阅读说明：本技术 一种微流道装置和高通量可编程多浓度药物的建立方法 (Micro-channel device and method for establishing high-flux programmable multi-concentration medicine ) 是由 黄璐 杨德钰 周建华 于 2021-07-30 设计创作，主要内容包括：本发明涉及微流控技术领域,尤其涉及一种微流道装置和高通量可编程多浓度药物的建立方法。本发明公开了一种微流控装置,该装置可实现含不同种类、不同剂量药物组合的微液滴的高通量制备。该装置可以产生多浓度并且有着较好重复性的稳定液滴,通过改变药物初始浓度和流量,可以获得一系列的药物种类及浓度组合。该装置具有微量、高效、高通量和自动化的优点,能较好地克服多孔板筛选系统的不足。该装置可广泛应用于基因分析、药物筛选、细胞培养等领域。(The invention relates to the technical field of microfluidics, in particular to a micro-channel device and a method for establishing high-flux programmable multi-concentration medicines. The invention discloses a microfluidic device which can realize high-throughput preparation of micro-droplets containing different types and different dosage of drug combinations. The device can generate stable liquid drops with multiple concentrations and good repeatability, and a series of drug types and concentration combinations can be obtained by changing the initial concentration and the flow of the drug. The device has the advantages of trace, high efficiency, high flux and automation, and can well overcome the defects of a porous plate screening system. The device can be widely applied to the fields of gene analysis, drug screening, cell culture and the like.)

1. A micro flow channel device, characterized by comprising: the water phase channel, the oil phase channel and the two-phase channel are communicated with each other;

the water phase channel is arranged at the upstream of the oil phase channel, and the outlet of the oil phase channel and the outlet of the water phase channel are communicated with the inlet of the two-phase channel;

the water phase channel comprises a first water phase channel, and the first water phase channel is used for introducing a medicament;

the aqueous phase channel and the oil phase channel are both liquid injection devices.

2. The micro flow channel device according to claim 1, wherein the number of the oil phase channels is an even number, and the oil phase channels are symmetrically arranged.

3. The micro flow channel device of claim 1, wherein the number of the first aqueous phase channels is at least two, and the drugs are introduced into the first aqueous phase channels differently.

4. The micro flow channel device according to claim 1, wherein the aqueous phase channel further comprises a second aqueous phase channel for passing water, the second aqueous phase channel is provided upstream of the oil phase channel, and an outlet of the second aqueous phase channel is communicated with an inlet of the two-phase channel.

5. A method for establishing a high-throughput programmable multi-concentration drug, which comprises the steps of using the micro flow channel device according to any one of claims 1 to 4, and comprising:

respectively introducing a first water phase and an oil phase into the first water phase channel and the oil phase channel, and respectively controlling the flow of the first water phase and the flow of the oil phase through a liquid injection device, so as to obtain drug droplets with different concentrations;

the first aqueous phase is a drug.

6. The method for establishing a drug delivery system according to claim 5, wherein the number of the first aqueous phase is at least two, and different drugs are introduced into different first aqueous phase channels.

7. The method according to claim 5, wherein the total flow rate of the aqueous phase is 60 to 100. mu.L/min.

8. The method according to claim 5, wherein the total flow rate of the oil phase is 100 to 200 μ L/min.

9. The method of claim 8, wherein the introducing the first aqueous phase and the oil phase further comprises introducing a second aqueous phase into the second aqueous phase channel, wherein the second aqueous phase is water.

10. A micro flow channel chip comprising a droplet collecting chip and the micro flow channel device according to any one of claims 1 to 4;

the outlet of the micro-channel device is communicated with the liquid drop collecting chip.

Technical Field

The invention relates to the technical field of microfluidics, in particular to a micro-channel device and a method for establishing high-flux programmable multi-concentration medicines.

Background

Drug screening as a key step in the development of new drugs refers to screening compounds (lead compounds) with biological activity or further research value from a plurality of natural products or artificially synthesized compounds, and the main mode is to screen a large number of compounds by adopting a proper drug action target. However, with the development of the subjects such as genomics, proteomics, metabonomics, combinatorial chemistry and the like, the drug molecule library is continuously expanded, and the drug action targets are more and more, so that the drug discovery range is gradually expanded, and the workload of drug screening is increased rapidly. In recent years, in order to accelerate the drug screening process, a high-throughput screening (HTS) technology is proposed, which is based on an experimental method at a molecular level or a cell level, and can realize simultaneous testing and analysis of thousands of reactions through an automatic operation system, a sensitive and rapid detection system and a data analysis system, thereby greatly improving the experimental scale and efficiency of drug screening.

At present, a developed and mature high-throughput drug screening system is mainly based on a multi-well plate, has the problems of single cell culture condition, long time consumption, large reagent consumption and the like, and is difficult to realize complex multi-concentration drug combination screening. In recent years, microfluidic droplet technology has been rapidly developed and widely applied to drug combination screening. The microfluidic droplet technology is a micro-nano technology which is used for separating and separating continuous fluid into droplets with discrete nano-scale volumes and the volumes below the discrete nano-scale volumes by utilizing the interaction between flow shearing force and surface tension in a micro-scale channel, usually a micro-machined channel structure, and controlling the droplets. Its advantages include: high sensitivity, high flux, automation and low cost. In addition, the method can establish a conveniently controlled microenvironment of the fluid and the particles. At present, the technology for processing small amounts of liquid (e.g., microliter/nanoliter droplets) based on microfluidic chips has become mature, and the preparation of droplets in microchannels has become increasingly important. In a microfluidic cell screening system, operations such as inoculation of cells, replacement or addition of culture solution, addition and washing of drugs, addition of cell staining reagents and the like are generally involved, and all the operations are realized through manipulation of microfluid.

Therefore, providing a high-efficiency and reliable cell high-throughput drug screening method is a technical problem to be solved urgently at present.

Disclosure of Invention

In view of the above, the present invention provides a micro flow channel device and a method for establishing a high-throughput programmable multi-concentration drug, wherein the micro flow channel device can form high-throughput multi-concentration drug droplets, provides a solution for generating high-throughput multi-concentration drug droplets, and can realize rapid and accurate multi-drug multi-concentration combination.

The specific technical scheme is as follows:

the present invention provides a micro flow channel device including: the water phase channel, the oil phase channel and the two-phase channel are communicated with each other;

the water phase channel is arranged at the upstream of the oil phase channel, and the outlet of the oil phase channel and the outlet of the water phase channel are communicated with the inlet of the two-phase channel;

the water phase channel comprises a first water phase channel, and the first water phase channel is used for introducing a medicament;

the aqueous phase channel and the oil phase channel are both liquid injection devices.

Preferably, the number of the oil phase channels is even, and the oil phase channels are symmetrically arranged.

Preferably, the number of the first water phase channels is at least two, and different medicines are introduced into different first water phase channels.

Preferably, the aqueous phase channel further includes a second aqueous phase channel for passing water, the second aqueous phase channel is disposed upstream of the oil phase channel, and an outlet of the second aqueous phase channel communicates with an inlet of the two-phase channel.

The invention also provides a method for establishing the high-flux programmable multi-concentration medicament, which adopts the micro-channel device and comprises the following steps:

respectively introducing a first water phase and an oil phase into the first water phase channel and the oil phase channel, and respectively controlling the flow of the first water phase and the flow of the oil phase through a liquid injection device, so as to obtain drug droplets with different concentrations;

the first aqueous phase is a drug.

Preferably, the number of the first aqueous phase is at least two, and different medicaments are introduced into different first aqueous phase channels.

Preferably, the total flow rate of the water phase is 60-100 mu L/min;

preferably, the total flow rate of the oil phase is 100-200 mu L/min.

Preferably, when the first aqueous phase and the oil phase are introduced, a second aqueous phase is introduced into the second aqueous phase channel, and the second aqueous phase is water.

The present invention also provides a micro flow channel chip comprising a droplet collecting chip and the micro flow channel device according to any one of claims 1 to 5;

the outlet of the micro-channel device is communicated with the liquid drop collecting chip.

According to the technical scheme, the invention has the following advantages:

the invention provides a micro-channel device which can realize high-throughput preparation of micro-droplets containing different types and different dosage of drug combinations. The device can generate stable liquid drops with multiple concentrations and good repeatability, and a series of drug types and concentration combinations can be obtained by changing the initial concentration and the flow of the drug. The device has the advantages of trace, high efficiency, high flux and automation, and can well overcome the defects of a porous plate screening system. The device can be widely applied to the fields of gene analysis, drug screening, cell culture and the like.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without inventive exercise.

FIG. 1 is a schematic view of a micro flow channel chip according to an embodiment of the present invention;

FIG. 2 is a diagram of a droplet collection chip and high-throughput micro-droplets according to an embodiment of the present invention;

FIG. 3 is a graph of fluorescence of three gradient concentrations of a single drug as a function of droplet position according to example 2 of the present invention;

FIG. 4 is a graph of fluorescence of five gradient concentrations of a single drug as a function of droplet position, as provided in example 3 of the present invention;

FIG. 5 is a graph showing the relationship between fluorescence and droplet position in a dual drug combination with three gradient concentrations provided in example 4 of the present invention;

FIG. 6 is a graph showing the relationship between fluorescence and droplet position in a five-gradient concentration mode for a two-drug combination in example 5 of the present invention;

the illustration is as follows:

1. a water phase channel; 2. an oil phase channel; 3. a droplet collection chip.

Detailed Description

In order to make the objects, features and advantages of the present invention more obvious and understandable, the technical solutions in the embodiments of the present invention will be clearly and completely described below, and it should be apparent that the embodiments described below are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The present invention provides a micro flow channel device, including: the water phase channel, the oil phase channel and the two-phase channel are communicated with each other;

the water phase channel is arranged at the upstream of the oil phase channel, and the outlet of the oil phase channel and the outlet of the water phase channel are communicated with the inlet of the two-phase channel.

In the invention, the outlet of the oil phase channel and the outlet of the water phase channel are communicated with the inlet of the two-phase channel through the silicone tube. The two-phase channel is preferably a silica gel tube connector.

The water phase channel comprises a first water phase channel, the first water phase channel is used for introducing the medicine, and the oil phase channel is used for introducing the oil phase;

the water phase channel and the oil phase channel are both liquid injection devices. The liquid injection device is used for providing an oil phase or an aqueous phase and providing a driving force to inject the oil phase or the aqueous phase into the two-phase channel, and the liquid injection device can control the perfusion rate of the aqueous phase or the oil phase. In the present invention, the liquid injection device is preferably composed of a syringe pump and a syringe.

In the invention, the medicine in the upstream first water phase channel and the oil phase in the downstream oil phase channel are converged in the two-phase channel, and the oil phase divides the continuous fluid medicine into water-in-oil discrete droplets at the flow field interface limited by the geometric structure of the two-phase channel under the action of flow shearing force and surface tension between different fluids. The invention can change the flow rate of the first water phase through the liquid injection device to obtain different concentrations of the medicine.

In the present invention, the number of the oil phase channels is at least 1. The arrangement of the oil phase channel is not particularly limited, and the oil phase channel is arranged at the downstream of the water phase channel. Preferably, when the number of the oil phase channels is an even number, the oil phase channels are symmetrically arranged at the side of the two-phase channels. In the embodiment of the invention, the number of the oil phase channels is 2, and the 2 oil phase channels are symmetrically arranged.

In the present invention, the number of the first aqueous phase channels is at least 1. When the number of the first aqueous phase channels is at least two, the drugs introduced into different first aqueous phase channels are different.

The first aqueous phase channel is not particularly limited in the present invention, and may be provided only upstream of the oil phase channel. Preferably, when the number of the first aqueous phase channels is even, the first aqueous phase channels can be symmetrically arranged on the side surfaces of the two-phase channels; when the number of the first water phase channels is odd, the even number of the first water phase channels are symmetrically arranged, and the rest first water phase channels are arranged on the opposite side of the outlet of the two-phase channel. In the embodiment of the invention, the number of the first water phase channels is 2, and the 2 first water phase channels are symmetrically arranged.

The water phase channel of the present invention further comprises a second water phase channel for passing water, the second water phase channel is disposed upstream of the oil phase channel, and an outlet of the second water phase channel is communicated with an inlet of the two-phase channel. Preferably, the second aqueous phase channel is disposed on the opposite side of the outlet of the two-phase channel, and the outlet of the second aqueous phase channel communicates with the inlet of the two-phase channel.

In the present invention, the water introduced into the second aqueous phase channel is preferably deionized water. The water in the second aqueous phase channel is used to dilute the drug in the first aqueous phase channel.

The invention also provides a method for establishing the high-flux programmable multi-concentration medicament, which adopts the micro-channel device and comprises the following steps:

respectively introducing a first water phase and an oil phase into the first water phase channel and the oil phase channel, and respectively controlling the flow rates of the first water phase and the oil phase through injection pumps so as to obtain drug droplets with different concentrations;

the first aqueous phase is the drug.

The principle of the high-throughput programmable multi-concentration drug establishing method is as follows:

after the flow rate of an injection pump in the liquid injection device is set, the upstream medicine solution and the downstream oil phase are converged in the two-phase channel, and the continuously flowing medicine is divided into water-in-oil discrete droplets by the oil phase at a flow field interface limited by the geometrical structure of the two-phase channel through the flow shearing force and the surface tension between different fluids. The dispersed droplets of the drug at different concentrations are obtained by varying the flow rate of the drug solution.

The establishment method of the high-flux programmable multi-concentration medicine provided by the invention has the advantages of simple operation, less medicine consumption, accuracy, controllability and good reproducibility, is convenient for generating quantitative medicine time and space gradients, and can be used for preparing high-flux medicine liquid drops.

In the invention, when the number of the first water phase channels is more than two, the initial concentrations of different drug solutions are set, and the flow rates of the different drug solutions are changed, so that the drug compositions with different concentrations are obtained.

In the invention, the method for calculating the concentration of a single drug in discrete droplets comprises the following steps: concentration of single drug solution/total flow of aqueous phase of the drug.

In the invention, the total flow of the water phase is 60-100 mu L/min, preferably 80 mu L/min; the total flow rate of the oil phase is 100 to 200. mu.L/min, preferably 150. mu.L/min. The aqueous and oil phases of the present invention can achieve uniform and stable droplets at the preferred flow rates described above.

In the invention, when the drug is used for cell experiments, the first water phase can be set with gradient concentration, so that the treatment and research of experimental samples are facilitated.

In the present invention, the oil phase is preferably liquid paraffin, silicone oil or fluorine oil, and is preferably liquid paraffin.

In the present invention, when the first aqueous phase and the oil phase are introduced, it is preferable to further introduce a second aqueous phase into the second aqueous phase channel, and the second aqueous phase is water. The water may further dilute the drug solution. Therefore, the present invention can also obtain different concentrations of dispersed drug droplets by adjusting the flow rate of the second aqueous phase.

The invention also provides a micro-channel chip, which comprises a liquid drop collecting chip and the micro-channel device;

the outlet of the micro-channel device is communicated with the droplet collecting chip.

In the invention, the micro-channel device is connected with the liquid drop collecting chip through the capillary silicone tube, the micro-channel device generates liquid drops, the liquid drops are led into the inlet of the liquid drop collecting chip through the capillary silicone tube, and then the liquid drops sequentially enter and are arranged in the liquid drop collecting chip.

The invention can realize the high-flux preparation of micro-droplets containing different types and different doses of drug combinations. Stable liquid drops with multiple concentration gradients and better repeatability are generated in the micro-liquid drop collecting chip, and a series of medicine types and concentration combinations can be obtained by changing the initial concentration and the flow of the medicine. The micro-fluidic control method has the advantages of trace quantity, high efficiency, high throughput and automation, and can well overcome the defects of a multi-plate screening system, such as: the problems of single cell culture condition, time and labor consumption, large reagent consumption and the like are solved, and an efficient and reliable technical means is provided for constructing a cell high-throughput drug screening system.

The scheme of the invention is further explained below.

Example 1

This example is a preparation of a droplet collecting chip, and the specific preparation steps are as follows:

step 1) designing a geometric channel template of the microfluidic chip by using AutoCAD, and etching the structure on an aluminum template by using a laser etching technology.

Step 2) Polydimethylsiloxane (PDMS) prepolymer and crosslinker were mixed in a 10: 1, pouring 50mL of the mixture onto a mold after uniformly stirring, vacuumizing for 30min to remove bubbles, and then placing on a hot plate at 80 ℃ for heating and curing.

Step 3) pour 10mL of PDMS prepolymer and crosslinker onto a transparent glass plate at a 10: 1, standing to form a PDMS thin layer, and then placing into a constant temperature box of 60 ℃ for curing.

Step 4) dropping a small drop of PDMS prepolymer and a crosslinking agent onto the cured glass plate in a ratio of 10: 1, uniformly coating the mixture on a glass plate by using a roller, stripping the cured PDMS from the template, tightly adhering the PDMS to the glass plate, standing for 10min, and standing on a hot plate at 120 ℃ for 1h for curing to finish the manufacture of a liquid drop collecting chip.

Example 2

This example is the preparation of a single drug triple concentration gradient droplet

Step 1) dissolving 2mg of rhodamine in 2mL of deionized water to prepare 1mg/mL of rhodamine mother liquor;

step 2) mixing 100 mu L of the mother liquor with 9.9mL of deionized water to dilute the mother liquor into 1/100, and preparing into 10 mu g/mL rhodamine diluent for later use;

step 3) respectively adding the liquid to be pushed into five syringes, wherein two syringes are filled with liquid paraffin to be used as oil phase, one syringe is filled with 10 mug/mL rhodamine diluent, the microfluidic device sequentially contains the rhodamine diluent (simulating a single medicine) from top to bottom, the remaining two syringes are filled with deionized water to be used as water phase, and the capillary silicone tube is connected with the syringes;

and 4) fixing the injector on an injection pump, and adjusting the perfusion speed, wherein the total perfusion speed of the oil phase is 150 mu L/min and the total perfusion speed of the water phase is 80 mu L/min, and under the condition, the liquid drops are uniform and stable on the chip and are not easy to adhere. Under the flow condition, the droplet generation area generates at least 65 stable droplets with uniform size per minute, and the time for filling the droplet collection chip with the droplets is about 4.5 min. Wherein, during three gradients, the flow of the inlet 1 (deionized water) is kept at 20 muL/min, the flow of the inlet 2 (rhodamine solution) is 10 muL/min, 30 muL/min and 50 muL/min in sequence, the flow of the corresponding inlet 3 (deionized water) is 50 muL/min, 30 muL/min and 10 muL/min in sequence, and the method for calculating the concentration of the rhodamine in the liquid drop comprises the following steps: concentration of rhodamine solution in syringe this syringe flow/total aqueous phase flow. Thereby obtaining three concentration gradients of rhodamine liquid drops.

Example 3

This example is the preparation of a single drug five concentration gradient droplet

Step 1) dissolving 2mg of rhodamine in 2mL of deionized water to prepare 1mg/mL of rhodamine mother liquor;

step 2) mixing 100 mu L of mother liquor with 9.9mL of deionized water to dilute the mother liquor into 1/100, and preparing into 10 mu g/mL rhodamine diluent for later use;

step 3) respectively adding the liquid to be pushed into five injectors, wherein two injectors are filled with liquid paraffin to be used as an oil phase, one injector is filled with 10 mu g/mL rhodamine diluent (simulating a single medicament), the other two injectors are filled with deionized water to be used as a water phase, and the capillary silicone tube is connected with the injectors;

and 4) fixing the injector on an injection pump, and adjusting the perfusion speed, wherein the total perfusion speed of the oil phase is 150 mu L/min, and the total perfusion speed of the water phase is 80 mu L/min. During five gradients, the flow of the inlet 1 (deionized water) is kept at 20 mu L/min, the flow of the inlet 2 (rhodamine solution) is 10 mu L/min, 20 mu L/min, 30 mu L/min, 40 mu L/min and 50 mu L/min in sequence, the flow of the corresponding inlet 3 (deionized water) is 50 mu L/min, 40 mu L/min, 30 mu L/min, 20 mu L/min and 10 mu L/min in sequence, and the method for calculating the concentration of rhodamine in the liquid drop comprises the following steps: concentration of rhodamine solution in the injector is equal to the flow of the injector/the total flow of the water phase, so that five concentration gradients of rhodamine liquid drops are obtained, and the liquid drops are collected into a micro-liquid drop collection chip.

Example 4

This example is the preparation of a dual drug combination triple concentration gradient droplet

Step 1) weighing 2mg of Hoechst33342 and dissolving in 1mL of deionized water to prepare 2mg/mL of Hoechst33342 mother solution; weighing 2mg of rhodamine 6G, and dissolving the rhodamine 6G in 1mL of deionized water to prepare 2mg/mL of rhodamine mother solution;

step 2) mixing 100 mu L of Hoechst33342 mother liquor with 9.9mL of deionized water, diluting the mother liquor to 1/100, and preparing 20 mu g/mL Hoechst33342 diluent for later use; mixing 100 mu L of rhodamine mother liquor with 9.9mL of deionized water to dilute the mother liquor into 1/100, and preparing 20 mu g/mL of rhodamine diluent for later use;

step 3) respectively adding the liquid to be pushed into five syringes, wherein two syringes are filled with liquid paraffin to be used as oil phase, one syringe is filled with 20 microgram/mL rhodamine diluent (simulating a first medicament), one syringe is filled with 20 microgram/mL Hoechst33342 diluent (simulating a second medicament), the other syringe is filled with deionized water to be used as water phase, and the capillary silicone tube is connected with the syringes;

and 4) fixing the injector on an injection pump, and adjusting the perfusion speed, wherein the total perfusion speed of the oil phase is 150 mu L/min, and the total perfusion speed of the water phase is 80 mu L/min. Under the flow condition, at least 65 stable and uniform-sized droplets are generated in the droplet generation area per minute, and the time for filling the collecting chip with the droplets is about 4.5 min. Wherein, during the three gradients, the flow of the inlet 1 (deionized water) is kept at 20 muL/min, the flow of the inlet 2 (rhodamine solution) is 10 muL/min, 30 muL/min and 50 muL/min in sequence, and the flow of the corresponding inlet 3(Hoechst33342 solution) is 50 muL/min, 30 muL/min and 10 muL/min in sequence. The method for calculating the concentration of rhodamine in liquid drops comprises the following steps: concentration of rhodamine solution in syringe this syringe flow/total flow of aqueous phase, method of calculation of Hoechst33342 concentration in droplets: concentration of Hoechst33342 solution in syringe this syringe flow/total flow of aqueous phase, resulting in three concentration gradients of dual drug droplets.

Example 5

This example is the preparation of a dual drug combination five concentration gradient droplet

Step 1) weighing 2mg of Hoechst33342 and dissolving in 1mL of deionized water to prepare 2mg/mL of Hoechst33342 mother solution; weighing 2mg of rhodamine 6G, and dissolving the rhodamine 6G in 1mL of deionized water to prepare 2mg/mL of rhodamine mother solution;

step 2) mixing 100 mu L of Hoechst33342 mother liquor with 9.9mL of deionized water, diluting the mother liquor to 1/100, and preparing 20 mu g/mL Hoechst33342 diluent for later use; mixing 100 mu L of rhodamine mother liquor with 9.9mL of deionized water to dilute the mother liquor into 1/100, and preparing 20 mu g/mL of rhodamine diluent for use;

step 3) respectively adding the liquid to be pushed into five injectors, wherein two injectors are filled with liquid paraffin to be used as an oil phase, one injector is filled with a rhodamine solution (simulating a first medicament), one injector is filled with a Hoechst33342 solution (simulating a second medicament), the other injector is filled with deionized water to be used as a water phase, and the capillary silicone tube is connected with the injectors;

and 4) fixing the injector on an injection pump, and adjusting the perfusion speed, wherein the total perfusion speed of the oil phase is 150 mu L/min, and the total perfusion speed of the water phase is 80 mu L/min. Under the flow condition, at least 65 stable and uniform-sized droplets are generated in the droplet generation area per minute, and the time for filling the collecting chip with the droplets is about 4.5 min. Wherein, the flow of the inlet 1 (deionized water) is kept at 20 muL/min during five gradients, the flow of the inlet 2 (rhodamine solution) is 10 muL/min, 20 muL/min, 30 muL/min, 40 muL/min and 50 muL/min in sequence, the flow of the corresponding inlet 3(Hoechst33342 solution) is 50 muL/min, 40 muL/min, 30 muL/min, 20 muL/min and 10 muL/min in sequence, and the rhodamine concentration calculation method in the liquid drop comprises the following steps: concentration of rhodamine solution in syringe this syringe flow/total flow of aqueous phase, method of calculation of Hoechst33342 concentration in droplets: concentration of Hoechst33342 solution in syringe this syringe flow/total flow of aqueous phase, resulting in five concentration gradients of dual drug droplets.

Example 6

High-flux fluorescence signal acquisition and data processing:

step 1) placing the micro-droplets of the embodiments 2-5 on an inverted fluorescence microscope stage after entering a droplet collection chip;

step 2) connecting all hardware, realizing the cooperative operation of a microscope objective table, an objective lens, a light source and a high-speed camera by using control software, and collecting a fluorescence signal diagram;

and 3) carrying out fluorescence analysis by using ImageJ to obtain a graph of the relation between the concentration and the fluorescence intensity.

As shown in FIG. 3, in example 2, the fluorescence intensity increased as the concentration of rhodamine increased.

As shown in FIG. 4, in example 3, the fluorescence intensity increased as the concentration of rhodamine increased.

As shown in FIG. 5, in example 4, the fluorescence intensity increased with the increase in the concentration of rhodamine, and the fluorescence intensity increased with the increase in the concentration of Hoechst 33342.

As shown in FIG. 6, in example 5, the fluorescence intensity increased with the increase in the concentration of rhodamine, and the fluorescence intensity increased with the increase in the concentration of Hoechst 33342.

The above-mentioned embodiments are only used for illustrating the technical solutions of the present invention, and not for limiting the same; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.