阅读说明：本技术 一种可调控多彩荧光发射的荧光素功能化聚二乙炔囊泡的制备方法及其分子逻辑门的构建 (Preparation method of fluorescein functionalized polydiacetylene vesicle capable of regulating and controlling multicolor fluorescence emission and construction of molecular logic gate of fluorescein ) 是由 杨光 侯娟 李光耀 纪捷 于 2019-09-27 设计创作，主要内容包括：一种可调控多彩荧光发射的荧光素功能化聚二乙炔囊泡的制备方法及其分子逻辑门的构建,属于有机共轭聚合物波组装技术领域。本发明为了解决现有荧光调控的功能化基材制备条件繁琐,pH控制要求较高,且功能化基材灵敏度小,受外界干扰较大等问题。本发明将二乙炔和荧光素基团相互结合,制得聚二乙炔复合囊泡,该复合囊泡基于聚二乙炔在外界刺激下红相和蓝相互相转变、荧光素基团对外界pH值变化响应的作用,来实现多重刺激聚二乙炔和荧光素基团二者颜色以及荧光发射变化调控的多彩荧光发射方法。本发明的复合囊泡制备过程简单,具有优异的荧光可调控性能,且荧光调控操作过程简便,可操作性强,并且抗环境干扰能力强,体系响应灵敏、快速。(A preparation method of fluorescein functionalized polydiacetylene vesicles capable of regulating and controlling multicolor fluorescence emission and construction of a molecular logic gate thereof belong to the technical field of organic conjugated polymer wave assembly. The invention aims to solve the problems that the existing fluorescence-controlled functionalized substrate is complicated in preparation conditions, high in pH control requirement, small in functionalized substrate sensitivity, large in external interference and the like. According to the invention, diacetylene and a fluorescein group are combined with each other to prepare the polydiacetylene composite vesicle, and the multi-color fluorescence emission method for regulating and controlling the colors of the polydiacetylene and the fluorescein group and the fluorescence emission change is realized on the basis of the mutual transformation of a red phase and a blue phase of the polydiacetylene under the external stimulation and the response of the fluorescein group to the change of the external pH value. The composite vesicle has the advantages of simple preparation process, excellent fluorescence regulation and control performance, simple and convenient fluorescence regulation and control operation process, strong operability, strong environmental interference resistance and sensitive and quick system response.)

1. A preparation method of fluorescein functionalized polydiacetylene vesicles capable of regulating and controlling multicolor fluorescence emission is characterized by comprising the following steps: the method comprises the following operation steps:

step 1, preparing aminated Fluorescein (FDNS) by using carboxyfluorescein and ethylenediamine;

step 2, preparing white powder PCDA-NHS by using carboxyl group diacetylene monomer (PCDA) and N-hydroxysuccinimide (NHS);

step 3, dissolving the FDNS and the PCDA-NHS prepared in the step 1 and the step 2 in a DMF solvent, and stirring at constant temperature in a dark place to obtain a fluorescein substituted diacetylene monomer (F-PCDA);

step 4, dissolving PCDA and F-PCDA in absolute ethyl alcohol, adding deionized water, heating and ultrasonically treating, cooling in a dark condition, and standing at a constant temperature to obtain a composite vesicle;

and 5, carrying out illumination polymerization on the composite vesicle obtained in the step 4 under 254nm ultraviolet light to obtain a blue polydiacetylene composite vesicle system.

2. The method for preparing the fluorescein functionalized polydiacetylene vesicle with controllable multicolor fluorescence emission according to claim 1, which is characterized in that: the specific operation process of the step 1 is as follows: the mass ratio of the materials is 1:1.2 placing the carboxyfluorescein and the ethylenediamine into absolute ethyl alcohol, heating and refluxing for 12h at the temperature of 80 ℃, cooling to room temperature, filtering, washing for 2-3 times by using the absolute ethyl alcohol, and recrystallizing to obtain the FDNS.

3. The method for preparing the fluorescein functionalized polydiacetylene vesicle with controllable multicolor fluorescence emission according to claim 1, which is characterized in that: the specific operation process of the step 2 is as follows: dissolving PCDA in dichloromethane solution, then adding NHS dropwise into the solution, stirring at 30 ℃ in the dark for 4h, filtering, separating by column chromatography, and vacuum drying at 30 ℃ for 12h to obtain white powdery PCDA-NHS.

4. The method for preparing the fluorescein functionalized polydiacetylene vesicle with controllable multicolor fluorescence emission according to claim 3, wherein the method comprises the following steps: the quantity ratio of the PCDA to the NHS is 1: 1.2; the developing solvent in the column chromatographic separation is dichloromethane.

5. The method for preparing the fluorescein functionalized polydiacetylene vesicle with controllable multicolor fluorescence emission according to claim 1, which is characterized in that: the specific operation process of the step 3 is as follows: and (3) dissolving PCDA-NHS and FDNS with the mass ratio of 1:1 in a DMF solvent, and stirring for 24 hours at 30 ℃ in the dark to obtain F-PCDA.

6. The method for preparing the fluorescein functionalized polydiacetylene vesicle with controllable multicolor fluorescence emission according to claim 1, which is characterized in that: the specific operation process of the step 4 is as follows: the total amount of the substances is 5 multiplied by 10^-5And (2) dissolving PCDA and F-PCDA in a molar ratio of 10:1 in 1.5ml of absolute ethyl alcohol, adding 300ml of deionized water, heating to 65 ℃, carrying out ultrasonic treatment for 65min under the condition of 40% of ultrasonic power, cooling in a dark condition, and standing for 12h at 4 ℃ to obtain the composite vesicle.

7. The method for preparing the fluorescein functionalized polydiacetylene vesicle with controllable multicolor fluorescence emission according to claim 1, which is characterized in that: and (3) heating the blue polydiacetylene composite vesicle system prepared in the step (5) to 80 ℃ to obtain the fluorescent red-phase polydiacetylene composite vesicle, wherein the system generates single red fluorescence emission.

8. The method for preparing the fluorescein functionalized polydiacetylene vesicle with controllable multicolor fluorescence emission according to claim 1, which is characterized in that: adding L-lysine and Cu into the blue polydiacetylene composite vesicle system prepared in the step 5²⁺When the pH value of the system rises, the fluorescein group responds, the polydiacetylene composite vesicle is in a blue phase, and the system generates green fluorescence emission; then heating to 80 ℃, wherein the polydiacetylene composite vesicle is changed into a red phase, and the two kinds of fluorescence are mixed to generate yellow fluorescence reflection; then adding H⁺The fluorescence of the ion and the fluorescein is rapidly quenched under the acidic condition, and the system returns to red fluorescence emission.

9. The method for preparing the fluorescein functionalized polydiacetylene vesicle with controllable multicolor fluorescence emission according to claim 1, which is characterized in that: adding L-lysine and Cu into the blue polydiacetylene composite vesicle system prepared in the step 5²⁺The ionic polydiacetylene complex vesicle is in a blue phase, and a fluorescein group responds to the ionic polydiacetylene complex vesicle, so that a system generates green emission; then addWhen the temperature is heated to 80 ℃, the polydiacetylene composite vesicle is changed into a red phase, and the two kinds of fluorescence are mixed, so that a system generates yellow fluorescence reflection; then adding Trinitrophenol (TNP), quenching red-phase fluorescence of the polydiacetylene complex vesicle by the TNP, and returning the system to green fluorescence emission; then adding H⁺Ionic and absolute ethanol, the system returns to red fluorescence emission.

10. The construction of the molecular logic gate of the controllable multicolor fluorescence emission fluorescein functionalized polydiacetylene vesicle according to claim 1, is characterized in that: the molecular logic gate comprises an OR logic gate and an INHIBIT logic gate;

the OR logic gate defines that an input signal is L-amino acid OR heating, namely the L-amino acid exists OR the heating is 1; absence of L-amino acid or heating to 0; taking the fluorescence color emitted by the polydiacetylene composite vesicle system as an output end, namely the yellow fluorescence emission of the system is 1, and otherwise, the yellow fluorescence emission is 0; defining the polydiacetylene vesicles to be in a blue phase state and defining green fluorescence emission of the system as an initial state;

the INHIBIT logic gate defines the input signal as H⁺Ions or OH^-Ions, i.e. presence of H⁺Ions or OH^-The ion is 1; absence of H⁺Ions or OH^-The ion is 0; taking the fluorescence color emitted by the system of the polydiacetylene composite vesicle as an output end, and taking the fluorescence color emitted by the system of the polydiacetylene composite vesicle as an output end, namely the yellow fluorescence emission of the system is 1, otherwise, the yellow fluorescence emission is 0; and defines polydiacetylene vesicles as the red phase and the system as the initial state of green fluorescence emission.

Technical Field

The invention relates to a preparation method of a fluorescein functionalized polydiacetylene vesicle capable of regulating and controlling multicolor fluorescence emission and construction of a molecular logic gate, belonging to the technical field of organic conjugated polymer wave assembly.

Background

The multiple stimulus response fluorescent polymer has the advantages of better fluorescence emission, convenient preparation, stable performance and the like, and has wider prospects in the aspects of biosensors, display materials, logic switches, information storage and the like.

Polydiacetylene can undergo visible and obvious color change under the stimulation of a plurality of external environments, such as temperature, pH value, chemical reagents, stress action and the like, and the typical color change is that blue is changed into red. However, most of the current fluorescence emission regulation and control depends on methods such as light, heat, electricity, chemical substances and the like, and the preparation conditions of the current fluorescence regulation and control functionalized substrate are complicated, the pH control requirement is higher, the sensitivity of the functionalized substrate is low, and the external interference is larger.

Disclosure of Invention

The invention provides a preparation method of a fluorescein functionalized polydiacetylene vesicle capable of regulating and controlling colorful fluorescence emission and construction of a molecular logic gate, aiming at solving the problems that the existing fluorescence-regulated functionalized substrate is complicated in preparation conditions, high in pH control requirement, small in functionalized substrate sensitivity, large in interference from the outside and the like.

The technical scheme of the invention is as follows:

a preparation method of fluorescein functionalized polydiacetylene vesicles capable of regulating and controlling multicolor fluorescence emission comprises the following specific operation steps:

step 1, preparing aminated Fluorescein (FDNS) by using carboxyfluorescein and ethylenediamine;

step 2, preparing white powder (PCDA-NHS) by using carboxyl group diacetylene monomer (PCDA) and N-hydroxysuccinimide (NHS);

step 3, dissolving FDNS and PCDA-NHS respectively obtained in the step 1 and the step 2 in a DMF solvent, and stirring at constant temperature in a dark place to obtain a fluorescein substituted diacetylene monomer (F-PCDA);

step 4, dissolving PCDA and F-PCDA in absolute ethyl alcohol, adding deionized water, heating and ultrasonically treating, cooling in a dark condition, and standing at a constant temperature to obtain a composite vesicle;

and 5, carrying out illumination polymerization on the composite vesicle obtained in the step 4 under 254nm ultraviolet light to obtain a blue polydiacetylene composite vesicle system.

The specific operation process of the step 1 is as follows: the mass ratio of the materials is 1:1.2 placing the carboxyfluorescein and the ethylenediamine into absolute ethyl alcohol, heating and refluxing for 12h at the temperature of 80 ℃, cooling to room temperature, filtering, washing for 2-3 times by using the absolute ethyl alcohol, and recrystallizing to obtain the FDNS.

The specific operation process of the step 2 is as follows: dissolving PCDA in dichloromethane solution, then adding NHS dropwise into the solution, stirring at 30 ℃ in the dark for 4h, filtering, separating by column chromatography, and vacuum drying at 30 ℃ for 12h to obtain white powdery PCDA-NHS.

The quantity ratio of the PCDA to the NHS is 1: 1.2; the developing solvent in the column chromatographic separation is dichloromethane.

The specific operation process of the step 3 is as follows: and (3) dissolving PCDA-NHS and FDNS with the mass ratio of 1:1 in a DMF solvent, and stirring for 24 hours at 30 ℃ in the dark to obtain F-PCDA.

The specific operation process of the step 4 is as follows: the total amount of the materials is 5 multiplied by 10^-5And (2) dissolving PCDA and F-PCDA in a molar ratio of 10:1 in 1.5ml of absolute ethyl alcohol, adding 300ml of deionized water, heating to 65 ℃, carrying out ultrasonic treatment for 65min under the condition of 40% of ultrasonic power, cooling in a dark condition, and standing for 12h at 4 ℃ to obtain the composite vesicle.

And (3) heating the blue polydiacetylene composite vesicle system prepared in the step (5) to 80 ℃ to obtain the fluorescent red-phase polydiacetylene composite vesicle, wherein the system generates single red fluorescence emission.

Adding L-lysine and Cu into the blue polydiacetylene composite vesicle system prepared in the step 5²⁺Ions enable the pH value of the system to rise, fluorescein groups respond, and the polydiacetylene complex vesicles are in a blue phase, so that the system generates green fluorescence emission; then addWhen the temperature is heated to 80 ℃, the polydiacetylene composite vesicle is changed into a red phase, and the two kinds of fluorescence are mixed, so that a system generates yellow fluorescence reflection; then adding H⁺The fluorescence of the ion and the fluorescein is rapidly quenched under the acidic condition, and the system returns to red fluorescence emission.

Adding L-lysine and Cu into the blue polydiacetylene composite vesicle system prepared in the step 5²⁺The ionic polydiacetylene complex vesicle is in a blue phase, and a fluorescein group responds to the ionic polydiacetylene complex vesicle, so that a system generates green emission; then heating to 80 ℃, wherein the polydiacetylene composite vesicle is changed into a red phase, and the two kinds of fluorescence are mixed to generate yellow fluorescence reflection; then adding Trinitrophenol (TNP), quenching red-phase fluorescence of the polydiacetylene complex vesicle by the TNP, and returning the system to green fluorescence emission; then adding H⁺Ionic and absolute ethanol, the system returns to red fluorescence emission.

Constructing a molecular logic gate of the fluorescein functionalized polydiacetylene vesicle based on controllable multicolor fluorescence emission, wherein the OR logic gate defines that an input signal is L-amino acid OR heating, namely the L-amino acid exists OR the heating is 1; absence of L-amino acid or heating to 0; taking the fluorescence color emitted by the polydiacetylene composite vesicle system as an output end, namely the yellow fluorescence emission of the system is 1, and otherwise, the yellow fluorescence emission is 0; defining the polydiacetylene vesicles to be in a blue phase state and defining green fluorescence emission of the system as an initial state; the INHIBIT logic gate defines the input signal as H⁺Ions or OH^-Ions, i.e. presence of H⁺Ions or OH^-The ion is 1; absence of H⁺Ions or OH^-The ion is 0; taking the fluorescence color emitted by the system of the polydiacetylene composite vesicle as an output end, and taking the fluorescence color emitted by the system of the polydiacetylene composite vesicle as an output end, namely the yellow fluorescence emission of the system is 1, otherwise, the yellow fluorescence emission is 0; and defines polydiacetylene vesicles as the red phase and the system as the initial state of green fluorescence emission.

The invention has the following beneficial effects: according to the method, diacetylene and a fluorescein group are combined with each other to prepare the polydiacetylene composite vesicle, and the polydiacetylene composite vesicle realizes the multi-stimulus-regulated colorful fluorescence emission method based on the mutual transformation of a red phase and a blue phase of the polydiacetylene under the external stimulus and the color and fluorescence emission change of the polydiacetylene and the fluorescein group under the action of the change response of the fluorescein group to the external pH value. The polydiacetylene composite vesicle is simple in preparation process, has excellent fluorescence regulation and control performance, is simple and convenient in fluorescence regulation and control operation process, strong in operability, strong in environmental interference resistance and sensitive and quick in system response, can be used for preparing a multi-stimulation regulation and control device, and has good use value in the aspects of intelligent display material preparation, optical storage and the like.

Drawings

FIG. 1 is a synthesis formula of FDNS according to the present invention;

FIG. 2 is a nuclear magnetic hydrogen spectrum of FDNS;

FIG. 3 is a fluorescence spectrum characterization of FDNS;

FIG. 4 is a synthetic formula of F-PCDA;

FIG. 5 is a nuclear magnetic hydrogen spectrum of F-PCDA;

FIG. 6 is a schematic diagram of preparation of polydiacetylene complex vesicles;

FIG. 7 is a schematic view of the multi-color fluorescence control of polydiacetylene complex vesicles;

FIG. 8 is an OR logic gate construction;

FIG. 9 shows the INHIBIT logic gate construction.

Detailed Description

The experimental procedures used in the following examples are conventional unless otherwise specified.