CN111632561A

CN111632561A - Method for preparing multi-surface anisotropic coding microspheres based on gas-assisted method and application thereof

Info

Publication number: CN111632561A
Application number: CN202010487232.XA
Authority: CN
Inventors: 黄超伯; 曲清莉; 张建; 陈龙; 唐国胜
Original assignee: Nanjing Forestry University
Current assignee: Nanjing Forestry University
Priority date: 2020-06-01
Filing date: 2020-06-01
Publication date: 2020-09-08

Abstract

The invention discloses a novel method for preparing a polyhedral anisotropic encoded microsphere in a green and nontoxic way. The method adopts gas shearing force to form liquid drops, forms microspheres through ion crosslinking and solidification, and programs and adjusts the color of each chamber in the ten-sided microspheres by injecting sodium alginate solution containing different fluorescent nanoparticles into a coaxial multi-channel nozzle system to form a large amount of codes, so that the method can be effectively applied to high-throughput biological detection, multiple information storage, anti-counterfeiting and the like. More importantly, the preparation process of the method does not contain toxic, harmful or unnecessary components such as organic solvent, photoinitiator, surfactant and the like in the whole process, and can be applied to the fields of food and medicine.

Description

Method for preparing multi-surface anisotropic coding microspheres based on gas-assisted method and application thereof

Technical Field

The invention belongs to the technical field of application of biological materials, and particularly relates to a method for preparing multi-surface anisotropic coded microspheres based on a gas-assisted method and application thereof.

Background

In recent years, the application of coding technology in the fields of medicine and biology, etc. has attracted extensive attention, especially in high-throughput biological detection, multiple information storage and anti-counterfeiting. Several methods for manufacturing coded particles or fibers have been developed and optimized to date, including photobleaching techniques, as well as photonic, electrical and biomolecular coding techniques, among others. From the perspective of encoding information and decoding methods, it can be divided into optical encoding, electronic encoding, patterned encoding, etc., and optical encoding is the most common encoding method among them because of its simpler preparation and decoding.

In recent years, based on the continuous development of anisotropic encoding microcarrier technology, carriers with various shapes such as a rod shape, a sheet shape, a fiber shape, a capsule shape and the like are proved to have certain encoding capacity, but compared with other technical means, the anisotropic microspheres can achieve larger encoding amount and more flexible application mode. However, most anisotropic encoded microspheres are inevitably prepared using photoinitiators, cross-linking agents, surfactants and/or polymers that are not approved for use in food and drug products, etc., and may be adversely affected when it is desired to bring the encoded microspheres into direct contact with sensitive biomolecules or cells, particularly in drug and food labeling. Due to toxicology screening and formula compatibility testing required by a pharmaceutical preparation, the development of a time-saving, labor-saving, economical and efficient preparation technology of the bio-friendly coding microspheres is a core problem for expanding the application range of the coding microspheres.

Disclosure of Invention

The purpose of the invention is as follows: aiming at the defects in the prior art, the invention adopts a gas-assisted method to prepare the multi-surface anisotropic coding microsphere. The whole preparation process is green and environment-friendly, the coding amount of the microspheres is effectively enlarged, the use process is convenient and flexible, the coding information is easy to read, and the method has industrial production feasibility.

The technical scheme is as follows: in order to achieve the purpose of the invention, the invention adopts the technical scheme that:

a method for preparing multi-faceted heterodromous coded microspheres by a high-flux one-step gas-assisted method comprises the following steps: first, a specific coaxial multi-channel nozzle system is prepared, which consists of a supporting needle, a housing needle and several channel needles. And secondly, preparing an aqueous solution of sodium alginate and calcium chloride, uniformly dispersing the red, green and blue fluorescent nanoparticles in the sodium alginate solution, and easily constructing up to seven colors by adjusting the proportion of the fluorescent nanoparticles with different colors. And finally injecting the sodium alginate solutions of the fluorescent nanoparticles with different colors and proportions into different channels of the nozzle system, and obtaining the multi-surface anisotropic microspheres with corresponding color codes through a gas-assisted preparation device. To facilitate reading and defining the starting point of the code, magnetic nanoparticles are additionally integrated into one cavity of the microsphere. The microspheres are the multi-faceted heterodromous encoded microspheres.

In the above step, the concentration of sodium alginate is 2% -4% (w/v).

In the above step, the concentration of calcium chloride is 1-5% (w/v).

In the above steps, the number of the channel needles in the coaxial multi-channel nozzle system can be 4-10.

Has the advantages that: compared with the prior art, the multi-surface heterodromous coding microsphere without using any organic reagent or other unnecessary harmful components is obtained on the premise of not influencing the coding amount and the preparation efficiency. And the microspheres can stably store information for a long time after being verified.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without inventive exercise. Wherein:

FIG. 1 is a schematic diagram of the preparation and encoding of a multi-faceted heterodromous encoded microsphere.

FIG. 2 is a diagram of an apparatus for preparing a multi-faceted heterodromous encoded microsphere. (A) The general device picture, (B) the coaxial multi-channel nozzle system, and (C) the obtained multi-surface anisotropic microspheres.

FIG. 3 shows seven colors of microspheres obtained by different color ratios. (A) The three primary colors are superposed to form seven colors, (B) fluorescent microspheres with different colors, and (C) fluorescent microspheres with seven colors.

FIG. 4 is a fluorescence image of a multi-faceted heterodromous encoded microsphere.

FIG. 5 is a particle size statistic of a multi-faceted heterodromous encoded microsphere. (A) The particle size distribution of the microspheres under the optimal condition, and (B) the particle size of the microspheres prepared by different air flow speeds is changed.

Fig. 6 shows that the microspheres with different particle sizes expand the information storage capacity. (A) Schematic diagram, and (B) object diagram.

FIG. 7 is a graph of the change in fluorescence intensity of microspheres over time. (A) Red fluorescence, (B) green fluorescence, (C) blue fluorescence.

Detailed Description

In order that the above objects, features and advantages of the present invention may be better understood, particular embodiments of the invention will be described below in conjunction with specific embodiments.

The structures, proportions, and dimensions shown in the drawings and described in the specification are for understanding and reading the present disclosure, and are not intended to limit the scope of the present disclosure, which is defined in the claims, and are not essential to the skilled in the art. In addition, the terms "upper", "lower", "front", "rear" and "middle" used in the present specification are for clarity of description, and are not intended to limit the scope of the present invention, and the relative relationship between the terms and the relative positions may be changed or adjusted without substantial technical changes.

Furthermore, reference herein to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one implementation of the invention. The appearances of the phrase "in one embodiment" in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments.

Example 1

3 dried screw bottles of 50mL are taken, added with the prepared 2% sodium alginate solution, and then respectively added with the red fluorescent nanoparticles, the green fluorescent nanoparticles and the ferroferric oxide magnetic nanoparticles, and evenly mixed. Adding a sodium alginate solution containing magnetic nanoparticles into any channel, injecting the sodium alginate solution containing red fluorescent nanoparticles and the sodium alginate solution containing green fluorescent nanoparticles into a coaxial ten-channel nozzle system through an injector according to an alternating sequence, introducing nitrogen at the flow rate of 6L/min to form ten-sided anisotropic liquid drops with red and green alternated, and curing the liquid drops through a 2% calcium chloride receiving bath to form microspheres, as shown in figure 2.

Example 2

Adding the prepared 2% sodium alginate solution into a series of 50mL dry screw bottles, respectively adding one or more red, green and blue fluorescent nanoparticles according to the required color, and uniformly mixing. And (3) passing the mixed solution series through a single-channel nozzle system, and preparing the microspheres with corresponding colors by adopting a gas-assisted method. As shown in fig. 3, the microspheres added with the green and blue fluorescent nanoparticles simultaneously appear cyan, the microspheres added with the green and red fluorescent nanoparticles simultaneously appear yellow, the microspheres added with the red and blue fluorescent nanoparticles simultaneously appear pink, the microspheres added with the red, green and blue fluorescent nanoparticles simultaneously appear white, and the original red, green and blue, the 7 colors which can be easily distinguished by naked eyes can be easily obtained, while the microspheres not added with any fluorescent nanoparticles appear black.

Injecting the corresponding solution into a coaxial multi-channel nozzle system through an injector according to a set sequence (black-green-red-green-blue-yellow-white-yellow-blue-red), and introducing nitrogen gas with the flow rate of 6L/min to enable the multi-surface anisotropic coding microspheres to fall into a calcium chloride receiving bath for solidification, so as to obtain the multi-surface anisotropic coding microspheres, wherein the multi-surface anisotropic coding microspheres are shown in figure 4.

Example 3

The average particle size of the multifaceted heterodromous encoded microspheres under the most suitable preparation conditions (nitrogen flow rate of 6L/min) was 665.48 μm, standard deviation was 11.41, and coefficient of variation was 1.71% as shown in FIG. 5(A) by statistical analysis of a large number of encoded microspheres.

By changing the gas flow speed of nitrogen required for preparing the microspheres, the particle size of the obtained multi-faceted heterodromous coded microspheres is changed. In the course of increasing the flow rate from 1L/min to 6L/min, the particle size of the microspheres decreased from 2000 μm to 500. mu.m, as shown in FIG. 5 (B).

By using the multi-surface anisotropic coded microspheres with different sizes, the information storage capacity can be effectively expanded, and as shown in fig. 6(a), under different air flow rates, ten-surface anisotropic coded microspheres with two different barcodes can be obtained. The prepared microsphere barcodes and corresponding decoding information are shown in fig. 6 (B). Thus, the microspheres with different particle sizes can be used together with the multi-surface microspheres with different color sequences, so that the coding amount of the multi-surface anisotropic coding microspheres is obviously improved.

In order to evaluate the feasibility of the multi-faceted heterodromous encoded microsphere in practical applications, the stability of its fluorescence intensity under conventional conditions was examined. As shown in FIG. 7, the fluorescence intensity of all three primary colors of the multi-faceted heterodromous encoded microsphere remains substantially unchanged within 120 days of evaluation, which meets the requirements of practical application.

It should be noted that the above-mentioned embodiments are only used for illustrating the technical solutions of the present invention and not for limiting, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that several modifications and amendments can be made without departing from the spirit and scope of the technical solutions of the present invention, which should be covered by the claims of the present invention.

Claims

1. A method for preparing multi-surface anisotropic coded microspheres based on a gas-assisted method is characterized by comprising the following steps: the method comprises the following steps:

the method comprises the following steps: preparing a calcium chloride solution and a sodium alginate solution containing fluorescent nanoparticles with three different colors and proportions;

step two: preparing a specific coaxial multi-channel nozzle system;

step three: preparing the multi-surface anisotropic coding microspheres by a gas-assisted method;

step four: and decoding the information of the multi-surface anisotropic coded microspheres.

2. The method of claim 1, wherein the method comprises: step one, preparing a calcium chloride solution with the concentration of 1-5% (w/v).

3. The method of claim 1, wherein the method comprises: step one, preparing a sodium alginate solution containing fluorescent nanoparticles with three different colors and proportions, wherein the concentration of the sodium alginate solution is 2-4% (w/v).

4. The fluorescent nanoparticle of claim 3, wherein: including three different colors of red, green and blue.

5. The method of claim 1, wherein the method comprises: the specific coaxial multi-channel nozzle system in the step two consists of a supporting needle head, a shell needle head and a plurality of channel needle heads.

6. The coaxial multi-channel nozzle system of claim 5, wherein: the number of the passage needles can be 4-10.

7. The method of claim 1, wherein the method comprises: and step three, the gas-assisted method is to form liquid drops by adopting gas shearing force, and form microspheres by ion crosslinking and solidification to obtain the multi-surface anisotropic coding microspheres.

8. The multifaceted anisotropic encoded microsphere prepared by the method for preparing a multifaceted anisotropic encoded microsphere according to any one of claims 1 to 7, wherein: the preparation process of the multi-face anisotropic coding microspheres is green and nontoxic in the whole process, different permutation and combination of seven colors are constructed through three primary colors, a large number of codes are formed, and the multi-face anisotropic coding microspheres are effectively applied to high-throughput biological detection, multiple information storage, anti-counterfeiting and the like.