CN110407163B - Integrated microfluidic chip for synthesizing composite droplet pair filled hydrogel microfiber and application - Google Patents
Integrated microfluidic chip for synthesizing composite droplet pair filled hydrogel microfiber and application Download PDFInfo
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Abstract
Integrated micro-device for synthesizing composite liquid drop pair filled hydrogel microfiberA fluidic chip and application relate to the technical field of microfluidics. The invention aims to solve the problem that the micro fiber filled with the liquid drop can not realize the controllable release of the internal encapsulation due to the solid structure of the existing single emulsion oil drop. A method; w is to bem1、Wm2、WcAnd WoContinuously injecting four fluids from corresponding inlets respectively, wherein the four fluids form solid microfibers at an outlet at the right end of the circular capillary tube c; after the solid microfiber is formed for 5-10 min, W is addedi1、Wi2、Om1And Om2Continuously injecting from corresponding inlets respectively, and obtaining the hydrogel microfiber filled with the composite liquid drops at an outlet at the right end of the circular capillary tube c. The invention can obtain an integrated micro-fluidic chip for synthesizing the hydrogel microfiber filled with the composite liquid drop pairs and application thereof.
Description
Technical Field
The invention relates to the technical field of microfluidics, in particular to an integrated microfluidic chip for synthesizing hydrogel microfibers filled with composite droplet pairs and application thereof.
Background
The microcarrier for co-encapsulation and controllable release of multiple functional substances has unique advantages and wide application prospects in the aspects of drug delivery, tissue regeneration, enzyme catalytic reaction and the like. The cavities of the multi-cavity microcarrier are separated by the shell material and are mutually independent, so that the co-encapsulation of multiple substances is facilitated, and the cross contamination among encapsulated substances is effectively avoided, which has important significance for enhancing the synergistic effect among multiple medicines in the aspects of biomedicine such as combined cancer chemotherapy, tissue regeneration, disease diagnosis and the like. With microfluidic technology, although many microcapsules with multi-cavity structures are generated for simultaneous and sequential release of multiple substances, the nested structure inside and outside the microcapsules makes it still a technical problem to achieve selective release of multiple encapsulants as required. Therefore, it is necessary to prepare a novel microcarrier that can realize parallel independent encapsulation and selective controlled release of multiple substances.
Droplet-filled microfibers, in which the droplets are regularly distributed within the fiber as discrete micro-cavities that can be effectively used for the independent encapsulation of functional substances, have attracted considerable attention in recent years by scholars both at home and abroad as an alternative microcarrier. To date, the use of microfluidic technology to produce oil droplet-encapsulated microfibers has been extensively studied and successfully applied to the encapsulation of various cells and functional particles. The solid structure of the oil droplets of the single emulsion makes such fibers incapable of achieving controlled release of the internal encapsulates.
Disclosure of Invention
The invention aims to solve the problem that microfiber filled in a liquid drop cannot realize controllable release of an internal packaging material due to the solid structure of the existing single-emulsion oil drop, and provides an integrated microfluidic chip for synthesizing composite liquid drop to the filled hydrogel microfiber and application thereof.
An integrated microfluidic chip comprises two droplet generation modules and a microfiber spinning module, wherein the two droplet generation modules are horizontally and symmetrically arranged, and the microfiber spinning module is arranged on the right side of the two droplet generation modules;
the liquid drop generation module is composed of a circular capillary tube a, a circular capillary tube b and a square capillary tube a, one end of the circular capillary tube a and one end of the circular capillary tube b are both in a conical structure, the inner diameter of a conical structure port of the circular capillary tube a is smaller than that of a conical structure port of the circular capillary tube b, the conical structure end of the circular capillary tube a extends into the square capillary tube a from the left end of the square capillary tube a, the conical structure end of the circular capillary tube b extends into the square capillary tube a from the right end of the square capillary tube a, the outer walls of the circular capillary tube a and the circular capillary tube b are both bonded with the inner wall of the bottom surface of the square capillary tube a in a non-sealing mode, the circular capillary tube a and the circular capillary tube b are arranged coaxially, and the axial distance L between the conical structure port of the circular capillary tube a and the conical structure port of the circular capillary tube b is larger than that the conical structure port of the square capillary tube a is smaller than that of the circular capillary tube b, the conical structure end of the circular capillary tube a is arranged between the circular capillary tube a and the conical structure end of the circular capillary tube b, the circular capillary tube a and the conical structure end of the circular capillary tube b is arranged coaxially1130-160 μm;
the microfiber spinning module is composed of a round capillary tube c and a square capillary tube b, the right end of the round capillary tube c extends into the square capillary tube b from the left end of the square capillary tube b, the outer wall of the round capillary tube c is in non-sealing bonding with the inner wall of the bottom surface of the square capillary tube b, the right ends of the round capillary tubes b of the two liquid drop generating modules extend into the round capillary tube c, the outer wall of the round capillary tube b is in non-sealing bonding with the inner wall of the round capillary tube c, and the distance L between the port at the right end of the round capillary tube b and the port at the right end of the round capillary tube c2Is 70-120 mu m.
A preparation method of an integrated microfluidic chip comprises the following steps:
firstly, horizontally and symmetrically placing two square capillary tubes a on the left side of a glass sheet, and bonding the outer walls of the two square capillary tubes a with the surface of the glass sheet; after the bonding is finished, the conical structure ends of the two circular capillary tubes a extend into the square capillary tubes a from the left ends of the two square capillary tubes a respectively, the outer walls of the two circular capillary tubes a are bonded with the inner walls of the square capillary tubes a in a non-sealing mode, then the conical structure ends of the two circular capillary tubes b extend into the square capillary tubes a from the right ends of the square capillary tubes a respectively, the two circular capillary tubes a and the two circular capillary tubes b are arranged to be coaxial respectively, and the axial distance L between the conical structure ports of the circular capillary tubes a and the conical structure ports of the circular capillary tubes b is ensured1The outer walls of the two round capillaries b are bonded with the inner wall of the square capillary a in a non-sealing way, wherein the outer wall is 130-160 mu m;
secondly, the right ends of the circular capillaries b of the two liquid drop generating modules extend into the circular capillary c, so that the distance L between the port at the right end of the circular capillary b and the port at the right end of the circular capillary c is ensured2The diameter is 70-120 mu m, and the outer walls of the two circular capillaries b are bonded with the inner wall of the circular capillary c in a non-sealing way; after the bonding is finished, sleeving the square capillary tube b on the circular capillary tube c, and bonding the outer wall of the circular capillary tube c with the inner wall of the bottom surface of the square capillary tube b in a non-sealing manner; and after the bonding is finished, bonding the outer wall of the square capillary tube b with the surface of the glass sheet to finish the preparation of the integrated microfluidic chip.
The method for synthesizing the hydrogel microfiber filled with the composite droplet pair by using the integrated microfluidic chip is completed by the following steps:
(1) respectively loading eight fluids into eight sample injectors, respectively fixing the eight fluids on eight injection pumps, and respectively and hermetically connecting the sample injectors loaded with the eight fluids with an inlet a, an inlet b, an inlet c, an inlet d, an inlet e, an inlet f, an inlet h and an inlet i which are arranged on the integrated microfluidic chip;
(2) w is injected by a syringe pumpm1、Wm2、WcAnd WoThe four fluids are continuously injected from the corresponding inlets a, b, c and d respectively, Wm1、Wm2、WcAnd WoThe outlets of the four fluids at the right end of the circular capillary c form a solid microfiber, Wm1And Wm2The flow rate of (A) is 2.3 mL/h-3 mL/h, WcThe flow rate of (A) is 6mL/h to 8mL/h, WoThe flow rate of (A) is 30 mL/h-40 mL/h;
(3) w to be used in step (2)m1、Wm2、WcAnd WoForming solid microfiber at the outlet of the right end of the circular capillary for 5-10 min, and pumping W with a syringe pumpi1、Wi2、Om1And Om2Continuously injecting the mixture from the corresponding inlet e, inlet f, inlet h and inlet i respectively to obtain hydrogel microfibers filled with composite liquid drops at the outlet at the right end of the circular capillary tube c; wi1And Wi2The flow rate of the mixed solution is 700 mu L/h-900 mu L/h, Om1And Om2The flow rate of the flow is 400 mu L/h-600 mu L/h.
The principle of the invention is as follows:
the invention relates to an integrated microfluidic chip for synthesizing composite liquid drop pair filled hydrogel microfibers and application thereof, which is used for sequentially integrating two technologies of composite liquid drop generation and fiber spinning based on a microfluidic technology. Two processes are mainly involved: the generation of two composite droplets based on the flow focusing method and the synthesis of microfibers based on the co-axial flow method. First of all two identical glass capillary drop generation modules in the chip and the fluid flow rate regulation controlled by the syringe pump ensure the simultaneous generation of two drops in both modules, during which Wi1And Wi2，Wm1And Wm2And Om1And Om2The three sets of fluid flow rates remain the same, respectively; second, the distance L between the circular capillary b and the circular capillary c2And a fluid W introduced from an inlet ccThe two synchronously generated liquid drops are ensured to be orderly matched and arranged after meeting in the circular capillary tube c; finally at the outlet of the circular capillary tube c due to WoAnd WcChemical cross-linking reaction between two fluidsIt is such that the formation of the microfibers simultaneously wraps the side-by-side pairs of composite droplets inside the fibers, resulting in hydrogel microfibers filled with the pairs of composite droplets.
The invention has the advantages that:
1. the invention relates to an integrated microfluidic chip for synthesizing composite liquid drop pairs filled with hydrogel microfibers and application thereof, wherein two composite liquid drop pairs arranged side by side in the microfibers can be used as independent chambers for co-packaging two functional materials, and the mutual independence of the two liquid drops effectively avoids cross contamination between the two packaged functional materials and between the packaging material and the external environment;
2. the external hydrogel fiber matrix has good mechanical properties and visualization characteristics, so that the side-by-side liquid drop pairs wrapped inside have the characteristic of easy operation;
3. by means of a fluid Om1And Om2For example, using temperature or pH sensitive materials (waxy materials and paraffinic materials, etc.) as Om1And Om2The two types of liquid drops wrapped in the fibers can be endowed with corresponding temperature or pH value responsiveness, and the shell membrane of the liquid drops can be cracked by changing the temperature or the pH value of the external environment, so that the controllable release of the internal packaging material is realized.
The invention can obtain an integrated micro-fluidic chip for synthesizing the hydrogel microfiber filled with the composite liquid drop pairs and application thereof.
Drawings
FIG. 1 is a diagram of an integrated microfluidic chip disposed on a glass plate according to an embodiment;
FIG. 2 is a schematic view of I in FIG. 1 enlarged twice;
FIG. 3 is a schematic view of II of FIG. 1 at a magnification of two times;
FIG. 4 is a schematic diagram of a droplet generation module according to a first embodiment;
FIG. 5 is a schematic view of a microfiber spinning module according to one embodiment;
FIGS. 6 and 7 are the synthesis process of the composite droplet pair filled hydrogel microfiber in the third embodiment;
figure 8 is a composite drop pair filled hydrogel microfiber of example three.
Wherein, 1, glass sheet; 2. a circular capillary tube a; 3. an inlet h; 4. a square capillary tube a; 5. a circular capillary tube b; 6. an inlet a; 7. a circular capillary tube c; 8. a square capillary tube b; 9. an inlet d; 10. an inlet c; 11. an inlet b; 12. an inlet i; 13. an inlet e; 14. and an inlet f.
Detailed Description
The first embodiment is as follows: the integrated microfluidic chip comprises two droplet generation modules and a microfiber spinning module, wherein the two droplet generation modules are horizontally and symmetrically arranged, and the microfiber spinning module is arranged on the right side of the two droplet generation modules;
the liquid drop generation module comprises a circular capillary tube a2, a circular capillary tube b5 and a square capillary tube a4, wherein one end of each of the circular capillary tube a2 and the circular capillary tube b5 is of a tapered structure, the inner diameter of a tapered structure port of the circular capillary tube a2 is smaller than that of a tapered structure port of the circular capillary tube b5, the tapered structure end of the circular capillary tube a2 extends into the square capillary tube a4 from the left end of the square capillary tube a4, the tapered structure end of the circular capillary tube b5 extends into the square capillary tube a4 from the right end of the square capillary tube a4, the outer walls of the circular capillary tube a2 and the circular capillary tube b5 are in non-sealing adhesion with the inner wall of the bottom face of the square capillary tube a4, the circular capillary tube a2 and the circular capillary tube b5 are coaxially arranged, and the axial distance L between the tapered structure port of the circular capillary tube a2 and the tapered structure port of the circular capillary tube b5 is L1130-160 μm;
the microfiber spinning module consists of a round capillary tube c7 and a square capillary tube b8, the right end of a round capillary tube c7 extends into the square capillary tube b8 from the left end of a square capillary tube b8, the outer wall of a round capillary tube c7 is in non-sealing bonding with the inner wall of the bottom surface of the square capillary tube b8, the right ends of round capillary tubes b5 of the two liquid drop generation modules extend into the round capillary tube c7, the outer wall of a round capillary tube b5 is in non-sealing bonding with the inner wall of a round capillary tube c7, and the distance L between the port at the right end of a round capillary tube b5 and the port at the right end of a round capillary tube c72Is 70-120 mu m.
The second embodiment is as follows: the present embodiment differs from the present embodiment in that: the integrated microfluidic chip is horizontally arranged on the glass sheet 1, and the square capillary a4 and the square capillary b8 of the integrated microfluidic chip are bonded with the surface of the glass sheet 1; the inner diameter of the conical structure port of the circular capillary tube a2 is 40-60 μm, and the inner diameter of the conical structure port of the circular capillary tube b5 is 160-180 μm.
Other steps are the same as those in the first embodiment.
The third concrete implementation mode: the first or second differences from the present embodiment are as follows: a preparation method of an integrated microfluidic chip comprises the following steps:
firstly, horizontally and symmetrically placing two square capillary tubes a4 on the left side of a glass sheet 1, and bonding the outer walls of the two square capillary tubes a4 with the surface of the glass sheet 1; after the bonding is finished, the tapered structure ends of the two round capillary tubes a2 are respectively extended into the square capillary tube a4 from the left end of the two square capillary tubes a4, the outer walls of the two round capillary tubes a2 are bonded with the inner wall of the square capillary tube a4 in a non-sealing mode, then the tapered structure ends of the two round capillary tubes b5 are respectively extended into the square capillary tube a4 from the right end of the square capillary tube a4, the two round capillary tubes a2 and the two round capillary tubes b5 are respectively arranged to be coaxial, and the axial distance L between the tapered structure port of the round capillary tube a2 and the tapered structure port of the round capillary tube b5 is ensured1The diameter is 130-160 mu m, and then the outer walls of the two round capillaries b5 are bonded with the inner wall of the square capillary a4 in a non-sealing way;
secondly, the right ends of the circular capillary tubes b5 of the two liquid drop generating modules extend into the circular capillary tube c7, so that the distance L between the port at the right end of the circular capillary tube b5 and the port at the right end of the circular capillary tube c7 is ensured270-120 μm, and bonding the outer walls of the two round capillaries b5 and the inner wall of the round capillary c7 in a non-sealing manner; after the bonding is finished, sleeving the square capillary tube b8 on the circular capillary tube c7, and bonding the outer wall of the circular capillary tube c7 and the inner wall of the bottom surface of the square capillary tube b8 in a non-sealing manner; after the bonding is completed, the outer wall of the square capillary b8 is bonded with the surface of the glass sheet 1, and the preparation of the integrated microfluidic chip is completed.
The other steps are the same as those in the first or second embodiment.
The fourth concrete implementation mode: the difference between this embodiment and one of the first to third embodiments is as follows: the round capillary a2 in step one is subjected to hydrophobic treatment before use, and the hydrophobic treatment steps are as follows: under the condition of continuously introducing nitrogen into the circular capillary tube a2, placing the circular capillary tube a2 in a soaking solution for soaking for 1 min-2 min, and then drying the soaked circular capillary tube a2 at the temperature of 160-180 ℃ for 30 min-40 min; the soaking solution is prepared from trichlorooctadecylsilane and a toluene solution, wherein the volume ratio of the trichlorooctadecylsilane to the toluene solution is 1: 49-99.
The other steps are the same as those in one of the first to third embodiments.
The fifth concrete implementation mode: the difference between this embodiment and one of the first to fourth embodiments is: the outer walls of the two round capillaries a2 and the inner wall of the square capillary a4 are bonded in a non-sealing mode, the outer walls of the two round capillaries b5 and the inner wall of the square capillary a4 are bonded in a non-sealing mode, the outer walls of the two round capillaries b5 and the inner wall of the round capillary c7 are bonded in a non-sealing mode, the outer walls of the two round capillaries b 7 and the inner wall of the bottom face of the square capillary b8 are bonded in a non-sealing mode, AB glue is used for bonding, and the outer walls of the two square capillaries a4 and the surface of the glass sheet 1 and the outer walls of the square capillaries b8 and the surface of the glass sheet 1 are bonded in a shadowless glue mode.
The other steps are the same as those in one of the first to fourth embodiments.
The sixth specific implementation mode: the difference between this embodiment and one of the first to fifth embodiments is as follows: the method for synthesizing the hydrogel microfiber filled with the composite droplet pair by using the integrated microfluidic chip is completed by the following steps:
(1) respectively loading eight kinds of fluids into eight sample injectors, respectively fixing the eight kinds of fluids on eight injection pumps, and then respectively and hermetically connecting the sample injectors loaded with the eight kinds of fluids with an inlet a6, an inlet b11, an inlet c10, an inlet d9, an inlet e13, an inlet f14, an inlet h3 and an inlet i12 which are arranged on the integrated microfluidic chip;
(2) w is injected by a syringe pumpm1、Wm2、WcAnd WoThe four fluids are continuously injected from the corresponding inlets a6, b11, c10 and d9 respectively, and W ism1、Wm2、WcAnd WoThe outlets of the four fluids at the right end of the circular capillary c7 form a solid microfiber, Wm1And Wm2The flow rate of (A) is 2.3 mL/h-3 mL/h, WcThe flow rate of (A) is 6mL/h to 8mL/h, WoThe flow rate of (A) is 30 mL/h-40 mL/h;
(3) w to be in step (2)m1、Wm2、WcAnd WoForming solid microfibers from outlets of the four fluids at the right end of the circular capillary c7 for 5-10 min, and then pumping W by using a syringe pumpi1、Wi2、Om1And Om2Continuously injecting the mixture from the corresponding inlet e13, inlet f14, inlet h3 and inlet i12 respectively, and obtaining hydrogel microfibers filled with composite liquid drops at an outlet at the right end of the circular capillary c 7; w is a group ofi1And Wi2The flow rate of the mixed solution is 700 mu L/h-900 mu L/h, Om1And Om2The flow rate of the flow is 400 mu L/h-600 mu L/h.
The other steps are the same as those in one of the first to fifth embodiments.
The principle of the present embodiment:
the method for synthesizing the hydrogel microfiber filled with the composite droplet pair by using the integrated microfluidic chip is a method for sequentially integrating two technologies of composite droplet generation and fiber spinning based on the microfluidic technology. Two processes are mainly involved: the generation of two composite droplets based on the flow focusing method and the synthesis of microfibers based on the co-axial flow method. First of all two identical glass capillary drop generation modules in the chip and the fluid flow rate regulation controlled by the syringe pump ensure the simultaneous generation of two drops in both modules, during which Wi1And Wi2，Wm1And Wm2And Om1And Om2The three sets of fluid flow rates remain the same, respectively; second, distance L between circular capillary b5 and circular capillary c72And a fluid W introduced from an inlet c10cEnsure thatTwo synchronously generated liquid drops are orderly matched and arranged after meeting in a circular capillary tube c 7; finally at the outlet of the circular capillary tube c7 due to WoAnd WcAnd (3) carrying out chemical crosslinking reaction between the two fluids, so that the side-by-side compound droplet pairs are wrapped in the fibers while the microfibers are generated, and obtaining the hydrogel microfibers filled with the compound droplet pairs.
The advantages of this embodiment:
1. in the embodiment, the method for synthesizing the hydrogel microfiber filled with the composite droplet pairs by using the integrated microfluidic chip is utilized, two composite droplet pairs arranged side by side in the microfiber can be used as independent chambers for co-packaging two functional materials, and the mutual independence of the two droplets effectively avoids cross contamination between the two packaged functional materials and between the packaging material and the external environment;
2. the external hydrogel fiber matrix has good mechanical properties and visualization characteristics, so that the side-by-side liquid drop pairs wrapped inside have the characteristic of easy operation;
3. by means of a fluid Om1And Om2For example, using temperature or pH sensitive materials (waxy materials and paraffinic materials, etc.) as Om1And Om2The two types of liquid drops wrapped in the fibers can be endowed with corresponding temperature or pH value responsiveness, and the shell membrane of the liquid drops can be cracked by changing the temperature or the pH value of the external environment, so that the controllable release of the internal packaging material is realized.
The seventh embodiment: the difference between this embodiment and one of the first to sixth embodiments is: w isi1And Wi2The preparation steps are as follows:
1)Wi1the preparation of (1): dissolving polyvinyl alcohol in deionized water, stirring for 10-12 h at 70-85 ℃, and filtering by a 0.80 mu m filter to obtain Wi1The volume ratio of the polyvinyl alcohol to the deionized water is 1: 49-99 parts;
2)Wi2the preparation of (1): w obtained in step 1)i1Adding methylene blue, and filtering by 0.45 μmFiltering with a filter to obtain Wi2Methylene blue and Wi1Is 1: 499-999 percent.
The other steps are the same as those in one of the first to sixth embodiments.
The specific implementation mode is eight: the difference between this embodiment and one of the first to seventh embodiments is: said O ism1And Om2Is the same fluid, and is dimethyl silicone oil with the viscosity of 150 cst-300 cst.
The other steps are the same as those in one of the first to seventh embodiments.
The specific implementation method nine: the difference between this embodiment and the first to eighth embodiments is: w ism1、Wm2And WcIs the same fluid, is a mixed solution A, and has the following preparation steps: sequentially dissolving sodium alginate and polyvinyl alcohol in deionized water, placing the mixture on a magnetic stirrer at room temperature, stirring the mixture for 12 to 14 hours at a stirring speed of 500 to 600r/min, and then filtering the mixture through a 0.80 mu m filter to obtain a mixed solution A, wherein the volume ratio of the sodium alginate to the deionized water is 1: 99-166, wherein the volume ratio of the polyvinyl alcohol to the deionized water is 1: 49 to 99 parts.
The other steps are the same as those in the first to eighth embodiments.
The detailed implementation mode is ten: the difference between this embodiment and one of the first to ninth embodiments is as follows: w is as describedoThe preparation steps are as follows: dissolving calcium chloride in deionized water, manually stirring for 3-7 min at room temperature to obtain calcium chloride solution, filtering the calcium chloride solution through a 0.45-micron filter, and mixing the filtered calcium chloride solution with pure glycerol to obtain WoThe ratio of the mass of calcium chloride to the volume of deionized water is 1 g: 32 mL-49 mL, and the volume ratio of the calcium chloride solution to the pure glycerol is 1: 4.
the other steps are the same as those in one of the first to ninth embodiments.
The following examples were used to demonstrate the beneficial effects of the present invention:
the first embodiment is as follows: as shown in fig. 1 to 5, an integrated microfluidic chip comprises two droplet generation modules and a microfiber spinning module, wherein the integrated microfluidic chip is horizontally arranged on a glass sheet 1, and the microfiber spinning module is arranged on the right side of the two droplet generation modules on the glass sheet 1;
the liquid drop generation module comprises a circular capillary tube a2, a circular capillary tube b5 and a square capillary tube a4, the surface of the square capillary tube a4 and the surface of the glass sheet 1 are bonded by adopting shadowless glue, one end of the circular capillary tube a2 and one end of the circular capillary tube b5 are both in a conical structure, the inner diameter of a conical structure port of the circular capillary tube a2 is 40-60 mu m, the inner diameter of a conical structure port of the circular capillary tube b5 is 160-180 mu m, the conical structure end of the circular capillary tube a2 extends into the square capillary tube a4 from the left end of the square capillary tube a4, the tapered end of circular capillary b5 extends from the right end of square capillary a4 into square capillary a4, and the outer walls of the round capillary tube a2 and the round capillary tube b5 are in non-sealing bonding with the inner wall of the bottom surface of the square capillary tube a4 by AB glue, the round capillary tube a2 and the round capillary tube b5 are coaxially arranged, and the axial distance L between the tapered structure port of the round capillary tube a2 and the tapered structure port of the round capillary tube b5 is shorter than that L.1130-160 μm;
the microfiber spinning module comprises a circular capillary c7 and a square capillary b8, the outer wall of the square capillary b8 is bonded with the surface of the glass sheet 1 through shadowless glue, the right end of the circular capillary c7 extends into the square capillary b8 from the left end of the square capillary b8, the outer wall of the circular capillary c7 is bonded with the inner wall of the bottom surface of the square capillary b8 through AB glue in a non-sealing mode, the right ends of the circular capillaries b5 of the two liquid drop generation modules extend into the circular capillary c7, the outer wall of the circular capillary b5 is bonded with the inner wall of the circular capillary c7 through AB glue in a non-sealing mode, and the distance L between the port of the right end of the circular capillary b5 and the port of the right end of the circular capillary c7 is L2Is 70-120 mu m.
Example two: a preparation method of an integrated microfluidic chip comprises the following steps:
firstly, the round capillary a2 is subjected to hydrophobic treatment before use, and the hydrophobic treatment steps are as follows: the solution is prevented from contacting the inner wall of the capillary, so that the circular capillary a2 is soaked in a soaking solution for 1min to 2min under the condition that nitrogen is continuously introduced into the circular capillary a2, the soaked circular capillary a2 is dried for 30min to 40min at the temperature of 160 ℃ to 180 ℃, the hydrophobic treatment of the circular capillary a2 is completed, the rest circular capillaries are not required to be treated, the soaking solution is prepared from trichlorooctadecylsilane and a toluene solution, and the volume ratio of the trichlorooctadecylsilane to the toluene solution is 1: 49-99 parts;
secondly, horizontally and symmetrically placing the two square capillary tubes a4 on the left side of the glass sheet 1, and bonding the outer walls of the two square capillary tubes a4 with the surface of the glass sheet 1 by adopting shadowless glue; after the bonding is finished, the conical structure ends of the two round capillary tubes a2 are respectively extended into the square capillary tube a4 from the left end of the two square capillary tubes a4, the outer walls of the two round capillary tubes a2 and the inner wall of the square capillary tube a4 are bonded in a non-sealing mode by AB glue, then the conical structure ends of the two round capillary tubes b5 are respectively extended into the square capillary tube a4 from the right end of the square capillary tube a4, the two round capillary tubes a2 and the two round capillary tubes b5 are respectively arranged to be coaxial, and the axial distance L between the conical structure port of the round capillary tube a2 and the conical structure port of the round capillary tube b5 is ensured1The thickness is 130-160 μm, and then the outer walls of the two round capillaries b5 and the inner wall of the square capillary a4 are bonded in a non-sealing way by AB glue;
thirdly, the right ends of the circular capillary tubes b5 of the two liquid drop generating modules extend into the circular capillary tube c7, and the distance L between the port at the right end of the circular capillary tube b5 and the port at the right end of the circular capillary tube c7 is ensured2The diameter of the capillary tube is 70-120 mu m, and the outer walls of the two circular capillary tubes b5 and the inner wall of the circular capillary tube c7 are bonded in a non-sealing way by AB glue; after the bonding is finished, sleeving the square capillary tube b8 on the circular capillary tube c7, and performing non-sealing bonding on the outer wall of the circular capillary tube c7 and the inner wall of the bottom surface of the square capillary tube b8 by using AB glue; and after the bonding is finished, the outer wall of the square capillary tube b8 is bonded with the surface of the glass sheet 1 by adopting shadowless glue, so that the preparation of the integrated microfluidic chip is finished.
Example three: as shown in fig. 6 to 8, a method for synthesizing hydrogel microfibers filled with composite droplet pairs by using an integrated microfluidic chip is completed according to the following steps:
(1) preparation of eight fluids:
Wi1the preparation of (1): dissolving polyvinyl alcohol in deionized water, stirring for 10-12 h at 70-85 ℃, and filtering by a 0.80 mu m filter to obtain Wi1The volume ratio of the polyvinyl alcohol to the deionized water is 1: 49-99 parts; wi2The preparation of (1): w obtained in step 1)i1Adding methylene blue, and filtering with 0.45 μm filter to obtain Wi2Methylene blue and Wi1Is 1: 499-999 parts; o ism1And Om2Is the same fluid, namely dimethyl silicone oil with the viscosity of 150 cst-300 cst; wm1、Wm2And WcIs the same fluid, is a mixed solution A, and has the following preparation steps: sequentially dissolving sodium alginate and polyvinyl alcohol in deionized water, placing the mixture on a magnetic stirrer at room temperature, stirring the mixture for 12 to 14 hours at a stirring speed of 500 to 600r/min, and then filtering the mixture through a 0.80 mu m filter to obtain a mixed solution A, wherein the volume ratio of the sodium alginate to the deionized water is 1: 99-166, wherein the volume ratio of the polyvinyl alcohol to the deionized water is 1: 49-99 parts; woThe preparation steps are as follows: dissolving calcium chloride in deionized water, manually stirring for 3-7 min at room temperature to obtain calcium chloride solution, filtering the calcium chloride solution through a 0.45-micron filter, and mixing the filtered calcium chloride solution with pure glycerol to obtain WoThe ratio of the mass of calcium chloride to the volume of deionized water was 1 g: 32 mL-49 mL, and the volume ratio of the calcium chloride solution to the pure glycerol is 1: 4;
(2) opening a computer, a microscope and a CCD camera which are connected with the microscope, observing whether the equipment runs normally, and then opening CellSens Entry image acquisition software; fixing the microfluidic chip on an objective table, adjusting the position and the focal length of the chip, and observing the scene on the objective table of the microscope in real time;
(3) respectively loading eight kinds of fluids into eight sample injectors, respectively fixing the eight kinds of fluids on eight injection pumps, and then respectively and hermetically connecting the sample injectors loaded with the eight kinds of fluids with an inlet a6, an inlet b11, an inlet c10, an inlet d9, an inlet e13, an inlet f14, an inlet h3 and an inlet i12 which are arranged on the integrated microfluidic chip;
(4) using a syringe pump to pump Wm1、Wm2、WcAnd WoThe four fluids are continuously injected from the corresponding inlets a6, b11, c10 and d9 respectively, and W ism1、Wm2、WcAnd WoThe outlets of the four fluids at the right end of the circular capillary c7 form a solid microfiber, Wm1And Wm2The flow rate of (A) is 2.3 mL/h-3 mL/h, WcThe flow rate of (A) is 6mL/h to 8mL/h, WoThe flow rate of (A) is 30 mL/h-40 mL/h;
(5) w to be used in the step (4)m1、Wm2、WcAnd WoForming solid microfibers from the outlets of the four fluids at the right end of the circular capillary c7 for 5-10 min, and pumping W by using a syringe pumpi1、Wi2、Om1And Om2Continuously injecting from the corresponding inlets e13, f14, h3 and i12 respectively, Wi1And Wi2The flow rate of the mixed solution is 700 mu L/h-900 mu L/h, Om1And Om2The flow rate of the flow is 400 mu L/h-600 mu L/h.
As shown in FIG. 5, due to the interaction of the three immiscible fluids within each droplet generation module, two composite droplets are generated simultaneously within both droplet generation modules and follow Wm1、Wm2Flows downstream along two circular capillaries b5, respectively, and then two droplets flowing into circular capillary c7 meet and meet at WcAre arranged side by side in the form of pairs of droplets. Finally encapsulated in pairs in hydrogel microfibers at the outlet of circular capillary c7, resulting in hydrogel microfibers filled with composite droplet pairs. Fig. 6 shows the composite droplet pair filled calcium alginate microfibers prepared under an optical microscope.
Claims (10)
1. An integrated microfluidic chip, characterized in that it: the device comprises two liquid drop generating modules and a microfiber spinning module, wherein the two liquid drop generating modules are horizontally and symmetrically arranged, and the microfiber spinning module is arranged on the right side of the two liquid drop generating modules;
the liquid drop generation module is composed of a round capillary tube a (2), a round capillary tube b (5) and a square capillary tube a (4), one ends of the round capillary tube a (2) and the round capillary tube b (5) are both in a conical structure, the inner diameter of a conical structure port of the round capillary tube a (2) is smaller than that of a conical structure port of the round capillary tube b (5), the conical structure end of the round capillary tube a (2) extends into the square capillary tube a (4) from the left end of the square capillary tube a (4), the conical structure end of the round capillary tube b (5) extends into the square capillary tube a (4) from the right end of the square capillary tube a (4), the outer walls of the round capillary tube a (2) and the round capillary tube b (5) are both bonded with the inner wall of the bottom surface of the square capillary tube a (4) in a non-sealing mode, and the round capillary tube a (2) and the round capillary tube b (5) are arranged coaxially, axial distance L between conical structure port of circular capillary a (2) and conical structure port of circular capillary b (5)1130 to 160 μm;
the microfiber spinning module comprises a round capillary tube c (7) and a square capillary tube b (8), the right end of the round capillary tube c (7) extends into the square capillary tube b (8) from the left end of the square capillary tube b (8), the outer wall of the round capillary tube c (7) is in non-sealing bonding with the inner wall of the bottom surface of the square capillary tube b (8), the right ends of the round capillary tubes b (5) of the two liquid drop generation modules extend into the round capillary tube c (7), the outer wall of the round capillary tube b (5) is in non-sealing bonding with the inner wall of the round capillary tube c (7), and the distance L between the port at the right end of the round capillary tube b (5) and the port at the right end of the round capillary tube c (7)2Is 70 μm to 120 μm.
2. The integrated microfluidic chip according to claim 1, wherein the integrated microfluidic chip is horizontally arranged on the glass sheet (1), and the square capillary a (4) and the square capillary b (8) of the integrated microfluidic chip are bonded with the surface of the glass sheet (1); the inner diameter of the conical structure port of the circular capillary tube a (2) is 40-60 mu m, and the inner diameter of the conical structure port of the circular capillary tube b (5) is 160-180 mu m.
3. A preparation method of an integrated microfluidic chip is characterized by comprising the following steps:
firstly, horizontally and symmetrically placing two square capillary tubes a (4) on the left side of a glass sheet (1), and bonding the outer walls of the two square capillary tubes a (4) with the surface of the glass sheet (1); after the bonding is finished, the conical structure ends of the two circular capillaries a (2) respectively extend into the square capillaries a (4) from the left ends of the two square capillaries a (4), the outer walls of the two circular capillaries a (2) are bonded with the inner walls of the square capillaries a (4) in a non-sealing manner, then the conical structure ends of the two circular capillaries b (5) respectively extend into the square capillaries a (4) from the right ends of the square capillaries a (4), the two circular capillaries a (2) and the two circular capillaries b (5) are arranged to be coaxial respectively, and the axial distance L between the conical structure port of the circular capillary a (2) and the conical structure port of the circular capillary b (5) is ensured1The thickness of the capillary tube is 130-160 micrometers, and then the outer walls of the two round capillary tubes b (5) are bonded with the inner wall of the square capillary tube a (4) in a non-sealing mode;
secondly, the right ends of the circular capillaries b (5) of the two liquid drop generating modules extend into the circular capillary c (7), so that the distance L between the port at the right end of the circular capillary b (5) and the port at the right end of the circular capillary c (7) is ensured2The diameter of the capillary tube is 70-120 mu m, and the outer walls of the two circular capillary tubes b (5) are bonded with the inner wall of the circular capillary tube c (7) in a non-sealing manner; after the bonding is finished, sleeving the square capillary tube b (8) on the circular capillary tube c (7), and bonding the outer wall of the circular capillary tube c (7) with the inner wall of the bottom surface of the square capillary tube b (8) in a non-sealing manner; and after the bonding is finished, bonding the outer wall of the square capillary tube b (8) with the surface of the glass sheet (1) to finish the preparation of the integrated microfluidic chip.
4. The method for preparing an integrated microfluidic chip according to claim 3, wherein the circular capillary a (2) in the first step is subjected to a hydrophobic treatment before use, and the hydrophobic treatment comprises the following steps: under the condition that nitrogen is continuously introduced into the circular capillary tube a (2), the circular capillary tube a (2) is placed in a soaking solution to be soaked for 1 min-2 min, and then the soaked circular capillary tube a (2) is dried for 30 min-40 min at the temperature of 160-180 ℃; the soaking solution is prepared from trichlorooctadecylsilane and a toluene solution, wherein the volume ratio of the trichlorooctadecylsilane to the toluene solution is 1: 49-99.
5. The method for preparing an integrated microfluidic chip according to claim 3, wherein the non-sealing bonding of the outer walls of the two circular capillaries a (2) and the inner wall of the square capillary a (4), the non-sealing bonding of the outer walls of the two circular capillaries b (5) and the inner wall of the circular capillary c (7), and the non-sealing bonding of the outer walls of the circular capillaries c (7) and the inner wall of the bottom surface of the square capillary b (8) are performed by AB glue, the outer walls of the two square capillaries a (4) are bonded with the surface of the glass sheet (1), and the outer walls of the square capillaries b (8) are bonded with the surface of the glass sheet (1) by adopting shadowless glue.
6. The method for synthesizing composite droplet pair filled hydrogel microfibers by using the integrated microfluidic chip as claimed in claim 1 or 2, wherein the method is completed by the following steps:
(1) respectively loading eight kinds of fluids into eight sample injectors, respectively fixing the eight kinds of fluids on eight injection pumps, and respectively and hermetically connecting the sample injectors loaded with the eight kinds of fluids with an inlet a (6), an inlet b (11), an inlet c (10), an inlet d (9), an inlet e (13), an inlet f (14), an inlet h (3) and an inlet i (12) which are arranged on the integrated microfluidic chip;
(2) using a syringe pump to pump Wm1、Wm2、WcAnd WoThe four fluids are continuously injected from the corresponding inlets a (6), b (11), c (10) and d (9), Wm1、Wm2、WcAnd WoThe outlets of the four fluids at the right end of the round capillary c (7) form a solid microfiber, Wm1And Wm2The flow rate of (a) is 2.3mL/h to 3mL/h, WcThe flow rate of (A) is 6mL/h to 8mL/h, WoThe flow rate of (a) is 30-40 mL/h;
(3) w to be used in step (2)m1、Wm2、WcAnd WoForming solid microfibers at the outlet of the right end of the circular capillary c (7) by the four fluids for 5-10 min, and then pumping W by using a syringe pumpi1、Wi2、Om1And Om2Continuously injecting four fluids from an inlet e (13), an inlet f (14), an inlet h (3) and an inlet i (12) which correspond to each other, and obtaining hydrogel microfibers filled with composite droplet pairs at an outlet at the right end of the circular capillary tube c (7); wi1And Wi2The flow rate of the first and second flow paths is 700 mu L/h to 900 mu L/h, Om1And Om2The flow rate of (a) is 400 to 600 mu L/h.
7. The method of claim 6, wherein W isi1And Wi2The preparation steps are as follows:
1）Wi1the preparation of (1): dissolving polyvinyl alcohol in deionized water, stirring for 10-12 h at 70-85 ℃, and filtering through a 0.80-micron filter to obtain Wi1The volume ratio of the polyvinyl alcohol to the deionized water is 1: 49-99 parts;
2）Wi2the preparation of (1): w obtained in step 1)i1Adding methylene blue, and filtering with 0.45 μm filter to obtain Wi2Methylene blue and Wi1Is 1: 499-999.
8. The method of claim 6, wherein said O is selected from the group consisting ofm1And Om2The silicone oil is the same fluid and is dimethyl silicone oil with the viscosity of 150 cst-300 cst.
9. The method of claim 6, wherein W ism1、Wm2And WcIs the same fluid, is a mixed solution A, and has the following preparation steps: sequentially dissolving sodium alginate and polyvinyl alcohol in deionized water, placing the mixture on a magnetic stirrer at room temperature, stirring the mixture for 12 to 14 hours at a stirring speed of 500 to 600r/min, and filtering the mixture through a 0.80-micron filter to obtain a mixed solution A, wherein the volume ratio of the sodium alginate to the deionized water is 1: 99-166, wherein the volume ratio of the polyvinyl alcohol to the deionized water is 1: 49-99.
10. The method of claim 6, wherein W is selected from the group consisting ofoThe preparation steps are as follows: dissolving calcium chloride in deionized water, manually stirring for 3-7 min at room temperature to obtain a calcium chloride solution, filtering the calcium chloride solution through a 0.45-micrometer filter, and mixing the filtered calcium chloride solution with pure glycerol to obtain WoThe ratio of the mass of calcium chloride to the volume of deionized water is 1 g: 32-49 mL, wherein the volume ratio of the calcium chloride solution to the pure glycerol is 1: 4.
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