CN112675798B

CN112675798B - Microfluidic reaction device and microfluidic reaction driving method

Info

Publication number: CN112675798B
Application number: CN202011464345.4A
Authority: CN
Inventors: 林柏全; 席克瑞; 李伟; 欧阳珺婷; 粟平; 秦锋
Original assignee: Shanghai Tianma Microelectronics Co Ltd
Current assignee: Shanghai Tianma Microelectronics Co Ltd
Priority date: 2020-12-14
Filing date: 2020-12-14
Publication date: 2022-11-08
Anticipated expiration: 2040-12-14
Also published as: CN112675798A

Abstract

The embodiment of the invention provides a microfluid reaction device and a microfluid reaction driving method, wherein the microfluid reaction device comprises: the micro-fluid device comprises a first substrate and a second substrate which are arranged at intervals along the thickness direction of a micro-fluid; the first electrode layer is arranged on one side, facing the second substrate, of the first substrate and comprises a plurality of first electrodes; the first hydrophobic layer is arranged on one side, away from the first substrate, of the first electrode layer; the second hydrophobic layer is arranged on one side, facing the first hydrophobic layer, of the second substrate, the second hydrophobic layer and the first hydrophobic layer are distributed at intervals, and an accommodating space for accommodating liquid drops is formed between the first hydrophobic layer and the second hydrophobic layer; the second electrode layer is arranged on one side, facing the second hydrophobic layer, of the second substrate and comprises a plurality of second electrodes which are arranged independently, and each second electrode is respectively positioned in each temperature zone; and the heating layer is arranged between the second electrode layer and the second substrate, the heating substrate comprises a plurality of heating electrodes, and each heating electrode is respectively positioned in each temperature area.

Description

Microfluidic reaction device and microfluidic reaction driving method

Technical Field

The invention relates to the technical field of microfluid reaction equipment, in particular to a microfluid reaction device and a microfluid reaction driving method.

Background

Droplet microfluidics (droplet microfluidics) is a relatively new but rapidly developing field. Droplet microfluidic reaction devices typically employ electrowetting, electrophoresis, dielectrophoresis, and like techniques to manipulate droplet flow. Can be used in the fields of medical diagnosis, cancer screening, drug discovery, food safety inspection, environmental monitoring, forensic analysis and the like. Has the advantages of low cost, automation, low energy consumption and the like.

At present, when the droplet microfluidic reaction device is used for droplet experiments, when the droplets need to react at different temperatures, the microfluidic reaction device changes the temperature of the position where the droplet is located through temperature conversion. The temperature transformation is seriously influenced by the environment, and when the difference between the environment temperature and the target temperature is large, the time consumption of temperature transformation is long, and the experiment efficiency is seriously influenced.

Therefore, a new microfluidic reaction device and a driving method thereof are needed.

Disclosure of Invention

The embodiment of the invention provides a microfluid reaction device and a microfluid reaction driving method, and aims to solve the problem that the time consumption is long when a microfluid reaction device is used for carrying out experiments.

Embodiments of the first aspect of the invention provide a microfluidic reaction device having a plurality of temperature zones, the microfluidic reaction device comprising: the micro-fluid device comprises a first substrate and a second substrate which are arranged at intervals along the thickness direction of a micro-fluid; the first electrode layer is arranged on one side, facing the second substrate, of the first substrate and comprises a plurality of first electrodes; the first hydrophobic layer is arranged on one side, away from the first substrate, of the first electrode layer; the second hydrophobic layer is arranged on one side, facing the first hydrophobic layer, of the second substrate, the second hydrophobic layer and the first hydrophobic layer are distributed at intervals, and an accommodating space for accommodating liquid drops is formed between the first hydrophobic layer and the second hydrophobic layer; the second electrode layer is arranged on one side, facing the second hydrophobic layer, of the second substrate and comprises a plurality of second electrodes which are arranged independently, and each second electrode is located in each temperature zone; and the heating layer is arranged between the second electrode layer and the second substrate and comprises a plurality of heating electrodes, and each heating electrode is respectively positioned in each temperature area.

Embodiments of the second aspect of the present invention further provide a microfluidic reaction driving method using the microfluidic reaction device according to any one of the embodiments of the first aspect, the method including:

mixing the droplets and the reagent to form mixed droplets;

driving the mixed liquid drop to move to the central area of the preset temperature area;

the temperature zone where the mixed liquid drop is located is heated to the reaction temperature through the heating electrode, so that the liquid drop in the mixed liquid drop and the reagent can react with each other.

In another embodiment of the second aspect of the present invention, there is provided a microfluidic reaction driving method using the microfluidic reaction device of any one of the embodiments of the first aspect, the method including:

driving the liquid drops and the reagent to respectively move to a central area of a preset temperature area in sequence so as to mix the liquid drops and the reagent in the central area;

before or after the step of driving the liquid drop and the reagent to respectively move to the central area of the preset temperature area in sequence, the method further comprises the following steps: the preset temperature zone is heated to a reaction temperature by a heating electrode so that the liquid drop and the reagent located in the central zone can react with each other.

In a further embodiment of the second aspect of the present invention, there is provided a microfluidic reaction driving method using the microfluidic reaction device according to any one of the first aspect of the present invention, in which a reagent is embedded in a central region of each temperature zone, the method including:

driving the liquid drop to move to the central area of the preset temperature area;

before or after the step of driving the liquid drop to move to the central area of the preset temperature area, the method further comprises the following steps: the preset temperature zone is heated to a reaction temperature by a heating electrode so that the liquid drop and the reagent located in the central zone can react with each other.

In the microfluidic reaction device provided by the embodiment of the invention, the microfluidic reaction device comprises a first substrate, a second substrate, a first electrode layer, a first hydrophobic layer, a second electrode layer and a heating layer, wherein the first electrode layer, the first hydrophobic layer, the second electrode layer and the heating layer are positioned between the first substrate and the second substrate. Wherein, an accommodating space is formed between the first hydrophobic layer and the second hydrophobic layer, so that the liquid drop can be positioned in the accommodating space and move in the accommodating space. When the microfluidic reaction device is used for a droplet experiment, the temperature of the temperature zone can be fixed, and the droplet moves between different temperature zones by the interaction of the first electrode and the second electrode and by utilizing the electrowetting principle, so that the purpose of reacting the droplet at different temperatures is realized. The heating electrodes of the heating layer are respectively positioned in each temperature area, so that the heating electrodes can be heated in each temperature area, mutual heating among the temperature areas cannot be influenced, the same temperature area is not required to be subjected to temperature conversion, the influence of the environment temperature on a liquid drop reaction experiment can be improved, the time consumption of the temperature conversion is saved, and the efficiency of the liquid drop reaction is improved.

In addition, the second electrode layer comprises a plurality of second electrodes which are arranged independently, each second electrode is located in each temperature zone, the second electrodes in different temperature zones are independent, heat conduction between the second electrodes corresponding to different temperature zones is avoided, and mutual influence among different temperature zones can be further improved.

Drawings

Other features, objects and advantages of the invention will become apparent from the following detailed description of non-limiting embodiments with reference to the accompanying drawings in which like or similar reference characters refer to the same or similar parts.

FIG. 1 is a schematic top view of a microfluidic reaction device provided in an embodiment of the first aspect of the present invention;

FIG. 2 isbase:Sub>A cross-sectional view taken at A-A of FIG. 1;

FIG. 3 is a cross-sectional view taken at B-B of FIG. 1;

FIG. 4 is a schematic diagram of a driving circuit in a microfluidic reaction device according to an embodiment of the first aspect of the present invention;

FIG. 5 is a schematic diagram of a partial layer structure of a microfluidic reaction device according to an embodiment of the first aspect of the present invention.

FIG. 6 isbase:Sub>A cross-sectional view of FIG. 1 at A-A in another embodiment;

FIG. 7 is a schematic diagram of another part of the layer structure of a microfluidic reaction device according to an embodiment of the first aspect of the present invention.

FIG. 8 is a schematic diagram of the location of a temperature zone in a microfluidic reaction device according to an embodiment of the first aspect of the present invention.

FIG. 9 is a schematic diagram of the location of a temperature zone in a microfluidic reaction device according to another embodiment of the first aspect of the present invention.

FIGS. 10 to 15 are schematic views showing the procedure of conducting an assay using a microfluidic device according to an embodiment of the first aspect of the present invention;

FIG. 16 is a schematic flow chart of a microfluidic reaction driving method according to an embodiment of the second aspect of the present invention;

FIG. 17 is a schematic flow chart of a microfluidic reaction driving method according to another embodiment of the second aspect of the present invention

Fig. 18 is a schematic flow chart of a microfluidic reaction driving method according to another embodiment of the second aspect of the present invention.

Description of reference numerals:

10. a microfluidic reaction device; TA1, a first temperature zone; TA2, a second temperature zone; TA3, a third temperature zone;

100. a first substrate; 110. a thin film transistor; 110a, a switch tube; 110b, a drive tube; 111. a gate electrode; 112. a source electrode; 113. a drain electrode; 120. an organic layer; 130. a temperature measuring unit; 131. a first temperature measuring electrode; 132. a bismuth telluride layer; 133. a second temperature measuring electrode; 134. a first signal line; 135. a second signal line; 140. a first electrode layer; 141. a first electrode; 150. a first dielectric layer; 160. a first hydrophobic layer; 170. a data line; 180. scanning lines;

200. a second substrate; 210. a heating layer; 211. heating the electrode; 220. a second electrode layer; 221. a second electrode; 230. a second dielectric layer; 240. a second hydrophobic layer;

300. an accommodating space;

400. a packaging layer;

510. a first insulating layer; 520. a second insulating layer; 530. a third insulating layer; 540. a fourth insulating layer;

20. a droplet.

Detailed Description

Features and exemplary embodiments of various aspects of the present invention will be described in detail below. In the following detailed description, numerous specific details are set forth in order to provide a thorough understanding of the invention. It will be apparent, however, to one skilled in the art that the present invention may be practiced without some of these specific details. The following description of the embodiments is merely intended to provide a better understanding of the present invention by illustrating examples of the present invention. In the drawings and the following description, at least some well-known structures and techniques have not been shown in detail in order to avoid unnecessarily obscuring the present invention; also, the dimensions of some of the structures may be exaggerated for clarity. Furthermore, the described features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

In the description of the present invention, it is to be noted that, unless otherwise specified, "a plurality" means two or more; the terms "upper," "lower," "left," "right," "inner," "outer," and the like, as used herein, refer to an orientation or positional relationship indicated for convenience in describing the invention and to simplify description, but do not indicate or imply that the referenced device or element must have a particular orientation, be constructed and operated in a particular orientation, and thus should not be construed as limiting the invention. Furthermore, the terms "first," "second," and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.

The directional terms appearing in the following description are intended to be illustrative in all directions, and are not intended to limit the specific construction of embodiments of the present invention. In the description of the present invention, it should also be noted that, unless otherwise explicitly stated or limited, the terms "mounted" and "connected" are to be construed broadly, e.g., as being fixed or detachable or integrally connected; can be directly connected or indirectly connected. The specific meaning of the above terms in the present invention can be understood as appropriate to those of ordinary skill in the art.

At present, a droplet microfluidic reaction device includes a droplet accommodating portion and two electrodes located at upper and lower sides of the droplet accommodating portion, one of the two electrodes is a control electrode and the other is a common electrode. The common electrode is usually provided over the entire surface. In order to enable the liquid drops to react at different temperatures, the microfluidic reaction device is provided with a plurality of different temperature areas, and the liquid drop microfluidic reaction device drives the liquid drops to move between the different temperature areas through two electrodes.

The inventor finds that there is an interaction between different temperature zones, and it is difficult to ensure that the temperature in the temperature zones is stably at the preset temperature. The inventor further researches and discovers that due to the fact that the common electrode is arranged on the whole surface, heat conduction is conducted between different temperature areas through the common electrode, and mutual influence exists between the different temperature areas.

The present invention is proposed to solve the above-mentioned technical problems. For better understanding of the present invention, the microfluidic reaction device and the microfluidic reaction driving method according to the embodiment of the present invention will be described in detail below with reference to fig. 1 to 15.

Referring to fig. 1 and 2, fig. 1 is a schematic top view of a microfluidic reaction device 10 according to a first embodiment of the present invention. Fig. 2 isbase:Sub>A cross-sectional view atbase:Sub>A-base:Sub>A in fig. 1.

The microfluidic reaction device 10 has a plurality of temperature zones, and the microfluidic reaction device 10 includes: a first substrate 100 and a second substrate 200 disposed at an interval in a thickness direction of the microfluidic (Z direction in fig. 1); a first electrode layer 140 disposed on a side of the first substrate 100 facing the second substrate 200, the first electrode layer 140 including a plurality of first electrodes 141; a first hydrophobic layer 160 disposed on a side of the first electrode layer 140 away from the first substrate 100; the second hydrophobic layer 240 is disposed on one side of the second substrate 200 facing the first hydrophobic layer 160, and the second hydrophobic layer 240 and the first hydrophobic layer 160 are distributed at intervals, so that a receiving space 300 for receiving the droplet 20 is formed between the first hydrophobic layer 160 and the second hydrophobic layer 240; a second electrode layer 220 disposed on a side of the second substrate 200 facing the second water-repellent layer 240, wherein the second electrode layer 220 includes a plurality of second electrodes 221 independently disposed, and each second electrode 221 is located in each temperature zone; and a heating layer 210 disposed between the second electrode layer 220 and the second substrate 200, wherein the heating layer 210 includes a plurality of heating electrodes 211, and each heating electrode 211 is located in each temperature zone.

In the microfluidic reaction device 10 provided in the embodiment of the present invention, the microfluidic reaction device 10 includes a first substrate 100, and a first electrode layer 140 and a first water-repellent layer 160 disposed on the first substrate 100, a second substrate 200, and a heating layer 210, a second electrode layer 220, and a second water-repellent layer 240 disposed on the second substrate 200. The first substrate 100 and the second substrate 200 are butt-joined such that the first electrode layer 140, the first water-repellent layer 160, the heating layer 210, the second electrode layer 220, and the second water-repellent layer 240 are located between the first substrate 100 and the second substrate 200. Wherein the first water-repellent layer 160 and the second water-repellent layer 240 form an accommodating space 300 therebetween so that the droplet 20 can be positioned in the accommodating space 300 and move within the accommodating space 300.

When the microfluidic reaction device 10 of the embodiment of the present invention is used for a droplet experiment, the temperature of each temperature zone may be fixed, and the droplet 20 moves between different temperature zones by the interaction between the first electrode 141 and the second electrode 221 and using the electrowetting principle, so as to achieve the purpose of performing a reaction on the droplet 20 at different temperatures. The plurality of heating electrodes 211 of the heating layer 210 are respectively located in each temperature zone, so that the heating electrodes 211 can be heated in each temperature zone, mutual heating between each temperature zone does not affect each other, temperature conversion in the same temperature zone is not required, the influence of the environmental temperature on the droplet reaction experiment can be improved, the time consumption of temperature conversion is saved, and the efficiency of droplet reaction is improved.

The heating electrode 211 may be disposed in various ways, for example, according to the temperature requirements of different temperature zones, the electrode plate size (for example, the plate width) of the heating electrode 211 in each temperature zone and/or the magnitude of the signal current may be changed, and the heating between the temperature zones may not affect each other.

In a droplet experiment, the speed of the droplet 20 can reach 80mm/s, and the distance between two adjacent temperature zones is usually 100mm, so that the droplet can move from one temperature zone to the next temperature zone within the time of 1s to 2s. In the prior art, when the temperature conversion is utilized to realize the reaction of the liquid drops at different temperatures in the same temperature region, on one hand, the influence of the environmental temperature on the temperature conversion is large, so that the experimental efficiency cannot be ensured. On the other hand, when the temperature-drop treatment is required for the temperature-drop treatment, the temperature-drop speed is usually very slow, and it may take 2min or even longer to realize the temperature conversion. Therefore, in the embodiment of the present application, the microfluidic reaction device 10 includes a plurality of temperatures, and the droplet 20 is moved to react at different temperatures, so that the efficiency of the droplet experiment can be effectively improved.

In addition, the second electrode layer 220 includes a plurality of second electrodes 221 that are disposed independently of each other, each second electrode 221 is located in each temperature zone, different second electrodes 211 are independent of each other, and adjacent second electrodes 221 do not conduct heat with each other, which can further improve the mutual influence between different temperature zones.

With continued reference to fig. 1, in some alternative embodiments, the microfluidic reaction device 10 includes 5 temperature zones and 5 second electrodes 221 and 5 heating electrodes 211 respectively located in each temperature zone. In other embodiments, the microfluidic reaction device 10 may further include 2, 3, and 4 isothermal zones, and correspondingly, the number of the second electrodes 221 may also be 2, 3, or 4, and the number of the heating electrodes 211 may also be 2, 3, or 4, as long as there are a plurality of temperature zones, the number of the second electrodes 221, and the number of the heating electrodes 211, and each of the second electrodes 221 and each of the heating electrodes 211 is located in each temperature zone. The plurality of heating electrodes 211 located in the same temperature zone may be independent from each other or may be connected to each other. When mutually independent, each heating electrode 211 comprises a heating signal terminal for connection with a driving module; when interconnected, a temperature zone has only two heating signal terminals.

Optionally, the heating electrode 211 is formed to extend along the bending path, so as to increase the length of the heating electrode 211 in the temperature zone, and improve the heating effect of the heating electrode 211.

Optionally, when the dielectric constant of the first hydrophobic layer 160 is insufficient, resulting in insufficient driving force for the droplet 20, or when the dielectric strength of the first hydrophobic layer 160 is insufficient, such that the first hydrophobic layer 160 is easily broken down, in some optional embodiments, a first dielectric layer 150 is further disposed between the first hydrophobic layer 160 and the first electrode layer 140. The dielectric constant and/or dielectric strength of first hydrophobic layer 160 can be increased by providing first dielectric layer 150.

Alternatively, when the dielectric constant of the second hydrophobic layer 240 is insufficient, resulting in insufficient driving force for the droplet 20, or when the dielectric strength of the second hydrophobic layer 240 is insufficient, such that the second hydrophobic layer 240 is easily broken down, in some alternative embodiments, the second electrode layer 220 is disposed between the heating layer 210 and the second hydrophobic layer 240. A second dielectric layer 230 is also disposed between the second hydrophobic layer 240 and the second electrode layer 220. The dielectric constant and/or the dielectric strength of the second hydrophobic layer 240 can be improved by the second dielectric layer 230.

Referring to fig. 3, fig. 3 is a cross-sectional view taken along line B-B of fig. 1.

In some optional embodiments, the microfluidic reaction device 10 further includes an encapsulation layer 400, which is encapsulated between the first hydrophobic layer 160 and the second hydrophobic layer 240, so that the first hydrophobic layer 160, the second hydrophobic layer 240, and the encapsulation layer 400 enclose a relatively closed accommodating space 300, and leakage of the droplet 20 is avoided. Optionally, a first insulating layer 510 is disposed between the heating layer 210 and the second electrode layer 220 to ensure that the heating electrode 211 in the heating layer 210 and the second electrode 221 in the second electrode layer 220 are disposed in an insulating manner.

The first substrate 100 and the second substrate 200 are not limited to be disposed, and for example, the first substrate 100 and/or the second substrate 200 may be a glass substrate or a flexible substrate.

There are various ways to drive the droplet 20 to move in the containing space 300 through the first electrode 141 and the second electrode 221, and in some alternative embodiments, the microfluidic reaction device 10 further includes a thin film transistor 110 connected to the first electrode 141 and used for controlling the operating state of the first electrode 141.

In these alternative embodiments, on one hand, the driving of the first electrode 141 by the thin film transistor 110 can ensure the control accuracy of the microfluidic reaction device 10, so that the microfluidic reaction device 10 has the advantages of high throughput and high accuracy. On the other hand, the thin film transistor 110 is connected to the first electrode 141, that is, the first electrode 141 is a control electrode, the first electrode 141 is located on one side of the first hydrophobic layer 160 and the second hydrophobic layer 240 away from the heating layer 210, so that the distance between the first electrode 141, the thin film transistor 110 and the heating layer 210 can be increased, the risk of the change of the thermal characteristics of the thin film transistor 110 is reduced, and the long-term normal and stable operation of the thin film transistor 110 is ensured.

Referring to fig. 4, fig. 4 is a schematic diagram of a driving circuit structure of a microfluidic reaction device 10 according to an embodiment of the present invention.

Optionally, the first substrate 100 is further provided with a scan line 180 and a data line 170; the gate 111 of the thin film transistor 110 is connected to the scan line 180, the drain 113 of the thin film transistor 110 is connected to the data line 170, and the source 113 of the thin film transistor 110 is connected to the first electrode 141, so that a control signal is transmitted to the first electrode 141 through the source 113 of the thin film transistor 110.

Optionally, the scanning lines 180 are multiple, and the multiple scanning lines 180 are distributed at intervals along the Y direction (the second direction); the data lines 170 are spaced apart from each other in the X direction (first direction), and the scan lines 180 intersect the data lines 170 to form intersections. When current exists on a certain data line 170 and a certain scanning line 180, the thin film transistor 110 at the position of the intersection of the data line 170 and the scanning line 180 receives a control signal, so as to drive the droplet 20 at the position of the intersection to move. Accurate movement of the droplet 20 can be achieved through the plurality of data lines 170 and the plurality of scan lines 180.

With continued reference to fig. 2, in order to obtain the temperatures of different temperature zones or locations of the droplets 20 in time, in some alternative embodiments, the microfluidic reaction device 10 further comprises: the temperature measuring unit 130 is located on one side of the first electrode layer 140 facing the first substrate 100. The temperature measuring unit 130 is arranged to obtain the actual temperature in the microfluidic reaction device 10 in real time, so that the microfluidic reaction device 10 can control the heating temperature of the heating layer 210 according to the actual temperature, the droplet 20 is guaranteed to react within a preset temperature range, and the accuracy of the microfluidic reaction experiment is improved.

The temperature measuring unit 130 can be disposed in various manners, and the temperature measuring unit 130 can be a temperature sensor, for example, the temperature measuring unit 130 is a thermistor.

In some optional embodiments, the thermometry unit 130 comprises: a first temperature measuring electrode 131; the bismuth telluride layer 132 is positioned on one side of the first temperature measuring electrode 131, which is far away from the first electrode layer 140; and the second temperature measuring electrode 133 is positioned on the side of the bismuth telluride layer 132, which is far away from the first temperature measuring electrode 131, and the first temperature measuring electrode 131 and the second temperature measuring electrode 133 are mutually connected through the bismuth telluride layer 132.

In these alternative embodiments, the temperature measuring unit 130 comprises a first temperature measuring electrode 131, a second temperature measuring electrode 133 and a bismuth telluride layer 132 between the first temperature measuring electrode 131 and the second temperature measuring electrode 133, and the temperature of the position where the temperature measuring unit 130 is located can be obtained by measuring the potential difference between the first temperature measuring electrode 131 and the second temperature measuring electrode 133 by using the seebeck effect. The temperature is measured by using the seebeck effect, so that the measurement range of the temperature measuring unit 130 can be expanded and the measurement accuracy can be improved.

Optionally, a second insulating layer 520 is disposed between the first electrode layer 140 and the first thermometric electrode 131 to ensure that the first electrode 141 in the first electrode layer 140 and the first thermometric electrode 131 are insulated from each other.

The second temperature measuring electrode 133 can be disposed at various positions, for example, the second temperature measuring electrode 133 is disposed on the side of the thin film transistor 110 where the source and drain electrodes 113 face the first electrode layer 140, or the second temperature measuring electrode 133 is disposed on the side of the thin film transistor 110 where the source and drain

electrodes

112 and 113 face away from the first electrode layer 140.

Optionally, in some embodiments, the second thermometric electrode 133 is disposed on the same layer as the source 112 and the drain 113 of the thin film transistor 110. In the preparation process of the microfluidic reaction device 10, the source 112, the drain 113 and the second temperature measuring electrode 133 of the thin film transistor 110 can be prepared and molded in the same process step, so that the preparation process of the microfluidic reaction device 10 can be simplified, and the preparation efficiency of the microfluidic reaction device 10 can be improved.

In addition, when the microfluidic reaction device 10 includes the data line 170 and the scan line 180, the data line 170 or the scan line 180 is disposed on the same layer as the source electrode 112 and the drain electrode 113 of the thin film transistor 110. For example, the data line 170 is disposed on the same layer as the source electrode 112 and the drain electrode 113 of the thin film transistor 110. So that the second thermometric electrode 133 and the data line 170 are disposed on the same layer.

The relative position arrangement between the first thermometric electrode 131 and the second thermometric electrode 133 is various, and optionally, as long as the orthographic projection of the first thermometric electrode 131 on the first substrate 100 and the orthographic projection of the second thermometric electrode 133 on the first substrate 100 are at least partially overlapped, the bismuth telluride layer 132 extends along the Z direction and can be connected between the first thermometric electrode 131 and the second thermometric electrode 133.

In the temperature measurement unit 130, the overlapping area of the orthographic projection of the first temperature measurement electrode 131 on the first substrate 100 and the orthographic projection of the second temperature measurement electrode 133 on the first substrate 100 determines the temperature measurement area of the temperature measurement unit 130.

The temperature measuring unit 130 can be disposed at various positions, for example, the temperature measuring unit 130 is disposed at different positions in the microfluidic reaction device 10, so as to obtain the temperature at different positions in the microfluidic reaction device 10. Optionally, a temperature measuring unit 130 is disposed in each temperature zone to obtain the actual temperature of each temperature zone in real time.

The temperature measuring unit 130 can be arranged in various ways in the temperature zone, for example, the temperature measuring unit 130 is arranged at different positions in the temperature zone. Alternatively, when the liquid droplet 20 reacts in the central region of the temperature zone, the temperature measuring unit 130 is disposed in the central region of the temperature zone to be able to acquire the actual temperature of the central region of the temperature zone in real time, that is, the actual temperature of the liquid droplet 20 when reacting.

When the temperature measuring unit 130 is disposed in each temperature zone, one temperature measuring unit 130 may be disposed in each temperature zone, and the actual temperature of the temperature zone is obtained through one temperature measuring unit 130.

In other embodiments, two or more temperature measuring units 130 are disposed in each temperature zone. More than two temperature measuring units 130 in the same temperature zone can be independently arranged, and the temperatures of different positions of the temperature zone can be obtained through a plurality of temperature measuring units 130.

Referring to fig. 1, the temperature measuring unit 130 is disposed on a side of the first electrode 141 facing the thin film transistor 110, the first electrode 141 is connected to the source 112 of the thin film transistor 110, a via hole is disposed on the second insulating layer 520 between the first electrode 141 and the thin film transistor 110, and a connecting wire is connected between the source 112 and the first electrode 141 via the via hole.

The following description will be given taking an example in which one temperature measuring unit 130 is provided in each temperature zone.

When one temperature measuring unit 130 is arranged in each temperature zone, the number of the first temperature measuring electrodes 131 and the second temperature measuring electrodes 133 of the temperature measuring unit 130 is multiple.

Referring to fig. 5, fig. 5 is a schematic diagram of a partial layer structure of a microfluidic reaction device 10 according to an embodiment of the present invention. In fig. 5, in order to better show the structure of the first thermometric electrode 131 and the second thermometric electrode 133, the size of the second thermometric electrode 133 is smaller than that of the first thermometric electrode 131. In other embodiments, the first thermometric electrode 131 and the second thermometric electrode 133 in the thermometric unit 130 may be the same size.

Optionally, the number of the first temperature measuring electrodes 131 and the number of the second temperature measuring electrodes 133 are multiple, each of the first temperature measuring electrodes 131 and each of the second temperature measuring electrodes 133 are respectively disposed correspondingly, the multiple first temperature measuring electrodes 131 are connected to each other and/or the multiple second temperature measuring electrodes 133 are connected to each other, and the temperature measuring unit 130 includes the multiple first temperature measuring electrodes 131, so that the temperature measuring area of the temperature measuring unit 130 can be enlarged.

Wherein, the corresponding arrangement of each first thermometric electrode 131 and each second thermometric electrode 133 respectively means that: the orthographic projection of each first thermometric electrode 131 on the first substrate 100 and the orthographic projection of each second thermometric electrode 133 on the first substrate 100 are at least partially overlapped. As shown in FIG. 5, the orthographic projection of each second thermometric electrode 133 on the first substrate 100 is located within the orthographic projection of each first thermometric electrode 131 on the first substrate 100.

In other embodiments, the orthographic projection of each first thermometric electrode 131 on the first substrate 100 can also completely overlap the orthographic projection of the second thermometric electrode 133 on the first substrate 100, so as to reduce the distribution area of the thermometric units 130.

When the first temperature measuring electrodes 131 and the second temperature measuring electrodes 133 are both multiple, the orthographic projections of the single first temperature measuring electrode 131 on the first substrate 100 are respectively overlapped with the orthographic projections of the single first electrode 141 on the first substrate 100, and each first temperature measuring electrode 131 and the connecting lead are arranged in a staggered manner, so that the first temperature measuring electrode 131, the connecting lead and the first electrode 141 are mutually insulated.

In order to make the first thermometric electrode 131 and the connecting lead arranged in a staggered manner, as shown in fig. 5, optionally, the orthographic projection of the first thermometric electrode 131 on the first substrate 100 is located within the orthographic projection of the first electrode 141 on the first substrate 100. In other embodiments, the orthographic projection of the first thermometric electrode 131 on the first substrate 100 may also partially overlap the orthographic projection of the first electrode 141 on the first substrate 100.

When the number of the first temperature measuring electrode 131 and the second temperature measuring electrode 133 is plural, the number of the bismuth telluride layers 132 is plural, and there may be one bismuth telluride layer 132, and one bismuth telluride layer 132 is connected between the first temperature measuring electrode 131 and the second temperature measuring electrode 133 which are oppositely arranged in the Z direction. Or the number of the bismuth telluride layers 132 is plural, and each bismuth telluride layer 132 is connected between the same set of the first temperature measuring electrode 131 and the second temperature measuring electrode 133 which are opposite to each other in the Z direction.

Optionally, two adjacent first temperature measurement electrodes 131 are connected to each other through a first signal line 134, and the first signal line 134 is insulated from the connection lead, so that the plurality of first temperature measurement electrodes 131 are interconnected to form one temperature measurement electrode through the first signal line 134, thereby forming one temperature measurement unit 130. The mutual insulation of the first signal line 133 and the first electrode 141 can avoid a short circuit, ensure the normal operation of the temperature measuring unit 130, and avoid the temperature measuring unit 130 from influencing the control of the thin film transistor 110 on the first electrode 141.

Optionally, when the temperature measuring unit 130 includes more than two first temperature measuring electrodes 131, the more than two first temperature measuring electrodes 131 are disposed corresponding to the adjacent first electrodes 141, so that a distance between the two adjacent first temperature measuring electrodes 141 can be reduced, a distribution area of the temperature measuring unit 130 can be reduced, and an extending distance of the first signal line 134 can be reduced.

For example, referring to fig. 5, when four first temperature measuring electrodes 131 are correspondingly disposed in the temperature zone, the four first temperature measuring electrodes 131 are disposed corresponding to the four first electrodes 141, the four first electrodes 141 are arranged in two rows and two columns along the X direction and the Y direction, and two of the four first electrodes 141 are disposed adjacent to each other, so as to reduce the distance between the four first temperature measuring electrodes 131. In other embodiments, the number of the first thermometric electrodes 131 can also be 2, 3 or 5, etc.

When the plurality of first thermometric electrodes 131 are connected to each other through the first signal line 134, the plurality of second thermometric electrodes 144 may be disposed independently of each other. Alternatively, referring to fig. 5, the second thermometric electrodes 144 are connected to each other through the second signal line 135.

When the second thermometric electrodes 133 and the data lines 170 are disposed on the same layer, the data lines 170 generally extend in the Y direction, and in order to ensure the mutual insulation between the second thermometric electrodes 133 and the data lines 170, in the same temperature region, the second thermometric electrodes 133 in the same column are connected to each other through the second signal lines 135, the number of the second signal lines 135 is two or more, and the two or more second signal lines 135 are connected to each other. In this way, it is possible to connect the plurality of second thermometric electrodes 133 through the two or more second signal lines 135 and to prevent the second thermometric electrodes 133 from crossing the data lines 170.

Alternatively, as shown in fig. 5, the second signal line 135 extends to the edge of the microfluidic reaction device 10, and two or more second signal lines 135 cross over one end of the data line 170 and are connected to each other. In still other alternative embodiments, the number of the first thermometric electrodes 131 in the thermometric unit 130 is one, and one first thermometric electrode 131 may be disposed corresponding to one first electrode 141, that is, an orthographic projection of the first thermometric electrode 131 on the first substrate 100 at least partially overlaps an orthographic projection of a single first electrode 141 on the first substrate 100, and the first thermometric electrode 131 and the connecting lead are insulated from each other.

Referring also to fig. 6, fig. 6 isbase:Sub>A cross-sectional view of fig. 1 atbase:Sub>A-base:Sub>A in another embodiment. Fig. 6 is different from fig. 2 in that the first thermometric electrode 131 has a large area.

Optionally, an orthographic projection of the first temperature measuring electrode 131 on the first substrate 100 overlaps with an orthographic projection of more than two first electrodes 141 on the first substrate 100, a first yielding hole is formed in the first temperature measuring electrode 131, and the connecting wire is connected between the first electrode 141 and the thin film transistor 110 through the first yielding hole. In these alternative embodiments, the area of the first thermometric electrode 131 is larger, which can increase the detection area of the thermometric unit 130.

Referring to fig. 7, in the above embodiment, the number of the second thermometric electrodes 133 may be one, and the orthographic projection of one second thermometric electrode 133 on the first substrate 100 is overlapped with the orthographic projection of the first thermometric electrode 131 on the first substrate 100. When the second temperature measuring electrode 133, the data line 170, the source electrode 112, and the drain electrode 113 are disposed on the same layer, the second temperature measuring electrode 133 is provided with a second recess, and the data line 170, the source electrode 112, and the drain electrode 113 are disposed in the second recess, so that the second temperature measuring electrode 133, the data line 170, the source electrode 112, and the drain electrode 113 can be insulated from each other.

Alternatively, referring to fig. 7, in order to better show the structure of the second temperature measuring electrode 133, the first temperature measuring electrode 131 is omitted in fig. 7.

The number of the second thermometric electrodes 133 may be two or more, and the orthographic projections of the two or more second thermometric electrodes 133 on the first substrate 100 may be located within the orthographic projection of the first thermometric electrode 131 on the first substrate 100. Each second temperature measuring electrode 133 is located between two adjacent data lines 170, and each second temperature measuring electrode 133 is disposed in a staggered manner with respect to the source electrode 112 and the drain electrode 113, so as to ensure that the second temperature measuring electrode 133 is insulated from the data lines 170, the source electrode 112 and the drain electrode 113.

In some alternative embodiments, the microfluidic reaction device 10 further comprises: the organic layer 120 is disposed between the first temperature measuring electrode 131 and the second temperature measuring electrode 133, a first via hole (not shown) is formed through the organic layer 120, and the bismuth telluride layer 132 is connected between the first temperature measuring electrode 131 and the second temperature measuring electrode 133 through the first via hole. Referring to fig. 1, in fig. 1, a portion of the bismuth telluride layer 132 penetrating through the organic layer 120 is a first via.

In these alternative embodiments, by providing the organic layer 120, a first via hole is provided on the organic layer 120, and the bismuth telluride layer 132 is connected to the first thermometric electrode 131 and the second electrode 221 via the first via hole. Through the thickness of the organic layer 120, the distance between the first temperature measuring electrode 131 and the second temperature measuring electrode 133 can be adjusted, and the extension length of the bismuth telluride layer 132 in the Z direction can be adjusted, so that the temperature measuring unit 130 can meet the temperature measuring requirement. In addition, the organic layer 120 can provide a heat insulation effect to prevent the heating electrode 211 from heating the thin film transistor 110.

Optionally, the thickness of the organic layer 120 is, for example, 2 μm to 8 μm. When the thickness of the organic layer 120 is within the above range, it is possible to prevent both the thickness of the microfluidic reaction device 10 from being excessively large due to the excessive thickness of the organic layer 120 and the size of the microfluidic reaction device 10 from being excessively large; the problem that the extension length of the bismuth telluride layer 132 in the Z direction is insufficient due to the over-small thickness of the organic layer 120, so that the temperature measuring unit 130 cannot meet the temperature measuring requirement, can also be avoided.

Optionally, the organic layer 120 is disposed on a side of the second insulating layer 520 facing away from the first electrode layer 140. A third insulating layer 530 is further disposed between the source electrode 112, the gate electrode 111, and the second thermometric electrode 133, and the organic layer 120.

Optionally, the thin film transistor 110 further includes a gate 111, and the gate 111 is located on a side of the source 112 and the drain 113 facing away from the first electrode layer 140. Optionally, a fourth insulating layer 540 is further disposed between the gate 111 and the source and drain

electrodes

112 and 113.

There are various ways of setting the temperature zones in the microfluidic reaction device 10, for example, when the droplets 20 need to react at two different temperatures, more than two temperature zones with different temperatures may be set in the microfluidic reaction device 10. Or when the droplets 20 need to react at three different temperatures, more than three temperature zones of different temperatures may be provided in the microfluidic reaction device 10.

Referring to fig. 8, fig. 8 is a schematic diagram illustrating a location of a temperature zone in a microfluidic reaction device 10 according to an embodiment of the present invention.

According to the microfluidic reaction device 10 provided in the embodiment of the present invention, the plurality of temperature regions of the microfluidic reaction device 10 include the initial temperature region TA1, the end temperature region TA3, and one or more intermediate temperature regions TA2 located between the initial temperature region TA1 and the end temperature region TA3, the initial temperature region TA1, the intermediate temperature region TA2, and the end temperature region TA3 are distributed along a predetermined annular path, the number of the initial temperature regions TA1 and the end temperature regions TA3 is one or more, and at least one of the initial temperature regions TA1 and at least one of the end temperature regions TA3 are partially or completely overlapped, so that the droplets 20 can continue to circularly move to the initial temperature region TA1 from the end temperature region TA3 after moving from the initial temperature region TA1 to the end temperature region TA3.

The initial temperature zone TA1 is a temperature zone in which the droplet 20 undergoes the first reaction, and the final temperature zone is a temperature zone in which the droplet 20 undergoes the last reaction. For example, when the microfluidic reaction device 10 is used to perform a Polymerase Chain Reaction (PCR) experiment, the droplets 20 need to react at about 95 ℃, about 60 ℃, and about 72 ℃ in that order. The droplets 20 need to undergo a first reaction at around 95 c and a final reaction at around 72 c. The initial temperature zone TA1 may be a temperature zone having a temperature of about 95 deg.c, the intermediate temperature zone TA2 may be a temperature zone having a temperature of about 60 deg.c, and the finishing temperature zone TA3 may be a temperature zone having a temperature of about 72 deg.c.

The predetermined circular path is, for example, a moving path of the droplet 20, and the droplet 20 moves along the predetermined circular path. The predetermined circular path may be triangular, quadrangular or curved as long as the droplet 20 moves along the predetermined circular path, and then continuously moves circularly from the initial temperature zone TA1 to the ending temperature zone TA3 after moving from the initial temperature zone TA1 to the ending temperature zone TA3.

In these alternative embodiments, at least one of the initial temperature zones TA1 and at least one of the end temperature zones TA3 partially or completely overlap. Therefore, when the liquid drop 20 moves along the preset annular path, after the liquid drop moves from the initial temperature area TA1 to the ending temperature area TA3, the liquid drop can continuously and circularly move from the ending temperature area TA3 to the initial temperature area TA1, so that the circular reaction of the liquid drop 20 is realized, and the consumption of samples or reagents used in the experiment can be effectively reduced.

When the droplet 20 circulates through the initial temperature zone TA1 a plurality of times on the preset loop path, the initial temperature zone TA1 may be the initial temperature zone TA1 at the same position or the initial temperature zones TA1 at different positions. When the droplet 20 circulates through the ending temperature region TA3 a plurality of times on the predetermined circular path, the ending temperature region TA3 may be the ending temperature region TA3 at the same position or the ending temperature region TA3 at different positions.

In some optional embodiments, the number of the initial temperature zone TA1, the intermediate temperature zone TA2, and the ending temperature zone TA3 is one, and the initial temperature zone TA1 and the ending temperature zone TA3 are located on at least one side of the intermediate temperature zone TA2 in the X direction; the intermediate temperature zone TA2 and the initial temperature zone TA1 at least partially overlap in the X direction so that the droplet 20 can move between the initial temperature zone TA1 and the intermediate temperature zone TA2 when moving in the X direction; the intermediate temperature region TA2 and the end temperature region TA3 at least partially overlap in the X direction so that the liquid droplet 20 can move between the intermediate temperature region TA2 and the end temperature region TA3 when moving in the X direction.

In these alternative embodiments, the intermediate temperature zone TA2 and the initial temperature zone TA1 at least partially overlap in the X direction by: at least part of the intermediate temperature area TA2 and the initial temperature area TA1 are sequentially distributed along the X direction, so that the liquid drop 20 can move between the intermediate temperature area TA2 and the initial temperature area TA1 when moving along the X direction, the liquid drop 20 does not need to move in a curve, and the moving mode of the liquid drop 20 can be simplified. Similarly, the intermediate temperature zone TA2 and the end temperature zone TA3 at least partially overlap in the X direction: at least part of the intermediate temperature area TA2 and the ending temperature area TA3 are distributed in sequence along the X direction, so that the liquid drop 20 can move between the intermediate temperature area TA2 and the ending temperature area TA3 when moving along the X direction, the liquid drop 20 does not need to move in a curve, and the moving mode of the liquid drop 20 can be simplified.

Optionally, the initial temperature zone TA1 and the ending temperature zone TA3 located on the same side of the intermediate temperature zone TA2 in the X direction are distributed along the Y direction, so that the droplet 20 can move between the ending temperature zone TA3 and the initial temperature zone TA1 when moving along the Y direction. The initial temperature zone TA1 and the ending temperature zone TA3 move along the Y direction, so that the droplet 20 can move between the initial temperature zone TA1 and the ending temperature zone TA3 when moving along the Y direction, the droplet 20 does not need to perform curvilinear motion, and the moving mode of the droplet 20 can be simplified.

The size and number of the initial temperature zone TA1, the intermediate temperature zone TA2, and the ending temperature zone TA3 may be set in various manners, for example, in some alternative embodiments, as shown in fig. 5, the areas and shapes of the initial temperature zone TA1, the intermediate temperature zone TA2, and the ending temperature zone TA3 are the same. The number of the initial temperature area TA1, the intermediate temperature area TA2 and the ending temperature area TA3 is respectively 1, and the initial temperature area TA1, the intermediate temperature area TA2 and the ending temperature area TA3 are arranged according to a 'Ping' shape figure.

The middle region of the temperature zone is typically a temperature stable region, and the droplet 20 is typically located in the middle region of the temperature zone so that the droplet 20 can be uniformly heated. In these alternative embodiments described above, the droplet 20 may move in the X direction from the central region of the initial temperature zone TA1 to the intermediate temperature zone TA2, and then the droplet 20 moves in the Y direction to the central region of the intermediate region TA 2. The liquid drop 20 can move from the intermediate temperature area TA2 to the ending temperature area TA3 continuously along the Y direction and can move back to the initial temperature area TA1 continuously along the Y direction, and the circulation experiment of the liquid drop 20 is realized repeatedly. The liquid droplets 20 circulate on a preset loop path through a plurality of initial temperature zones TA1, and these initial temperature zones TA1 are initial temperature zones TA1 at the same position.

Referring to fig. 9, fig. 9 is a schematic diagram illustrating a location of a temperature zone in a microfluidic reaction device 10 according to another embodiment of the first aspect of the present invention.

According to the microfluidic reaction device 10 provided in another embodiment of the present invention, the initial temperature zone TA1 and the final temperature zone TA3 are both two; the two initial temperature zones TA1 are respectively arranged at two sides of the middle temperature zone TA2 in the X direction, and the two initial temperature zones TA1 are distributed in a staggered manner in the X direction; the two ending temperature areas TA3 are respectively arranged at two sides of the middle temperature area TA2 in the X direction, and the two ending temperature areas TA3 are distributed in a staggered manner in the X direction; the initial temperature zone TA1 and the end temperature zone TA3 located on both sides of the intermediate temperature zone TA2 in the X direction are distributed in the Y direction.

In these alternative embodiments, when the droplet 20 moves along the predetermined circular path, the droplet 20 only needs to move along the X direction to move from the central region of the initial temperature zone TA1 to the central region of the intermediate temperature zone TA2, the droplet 20 continues to move along the X direction to move from the central region of the intermediate temperature zone TA2 to the central region of the ending temperature zone TA3, the droplet 20 continues to move along the Y direction to move from the central region of the ending temperature zone TA3 to the central region of the next initial temperature zone TA1, then the droplet 20 continues to move along the X direction to move to the central region of the next intermediate temperature zone TA2, continues to move along the X direction to move to the central region of the next ending temperature zone TA3, and then continues to move along the Y direction to return to the original initial temperature zone TA1. Further, in these alternative embodiments, when the liquid droplet 20 circulates through the initial temperature zone TA1 a plurality of times on the preset loop path, the initial temperature zone TA1 may be an initial temperature zone TA1 of different positions.

In these embodiments of the present invention, the droplet 20 only needs to move in one direction to move from the central region of one temperature zone to the central region of the next adjacent temperature zone, which can further simplify the moving manner of the droplet 20. .

Optionally, the area of the intermediate temperature zone TA2 is larger, and the extension size of the intermediate temperature zone TA2 in the Y direction is equal to the sum of the extension sizes of the initial temperature zone TA1 and the end temperature zone TA3 in the Y direction. The extension of the intermediate temperature zone TA2 in the X direction is equal to the extension of the initial temperature zone TA1 and the end temperature zone TA3 in the X direction. Alternatively, the initial temperature zone TA1 and the end temperature zone TA3 have the same extension in the Y direction.

In some optional embodiments, the central area of each temperature zone may also be embedded with a reagent. For example, when the microfluidic reaction device 10 is used to perform a Polymerase Chain Reaction (PCR) experiment, reagents such as primers, bases, and polymerase may be embedded in the central region of each temperature zone.

In these alternative embodiments, reagents are embedded in the temperature zones, and when the microfluidic reaction device 10 is used, it is only necessary to drive the droplets 20 containing the sample to be amplified without driving the reagents to move to the central zone, thereby improving the experimental efficiency. In addition, the reagent is embedded in the central area of each temperature zone, so that the liquid drop 20 can react with the reagent in the central area, the influence of temperature deviation existing in the boundary area on an experimental result can be improved, and the accuracy of the experimental result is improved.

In some alternative embodiments, the microfluidic reaction device 10 further comprises a liquid inlet, the liquid inlet being in communication with the accommodation space 300 such that the liquid droplets 20 can enter the accommodation space 300 through the liquid inlet. The liquid inlet is disposed at a plurality of positions, for example, the liquid inlet may be disposed at an edge of one of the temperature zones, for example, the liquid inlet is disposed at an edge of the first temperature zone TA1, so that the liquid droplets 20 can enter the first temperature zone TA1 from the liquid inlet for reaction. The liquid inlet is, for example, disposed on the package layer 400, and a through hole communicating with the accommodating space 300 is disposed on the package layer 400 as the liquid inlet.

In still other embodiments, the liquid inlet may also be used as a liquid outlet, so that the reacted liquid droplets 20 flow out of the liquid inlet. Alternatively, the microfluidic reaction device 10 further comprises a liquid outlet, which is communicated with the accommodating space 300, so that the reacted liquid droplets 20 can flow out from the liquid outlet.

The following description will discuss a method of using the microfluidic reaction device 10, by taking as an example the microfluidic reaction device 10 shown in FIGS. 2 and 9, for performing a PCR experiment. Reagents are not embedded in each temperature zone, and the droplet 20 contains a sample to be amplified (e.g., a specific DNA fragment), primers, bases, polymerase, and the like.

Referring to fig. 10, for example, the starting position of the droplet 20 is a first initial temperature zone TA1 (an upper left initial temperature zone TA1 in fig. 10), a liquid inlet is disposed on the first initial temperature zone TA1, the droplet 20 enters the initial temperature zone TA1 from the liquid inlet, and the droplet 20 reacts in the initial temperature zone TA1 (e.g., a central region of the initial temperature zone TA 1), for example, the temperature of the initial temperature zone TA1 is about 95 ℃, and double strands of DNA are unwound into single strands in the initial temperature zone TA1.

Then, the first electrode 141 is controlled by the thin film transistor 110, and the first electrode 141 and the second electrode 221 cooperate with each other to drive the droplet 20 to move in the direction indicated by the arrow in fig. 10, so that the droplet 20 moves from the central region of the initial temperature zone TA1 to the central region of the intermediate temperature zone TA2, as shown in fig. 11. The single strand is allowed to react in the central region of the intermediate temperature zone TA 2. For example, the temperature of the intermediate temperature region TA2 is about 60 ℃, and the single strand is bonded with the primer in the intermediate temperature region TA2 according to the base complementary pairing principle.

Next, the first electrode 141 is controlled by the thin film transistor 110, and the first electrode 141 and the second electrode 221 cooperate with each other to drive the droplet 20 to move in the direction indicated by the arrow in fig. 11, so that the droplet 20 moves from the central region of the intermediate temperature zone TA2 to the central region of the end temperature zone TA3, as shown in fig. 12. The paired single chains are reacted in the central area of the ending temperature zone TA3. For example, the finishing temperature zone TA3 is about 72 ℃ and the base forms a complementary pair with the single strand under the action of polymerase to form a double strand. The first round of amplification of the DNA fragment was completed.

When the DNA fragments obtained after the reaction of the set of initial temperature zone TA1, intermediate temperature zone TA2 and ending temperature zone TA3 are too small to meet the use requirement, the first electrode 141 is controlled by the thin film transistor 110, and the first electrode 141 and the second electrode 221 cooperate with each other to drive the droplet 20 to move along the arrow direction shown in fig. 12, so that the droplet 20 moves from the central area of the ending temperature zone TA3 to the central area of the next initial temperature zone TA1 (the lower right initial temperature zone TA1 in fig. 13), as shown in fig. 13. The droplet 20 is subjected to the next round of unwinding reaction in the next initial temperature zone TA1. The droplet 20 then moves to the central area of the intermediate temperature zone TA2 along the arrow shown in fig. 13, as shown in fig. 14. After the reaction in the central region of the intermediate temperature region TA2 is completed, the droplet 20 moves to the central region of the end temperature region TA3 along the arrow shown in fig. 14, as shown in fig. 15. When the droplet 20 still fails to satisfy the requirement after two rounds of reactions, the droplet 20 may move to the central region of the first initial temperature region TA1 in the direction indicated by the arrow in fig. 15, and continue the next round of reactions. The above steps are repeated until the reaction product meets the requirement.

When the liquid drops 20 are evaporated after multiple rounds of reaction, the liquid drops 20 can be supplemented through the liquid inlet, so that the liquid drops 20 and the original liquid drops 20 are mixed and then continue to react.

Referring to fig. 16, fig. 16 is a schematic flow chart of a microfluidic driving method according to a second embodiment of the present invention.

According to an embodiment of the second aspect of the present invention, a microfluidic reaction driving method is provided, which uses any one of the microfluidic reaction devices 10 provided in the embodiments of the first aspect. The microfluidic reaction driving method includes:

step S01: the droplets and the reagents are mixed to form mixed droplets.

Step S02: and driving the mixed liquid drop to move to the central area of the preset temperature area.

Step S03: the temperature zone where the mixed liquid drop is located is heated to the reaction temperature through the heating electrode, so that the liquid drop in the mixed liquid drop and the reagent can react with each other.

In the microfluidic reaction driving method provided by the embodiment of the present invention, the droplets 20 and the reagents are first mixed. For example, when the microfluidic reaction driving method provided by the embodiment of the present invention is used in a PCR experiment, the droplet 20 includes a sample to be amplified, and the reagents include primers, bases, and polymerase. And then driving the mixed liquid drop to move to the central area of the preset temperature area. The preset temperature zone may be a temperature zone in which the mixed liquid droplet is to be reacted. Finally, the temperature zone of the mixed droplet is heated to the reaction temperature by the heating electrode 211, for example, when the mixed droplet is to be despin, the preset temperature zone is the first temperature zone TA1, the temperature of the first temperature zone TA1 is heated to about 95 ℃ by the heating electrode 211, and the sample to be amplified is despin to be a single chain in the first temperature zone TA1.

In the microfluidic reaction driving method provided by the embodiment of the invention, in step S02, the mixed droplet is driven to the central region of the preset temperature zone, so that the mixed droplet reacts in the central region but not in the boundary region. In the microfluidic reaction device 10, when a plurality of temperature zones are disposed adjacent to each other or a plurality of temperature zones are disposed at intervals, the temperature zones may have temperature deviations from the regions around the temperature zones, resulting in possible temperature deviations in the boundary regions of the temperature zones. According to the method provided by the embodiment of the invention, the mixed liquid drop reacts in the central area instead of the boundary area, so that the influence of the temperature deviation on the reaction result can be avoided, and the accuracy of the reaction result is improved.

In addition, in the embodiment of the present invention, the heating electrode 211 is used to heat the preset temperature region after the mixed liquid droplet moves to the central region, so as to prevent the actual temperature of the temperature region from reaching the reaction temperature before the mixed liquid droplet moves to the central region, so that the liquid droplet 20 will react before moving to the central region, which affects the accuracy of the reaction result.

Referring to fig. 17, fig. 17 is a flow chart illustrating a microfluidic driving method according to another embodiment of the second aspect of the present invention, which uses the microfluidic reaction device 10 according to any of the first aspect of the present invention. The microfluidic reaction driving method includes:

step S01': and heating the preset temperature zone to the reaction temperature through the heating electrode.

Step S02': and driving the liquid drops and the reagent to respectively move to the central area of the preset temperature area in sequence so as to mix the liquid drops and the reagent in the central area.

In the embodiment of the present invention, step S01 'is performed before step S02', and the temperature zone starts to be heated when the droplet 20 does not run to the central region of the preset temperature zone, so that the efficiency of the microfluidic reaction experiment can be improved. The droplets 20 and the reagents are mixed in the central region so that the droplets 20 and the reagents located in the central region can react with each other. The temperature of the central region is closer to the preset temperature, and thus the reaction efficiency can be improved by the step S01'.

Alternatively, in other embodiments, step S02 'may also be performed after step S01',

in the microfluidic reaction driving method provided by the embodiment of the invention, the droplet 20 and the reagent move to the central area of the preset temperature area in sequence, namely, the droplet 20 and the reagent are mixed in the central area to form a mixed droplet. Then, since the droplet 20 and the reagent do not react with each other before moving to the central region even if the temperature reaches the reaction temperature. Therefore, the preset temperature zone can be heated before or after step S01' and heated to the reaction temperature. In the embodiment of the present invention, the droplets 20 and the reagents are mixed in the central region, and the influence of the temperature deviation of the boundary region on the reaction experiment can be avoided.

Referring to fig. 18, fig. 18 is a flow chart of a microfluidic reaction driving method according to another embodiment of the second aspect of the present invention, which uses the microfluidic reaction device 10 according to any of the first aspect of the present invention. The central area of each temperature zone is embedded with a reagent. The microfluidic reaction driving method includes:

step S01': and embedding a reagent in the central area of the preset temperature zone.

Step S02': the preset temperature zone is heated to the reaction temperature by the heating electrode 211.

Step S03': and driving the liquid drop to move to the central area of the preset temperature zone.

In the embodiment of the invention, step S01 "is performed before step S02", that is, the reagent is embedded in the central region of the preset temperature zone in advance, and during the reaction process, the reagent does not need to be driven to move, so that the experimental efficiency can be improved. When the liquid drop 20 does not run to the central area of the preset temperature area, the temperature area is heated, and the efficiency of the microfluid reaction experiment can be improved. The reagent is embedded in the central region, the droplet 20 and the reagent are mixed in the central region, and the droplet 20 and the reagent react with each other in the central region. The temperature of the central region is closer to the preset temperature, and thus the reaction efficiency can be improved by the step S01'.

Optionally, in some other embodiments, step S03 "may also be performed after step S01".

In the microfluidic reaction driving method provided in the embodiment of the present invention, since the reagent is embedded in the central region of the temperature zone, the droplet 20 and the reagent cannot react with each other even if the temperature of the preset temperature zone reaches the reaction temperature before the droplet 20 moves to the central region of the preset temperature zone. Therefore, the preset temperature zone can be heated before or after step S01 ″ and heated to the reaction temperature. In the embodiment of the invention, the reagent is embedded in the central area, the liquid drop 20 and the reagent react with each other in the central area, and the influence of the temperature deviation of the boundary area on the reaction experiment can be avoided.

While the application has been described with reference to a preferred embodiment, various modifications may be made and equivalents may be substituted for elements thereof without departing from the scope of the application. In particular, the technical features mentioned in the embodiments can be combined in any way as long as there is no structural conflict. The present application is not intended to be limited to the particular embodiments disclosed herein but is to cover all embodiments that may fall within the scope of the appended claims.

Claims

1. A microfluidic reaction device having a plurality of temperature zones, the microfluidic reaction device comprising:

the first substrate and the second substrate are arranged at intervals along the thickness direction of the microfluid;

the first electrode layer is arranged on one side, facing the second substrate, of the first substrate and comprises a plurality of first electrodes;

the first hydrophobic layer is arranged on one side, away from the first substrate, of the first electrode layer;

the second hydrophobic layer is arranged on one side, facing the first hydrophobic layer, of the second substrate, the second hydrophobic layer and the first hydrophobic layer are distributed at intervals, and an accommodating space for accommodating liquid drops is formed between the first hydrophobic layer and the second hydrophobic layer;

the second electrode layer is arranged on one side, facing the second hydrophobic layer, of the second substrate and comprises a plurality of second electrodes which are arranged independently, and each second electrode is respectively positioned in each temperature zone;

the zone of heating, set up in the second electrode layer with between the second base plate, the zone of heating includes heating electrode, heating electrode is a plurality of, each heating electrode is located each respectively the warm-area.

2. The microfluidic reaction device of claim 1, further comprising: and the thin film transistor is connected to the first electrode through a connecting wire and is used for controlling the working state of the first electrode.

3. The microfluidic reaction device according to claim 2,

the first substrate is also provided with a scanning line and a data line;

the grid electrode of the thin film transistor is connected with the scanning line, the drain electrode of the thin film transistor is connected with the data line, and the source electrode of the thin film transistor is connected with the first electrode.

4. The microfluidic reaction device of claim 2, further comprising: and the temperature measuring unit is positioned on one side of the first electrode layer facing the first substrate.

5. The microfluidic reaction device according to claim 4, wherein the thermometry unit comprises:

a first temperature measuring electrode;

the bismuth telluride layer is positioned on one side, away from the first electrode layer, of the first temperature measuring electrode;

and the second temperature measuring electrode is positioned on one side of the bismuth telluride layer, which is far away from the first temperature measuring electrode, and the first temperature measuring electrode and the second temperature measuring electrode are mutually connected through the bismuth telluride layer.

6. The microfluidic reaction device according to claim 5,

one temperature measuring unit is arranged in each temperature zone;

or two or more mutually independent temperature measuring units are arranged in each temperature zone.

7. The microfluidic reaction device according to claim 6, wherein the second thermometric electrode is disposed in the same layer as the source and drain of the thin film transistor.

8. The microfluidic reaction device according to claim 7, wherein the first thermometric electrodes and the second thermometric electrodes are multiple, each first thermometric electrode and each second thermometric electrode are respectively disposed correspondingly, and the multiple first thermometric electrodes are connected to each other and/or the multiple second thermometric electrodes are connected to each other;

the orthographic projection of the single first temperature measuring electrode on the first substrate is overlapped with the orthographic projection of the single first temperature measuring electrode on the first substrate, and the first temperature measuring electrodes and the connecting lead are arranged in a staggered mode.

9. The microfluidic reaction device according to claim 8,

the plurality of first temperature measuring electrodes are connected with one another through first signal lines;

and/or in the same temperature zone, the second temperature measuring electrodes in the same column are connected with each other through second signal lines, the number of the second signal lines is more than two, and the more than two second signal lines are connected with each other.

10. The microfluidic reaction device according to claim 7,

the temperature measuring unit comprises one first temperature measuring electrode, the orthographic projection of the first temperature measuring electrode on the first substrate and the orthographic projection of more than two first electrodes on the first substrate are mutually overlapped, a first abdicating hole is formed in the first temperature measuring electrode, and the connecting lead is connected between the first electrode and the thin film transistor through the first abdicating hole;

the number of the second temperature measuring electrodes is one, a second yielding hole is formed in each second temperature measuring electrode, and the data line, the source electrode and the drain electrode are arranged in the second yielding hole; or, the number of the second temperature measuring electrodes may be more than two, each of the second temperature measuring electrodes is located between two adjacent data lines, and the second temperature measuring electrodes are arranged in a staggered manner with the source and the drain.

11. The microfluidic reaction device according to claim 5, further comprising: the organic layer is arranged between the first temperature measuring electrode and the second temperature measuring electrode, a first through hole penetrates through the organic layer, and the bismuth telluride layer is connected between the first temperature measuring electrode and the second temperature measuring electrode through the first through hole.

12. The microfluidic reaction device according to claim 11, wherein the organic layer has a thickness of 2 μm to 8 μm.

13. The microfluidic reaction device according to claim 1, wherein the plurality of temperature zones include an initial temperature zone, an end temperature zone, and one or more intermediate temperature zones located between the initial temperature zone and the end temperature zone, the initial temperature zone, the intermediate temperature zone, and the end temperature zone are distributed along a predetermined circular path, the number of the initial temperature zone and the end temperature zone is one or more, and at least one of the initial temperature zone and at least one of the end temperature zone are partially or completely overlapped, so that the droplets can continue to move cyclically from the end temperature zone to the initial temperature zone after moving from the initial temperature zone to the end temperature zone.

14. The microfluidic reaction device according to claim 13, wherein the number of the initial temperature zone, the intermediate temperature zone and the finishing temperature zone is one,

the initial temperature zone and the finishing temperature zone are positioned on at least one side of the intermediate temperature zone in the first direction;

the intermediate temperature zone and the initial temperature zone at least partially overlap in the first direction to enable the droplet to move between the initial temperature zone and the intermediate temperature zone as the droplet moves in the first direction;

the intermediate temperature zone and the ending temperature zone at least partially overlap in the first direction to enable movement between the intermediate temperature zone and the ending temperature zone when the droplet moves in the first direction.

15. The microfluidic reaction device according to claim 14, wherein the initial temperature zone and the ending temperature zone located on the same side of the intermediate temperature zone in the first direction are distributed in a second direction so that the droplet can move between the ending temperature zone and the initial temperature zone while moving in the second direction.

16. The microfluidic reaction device according to claim 13,

the initial temperature zone and the finishing temperature zone are both two;

the two initial temperature zones are respectively arranged on two sides of the intermediate temperature zone in a first direction, and the two initial temperature zones are distributed in a staggered manner in the first direction;

the two ending temperature areas are respectively arranged on two sides of the intermediate temperature area in the first direction, and the two ending temperature areas are distributed in a staggered manner in the first direction;

the initial temperature zone and the finishing temperature zone which are positioned at two sides of the intermediate temperature zone in the first direction are distributed along a second direction.

17. A microfluidic reaction driving method using the microfluidic reaction device according to any one of claims 1 to 16, the method comprising:

mixing the droplets and the reagent to form mixed droplets;

driving the mixed liquid dropping liquid to move to a central area of a preset temperature area;

and heating a temperature zone where the mixed liquid drop is located to a reaction temperature through the heating electrode, so that the liquid drop in the mixed liquid drop and the reagent can react with each other.

18. A microfluidic reaction driving method using the microfluidic reaction device according to any one of claims 1 to 16, the method comprising:

before or after the step of driving the liquid drop and the reagent to respectively move to the central area of the preset temperature area in sequence, the method further comprises the following steps: heating the preset temperature zone to a reaction temperature by the heating electrode so that the liquid drop and the reagent located in the central zone can react with each other.

19. A microfluidic reaction driving method using the microfluidic reaction device according to any one of claims 1 to 16, wherein a reagent is embedded in a central region of each of the temperature zones, the method comprising:

before or after the step of driving the liquid drop to move to the central area of the preset temperature area, the method further comprises the following steps: heating the preset temperature zone to a reaction temperature by the heating electrode so that the liquid drop and the reagent located in the central zone can react with each other.