CN211385058U - Chaotic convection mixing device - Google Patents
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- CN211385058U CN211385058U CN201922175005.9U CN201922175005U CN211385058U CN 211385058 U CN211385058 U CN 211385058U CN 201922175005 U CN201922175005 U CN 201922175005U CN 211385058 U CN211385058 U CN 211385058U
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Abstract
The utility model discloses a chaotic convection mixing device, which comprises an upper layer structure, a middle layer structure and a lower layer structure from top to bottom in sequence, wherein the upper layer structure is provided with a disturbance column through hole at a position corresponding to a reaction cavity of a microfluid chip in a penetrating way, an elastic membrane is arranged between the upper layer structure and the middle layer structure, the elastic membrane is provided with a disturbance column, and the disturbance column passes through the disturbance column through hole and is contacted with an elastic membrane coated outside the bottom of the reaction cavity; the upper layer structure and the middle layer structure are provided with an elastic membrane cavity in a matching way at the position corresponding to the reaction cavity, the elastic membrane cavity is provided with a middle layer exhaust port and at least one disturbance air inlet, and the lower layer structure is provided with a lower layer exhaust port at the position corresponding to the middle layer exhaust port; the middle layer structure is also provided with an air inlet and an air inlet channel, and the air inlet is communicated with the disturbance air inlet through the air inlet channel. Can be used with the integration of microfluid chip, its mixing is efficient, and the reaction is more abundant, can reduce cost, and it is more simple and convenient to operate.
Description
Technical Field
The utility model belongs to the technical field of medical equipment, especially, relate to a chaotic convection mixing device.
Background
Microfluidics is a technology applied across various disciplines including engineering, physics, chemistry, microtechnology, and biotechnology. Microfluidics involves the study of minute quantities of fluids and how to manipulate, control and use such small quantities of fluids in various microfluidic systems and devices, such as microfluidic detection chips. For example: microfluidic biochips (known as "lab-on-a-chip") are used in the field of molecular biology to integrate assay operations for purposes such as analyzing enzymes and DNA, detecting biochemical toxins and pathogens, diagnosing diseases, and the like.
Microfluidic chip (microfluidic chip) is a hot spot area for the development of current micro total Analysis Systems (miniaturedtotal Analysis Systems). The micro-fluid detection chip analysis takes a chip as an operation platform, simultaneously takes analytical chemistry as a basis, takes a micro-electromechanical processing technology as a support, takes a micro-pipeline network as a structural characteristic, takes life science as a main application object at present, and is the key point of the development of the field of the current micro total analysis system. Its goal is to integrate the functions of the entire laboratory, including sampling, dilution, reagent addition, reaction, separation, detection, etc., on a microchip. The microfluid detection chip is a main platform for realizing microfluid technology. The device is characterized mainly by the fact that its active structures (channels, chambers and other functional elements) containing the fluid are of micron-scale dimensions at least in one dimension. Due to the micro-scale structure, the fluid exhibits and develops specific properties therein that differ from those of the macro-scale. Thus developing unique assay-generated properties. The characteristics and development advantages of the microfluid detection chip are as follows: the microfluid detection chip has the characteristics of controllable liquid flow, extremely less consumption of samples and reagents, ten-fold or hundred-fold improvement of analysis speed and the like, can simultaneously analyze hundreds of samples in a few minutes or even shorter time, and can realize the whole processes of pretreatment and analysis of the samples on line. The application of the micro total analysis system aims to realize the ultimate goal of the micro total analysis system, namely a lab-on-a-chip, and the key application field of the current work development is the field of life science.
However, when the microfluid chip is used for detection at present, how to improve the uniform mixing effect of a sample in a closed narrow reaction cavity of the microfluid chip is an urgent problem to be solved, a mode of uniformly mixing magnetic beads is adopted at present, compared with the traditional separation method, the magnetic beads are used for separating complex components of biochemical samples, separation and enrichment can be carried out simultaneously, the separation speed and the enrichment efficiency are effectively improved, and meanwhile, the sensitivity of analysis and detection is greatly improved. The method can be used for separating and purifying target bodies in a sample by coating specific antibodies, receptors and the like on the surfaces of magnetic beads. Particularly, in-vitro diagnosis immunoassay, a reaction system is generally required to be uniformly mixed, so that the reaction is more sufficient; however, a large number of experiments prove that the mixing effect of uniformly mixing the magnetic beads still needs to be further improved.
Therefore, there is a need to develop a microfluid detection chip, which can be mixed with a high-efficiency mixing device, and has the advantages of high mixing efficiency, high accuracy of detection result, reduced cost, and simple operation.
SUMMERY OF THE UTILITY MODEL
The to-be-solved technical problem of the utility model is to provide a chaotic convection mixing device can realize high-efficient mixing, and can set up the use with microfluid chip integration, and its mixing is efficient, and the result rate of accuracy is high, and can reduce cost, operates more portably.
In order to solve the technical problem, the utility model discloses a technical scheme is: the chaotic convection blending device sequentially comprises an upper layer structure, a middle layer structure and a lower layer structure from top to bottom, wherein a disturbance column through hole penetrates through the upper layer structure at a position corresponding to the reaction cavity, an elastic membrane is arranged between the upper layer structure and the middle layer structure, a disturbance column is arranged on the elastic membrane, and the disturbance column penetrates through the disturbance column through hole to be in contact with an elastic membrane coated outside the bottom of the reaction cavity; the upper layer structure and the middle layer structure are provided with an elastic membrane cavity in a matching way at the position corresponding to the reaction cavity, the elastic membrane cavity is provided with a middle layer air outlet and at least one disturbance air inlet, and the lower layer structure is provided with a lower layer air outlet at the position corresponding to the middle layer air outlet; the middle layer structure is also provided with an air inlet and an air inlet channel, and the air inlet is communicated with the disturbance air inlet through the air inlet channel. By adopting the technical scheme, the microfluid chip and the chaotic convection blending device are integrally arranged, wherein the convection blending device is of a 3-layer structure, the material of the convection blending device is plastic as that of the microfluid chip, and the convection blending device is bonded or adhered with the microfluid chip into a whole after being integrally bonded for detection, namely the microfluid chip is provided with the blending device, no additional blending instrument is needed, and the use is convenient; when the chaotic convection blending device is used, an air source and a pulse airflow controller are required to be configured to act on the chaotic convection blending device to form an integral blending instrument, liquid to be blended forms chaotic convection through two alternative disturbance modes, the reaction time is shortened, the blending effect is better, and the detection result is more accurate. The disturbing column penetrates through the disturbing column through hole to be in contact with the elastic film at the bottom of the reaction cavity, so that the sample liquid in the reaction cavity is disturbed, and the liquid in the reaction cavity is uniformly mixed; wherein the diameter of the through hole of the perturbation column is larger than that of the perturbation column.
As a preferred technical solution of the present invention, the elastic membrane cavity includes an upper layer elastic membrane cavity disposed on the back side of the upper layer structure and a middle layer elastic membrane cavity disposed on the front side of the middle layer structure; the elastic diaphragm is arranged between the upper layer elastic diaphragm cavity and the middle layer elastic diaphragm cavity; the middle layer air outlet and the disturbance air inlet are arranged on the middle layer elastic membrane cavity. The upper layer structure and the middle layer structure are tightly bonded by the arrangement, the elastic membrane is ensured to be air-tight, and meanwhile, the elastic membrane can cover a disturbance air inlet on the middle layer elastic membrane cavity, so that two air flows are convected in the middle layer elastic membrane cavity to form two alternative motion modes, chaotic convection is formed, and then a sample liquid in a reaction cavity contacted with a disturbance column is disturbed by the disturbance column on the elastic membrane between the middle layer elastic membrane cavity and the upper layer elastic membrane cavity, so that the liquid in the reaction cavity is uniformly mixed.
As the utility model discloses a preferred technical scheme, the air inlet sets up the side of middle level structure, inlet channel sets up the back of middle level structure, inlet channel's one end with the air inlet is connected, inlet channel's the other end with the disturbance air inlet in middle level elastic membrane chamber is linked together. The arrangement can lead the air source to directly enter the middle layer elastic membrane cavity.
As the utility model discloses a preferred technical scheme, upper elastic diaphragm chamber is in the back of superstructure is equipped with upper exhaust passage, correspondingly, the structural middle level gas vent that is equipped with of middle level, the substructure is in the corresponding position department of middle level gas vent is equipped with lower floor's gas vent one, upper exhaust passage keeps away from the one end warp of upper elastic diaphragm chamber after superstructure, middle level structure and lower floor's structure cooperate to connect with middle level gas vent one is linked together.
As the utility model discloses an optimized technical scheme, the shape and the size in upper elastic diaphragm chamber with the shape and the size in reaction chamber are the same, the shape in middle level elastic diaphragm chamber with the shape in upper elastic diaphragm chamber is the same, just the size in middle level elastic diaphragm chamber is greater than the size in upper elastic diaphragm chamber, elastic diaphragm's size is not less than the size in middle level elastic diaphragm chamber.
As the utility model discloses a preferred technical scheme, the shape in elastic diaphragm chamber is olive shape, the quantity of disturbance post is 2, 2 the setting of disturbance post is in on the elastic diaphragm piece and the central line along olive shape's radial direction's both ends evenly distributed, correspondingly, the quantity of disturbance post through-hole is 2, establish superstructure with the corresponding position department of disturbance post.
As the utility model discloses a preferred technical scheme, upper strata exhaust passage extends to both sides along the olive radial direction of perpendicular to shape, makes middle level gas vent one sets up outside the middle level elastic membrane chamber.
As the utility model discloses a preferred technical scheme, the quantity of disturbance air inlet is 2, the setting of disturbance air inlet is in middle level elastic membrane intracavity and set up the both ends at the radial direction of olive shape. Such an arrangement may ensure an alternating perturbation of the flexible diaphragm by the inlet port.
As a preferred technical solution of the present invention, the number of the air inlets and the number of the air inlet channels are 2, 2 of the air inlets are respectively arranged on the side surface of the middle layer structure, and the distance between 2 of the air inlets is greater than the radial length of the middle layer elastic film cavity; the air inlet channel is L-shaped.
As the preferred technical scheme of the utility model, the middle level gas vent runs through the setting and is in the centre in the middle level elasticity membrane intracavity. So that the middle exhaust port is centered with respect to the perturbation column also on the elastomeric membrane.
The utility model discloses the technical problem who still solves provides a microfluid detects chip, can with chaos convection current mixing device integration setting, the cooperation mixing, the mixing is efficient, and the testing result rate of accuracy is high, and can reduce cost, operates more portably.
In order to solve the technical problem, the utility model discloses a technical scheme is: the microfluid chip used for detecting the cardiac troponin comprises a chip body, wherein the chip body sequentially comprises a lower chip, a middle chip and an upper chip from bottom to top; the chip body comprises a liquid filling port, a reaction cavity and a micro-channel, and the middle-layer chip and the lower-layer chip are matched to define a closed micro-channel and a reaction cavity; the liquid adding port is communicated with the reaction cavity through the micro-channel; at least one groove is arranged in the reaction cavity. By adopting the technical scheme, the grooves are formed in the reaction cavity and used for coating the antibodies, and the fluorescence labeled antibodies are fixed in the area outside the grooves in the reaction cavity, compared with the previous design, the positions of the coated antibodies are more concentrated, the fluorescence signals after reaction are more concentrated, the peak positions of fluorescence detection curves are more stable, the CV value of a detection result is smaller, the repeatability is good, and the detection accuracy is improved; the shape of reaction chamber wherein is olive shape, increases the reaction volume, makes the reaction more concentrated, then fluorescence signal is more concentrated, the detection of being convenient for, such reaction chamber also more is favorable to the mixing of sample antigen and peridium antibody in the reaction chamber, reduces the liquid dead volume at former design reaction chamber both ends.
The further improvement is that the bottom of the reaction cavity is provided with a chaotic convection blending device. The microfluid chip and the chaotic convection mixing device are integrally arranged, wherein the convection mixing device is of a 3-layer structure, the material of the convection mixing device is plastic as the microfluid chip, and the convection mixing device is bonded or adhered with the microfluid chip into a whole after being integrally bonded for detection, namely, the microfluid chip is provided with the mixing device, so that an additional matching mixing instrument is not needed, and the use is convenient.
As a preferred technical solution of the present invention, the reaction chamber includes an upper reaction chamber disposed on the back surface of the middle chip and a lower reaction chamber disposed on the lower chip in a penetrating manner, and the position of the upper reaction chamber on the back surface of the middle chip corresponds to the position of the lower reaction chamber on the lower chip; the bottom of the lower reaction chamber is covered on the back surface of the lower chip by an elastic film for sealing, and the groove is arranged in the upper reaction chamber. The fluorescence labeling antibody is fixed in the area outside the groove of the upper reaction chamber or on the elastic film of the lower reaction chamber on the front surface of the lower chip; the groove is arranged in the upper reaction chamber, so that the position of the coated antibody is more concentrated, the fluorescent signal after reaction is more concentrated, the repeatability is good, and the detection accuracy is improved; the bottom of the lower reaction chamber is covered and sealed by an elastic film, so that the transmission of blending energy is facilitated, and the high-efficiency blending device, namely a disturbance column cap of the chaotic convection blending device, is matched with the reaction chamber to enhance the blending effect.
As the utility model discloses an optimized technical scheme, the recess is inside sunken circular slot, the quantity of recess is 3, 3 the recess is in evenly distributed in the reaction chamber. Three circular grooves are formed in the reaction cavity, three antibodies can be coated, the chip product is a three-link card, three indexes of the same sample can be detected, the multi-index simultaneous detection of the sample is realized, and compared with the previous multi-index microfluidic chip, the multi-index microfluidic chip has the advantages that the production cost can be reduced, the experimental operation is simpler and more convenient, and the structure of a detection instrument used in a matched mode is simpler.
As a preferred technical solution of the present invention, the micro flow channel includes a reaction cavity input flow channel and a reaction cavity output flow channel, the reaction cavity input flow channel and the reaction cavity output flow channel are both disposed on the back side of the middle chip, the reaction cavity input flow channel is communicated with one end of the upper reaction chamber, and the reaction cavity output flow channel is communicated with the other end of the upper reaction chamber; the reaction cavity input flow channel is communicated with the liquid adding port; and an anti-backflow structure is arranged between the reaction cavity input flow channel and the liquid adding port. The backflow prevention structure is arranged, so that the sample liquid in the reaction cavity can normally shake within a range, and the sample liquid in the reaction cavity cannot flow in the front-back direction of the reaction cavity in the flow path. The reaction cavity input flow channel and the reaction cavity output flow channel are both arranged on the back surface of the middle chip, so that gas can be discharged, the reaction is ensured to be filled with sample liquid, and the gas can be prevented from occupying the upper part of the reaction cavity when the reaction cavity input flow channel and the reaction cavity output flow channel of the reaction cavity are arranged on the front surface of the lower chip, so that the sample liquid can flow out when the reaction cavity cannot be filled with the sample liquid; furthermore, such an arrangement may increase the depth of the reaction chamber.
As a preferred technical scheme of the utility model, the liquid filling opening is arranged on the upper chip in a penetrating way, a middle layer liquid filling through hole is arranged on the middle layer chip in a penetrating way at the corresponding position of the liquid filling opening, and a liquid filling input micro channel is arranged on the front surface of the lower chip at the corresponding position of the liquid filling opening; the liquid feeding input micro-channel is communicated with the liquid feeding port through the middle-layer liquid feeding through hole, and the reaction cavity input channel is communicated with the liquid feeding port through the liquid feeding input micro-channel. The filling opening is funnel-shaped on the upper chip, the opening of the filling opening is gradually reduced from the front surface of the upper chip to the back surface of the upper chip, and the sample to be detected is better added from the filling opening.
As the preferred technical scheme of the utility model, the backflow prevention structure comprises a first vertical flow passage, a second vertical flow passage and a backflow prevention connecting flow passage; the backflow-preventing connecting flow channel is arranged on the back face of the upper-layer chip, and the first vertical flow channel and the second vertical flow channel are arranged on the middle-layer chip in a penetrating mode; and the liquid adding input micro-channel is communicated with the reaction cavity input channel after sequentially passing through the vertical channel II, the backflow prevention connecting channel and the vertical channel I.
As a preferred technical solution of the present invention, the chip body further includes a waste liquid cavity, the waste liquid cavity is disposed on the lower chip, and a middle layer waste liquid cavity through hole is disposed on the middle chip at a position corresponding to the waste liquid cavity; the back of the upper chip is provided with a waste liquid cavity cover plate at a position corresponding to the waste liquid cavity, correspondingly, the front of the upper chip is provided with a waste liquid cavity exhaust hole, the front of the middle chip is provided with an auxiliary leakage-proof waste liquid cavity at the upper part of the waste liquid cavity, and the auxiliary leakage-proof waste liquid cavity is provided with a waste liquid cavity middle exhaust hole at a position corresponding to the waste liquid cavity exhaust hole in a penetrating manner.
As the utility model discloses an optimized technical scheme, the reaction chamber passes through in proper order reaction chamber output runner and waste liquid output miniflow way with the waste liquid chamber is linked together, waste liquid output miniflow way sets up the front of lower floor's chip, just waste liquid output miniflow way with be equipped with the conductive rubber valve between the reaction chamber output runner, the conductive rubber valve is including setting up upper conductive rubber valve structure on the upper chip is in with the setting the middle level conductive rubber valve structure of the corresponding position department of middle level chip.
The utility model discloses the technical problem who still solves provides an adopt microfluid chip to detect cardiac muscle troponin's method, and the immunoreaction mixing is efficient, and the result rate of accuracy is high, and can reduce cost, and it is more simple and convenient to operate.
In order to solve the above technical problem, the utility model adopts the technical scheme that the method for detecting cardiac troponin by using the microfluid chip specifically comprises the following steps:
(1) adding a quantitative sample plasma/serum from the liquid adding port by using a liquid transfer machine;
(2) the sample plasma/serum sequentially passes through the middle-layer liquid adding through hole, the liquid adding input micro-channel and the reaction cavity input channel to flow into the reaction cavity, the micro-fluid chip and the chaotic convection blending device are integrally designed, the chaotic convection blending device is started to blend the sample plasma/serum in the reaction cavity for 3-15 min, the sample antigen and the antibody in the reaction cavity carry out immunoreaction, and the chaotic convection blending device is stopped;
(3) pushing air into the liquid adding opening by adopting the liquid shifter, pushing liquid in the microfluidic chip to move forwards, and drying the reaction cavity and the flow channel;
(4) adding a cleaning solution into the liquid adding port until the reaction cavity is filled, starting the chaotic convection mixing device again, mixing for 1-3 min, cleaning, stopping the chaotic convection mixing device, and pushing air into the microfluidic chip by a liquid shifter to blow dry the microfluidic channel and the reaction cavity of the chip;
(5) repeating the step (4) and cleaning for 3-5 times;
(6) and carrying out fluorescence detection on the microfluidic chip to obtain a reaction fluorescence value.
By adopting the technical scheme, the sample plasma/serum is sucked by a liquid transfer device through manual sample introduction and is added into a liquid adding port, the sample enters a reaction cavity, the mixture is uniformly mixed by a chaotic convection uniformly mixing device, the immunoreaction is carried out, and the fluorescence signal value is detected after the steps of cleaning and the like; the sample reaction fluorescence value of the immune reaction carried out by uniformly mixing the mixture by the chaotic convection uniformly mixing device is high, and the immune reaction of the antibody and the antigen is more sufficient.
Drawings
The following is a more detailed description of embodiments of the present invention with reference to the accompanying drawings:
FIG. 1 is a schematic diagram of a three-dimensional structure of a microfluidic chip according to the present invention;
FIG. 2 is a schematic diagram of a three-layer explosion structure of the microfluidic chip of the present invention;
FIG. 3 is a schematic diagram of the front structure of the lower chip of the microfluidic chip of the present invention;
FIG. 4 is a schematic diagram of a back side structure of a lower chip of the microfluidic chip of the present invention;
FIG. 5 is a schematic diagram of the front side of the middle chip of the microfluidic chip of the present invention;
fig. 6 is a schematic diagram of a reverse structure of a middle layer chip of a microfluidic chip according to embodiment 1 of the present invention;
fig. 7 is a schematic diagram of a reverse structure of a middle layer chip of a microfluidic chip according to embodiment 2 of the present invention;
fig. 8 is a schematic structural diagram of the front side of the upper chip of the microfluidic chip according to the present invention;
fig. 9 is a schematic structural view of the upper chip of the microfluidic chip according to the present invention;
fig. 10 is a top three-layer explosion structure diagram of the chaotic convection blending device after the microfluidic chip is bonded with the chaotic convection blending device according to the present invention;
fig. 11 is a bottom three-layer explosion structure diagram of the chaotic convection blending device after the microfluidic chip is bonded with the chaotic convection blending device of the present invention;
fig. 12 is a perspective structure view of the chaos convection mixing device of the microfluidic chip according to the present invention;
fig. 13 is a top perspective view of the microfluidic chip of the present invention bonded to the chaotic convection mixer;
fig. 14 is a bottom perspective view of the microfluidic chip of the present invention after bonding with the chaotic convection mixer;
wherein: 1-lower chip; 2-middle layer chip; 3-upper chip; 4-a liquid adding port; 401-middle layer liquid adding through hole; 5-lower reaction chamber; 501-an upper reaction chamber; 502-a lower reaction chamber; 5021-elastic film; 503-grooves; 6-micro flow channel; 601-reaction chamber input runner; 602-a reaction chamber output flow channel; 603-liquid adding and inputting into a micro-channel; 604-waste liquid output micro flow channel; 7-chaotic convection mixing device; 701-upper layer structure; 702-a middle layer structure; 703-lower layer structure; 704-perturbation post vias; 705-elastic membrane cavity; 7051-upper elastic membrane chamber; 7052-lower elastic membrane cavity; 706-elastic membrane; 707-perturbation columns; 708-middle layer exhaust port; 709-turbulence inlet; 7010-lower vent; 7011-air intake; 7012 — an intake passage; 7013-upper exhaust passage; 7014-middle exhaust port one; 7015-lower exhaust port one; 8-backflow prevention structure; 9-waste liquid chamber; 901-middle layer waste liquid cavity through hole; 902-waste chamber cover plate; 903-a waste liquid cavity vent hole; 904-waste liquid anti-backflow structure; 905-a middle exhaust hole in the waste liquid cavity; 906-upper conductive rubber valve structure; 907-middle layer conductive rubber valve structure; 908-attached leak-proof waste liquid chamber.
Detailed Description
Example 1: as shown in fig. 1 to 6 and fig. 8 to 14, the microfluidic chip includes a chip body, and the chip body includes a lower chip 1, a middle chip 2, and an upper chip 3 in sequence from bottom to top; the chip body comprises a liquid adding port 4, a reaction cavity 5 and a micro-channel 6, and the middle-layer chip 2 and the upper-layer chip 3 are matched to define the closed micro-channel 6 and the reaction cavity 5; the liquid adding port 4 is communicated with the reaction cavity 5 through the micro-channel 6; a groove 503 is arranged in the reaction cavity 5; the groove 503 is an inward-concave circular groove, and the bottom of the reaction cavity 5 is provided with a chaotic convection blending device 7; the reaction cavity 5 comprises an upper reaction chamber 501 arranged on the back surface of the middle chip 2 and a lower reaction chamber 502 arranged on the lower chip 1 in a penetrating manner, and the position of the upper reaction chamber 501 on the back surface of the middle chip 2 corresponds to the position of the lower reaction chamber 502 on the lower chip 1; the bottom of the lower reaction chamber 502 is sealed by covering the elastic film 5021 on the back of the lower chip 1, and the groove 503 is arranged in the upper reaction chamber 501; the fluorescence labeled antibody is fixed on the region outside the groove 503 of the upper reaction chamber 501 or on the elastic film of the lower reaction chamber 502 of the front surface of the lower chip 1; the micro flow channel 6 comprises a reaction cavity input flow channel 601 and a reaction cavity output flow channel 602, the reaction cavity input flow channel 601 and the reaction cavity output flow channel 602 are both arranged on the back surface of the middle layer chip 2, the reaction cavity input flow channel 601 is communicated with one end of the upper reaction chamber 501, and the reaction cavity output flow channel 602 is communicated with the other end of the upper reaction chamber 501; the reaction cavity input runner 601 is communicated with the liquid adding port; an anti-backflow structure 8 is arranged between the reaction cavity input runner 601 and the liquid adding port 4; the upper chip 3 is provided with the liquid adding port 4 in a penetrating manner, the liquid adding port 4 is funnel-shaped on the upper chip 3, the opening of the liquid adding port 4 is gradually reduced from the front of the upper chip 3 to the back of the upper chip 3, the middle chip 2 is provided with a middle liquid adding through hole 401 in a penetrating manner at the corresponding position of the liquid adding port 4, and the front of the lower chip 1 is provided with a liquid adding input micro channel 603 at the corresponding position of the liquid adding port 4; the liquid adding input micro-channel 603 is communicated with the liquid adding port 4 through the middle layer liquid adding through hole 401, and the reaction cavity input channel 601 is communicated with the liquid adding port 4 through the liquid adding input micro-channel 603; the backflow prevention structure 8 comprises a first vertical flow passage, a second vertical flow passage and a backflow prevention connecting flow passage; the backflow-preventing connecting flow channel is arranged on the back surface of the upper chip 3, and the first vertical flow channel and the second vertical flow channel are both arranged on the middle chip 2 in a penetrating mode; the liquid adding input micro-channel 603 is communicated with the reaction cavity input channel 601 after sequentially passing through the vertical channel II, the anti-backflow connecting channel and the vertical channel I; the chip body further comprises a waste liquid cavity 9, the waste liquid cavity 9 is arranged on the lower chip 1, and a middle waste liquid cavity through hole 901 penetrates through the middle chip 2 at a position corresponding to the waste liquid cavity 9; a waste liquid cavity cover plate 902 is arranged on the back surface of the upper chip 3 at a position corresponding to the waste liquid cavity 9, correspondingly, a waste liquid cavity exhaust hole 903 is arranged on the front surface of the upper chip 3, and a waste liquid backflow prevention structure 904 is further arranged on the waste liquid output micro-channel 604; an auxiliary leakage-proof waste liquid cavity 908 is arranged on the upper part of the waste liquid cavity 9 on the front surface of the middle chip 2, and a waste liquid cavity middle layer exhaust hole 905 penetrates through the auxiliary leakage-proof waste liquid cavity 908 at a position corresponding to the waste liquid cavity exhaust hole 903; the reaction chamber 5 is communicated with the waste liquid chamber 9 through the reaction chamber output flow channel 602 and the waste liquid output micro flow channel 604 in sequence, the waste liquid output micro flow channel 604 is arranged on the front surface of the lower chip 1, and a conductive rubber valve is arranged between the waste liquid output micro flow channel 604 and the reaction chamber output flow channel 602, and the conductive rubber valve comprises an upper layer conductive rubber valve structure 906 arranged on the upper chip and a middle layer conductive rubber valve structure 907 arranged at a position corresponding to the middle chip; the chaotic convection blending device 7 is arranged on the back surface of the lower chip 1 and is integrated with the chip body; the chaotic convective uniform mixing device 7 sequentially comprises an upper layer structure 701, a middle layer structure 702 and a lower layer structure 703 from top to bottom, a disturbance column through hole 704 penetrates through the upper layer structure 701 at a position corresponding to the reaction cavity 5, an elastic membrane 706 is arranged between the upper layer structure 701 and the middle layer structure 702, a disturbance column 707 is arranged on the elastic membrane 706, and the disturbance column 707 penetrates through the disturbance column through hole 704 to be in contact with an elastic film 5021 coated outside the bottom of the reaction cavity 5; the upper layer structure 701 and the middle layer structure 702 are provided with an elastic membrane cavity 705 in a matching manner at a position corresponding to the reaction cavity 5, the elastic membrane cavity 705 is provided with a middle layer air outlet 708 and at least one disturbance air inlet 709, and the lower layer structure 703 is provided with a lower layer air outlet 7010 at a position corresponding to the middle layer air outlet 708; the middle layer structure 702 is further provided with an air inlet 7011 and an air inlet passage 7012, and the air inlet 7011 is communicated with the disturbance air inlet 709 through the air inlet passage 7012; the elastic film cavity 705 comprises an upper layer elastic film cavity 7051 arranged on the back side of the upper layer structure 701 and a middle layer elastic film cavity 7052 arranged on the front side of the middle layer structure 702; the elastic membrane 706 is arranged between the upper layer elastic membrane cavity 7051 and the middle layer elastic membrane cavity 7052; the middle layer air outlet 708 and the disturbance air inlet 709 are arranged on the middle layer elastic membrane cavity 7052; the air inlet 7011 is arranged on the side surface of the middle layer structure 702, the air inlet passage 7012 is arranged on the back surface of the middle layer structure 702, one end of the air inlet passage 7012 is connected with the air inlet 7011, and the other end of the air inlet passage 7012 is communicated with a disturbance air inlet 709 of the middle layer elastic membrane cavity 7052; the upper-layer elastic membrane cavity 7051 is provided with an upper-layer exhaust passage 7013 on the back of the upper-layer structure 701, correspondingly, the middle-layer structure 702 is provided with a middle-layer exhaust port 7014, the lower-layer structure 703 is provided with a lower-layer exhaust port 7015 at a position corresponding to the middle-layer exhaust port 7014, and one end of the upper-layer exhaust passage 7013, which is far away from the upper-layer elastic membrane cavity 7051, is communicated with the middle-layer exhaust port 7014 after being matched and connected with the upper-layer structure 701, the middle-layer structure 702 and the lower-layer structure 703; the shape and size of the upper elastic film cavity 7051 are the same as those of the reaction cavity 5, the shape of the middle elastic film cavity 7052 is the same as that of the upper elastic film cavity 7051, the size of the middle elastic film cavity 7052 is larger than that of the upper elastic film cavity 7051, and the size of the elastic membrane 706 is not smaller than that of the middle elastic film cavity 7052; the elastic membrane cavity 705 is shaped like an olive, the number of the perturbation columns 707 is 2, the 2 perturbation columns 707 are arranged on the elastic membrane 706 and are uniformly distributed on the center lines of the two ends of the elastic membrane along the radial direction of the olive, correspondingly, the number of the perturbation column through holes 704 is 2, and the perturbation column through holes are arranged at the positions of the upper layer structure 701 corresponding to the perturbation columns 707; the number of the upper layer exhaust passages 7013 is 2, and correspondingly, the number of the middle layer exhaust ports 7014 and the number of the lower layer exhaust ports 7015 are 2; the upper layer exhaust channel 7013 extends towards two sides along the radial direction perpendicular to the olive shape, so that 2 middle layer exhaust ports 7014 are arranged outside the middle layer elastic membrane cavity 7052; the number of the disturbance air inlets 709 is 2, and the disturbance air inlets 709 are arranged in the middle-layer elastic membrane cavity 7052 and at two ends of the olive shape in the radial direction; the number of the air inlets 7011 and the number of the air inlet passages 7012 are 2, 2 air inlets 7011 are respectively arranged on the side surface of the middle layer structure 702, and the distance between the 2 air inlets 7011 is greater than the radial length of the middle layer elastic film cavity 7052; the air inlet passage 7011 is L-shaped; the middle layer air outlet 708 penetrates through the middle part in the middle layer elastic membrane cavity 7052; such that the middle exhaust port 708 is centered with respect to the perturbation column 707 also on the elastomeric membrane 706.
Example 2: as shown in fig. 1 to 5 and fig. 7 to 14, the difference from embodiment 1 is that the number of the grooves is 3; specifically, the microfluidic chip comprises a chip body, wherein the chip body sequentially comprises a lower chip 1, a middle chip 2 and an upper chip 3 from bottom to top; the chip body comprises a liquid adding port 4, a reaction cavity 5 and a micro-channel 6, and the middle-layer chip 2 and the upper-layer chip 3 are matched to define the closed micro-channel 6 and the reaction cavity 5; the liquid adding port 4 is communicated with the reaction cavity 5 through the micro-channel 6; 3 grooves 503 are arranged in the reaction cavity 5; the grooves 503 are inwardly recessed circular grooves, and 3 grooves 503 are uniformly distributed in the upper reaction chamber 501; the bottom of the reaction cavity 5 is provided with a chaotic convection blending device 7; the reaction cavity 5 comprises an upper reaction chamber 501 arranged on the back surface of the middle chip 2 and a lower reaction chamber 502 arranged on the lower chip 1 in a penetrating manner, and the position of the upper reaction chamber 501 on the back surface of the middle chip 2 corresponds to the position of the lower reaction chamber 502 on the lower chip 1; the bottom of the lower reaction chamber 502 is sealed by covering the elastic film 5021 on the back of the lower chip 1, and the groove 503 is arranged in the upper reaction chamber 501; the fluorescence labeled antibody is fixed on the region outside the groove 503 of the upper reaction chamber 501 or on the elastic film of the lower reaction chamber 502 of the front surface of the lower chip 1; the micro flow channel 6 comprises a reaction cavity input flow channel 601 and a reaction cavity output flow channel 602, the reaction cavity input flow channel 601 and the reaction cavity output flow channel 602 are both arranged on the back surface of the middle layer chip 2, the reaction cavity input flow channel 601 is communicated with one end of the upper reaction chamber 501, and the reaction cavity output flow channel 602 is communicated with the other end of the upper reaction chamber 501; the reaction cavity input runner 601 is communicated with the liquid adding port; an anti-backflow structure 8 is arranged between the reaction cavity input runner 601 and the liquid adding port 4; the upper chip 3 is provided with the liquid adding port 4 in a penetrating manner, the liquid adding port 4 is funnel-shaped on the upper chip 3, the opening of the liquid adding port 4 is gradually reduced from the front of the upper chip 3 to the back of the upper chip 3, the middle chip 2 is provided with a middle liquid adding through hole 401 in a penetrating manner at the corresponding position of the liquid adding port 4, and the front of the lower chip 1 is provided with a liquid adding input micro channel 603 at the corresponding position of the liquid adding port 4; the liquid adding input micro-channel 603 is communicated with the liquid adding port 4 through the middle layer liquid adding through hole 401, and the reaction cavity input channel 601 is communicated with the liquid adding port 4 through the liquid adding input micro-channel 603; the backflow prevention structure 8 comprises a first vertical flow passage, a second vertical flow passage and a backflow prevention connecting flow passage; the backflow-preventing connecting flow channel is arranged on the back surface of the upper-layer chip 3, and the first vertical flow channel and the second vertical flow channel are both arranged on the middle-layer chip 2 in a penetrating mode; the liquid adding input micro-channel 603 is communicated with the reaction cavity input channel 601 after sequentially passing through the vertical channel II, the anti-backflow connecting channel and the vertical channel I; the chip body further comprises a waste liquid cavity 9, the waste liquid cavity 9 is arranged on the lower chip 1, and a middle waste liquid cavity through hole 901 penetrates through the middle chip 2 at a position corresponding to the waste liquid cavity 9; a waste liquid cavity cover plate 902 is arranged on the back surface of the upper chip 3 at a position corresponding to the waste liquid cavity 9, correspondingly, a waste liquid cavity exhaust hole 903 is arranged on the front surface of the upper chip 3, and a waste liquid backflow prevention structure 904 is further arranged on the waste liquid output micro-channel 604; an auxiliary leakage-proof waste liquid cavity 908 is arranged on the upper part of the waste liquid cavity 9 on the front surface of the middle chip 2, and a waste liquid cavity middle layer exhaust hole 905 penetrates through the auxiliary leakage-proof waste liquid cavity 908 at a position corresponding to the waste liquid cavity exhaust hole 903; the reaction chamber 5 is communicated with the waste liquid chamber 9 through the reaction chamber output flow channel 602 and the waste liquid output micro flow channel 604 in sequence, the waste liquid output micro flow channel 604 is arranged on the front surface of the lower chip 1, and a conductive rubber valve is arranged between the waste liquid output micro flow channel 604 and the reaction chamber output flow channel 602, and the conductive rubber valve comprises an upper layer conductive rubber valve structure 906 arranged on the upper chip and a middle layer conductive rubber valve structure 907 arranged at a position corresponding to the middle chip; the chaotic convection blending device 7 is arranged on the back surface of the lower chip 1 and is integrated with the chip body; the chaotic convective uniform mixing device 7 sequentially comprises an upper layer structure 701, a middle layer structure 702 and a lower layer structure 703 from top to bottom, a disturbance column through hole 704 penetrates through the upper layer structure 701 at a position corresponding to the reaction cavity 5, an elastic membrane 706 is arranged between the upper layer structure 701 and the middle layer structure 702, a disturbance column 707 is arranged on the elastic membrane 706, and the disturbance column 707 penetrates through the disturbance column through hole 704 to be in contact with an elastic film 5021 coated outside the bottom of the reaction cavity 5; the upper layer structure 701 and the middle layer structure 702 are provided with an elastic membrane cavity 705 in a matching manner at a position corresponding to the reaction cavity 5, the elastic membrane cavity 705 is provided with a middle layer air outlet 708 and at least one disturbance air inlet 709, and the lower layer structure 703 is provided with a lower layer air outlet 7010 at a position corresponding to the middle layer air outlet 708; the middle layer structure 702 is further provided with an air inlet 7011 and an air inlet passage 7012, and the air inlet 7011 is communicated with the disturbance air inlet 709 through the air inlet passage 7012; the elastic film cavity 705 comprises an upper layer elastic film cavity 7051 arranged on the back side of the upper layer structure 701 and a middle layer elastic film cavity 7052 arranged on the front side of the middle layer structure 702; the elastic membrane 706 is arranged between the upper layer elastic membrane cavity 7051 and the middle layer elastic membrane cavity 7052; the middle layer air outlet 708 and the disturbance air inlet 709 are arranged on the middle layer elastic membrane cavity 7052; the air inlet 7011 is arranged on the side surface of the middle layer structure 702, the air inlet passage 7012 is arranged on the back surface of the middle layer structure 702, one end of the air inlet passage 7012 is connected with the air inlet 7011, and the other end of the air inlet passage 7012 is communicated with a disturbance air inlet 709 of the middle layer elastic membrane cavity 7052; the upper-layer elastic membrane cavity 7051 is provided with an upper-layer exhaust passage 7013 on the back of the upper-layer structure 701, correspondingly, the middle-layer structure 702 is provided with a middle-layer exhaust port 7014, the lower-layer structure 703 is provided with a lower-layer exhaust port 7015 at a position corresponding to the middle-layer exhaust port 7014, and one end of the upper-layer exhaust passage 7013, which is far away from the upper-layer elastic membrane cavity 7051, is communicated with the middle-layer exhaust port 7014 after being matched and connected with the upper-layer structure 701, the middle-layer structure 702 and the lower-layer structure 703; the shape and size of the upper elastic film cavity 7051 are the same as those of the reaction cavity 5, the shape of the middle elastic film cavity 7052 is the same as that of the upper elastic film cavity 7051, the size of the middle elastic film cavity 7052 is larger than that of the upper elastic film cavity 7051, and the size of the elastic membrane 706 is not smaller than that of the middle elastic film cavity 7052; the elastic membrane cavity 705 is shaped like an olive, the number of the perturbation columns 707 is 2, the 2 perturbation columns 707 are arranged on the elastic membrane 706 and are uniformly distributed on the center lines of the two ends of the elastic membrane along the radial direction of the olive, correspondingly, the number of the perturbation column through holes 704 is 2, and the perturbation column through holes are arranged at the positions of the upper layer structure 701 corresponding to the perturbation columns 707; the number of the upper layer exhaust passages 7013 is 2, and correspondingly, the number of the middle layer exhaust ports 7014 and the number of the lower layer exhaust ports 7015 are 2; the upper layer exhaust channel 7013 extends towards two sides along the radial direction perpendicular to the olive shape, so that the first middle layer exhaust port 7014 is arranged outside the middle layer elastic membrane cavity 7052; the number of the disturbance air inlets 709 is 2, and the disturbance air inlets 709 are arranged in the middle-layer elastic membrane cavity 7052 and at two ends of the olive shape in the radial direction; the number of the air inlets 7011 and the number of the air inlet passages 7012 are 2, 2 air inlets 7011 are respectively arranged on the side surface of the middle layer structure 702, and the distance between the 2 air inlets 7011 is greater than the radial length of the middle layer elastic film cavity 7052; the air inlet passage 7011 is L-shaped; the middle layer air outlet 708 penetrates through the middle part in the middle layer elastic membrane cavity 7052; such that the middle exhaust port 708 is centered with respect to the perturbation column 707 also on the elastomeric membrane 706.
Example 3: as shown in fig. 10 to 12, the chaotic convection blending device 7 sequentially includes an upper layer structure 701, a middle layer structure 702 and a lower layer structure 703 from top to bottom, the upper layer structure 701 is provided with a disturbance column through hole 704 at a position corresponding to the reaction chamber 5, an elastic membrane 706 is provided between the upper layer structure 701 and the middle layer structure 702, the elastic membrane 706 is provided with a disturbance column 707, and the disturbance column 707 passes through the disturbance column through hole 704 and contacts with an elastic film 5021 coated outside the bottom of the reaction chamber 5; the upper layer structure 701 and the middle layer structure 702 are provided with an elastic membrane cavity 705 in a matching manner at a position corresponding to the reaction cavity 5, the elastic membrane cavity 705 is provided with a middle layer air outlet 708 and at least one disturbance air inlet 709, and the lower layer structure 703 is provided with a lower layer air outlet 7010 at a position corresponding to the middle layer air outlet 708; the middle layer structure 702 is further provided with an air inlet 7011 and an air inlet passage 7012, and the air inlet 7011 is communicated with the disturbance air inlet 709 through the air inlet passage 7012; the elastic film cavity 705 comprises an upper layer elastic film cavity 7051 arranged on the back side of the upper layer structure 701 and a middle layer elastic film cavity 7052 arranged on the front side of the middle layer structure 702; the elastic membrane 706 is arranged between the upper layer elastic membrane cavity 7051 and the middle layer elastic membrane cavity 7052; the middle layer air outlet 708 and the disturbance air inlet 709 are arranged on the middle layer elastic membrane cavity 7052; the air inlet 7011 is arranged on the side surface of the middle layer structure 702, the air inlet passage 7012 is arranged on the back surface of the middle layer structure 702, one end of the air inlet passage 7012 is connected with the air inlet 7011, and the other end of the air inlet passage 7012 is communicated with a disturbance air inlet 709 of the middle layer elastic membrane cavity 7052; the upper-layer elastic membrane cavity 7051 is provided with an upper-layer exhaust passage 7013 on the back of the upper-layer structure 701, correspondingly, the middle-layer structure 702 is provided with a middle-layer exhaust port 7014, the lower-layer structure 703 is provided with a lower-layer exhaust port 7015 at a position corresponding to the middle-layer exhaust port 7014, and one end of the upper-layer exhaust passage 7013, which is far away from the upper-layer elastic membrane cavity 7051, is communicated with the middle-layer exhaust port 7014 after being matched and connected with the upper-layer structure 701, the middle-layer structure 702 and the lower-layer structure 703; the shape and size of the upper elastic film cavity 7051 are the same as those of the reaction cavity 5, the shape of the middle elastic film cavity 7052 is the same as that of the upper elastic film cavity 7051, the size of the middle elastic film cavity 7052 is larger than that of the upper elastic film cavity 7051, and the size of the elastic membrane 706 is not smaller than that of the middle elastic film cavity 7052; the elastic membrane cavity 705 is shaped like an olive, the number of the perturbation columns 707 is 2, the 2 perturbation columns 707 are arranged on the elastic membrane 706 and are uniformly distributed on the center lines of the two ends of the elastic membrane along the radial direction of the olive, correspondingly, the number of the perturbation column through holes 704 is 2, and the perturbation column through holes are arranged at the positions of the upper layer structure 701 corresponding to the perturbation columns 707; the number of the upper layer exhaust passages 7013 is 2, and correspondingly, the number of the middle layer exhaust ports 7014 and the number of the lower layer exhaust ports 7015 are 2; the upper layer exhaust channel 7013 extends towards two sides along the radial direction perpendicular to the olive shape, so that the first middle layer exhaust port 7014 is arranged outside the middle layer elastic membrane cavity 7052; the number of the disturbance air inlets 709 is 2, and the disturbance air inlets 709 are arranged in the middle-layer elastic membrane cavity 7052 and at two ends of the olive shape in the radial direction; the number of the air inlets 7011 and the number of the air inlet passages 7012 are 2, 2 air inlets 7011 are respectively arranged on the side surface of the middle layer structure 702, and the distance between the 2 air inlets 7011 is greater than the radial length of the middle layer elastic film cavity 7052; the air inlet passage 7011 is L-shaped; the middle layer air outlet 708 penetrates through the middle part in the middle layer elastic membrane cavity 7052; such that the middle exhaust port 708 is centered with respect to the perturbation column 707 also on the elastomeric membrane 706.
Example 4: the method for detecting cardiac troponin by matching the micro-fluidic chip in the embodiment 1 with the chaotic convection blending device in the embodiment 3 specifically comprises the following steps:
(1) adding 200 μ L of sample plasma/serum from the addition port 4 with a pipette;
(2) the sample plasma/serum sequentially passes through the middle-layer liquid adding through hole 401, the liquid adding input micro-channel 603 and the reaction cavity input channel 601 to flow into the reaction cavity 5, the micro-fluid chip and the chaotic convection blending device 7 are integrally designed, the chaotic convection blending device 7 is started to blend the sample plasma/serum in the reaction cavity 5 for 10min, the sample antigen and the antibody in the reaction cavity 5 perform immunoreaction, and the chaotic convection blending device 7 is stopped;
(3) pushing air into a liquid adding port 4 of the microfluidic chip by using the liquid shifter, pushing liquid in the microfluidic chip to move forwards, and drying a reaction cavity 5 and a micro-channel 6;
(4) adding 200 mu L of cleaning solution into the liquid adding port 4 until the reaction cavity 5 is filled, starting the chaotic convection uniformly-mixing device 7 again, uniformly mixing for 1min, cleaning, stopping the chaotic convection uniformly-mixing device 7, and pushing air into the microfluidic chip by a liquid transfer device to blow dry the microfluidic channel 6 and the reaction cavity 5 of the chip;
(5) repeating the step (4) and cleaning for 3-5 times;
(6) and carrying out fluorescence detection on the microfluidic chip to obtain a reaction fluorescence value, and further detecting the content of the cardiac troponin index.
Example 5 (standing control example): the difference from the embodiment 4 is that in the step (2), a chaotic convection mixing device is not adopted for mixing, and the sample plasma/serum is added and kept standing for 10 min; in particular, the amount of the solvent to be used,
(1) adding 200 μ L of sample plasma/serum from the addition port 4 with a pipette;
(2) the sample plasma/serum sequentially flows into the reaction cavity 5 through the middle-layer liquid adding through hole 401, the liquid adding input micro-channel 603 and the reaction cavity input channel 601, and stands for 10min, so that the sample antigen and the antibody in the reaction cavity 5 perform immunoreaction;
(3) pushing air into a liquid adding port 4 of the microfluidic chip by using the liquid shifter, pushing liquid in the microfluidic chip to move forwards, and drying a reaction cavity 5 and a micro-channel 6;
(4) then adding 200 mu L of cleaning solution into the liquid adding port 4 until the reaction cavity 5 is filled, standing for 1min, cleaning, and pushing an air-blowing micro-channel 6 and the reaction cavity 5 into the microfluidic chip by a liquid shifter;
(5) repeating the step (4) and cleaning for 3-5 times;
(6) and carrying out fluorescence detection on the microfluidic chip to obtain a reaction fluorescence value, and further detecting the content of the cardiac troponin index.
Example 6: the difference from the embodiment 4 is that a rotor is arranged in the reaction cavity in advance, and the sample plasma/serum in the reaction cavity is uniformly mixed through the rotor; specifically, the method comprises the following steps:
(1) adding 200 μ L of sample plasma/serum from the addition port 4 with a pipette;
(2) the sample plasma/serum sequentially passes through the middle-layer liquid adding through hole 401, the liquid adding input micro-channel 603 and the reaction cavity input channel 601 to flow into the reaction cavity 5, the magnetic stirring device is started to uniformly mix the sample plasma/serum in the reaction cavity 5 for 10min, the sample antigen and the antibody in the reaction cavity 5 carry out immunoreaction, and the magnetic stirring device is stopped;
(3) pushing air into a liquid adding port 4 of the microfluidic chip by using the liquid shifter, pushing liquid in the microfluidic chip to move forwards, and drying a reaction cavity 5 and a micro-channel 6;
(4) adding 200 mu L of cleaning solution into the liquid adding opening 4 until the reaction cavity 5 is filled, starting the magnetic stirring device again, mixing uniformly for 1min, cleaning, stopping the magnetic stirring device, and pushing air into the microfluidic chip by a liquid shifter to blow dry the microfluidic channel 6 and the reaction cavity 5 of the chip;
(5) repeating the step (4) and cleaning for 3-5 times;
(6) and carrying out fluorescence detection on the microfluidic chip to obtain a reaction fluorescence value, and further detecting the content of the cardiac troponin index.
Example 7: the difference from the embodiment 4 lies in that the microfluidic chip is placed on a shaking table and the antibody and the sample in the reaction chamber are mixed uniformly by adopting a shaking table mixing mode, specifically:
(1) adding 200 μ L of sample plasma/serum from the addition port 4 with a pipette;
(2) the sample plasma/serum sequentially passes through the middle-layer liquid adding through hole 401, the liquid adding input micro-channel 603 and the reaction cavity input channel 601 to flow into the reaction cavity 5, the micro-fluid chip is placed into a shaking table and uniformly mixed for 10min, the sample antigen and the antibody in the reaction cavity 5 are subjected to immunoreaction, and the micro-fluid chip is taken out of the shaking table;
(3) pushing air into a liquid adding port 4 of the microfluidic chip by using the liquid shifter, pushing liquid in the microfluidic chip to move forwards, and drying a reaction cavity 5 and a micro-channel 6;
(4) adding 200 mu L of cleaning solution into the liquid adding port 4 until the reaction cavity 5 is filled, putting the mixture into a shaking table, uniformly mixing for 1min, cleaning, taking out the micro-liquid chip from the shaking table, and pushing air into the micro-liquid chip by a liquid transfer device to blow dry the micro-flow channel 6 and the reaction cavity 5 of the chip;
(5) repeating the step (4) and cleaning for 3-5 times;
(6) and carrying out fluorescence detection on the microfluidic chip to obtain a reaction fluorescence value, and further detecting the content of the cardiac troponin index.
The fluorescence values of the reaction obtained by mixing different sample concentrations in the methods of examples 4 to 7 are shown in Table 1.
Comparing the reaction fluorescence values of the standing control and the 3 mixing methods, the reaction fluorescence value of the chaotic convection mixing method is the highest, which shows that the mixing effect is better and the antibody-antigen immunoreaction is more sufficient.
TABLE 1 fluorescence values of the reactions of the different homogenization methods
The foregoing illustrates and describes the principles, general features, and advantages of the present invention. It will be understood by those skilled in the art that the present invention is not limited to the above embodiments, and that the embodiments and descriptions are illustrative only, and that various changes and modifications may be made without departing from the spirit and scope of the invention, for example, the layout structure of each chamber may be slightly modified, and that such changes and modifications are within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (10)
1. A chaotic convection blending device is characterized by comprising an upper layer structure, a middle layer structure and a lower layer structure from top to bottom in sequence, wherein a disturbance column through hole penetrates through the upper layer structure at a position corresponding to a reaction cavity of a microfluid chip, an elastic membrane is arranged between the upper layer structure and the middle layer structure, a disturbance column is arranged on the elastic membrane, and the disturbance column penetrates through the disturbance column through hole to be in contact with an elastic membrane coated outside the bottom of the reaction cavity; the upper layer structure and the middle layer structure are provided with an elastic membrane cavity in a matching way at the position corresponding to the reaction cavity, the elastic membrane cavity is provided with a middle layer air outlet and at least one disturbance air inlet, and the lower layer structure is provided with a lower layer air outlet at the position corresponding to the middle layer air outlet; the middle layer structure is also provided with an air inlet and an air inlet channel, and the air inlet is communicated with the disturbance air inlet through the air inlet channel.
2. The chaotic convective mixer of claim 1, wherein the elastic film cavities comprise an upper elastic film cavity disposed on the back of the upper structure and a middle elastic film cavity disposed on the front of the middle structure; the elastic diaphragm is arranged between the upper layer elastic diaphragm cavity and the middle layer elastic diaphragm cavity; the middle layer air outlet and the disturbance air inlet are arranged on the middle layer elastic membrane cavity.
3. The chaotic convective uniform mixing device of claim 2, wherein the air inlet is disposed at a side surface of the middle layer structure, the air inlet channel is disposed at a back surface of the middle layer structure, one end of the air inlet channel is connected to the air inlet, and the other end of the air inlet channel is communicated with a disturbance air inlet of the middle layer elastic membrane cavity.
4. The chaotic convective uniform mixing device according to claim 2, wherein the upper elastic membrane cavity is provided with an upper exhaust passage at a back surface of the upper structure, and correspondingly, the middle structure is provided with a first middle exhaust port, the lower structure is provided with a first lower exhaust port at a position corresponding to the first middle exhaust port, and one end of the upper exhaust passage away from the upper elastic membrane cavity is communicated with the first middle exhaust port after the upper structure, the middle structure and the lower structure are connected in a matching manner.
5. The chaotic convective uniform mixing device according to claim 3 or 4, wherein the shape and size of the upper elastic membrane cavity are the same as those of the reaction cavity, the shape of the middle elastic membrane cavity is the same as those of the upper elastic membrane cavity, the size of the middle elastic membrane cavity is larger than that of the upper elastic membrane cavity, and the size of the elastic membrane is not smaller than that of the middle elastic membrane cavity.
6. The chaotic convective uniform mixing device according to claim 5, wherein the elastic membrane cavity is shaped like an olive, the number of the disturbing columns is 2, 2 disturbing columns are arranged on the elastic membrane and are uniformly distributed along the central lines of the two ends of the olive in the radial direction, correspondingly, the number of the disturbing column through holes is 2, and the disturbing column through holes are arranged at the positions of the upper layer structure corresponding to the disturbing columns.
7. The chaotic convective mixer of claim 4, wherein the upper exhaust channel extends to both sides in a direction perpendicular to the olive-shaped radial direction, such that the first middle exhaust port is disposed outside the cavity of the middle elastic membrane.
8. The chaotic convective blending device of claim 6, wherein the number of the disturbance air inlets is 2, and the disturbance air inlets are disposed in the middle layer elastic membrane cavity and at two ends of the olive shape in the radial direction.
9. The chaotic convective uniform mixing device of claim 6, wherein the number of the air inlets and the air inlet channels is 2, 2 air inlets are respectively arranged on the side surface of the middle layer structure, and the distance between the 2 air inlets is greater than the radial length of the middle layer elastic membrane cavity; the air inlet channel is L-shaped.
10. The chaotic convective mixer of claim 6, wherein the middle air vent is disposed through the middle of the middle elastic membrane cavity.
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