourth auxiliary electrodes (254) can contact to the fourth electrodes (281). In these embodiments, the third auxiliary electrodes (252) contact with the third electrodes (271), and the fourth auxiliary electrodes (254) contact to the fourth electrodes (271), the area where electrons can pass through increases, and resistance decrease accordingly. The resistance decrease minimizes the voltage distortion transferred from the third (273) and the fourth contact (285) electrodes to the third (277) and the fourth contacting (287) electrodes, which allows one to measure the amount of hemoglobin more accurately.

EXAMPLE 3 FIGs 11-13 provides Example 3. Disclosed is a Layer I. The Layer I can be rectangular in profile and be 40 to 60 mm long, 6 mm wide, and 250 μιη and the microfluidic chip containing Layers I - V can be 1 mm thick. The Layer I may have: i) a first contacting electrode (313), a first connecting electrode (317), and a first wiring electrode (315) (together, a "first reference electrode");

ii) a second contacting electrode (323), a second connecting electrode (327), and a second wiring electrode (325) (together, a "first checking electrode"); and iii) a third contacting electrode (333), a third connecting electrode (337), and a third wiring electrode (335) (together, a "first working electrode").

The first contacting electrode (313) is connected to the first connecting electrode (317) by the first wiring electrode (315); the second contacting electrode (323) is connected to the second connecting electrode (327) by the second wiring electrode (325); the third contacting electrode (333) is connected to the third connecting electrode (337) by the third wiring electrode (335). Disposed together are the first contacting electrode (313), the second contacting electrode (323), and third contacting electrode (333). Further disposed together are the first connecting electrode (317), the second connecting electrode (327), and the third connecting electrode (337). Each of the first contacting electrode (313), the second contacting electrode (323), and third contacting electrode (333) can independently comprise a bend. The extremity of the first contacting electrode (313) can be bifurcated. The extremity of the third contacting electrode (333) can be bulbous. The bifurcations of the first contacting electrode (313) can at least partially envelop the extremity of the third contacting electrode (333).

Disclosed is Layer II that can separate a Layer I from a Layer III. Layer II can be 250 μιτι in depth. Disposed in Layer II is a void (331). The void (331) can be positioned over the first contacting electrode (313), the second contacting electrode (323), and the third contacting electrode (333). A fluid can partially or completely fill the void (331). The capacity of the void (331) or (351) can be in the range of 0.25 μί to 8 μί or preferably 0.25 μί to 6 μί, more preferably 0.5 to 5 μΙ and most preferably 0.5 μί to 2.5 μί. The capacity of the void (331) can be 2.5 μΙ. The void (331) can be positioned to ensure that a sufficient quantity of fluid reaches the first contacting electrode (313), the second contacting electrode (323), or independently the third contacting electrode (333). The void (331) can be positioned to ensure that a sufficient quantity of fluid reaches the first contacting electrode (313), the second contacting electrode (323), and the third contacting electrode (333). The void (331) can be present parallel or perpendicular to the longest axis of the Layer II. The void (331) can be 5.6 μιτι in depth. The void (331) can be exposed to atmospheric air.

Disclosed is a Layer III. The Layer I can be rectangular in profile and be 40 mm long, 6 mm wide, and 250 μιτι thick. The Layer III may have: i) a fourth contacting electrode (343), a fourth connecting electrode (347), and a fourth wiring electrode (345) (together, a "second reference electrode");

ii) a fifth contacting electrode (353), a fifth connecting electrode (357), and a fifth wiring electrode (355) (together, a "second checking electrode"); and iii) a sixth contacting electrode (363), a sixth connecting electrode (367), and a sixth wiring electrode (365) (together, a "second working electrode").

The fourth contacting electrode (343) is connected to the fourth connecting electrode (347) by the fourth wiring electrode (345); the fifth contacting electrode (353) is connected to the fifth connecting electrode (357) by the fifth wiring electrode (355); the sixth contacting electrode (363) is connected to the sixth connecting electrode (367) by the sixth wiring electrode (365). Disposed together are the fourth contacting electrode (343), the fifth contacting electrode (353), and sixth contacting electrode (363). Further disposed together are the fourth connecting electrode (347), the fifth connecting electrode (357), and the sixth connecting electrode (367). Each of the fourth contacting electrode (343), the fifth contacting electrode (353), and sixth contacting electrode (363) can independently comprise a bend. The extremity of the fourth contacting electrode (343) can be bifurcated. The extremity of the sixth contacting electrode (363) can be bulbous. The bifurcations of the fourth contacting electrode (343) can at least partially envelop the extremity of the sixth contacting electrode (363).

Disclosed is Layer IV that can separate a Layer III from a Layer V. Layer IV can be 250 μΙ in depth. Disposed in Layer IV is a void (351). The void (351) can be positioned over the fourth contacting electrode (343), the fifth contacting electrode (353), and the sixth contacting electrode (363). A fluid can partially or completely fill the void (351). The capacity of the void (351) can be in the range of 0.5 to 4 μΙ. The capacity of the void (351) can be 2.5 μΙ. The void (351) can be positioned to ensure that a sufficient quantity of fluid reaches the fourth contacting electrode (343), the fifth contacting electrode (353), or independently the sixth contacting electrode (363). The void (331) can be positioned to ensure that a sufficient quantity of fluid reaches the fourth contacting electrode (343), the fifth contacting electrode (353), and the sixth contacting electrode (363). The void (351) can be present parallel or perpendicular to the longest axis of the Layer II. The void (351) can be 5.6 μιη in depth. The void (351) can be exposed to atmospheric air.

Description of the Reference Numbers

Number Description

21 amplifier, first

23 converter, first

30 analysis unit, second

31 amplifier, second

33 converter, second

40 control unit

50 display unit

100 microfluidic chip

101 base layer

111 electrode, first

113 contacting electrode, second

115 wiring electrode, second

117 connecting electrode, second

121 electrode, second

123 contacting electrode, first

125 wiring electrode, first

127 connecting electrode, first

130 bonding layer, upper

131 detection part, first

140 protective layer, upper

141 opening, upper

143 vent hole, upper

150 bonding layer, lower

151 detection part, second

161 opening, lower

163 vent hole, lower

171 electrode, third

173 contacting electrode, third

175 wiring electrode, third

177 connecting electrode, third

181 electrode, fourth

183 contacting electrode, fourth

185 wiring electrode, fourth

187 connecting electrode, fourth

200 microfluidic chip

213 contacting electrode, second

215 wiring electrode, second

217 connecting electrode, second

223 contacting electrode, first

225 wiring electrode, first Number Description

227 connecting electrode, first

230 bonding layer, upper

231 detection part, first

234 auxiliary electrode, second

240 protective layer, upper

241 opening, upper

243 vent hole, upper

250 bonding layer, lower

251 detection part, second

252 auxiliary electrode, third

253 auxiliary electrode, third, connecting

254 auxiliary electrode, third, wiring

256 auxiliary electrode, fourth

257 auxiliary electrode, fourth, connecting

258 auxiliary electrode, fourth, wiring

261 opening, lower

263 vent hole, lower

271 electrode, third

273 contacting electrode, third

275 wiring electrode, third

277 connecting electrode, third

281 electrode, fourth

283 contacting electrode, fourth

285 wiring electrode, fourth

287 connecting electrode, fourth

311 electrode, first

313 contacting electrode, first

323 contacting electrode, second

333 contacting electrode, third

315 wiring electrode, first

325 wiring electrode, second

335 wiring electrode, third

317 connecting electrode, first

327 connecting electrode, second

337 connecting electrode, third

331 detection part

321 electrode, second

343 contacting electrode, first

353 contacting electrode, second

363 contacting electrode, third Number Description

345 wiring electrode, first

355 wiring electrode, second

365 wiring electrode, third

347 connecting electrode, first

357 connecting electrode, second

367 connecting electrode, third

351 detection part