TWI460429B - A microfluidic chip - Google Patents

Description

Microfluidic detection device

The present invention relates to a microfluidic detecting device, and more particularly to a microfluidic fake wine detecting device for detecting fake wine.

Methanol is a colorless and flammable liquid, also known as industrial alcohol. In addition to general industrial use, it is also used as a raw material for fake wine by unscrupulous traders. Methanol is very toxic, depending on the amount of human dose, it will cause different physiology. reaction. Methanol poisoning is generally caused by poisoning caused by eating fake wine, mainly causing retinal and optic neuropathy, and finally leading to optic atrophy and blindness. However, not only fake wines contain methanol, but general wines such as fruit-brewed wines may also contain excessive concentrations of methanol. Currently, common methanol detection methods include gas chromatography (GC) and potassium permanganate oxidation; Phase chromatography (GC) is an effective physical separation analysis method. It is necessary to set up a chromatography column and make a difference in the adsorption capacity of the components in the immiscible two phases (the stationary phase and the mobile phase). The flow rate is different, and the separation effect is achieved. In addition, a detection and recording system is required to calculate the component content. Potassium permanganate oxidation method: firstly set up a reaction device such as beaker, stirrer, etc., then add methanol and potassium permanganate to the beaker, stir the mixture to produce redox reaction in the beaker, and then add oxalic acid reagent to decolorize After the potassium permanganate is discolored, add a magenta (basic fuchsin) for color reaction, and then transfer the resulting product into the spectrophotometer sample tank for subsequent qualitative and quantitative analysis.

The Department of Health announces the use of spectrometers (spectrophotometers) with potassium permanganate oxidation to detect methanol in alcohol. Spectrophotometers, which are commonly used to analyze sample components, are analyzed by specific wavelengths. Typical spectrophotometers contain Five components: a light source can be ultraviolet or visible light, which is used to provide continuous incident light, a transparent sample cell, which is used to hold the sample, must use the material that can pass through the used light zone, generally made of quartz or smelting Preferably, the window of the sample well is completely perpendicular to the direction of the beam to reduce reflection losses, a wavelength selector, a photodetector, and a signal processing recorder. The sample tank of the prior art can only provide the sample of the sample to be tested. If the sample to be tested needs to be combined with some chemical reaction, it is necessary to set up a reaction device such as a beaker, a stirrer, etc., and obtain a sample of the sample to be tested from the reaction device. Then, the sample of the sample to be tested is placed in the sample cell of the spectrophotometer for subsequent component detection analysis.

In general, the above-mentioned conventional methanol detection method and spectrophotometer sample tank have the following disadvantages, for example, gas chromatography has the advantages of being fast and accurate, but the instrument and consumables are too expensive, so it is not suitable for development as a general folk test; Potassium manganate is a non-specific strong oxidant, which will cause the ethanol, pigment and sugar in the brewed wine to be oxidized by potassium permanganate. The resulting oxide will also be colored with magenta, which will interfere with the experimental results. To reduce the false positive of false positives, the sample of the sample to be tested needs to be subjected to complicated distillation treatment, so it is not particularly suitable for the detection of brewed wine. Furthermore, the sample of the sample to be tested after the reaction is moved into the sample tank by the outside, except that the sample to be tested may be contaminated by external contamination, which may affect the detection accuracy, and the required reaction dose is large, and the chemical is required. The waste disposal problem caused by the reaction is worth considering.

For the above reasons, it is necessary to further improve the above-mentioned conventional methanol detection method and in order to avoid the external contamination of the sample to be tested and reduce the required reaction dose, it is necessary to combine the new methanol test method with a microfluidic detection device.

In view of the above, the present invention improves the above-mentioned disadvantages by replacing potassium permanganate with a methanol oxidase (MOX) having a high specificity for methanol, and oxidizing methanol to formaldehyde in a microfluidic detection device in combination with magenta coloring. Then, the microfluidic detection device is directly placed in the spectrophotometer to detect the methanol content in the analyte.

SUMMARY OF THE INVENTION A primary object of the present invention is to provide a microfluidic device that can be tested with only a small amount of sample fluid, making it useful for rapid detection of fake wine.

A secondary object of the present invention is to provide a microfluidic detecting device, wherein the microfluidic device is provided with a transparent light transmitting hole portion, so that the microfluidic device can be directly inserted into the spectrophotometer sample tank for detecting methanol in the analyte. The content can avoid the external pollution of the analyte, affecting the qualitative and quantitative analysis results.

Still another object of the present invention is to provide a microfluidic detecting device, wherein a mixing tank in the microfluidic detecting device is designed to sufficiently mix the sample liquids to improve mixing uniformity and reaction efficiency.

Another object of the present invention is to provide a microfluidic device which is provided with a tortuous reaction channel so that the methanol to be detected can have a longer reaction time with the oxide to make the reaction more complete.

In order to achieve the foregoing object, the technical means utilized by the present invention and the effects achievable by the technical means include: a microfluidic detecting device comprising a feed layer plate, a mixed layer plate and a bus layer plate. The feed layer plate has a first surface and a second surface, and a plurality of through holes penetrating the feed layer plate are disposed between the first surface and the second surface, and the plurality of through holes of the feed layer plate comprise a The first injection port, a second injection port and a feeding port. The mixing layer has a first surface and a second surface, and the first surface of the mixed layer is combined with the second surface of the feeding layer, and a plurality of through-mixing is arranged between the first surface and the second surface of the mixed layer The through hole of the laminate comprises a first sample port, a second sample port, a perforation, a mixing tank and an analysis hole, the first sample port pair being located at the first injection port of the feed layer plate; The second sample port pair is located at the second injection port of the feed layer. The first surface of the mixed layer plate is provided with a feeding channel, at least one reaction channel and a feeding channel; the first end of the feeding channel is connected with the first sample port, and the second end is connected with the hole Connecting, the first end of the at least one reaction channel is connected to the mixing tank, the second end of the reaction channel is connected to the analysis hole, and the first end of the feeding channel corresponds to the feeding port of the feeding layer plate, The second end of the feed channel is connected to the analysis hole. The bussing plate has a first surface and a second surface, and the first surface of the busbar is combined with the second surface of the hybrid layer; the first surface has at least one first sample channel and at least one Two sample channels. The first end of the first sample channel corresponds to the perforation of the mixed layer plate, and the second end of the first sample channel corresponds to the mixing groove of the mixed layer plate. The first end of the second sample channel corresponds to the second sample port of the mixed layer, and the second end corresponds to the mixing tank of the mixed layer.

By injecting methanol from the first injection port of the feed layer, flowing through the feed channel of the mixed layer to the perforation, and then flowing into the mixed layer from the first sample channel of the manifold plate. a lower end of the mixing tank; an oxide may be injected from the second injection port of the feed layer, flowing through the second injection port of the mixed layer to the second sample channel of the bus layer, and then flowing into the mixed layer Mix the lower end of the tank. The methanol and the oxide mixture liquid undergoes a redox reaction in the at least one reaction channel to form formaldehyde, and the formaldehyde is finally collected in the analysis hole through the at least one reaction channel; and then the magenta is injected from the feed port. The magenta flows directly into the analysis through the feeding channel of the mixed layer to react the formaldehyde to the color. After the reaction is completed, the generated microfluidic detection device is collected in the protruding portion. When the analysis hole is made of a permeable material, the analysis hole provided in the protrusion can be directly placed into the sample slot of the spectrophotometer, and the absorption spectrum of the object to be tested is used to determine the qualitative characteristics of the fake wine. Quantitative analysis. According to the microfluidic detecting device of the invention, the microfluidic device can be detected only by a small amount of sample liquid, so that it has the function of rapidly detecting the fake wine.

The microfluidic detecting device of the present invention is provided with a light transmissive analysis hole portion, so that the analysis hole portion can be directly inserted into the sample slot of the spectrophotometer for detecting the content of methanol in the analyte to be tested. Qualitative and quantitative analysis can achieve the effect of avoiding the external contamination of the analyte and affecting the analysis results.

In the microfluidic detecting device of the present invention, the first sample channel and the second sample channel extend into the second end of the mixing tank in a manner opposite to each other, so that the sample liquid can be fully mixed and the mixing is improved. The uniformity makes the invention have the effect of improving the reaction efficiency.

In the microfluidic detecting device of the present invention, the microfluidic device is provided with a reaction channel surrounded by a meandering, so that methanol can have a longer reaction time with the oxide to achieve a more complete reaction.

The above and other objects, features and advantages of the present invention will become more <RTIgt; The microfluidic detecting device of the preferred embodiment of the present invention is composed of a plurality of laminates stacked on each other, and the plurality of laminates are preferably made of a light transmissive material. In the embodiment shown, the invention consists of a feed deck 1, a mixed laminate 2 and a busbar panel 3.

The feed layer 1 has a first surface 11 and a second surface 12. The feed layer 1 is further provided with a protrusion 13 which is cut downward by the first surface 11 to form a plurality of axial passages. The plurality of injection holes of the material layer plate 1 include a first injection port 14 , a second injection port 15 , at least one vent port 16 and a feeding port 17 . The methanol may be first selected by the first injection port 14 or the first One of the two injection ports 15 is injected, and then the oxide is injected from another injection port; at least one vent 16 is used for introducing gas; and the feed port 17 is for injecting magenta.

The mixed layer board 2 has a first surface 21 and a second surface 22. The mixed layer board 2 is further provided with a protrusion 23, and the first surface 21 of the mixed layer board 2 and the first layer 21 of the feed layer board 1 The two surfaces 12 are joined such that the feed layer 1 and the projections 13, 23 of the mixed layer 2 are overlapped with each other. The mixed layer plate 2 is provided with a plurality of through holes and a hollow opening extending axially through the mixed layer plate 2 from the first surface 21, and includes a first sample port 24 and a second sample port 25, at least A perforation 26, a mixing tank 27 and an analysis aperture 28. When the mixed layer plate 2 overlaps the feed layer plate 1, the first sample port 24 of the mixed layer plate 2 is located on the second side of the first injection port 14 of the feed layer plate 1; the mixed layer The second sample port 25 of the plate 2 is on the second side of the second injection port 15 of the feed deck 1.

The first surface 21 of the mixed layer 2 is further provided with a plurality of grooves including at least one feed channel 211, at least one vent channel 212, at least one reaction channel 213 and a feed channel 214. In the embodiment of the present embodiment, the feed channel 211 is drawn in two, the venting channel 212 is drawn in two, and the reaction channel 213 is drawn in two. The number of the at least one feed channel 211 is the same as the number of the at least one through hole 26, and the first end of each of the feed channels 211 is connected to the first sample port 24, and each of the feed channels 211 The second ends are respectively connected to one of the at least one through holes 26, so that the methanol injected from the first injection port 14 of the feed layer 1 can flow to the through holes 26 of the mixed layer 2. In order to enable the injected sample liquid to be quickly shunted and prevent the sample liquid from flowing back from the first injection port 14, the number of the feed channels 211 and the perforations 26 is preferably selected. The first ends of the at least one venting channel 212 respectively correspond to one of the vents 16 of the feed layer, and the second end of the venting channel 212 is connected to the analysis hole 28 such that the analysis hole 28 is at a relatively low pressure. The first end of the at least one reaction channel 213 is connected to the mixing tank 27, and the second end of the reaction channel 213 is connected to the analysis hole 28. The reaction channel 213 is preferably in a meandering manner to increase the flow path. For example, the reaction channel 213 is S-shaped, and thus, the reaction time can be increased to make the reaction more complete. The first end of the feeding channel 214 corresponds to the feeding port 17 of the feeding layer plate, and the second end of the feeding channel 214 is connected to the analysis hole 28 so that magenta can directly flow from the feeding port 17 to the Analyze the well 28.

The bussing board 3 has a first surface 31 and a second surface 32. The first surface 31 of the busbar 3 is combined with the second surface 22 of the hybrid layer 2. The busbar 3 is further provided with a convex surface. In the outlet portion 33, the projections 33 of the busbar plate 3 and the projections 13, 23 of the feed laminate 1 and the hybrid laminate 2 are overlapped with each other. Since the analysis hole 28 is a hollow opening in the mixed layer plate 2, the corresponding protruding portion 13 is obtained by sandwiching the mixed layer plate between the feeding layer plate 1 and the bus layer plate 3. 23, 33 is formed with a chamber for storing the mixed liquid, that is, the analysis hole 28. The first surface 31 of the bus bar 3 has a plurality of trenches including at least a first sample channel 34 and at least a second sample channel 35. The first end of the first sample channel 34 corresponds to the through hole 26 of the mixed layer plate 2, and the second end of the first sample channel 34 corresponds to the mixing groove 27 of the mixed layer plate 2, so that Methanol flowing to the perforation 26 can flow through the first sample channel 34 of the busbar plate to the mixing tank 27, and vortex mixing is formed by the lower end of the mixing tank 27 (i.e., the second surface 22 of the mixing layer 2). And rising to the upper end of the mixing tank 27; the first end of the second sample channel 35 corresponds to the second sample port 25 of the mixed layer plate 2, and the second end extends corresponding to the mixing of the mixed layer plate 2 The groove 27 allows the oxide to be injected from the second injection port 15 of the feed layer 1 , flows through the second sample port 25 of the mixed layer 2 to the second sample channel 35 of the bus layer 3, and From the second sample channel 35, it flows to the lower end of the mixing tank 27 of the mixed layer 2 (i.e., the second surface 22 of the mixed layer 2) to vortex the oxide with methanol. The first sample channel 34 and the second sample channel 35 are preferably formed in different lengths such that the flow rate of the methanol and the oxide is different, and thus the mixing of the methanol and the oxide can be more uniform.

Please refer to the second and third figures, which are the combination of the microfluidic detecting device according to the preferred embodiment of the present invention. The microfluidic detecting device is composed of three layers: a feed layer board 1, a mixed layer board 2 and a bus layer board 3. The laminates are stacked on each other, and the projections 13, 23, 33 of the three laminates are stacked toward the same side, so that the analysis holes 28 located in the mixed laminate 2 are fed by the feed laminate 1 and the bus layer. The plates 3 are co-interposed to form a chamber in which the mixed liquid can be stored. The first injection port 14 of the feed layer 1 corresponds to the first sample port 24 of the mixed layer plate 2; the second injection port 15 of the feed layer plate 1 and the second sample of the mixed layer plate 2 Corresponding to the mouth 25; at least one vent 16 of the feed deck 1 corresponds to the first end of at least one venting channel 212 of the mixed layer 2; the feed port 17 of the feed layer 1 is mixed with the The first end of the feed channel 214 of the laminate 2 corresponds. The second sample port 25 of the mixed layer plate 2 corresponds to the first end of the second sample channel 35 of the bus bar plate 3; the perforation 26 of the mixed layer plate 2 and the first sample of the bus bar plate 3 The first end of the channel 34 corresponds; the mixing groove 27 of the mixing layer 2 corresponds to the first sample channel 34 of the bus layer plate 3 and the second end of the second sample channel 35 of the bus layer plate.

Referring to the second and third figures, the microfluidic detecting device of the preferred embodiment of the present invention is applied to the detection of fake wine, and the sample liquids are fake wine (mainly containing methanol), oxide (MOX) and one. A coloring agent (for example: magenta). When methanol and oxide are separately injected into the mixing tank 27 of the mixed layer plate 2, a plurality of corresponding grooves 211, 34, 35 and a plurality of through holes are located in the mixed layer plate 2 and the bus layer plate 3. 14, 15, 24, 25, 26 make the methanol and oxide flow paths do not interact or cross each other. The mixing tank 27 is an accommodating space extending through the mixing layer 2, and the mixing tank 27 has an upper end (the first surface 21 of the mixed layer 2) and a lower end (the second surface 22 of the mixed layer 2) ). For example, methanol is injected from the first injection port 14 of the first surface 11 of the feed layer 1 and flows through the feed channel 211 of the mixed layer 2 to the perforation 26, followed by the confluence layer 3 A sample channel 34 flows into the lower end of the mixing tank 27 of the mixing layer 2; oxide can be injected from the second injection port 15 of the feed layer 1 and flows downward from the first surface 11 of the feed layer 1. Flowing through the second injection port 25 of the mixed layer plate 2 to the second sample channel 35 of the bus layer plate 3, and then flowing into the lower end of the mixing groove 27 of the mixed layer plate 2 (the second surface of the mixed layer plate 2) 22); at least one vent 16 of the feed layer 1 is ventilated by the venting channel 212 of the mixed layer 2, so that the analysis hole 28 of the mixed layer 2 is at a relatively low pressure because the analysis hole 28 is low pressure The methanol and oxide mixed liquid entering from the lower end of the mixing tank 27 is discharged from the upper end of the mixing tank 27, and the flow path length of the mixed liquid can be increased by the at least one reaction channel 213 surrounded by the meandering, thereby increasing the reaction. Time makes the reaction more complete. The methanol and oxide mixture liquid undergoes a redox reaction in the at least one reaction channel 213 to form formaldehyde, and the formaldehyde is finally collected in the analysis hole 28 via the at least one reaction channel 213; and then the feed port 17 is The magenta is injected, and the magenta flows directly into the analysis hole 28 through the addition channel 214 of the mixed layer 2 to react the formaldehyde to a color. The present invention can further inject hydrochloric acid into the feed port 17 after adding magenta to increase the color rendering effect.

Please refer to the fourth figure, which is a schematic diagram of each channel of the microfluidic detecting device of the preferred embodiment of the present invention. In the embodiment of the present embodiment, when the feeding channel 211 and the first sample are When the channel 34, the second sample channel 35, and the reaction channel 213 are each two and each has a different length, the flow rate of the methanol and the oxide flowing into the mixing tank 27 of the mixed layer 2 can be adjusted, and the inflow direction can be adjusted. Provide additional space for methanol and oxide by the length of the channel, except that the alcohol and oxide are not reversed from the injection port due to too much injection at one time and the methanol and oxide can be supplemented by the injection port through different directions. To the mixing tank 27. Methanol is injected from the first injection port 14 of the feed layer 1. Since the first injection port 14 corresponds to the two feed channels 211, a split is generated, and the methanol is fed from the mixed layer 2, respectively. The channels 211 each flow through the communicating perforations 26 to the first sample channel 34 of the busbar plate 3 and extend into the lower end of the mixing tank 27; the oxide is passed from the second injection port of the feed layer 1 15 is injected through the second sample port 25 of the mixed layer plate 2, and is split by the second sample channel 35, the oxide is respectively extended from the second sample channel 35 of the bus bar plate 3 and flows into the mixing tank 27 Lower end. Since the first sample channel 34 of the busbar layer 3 and the second sample channel 35 of the busbar layer 3 are respectively extended into the lower end of the mixing tank 27 by four different directions, and each of them is perpendicular to each other. The misalignment is opposite, and the two feed channels 211 and the second sample channels 35 are each formed in different lengths, or the first sample channel 34 and the second sample channel 35 are formed in different lengths, so that the injection is performed. Methanol and oxide can be vortexed and more uniformly mixed in the mixing tank 27 of the mixed layer 2 to improve mixing uniformity. The mixed methanol and oxide mixture flows out from the upper end of the mixing tank 27 of the mixed layer 2 due to the pressure, and then the mixed liquid is guided through at least one reaction channel 213 connected to the upper end of the mixing tank 27. To the analysis hole 28 (not shown).

In the microfluidic detecting device of the present invention, when the feeding layer plate 1, the mixed layer plate 2 and the mixed layer plate 2 are made of a light transmissive material, the microfluidic detecting device of the present invention can be convexly formed by the overlapping The outlets 13, 23, 33 are placed directly in the sample cell of the spectrophotometer, and an incident light source such as ultraviolet light or visible light is input to cause a sample of the characteristic absorption spectrum to be generated in the sample of the analysis hole 28, since each sample is It has its own unique absorption spectrum line, which is qualitatively analyzed according to the shape of the absorption spectrum line and the number of absorption peaks on the line, the wavelength corresponding to the peak and the relative height of the peak, to know the sample composition, and according to a certain characteristic peak. The height is proportional to the concentration of the substance for quantitative analysis to determine the sample content, which can be used to detect the efficacy of the fake wine.

With the microfluidic detecting device of the present invention, the sample detection can be consistent, so as to effectively reduce the contamination of the analyte and simplify the efficacy of the device. In addition, the microfluidic structure design of several through holes, a plurality of grooves, a mixing tank and a plurality of reaction channels can reduce the amount of sample liquid required for use, improve mixing uniformity, and make the reaction more complete. To achieve the effect of reducing pollution and improving the efficiency of the reaction.

While the invention has been described in connection with the preferred embodiments described above, it is not intended to limit the scope of the invention. The technical scope of the invention is protected, and therefore the scope of the invention is defined by the scope of the appended claims.

[this invention]

1. . . Feeding laminate

11. . . First surface

12. . . Second surface

13. . . Protrusion

14. . . First injection port

15. . . Second injection port

16. . . Vent

17. . . Feeding port

2. . . Mixed laminate

twenty one. . . First surface

211. . . Feed channel

212. . . Ventilation channel

213. . . Reaction channel

214. . . Feed channel

twenty two. . . Second surface

twenty three. . . Protrusion

twenty four. . . First sample port

25. . . Second sample port

26. . . perforation

27. . . Mixing tank

28. . . Analysis hole

3. . . Confluence layer

31. . . First surface

32. . . Second surface

33. . . Protrusion

34. . . First sample channel

35. . . Second sample channel

Figure 1 is an exploded perspective view of a microfluidic detection device in accordance with a preferred embodiment of the present invention.

Fig. 2 is a perspective view showing the combination of the microfluidic detecting device of the preferred embodiment of the present invention.

Fig. 3 is a view showing a combination of a mixed laminate and a busbar according to a preferred embodiment of the present invention.

Figure 4 is a schematic perspective view showing the arrangement of the fluid channels of the preferred embodiment of the present invention.

1. . . Feeding laminate

11. . . First surface

12. . . Second surface

13. . . Protrusion

14. . . First injection port

15. . . Second injection port

16. . . Vent

17. . . Feeding port

2. . . Mixed laminate

twenty one. . . First surface

211. . . Feed channel

212. . . Ventilation channel

213. . . Reaction channel

214. . . Feed channel

twenty two. . . Second surface

twenty three. . . Protrusion

twenty four. . . First sample port

25. . . Second sample port

26. . . perforation

27. . . Mixing tank

28. . . Analysis hole

3. . . Confluence layer

31. . . First surface

32. . . Second surface

33. . . Protrusion

34. . . First sample channel

341. . . First feed end

342. . . First feed end

35. . . Second sample channel

351. . . Second feed end

352. . . Second feed end

Claims (10)

A microfluidic detecting device comprises: a feeding layer plate having a first surface and a second surface, wherein the first surface and the second surface are provided with a plurality of through holes penetrating through the feeding layer, and the feeding The plurality of through holes of the layer plate comprise a first injection port, a second injection port and a feeding port; a mixed layer plate having a first surface and a second surface, the first surface of the mixed layer plate and the The second surface of the feed layer is combined, and the first surface and the second surface of the mixed layer are provided with a plurality of through holes penetrating the mixed layer, and the plurality of through holes of the mixed layer comprise a first sample a first sample port, two perforations, a mixing tank and an analysis hole, the first sample port pair is located at the first injection port of the feed layer plate, and the second sample port pair is located at the feed layer a second injection port of the plate; the first surface of the mixed layer plate is provided with two feeding channels, two reaction channels and a feeding channel, the length of the two feeding channels is different, and the two feeding channels are different One end is connected to the first sample port, and the second end of the two feed channel is respectively connected to the two perforations, the two reaction tanks The first end is connected to the mixing tank, and the second end of the two reaction channels are connected to the analysis hole, and the first end of the feeding channel corresponds to the feeding port of the feeding layer, and the feeding channel is The two ends are connected to the analysis hole; a bus bar having a first surface and a second surface, the first surface of the bus layer is combined with the second surface of the mixed layer; the first surface has the second a first channel of the first sample channel corresponding to two perforations of the mixed layer plate, and a second end of the two first sample channels respectively corresponding to the product channel and the second sample channel a mixing tank of the mixed layer; the second sample The length of the product channel is different, the first end of the second sample channel corresponds to the second sample port of the mixed layer plate, and the second ends of the second sample channels correspond to the mixed layer plate Mixing tank. A microfluidic detecting device comprises: a feeding layer plate having a first surface and a second surface, wherein the first surface and the second surface are provided with a plurality of through holes penetrating through the feeding layer, and the feeding The plurality of through holes of the layer plate comprise a first injection port, a second injection port and a feeding port; a mixed layer plate having a first surface and a second surface, the first surface of the mixed layer plate and the The second surface of the feed layer is combined, and the first surface and the second surface of the mixed layer are provided with a plurality of through holes penetrating the mixed layer, and the plurality of through holes of the mixed layer comprise a first sample a first sample port, two perforations, a mixing tank and an analysis hole, the first sample port pair is located at the first injection port of the feed layer plate, and the second sample port pair is located at the feed layer a second injection port; the first surface of the mixed layer plate is provided with two feed channels, two reaction channels and a feeding channel, and the first end of the two feeding channels is connected with the first sample port Connecting, the second ends of the two feed channels are respectively connected to the two perforations, and the first ends of the two reaction channels are connected to the mixing tank The second end of the two reaction channels is connected to the analysis hole, the first end of the feeding channel corresponds to the feeding port of the feeding layer, the second end of the feeding channel is connected to the analysis hole; a stratosphere The plate has a first surface and a second surface, and the first surface of the bus layer is combined with the second surface of the mixed layer; the first surface has two first sample channels and two second sample channels The lengths of the two first sample channels are different, and the first ends of the two first sample channels are respectively associated with the mixed layer Corresponding to the two perforations, the second ends of the two first sample channels correspond to the mixing grooves of the mixed layer; the lengths of the two second sample channels are different, and the first of the second sample channels is the first The ends all correspond to the second sample port of the mixed layer, and the second ends of the second sample channels correspond to the mixing grooves of the mixed layer. The microfluidic detection device according to claim 1 or 2, wherein the feed layer plate, the mixed layer plate and the bus layer plate are each provided with a protruding portion extending in the same direction and overlapping each other. . According to the microfluidic detecting device of the third aspect of the patent application, the analysis hole of the mixed layer plate is disposed at the protruding portion. According to the microfluidic detecting device according to claim 1 or 2, the feeding layer plate is made of a light transmitting material corresponding to the analysis hole portion. The microfluidic detection device according to claim 1 or 2, wherein the feed layer further comprises at least one vent, and the vent and the analysis hole are connected by at least one vent channel. The microfluidic detecting device according to item 6 of the patent application scope is provided with two venting ports and two venting channels. According to the microfluidic detection device of claim 1 or 2, the two first sample channels and the two second sample channels are extended from the lower end of the mixing tank into the mixing tank in a dislocated manner. . According to the microfluidic detecting device of claim 8, the length of each of the first sample channels is different from the length of each of the second sample channels. According to the microfluidic detecting device of claim 1 or 2, each of the reaction channels is formed in a meandering manner.