CN110907358A - Device and method for realizing light microflow microbubble cavity lead ion sensor - Google Patents
Device and method for realizing light microflow microbubble cavity lead ion sensor Download PDFInfo
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- CN110907358A CN110907358A CN201911250543.8A CN201911250543A CN110907358A CN 110907358 A CN110907358 A CN 110907358A CN 201911250543 A CN201911250543 A CN 201911250543A CN 110907358 A CN110907358 A CN 110907358A
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Abstract
The invention discloses a device and a method for realizing an optical microflow micro-bubble cavity lead ion sensor, wherein the device comprises a micro-bubble cavity, a micro-flow pump, an analyte container, a tunable laser controller, a tunable laser, an attenuation sheet, a polaroid, a fused cone optical fiber, a data acquisition card and a computer which are sequentially connected, and the computer is in signal connection with the data acquisition card and the tunable laser controller; the method realizes the lead ion detection with high sensitivity and high specificity by modifying GR-5 DNA enzyme on the inner wall of the micro-cavity. The implementation device and the implementation method are simple and flexible to operate, easy to implement and low in price.
Description
Technical Field
The invention relates to the technical field of realization of optical microfluidic micro-bubble cavity sensors, in particular to a device and a method for realizing an optical microfluidic micro-bubble cavity lead ion sensor.
Background
Lead is one of the earliest metals used by human beings and is also a heavy metal with strong toxicity, and exists in the earth crust in the form of a stable lead sulfide (namely galena) compound, and the content of the lead is only 0.0016%. As early as 5000 years ago, people could smelt lead from ore, but the amount of lead used and the exploitation of lead were very small at that time. With the rapid development of modern industry, lead pollution is rapidly increased, and the health and the natural environment of human beings are threatened. Automobile exhaust, home decoration materials, children toys, popcorn, cosmetics and other articles which people come into contact with daily contain lead. Lead is not easily biodegraded after entering the environment, and can finally cause the damage of human body functions, such as myasthenia, obnubilation, memory loss and syncope, even endanger life, especially children, through the continuous enrichment of biological chains. The nervous system of children is sensitive to lead and even if they are exposed to a low concentration of lead, they can cause severe irreversible damage to their nervous system. Lead can be discharged into the environment through various ways, lead pollution is wide from mining and transportation to application of lead ores, and the risk of the lead threatening the health of human beings is increased from air breathed by people to walking land to drinking water sources, wherein lead ions are one of the important forms of lead pollution. Accurate and efficient monitoring of the lead ion content, particularly monitoring of a water environment, is used for preventing further aggravation of lead ion pollution, and reduction of lead exposure of people is an extremely important step in the work of preventing and treating lead pollution. Therefore, relevant regulations are made for the lead ion degree of drinking water by various countries and environmental protection organizations, wherein the standard of China is not more than 48 nmol/L, and the maximum concentration specified by the United states environmental protection agency is 72 nmol/L.
The traditional methods for lead ion detection mainly comprise methods such as spectroscopy, mass spectrometry and the like. These analytical methods have high sensitivity but require bulky and expensive detection instruments, complicated and time-consuming pretreatment procedures, and require professional operation. These disadvantages make it impossible to meet the present-day demands solely by means of these conventional analytical methods, as the number of analytical samples to be tested increases today. For this reason, many efforts have been made by analysts, on the one hand, to combine traditional analysis methods with other techniques and new materials to improve detection efficiency; on the other hand, researchers have developed new technologies for rapid detection, and biosensors have attracted much attention because of their advantages such as simplicity, rapidity, and sensitivity. Deoxyribozymes (DNAzymes) are functional nucleic acids synthesized by in vitro molecular evolution technology, have the functions of identifying target molecules and catalyzing reactions, and have higher stability due to the lack of furanose ring 2-OH compared with RNA. Deoxyribozymes have the ability to catalyze a variety of reactions, such as ligation and hydrolysis of DNA, cleavage of DNA and RNA, and the like. Most of deoxyribozymes having RNA cleavage catalytic function require specific metal ions as cofactors, and thus have attracted much attention in the detection of heavy metal ions. In the presence of a cofactor, the deoxyribozyme exerts its catalytic activity for RNA cleavage, cleaving the substrate strand away from the cleavage site (rA). Compared with other enzymes, the deoxyribozyme has the advantages of simplicity, easiness in synthesis, stability, easiness in storage and the like. GR-5 DNAzyme, also an RNA splicing DNAzyme, was the first DNAzyme obtained by Breaker and Joyce et al in 1994 through in vitro molecular evolution techniques, and GR-5 DNAzyme has a higher specificity for lead ions than 8-17 DNAzyme and has recently been used in combination with fluorescence, electrochemical methods to develop sensors for detecting lead ions. Whispering Gallery Mode (WGM) optical microfluidic microcavity sensors enable lower detection limits because of their higher quality factor (Q) and smaller mode volume (V). In addition, the optical microfluidic micro-bubble cavity sensor is simple and convenient to prepare, low in price and naturally has the advantages of microfluidic channels and the like, so that the optical microfluidic micro-bubble cavity sensor is very suitable for detecting trace biochemical samples.
Disclosure of Invention
The invention aims to provide a device and a method for realizing a lead ion sensor with an optical microfluid micro-bubble cavity, so that the lead ion detection process has higher sensitivity and specificity, and the detection process is simpler to operate.
In order to solve the technical problem, the device for realizing the optical microflow microbubble cavity lead ion sensor comprises a microbubble cavity, a microflow pump, an analyte container, a tunable laser controller, a tunable laser, an attenuation sheet, a polaroid, a fused cone optical fiber, a data acquisition card and a computer which are sequentially connected, wherein the computer is in signal connection with the data acquisition card and the tunable laser controller; the two ends of the micro-bubble cavity are respectively connected with the micro-flow pump and the analyte container through the guide pipes, and the micro-flow pump pumps the analyte in the analyte container into the micro-bubble cavity; the fused cone optical fiber is arranged on one side of the micro-bubble cavity, one end of the fused cone optical fiber is connected with the output end of the polaroid, and the other end of the fused cone optical fiber is connected with the data acquisition card;
the computer sends a signal to the tunable laser controller to control the tunable laser to emit laser, the light sequentially passes through the attenuator and the polaroid to enter the fused-cone optical fiber, the fused-cone optical fiber couples the laser into the micro-bubble cavity, the laser which is continuously transmitted is received by the data acquisition card and is sent to the computer for processing, and the computer monitors the resonance wavelength of the received micro-bubble cavity to realize the real-time monitoring of the lead ions.
Furthermore, an acrylic box for fixing is arranged outside the micro-bubble cavity.
Further, the tunable laser is 1520 and 1630nm laser.
An implementation method of an optical microfluidic microbubble cavity lead ion sensor comprises the following steps:
s1: introducing piranha solution into the micro-bubble cavity to attach negative charges to the inner wall of the micro-bubble cavity;
s2: introducing polylysine into the micro-cavity to attach positive charges to the inner wall of the micro-cavity;
s3: introducing DNA polymerase chain into the micro-bubble cavity to attach the DNA polymerase chain to the inner wall of the micro-bubble cavity;
s4: introducing a substrate DNA chain into the microcavity, wherein the DNA chain and the substrate chain form a DNA double-chain structure, so that GR-5 DNA double chains are attached to the inner wall of the microcavity;
s5: introducing an analysis sample to be detected into the microbubble cavity;
s6: other metal ions are introduced into the microbubble cavity to test the specificity of the microbubble cavity.
Further, before the step S1, an ethanol aqueous solution with a refractive index gradient of five hundred thousandths is introduced into the micro-cavity, and the change of the resonance wavelength of the micro-cavity is monitored, so as to obtain the bulk refractive index sensitivity of the micro-cavity.
Further, the analysis sample to be detected in step S5 is a lead ion solution.
Further, the other metal ions in step S6 are eight metal ions, such as sodium, calcium, zinc, potassium, copper, iron, ferrous iron, and manganese, respectively.
By adopting the technical scheme, the invention has the following beneficial effects: by modifying GR-5 DNA enzyme on the inner wall of the light microflow micro-bubble cavity, the high-sensitivity lead ion detection is realized, the operation is simple and flexible, the realization is easy, and the price is low. In addition, the sensor can detect lead ions with high specificity by detecting different metal ions. The method has strong practicability.
Drawings
The invention is described in further detail below with reference to the following figures and embodiments:
FIG. 1 is a schematic view of an experimental apparatus according to the present invention;
FIG. 2 is a diagram illustrating the measurement result of the refractive index of the optical microfluidic microcavity sensor according to the present invention;
FIG. 3 is a schematic diagram of a specific modification process of the optical microfluidic microbubble cavity sensor according to the present invention;
FIG. 4 is a schematic diagram of the signal response of the optical microfluidic microbubble cavity sensor to lead ions according to the present invention;
fig. 5 is a schematic diagram of the response of the optical microfluidic microbubble cavity sensor to lead ion specific signals in the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be described in detail and completely with reference to the accompanying drawings.
As shown in fig. 1, the present invention provides an implementation apparatus of an optical microfluidic micro-cavity lead ion sensor, which includes a micro-cavity, a micro-flow pump, an analyte container, and a tunable laser controller, a tunable laser, an attenuator, a polarizer, a fused cone optical fiber, a data acquisition card and a computer, which are connected in sequence, wherein the computer is in signal connection with the data acquisition card and the tunable laser controller; the two ends of the micro-bubble cavity are respectively connected with the micro-flow pump and the analyte container through a catheter, an acrylic box for fixing is arranged outside the micro-bubble cavity, and the analyte in the analyte container is sucked into the micro-bubble cavity by the micro-flow pump; the fused cone optical fiber is arranged on one side of the micro-bubble cavity, one end of the fused cone optical fiber is connected with the output end of the polaroid, and the other end of the fused cone optical fiber is connected with the data acquisition card; the computer sends a signal to the tunable laser controller to control the tunable laser to emit laser, wherein the tunable laser is an 1520 + 1630nm laser, light sequentially enters the fused-cone optical fiber through the attenuator and the polarizing disc, the fused-cone optical fiber couples the light into the micro-bubble cavity, the laser continuously transmitted is received by the data acquisition card and sent to the computer for processing, and the computer monitors the received resonance wavelength of the micro-bubble cavity to realize the real-time monitoring of lead ions.
An implementation method of an optical microfluidic microbubble cavity lead ion sensor comprises the following steps:
s1: introducing piranha solution (piranha solution) into the micro-bubble cavity to make the inner wall of the micro-bubble cavity attach negative charges;
s2: introducing Polylysine (PLL) into the micro-bubble cavity to attach positive charges to the inner wall of the micro-cavity;
s3: introducing DNA polymerase chain into the micro-bubble cavity to attach the DNA polymerase chain to the inner wall of the micro-bubble cavity;
s4: introducing a substrate DNA chain into the microcavity, wherein the DNA chain and the substrate chain form a DNA double-chain structure, so that GR-5 DNA double chains are attached to the inner wall of the microcavity;
s5: introducing an analysis sample to be detected into the micro-bubble cavity, wherein the analysis sample to be detected is a lead ion solution;
s6: and introducing other metal ions into the micro-bubble cavity to test the specificity of the micro-bubble cavity, wherein the other metal ions are eight metal ions such as sodium, calcium, zinc, potassium, copper, iron, ferrous iron, manganese and the like.
Before S1, introducing an ethanol water solution with a refractive index gradient of five hundred thousandths into the micro-cavity, and monitoring the change of the resonance wavelength of the micro-cavity to obtain the refractive index sensitivity of the micro-cavity.
By modifying GR-5 DNA enzyme on the inner wall of the light microflow micro-bubble cavity, the high-sensitivity lead ion detection is realized, the operation is simple and flexible, the realization is easy, and the price is low. The sensor realizes the high specificity detection of lead ions by detecting the non-used metal ions. The method has strong practicability.
Before the inner wall of the microcavity is modified, the bulk refractive index of the microcavity needs to be tested. And (3) sequentially introducing an ethanol water solution with a refractive index gradient of five hundred thousandths into the micro-cavity, and monitoring the change of the resonance wavelength of the micro-cavity to obtain the sensitivity of the refractive index of the micro-cavity. The change in the microcavity resonance wavelength is shown in FIG. 2. The volume refractive index of the microbubble cavity was calculated to be 265.24 nm/RIU.
As shown in FIG. 3, the specific modification process of the inner wall of the micro-cavity is shown. Because the DNA chain can not be directly attached to the inner wall of the micro-cavity, the inner wall of the micro-cavity is firstly treated, negative charges are attached to the inner wall of the micro-cavity through the piranha solution, and positive charges are attached to the inner wall of the micro-cavity through polylysine. The amino group on the DNA strand has a negative charge and thus can adhere to the inner wall of the microcavity. And then introducing a DNA polymerase chain into the microcavity to be attached to the inner wall of the microcavity, and then introducing a substrate DNA chain into the inner wall of the microcavity, wherein the DNA polymerase chain and the substrate chain form a DNA double-chain structure, so that GR-5 DNA double chains are attached to the inner wall of the microcavity.
The lead ion solution was then tested using the treated lumen. Figure 4 shows the resonant wavelength of the lumen of the microbubbles as a function of time after lead ions have passed into the lumen of the microbubbles. It can be seen that the resonant wavelength first blue-shifts as the lead ions pass through the microbubble cavity and then gradually tends to stabilize. This result confirms the feasibility of the sensing device.
FIG. 5 shows the results of a test of the specificity of the microvesicle lumen for lead ions. Except for lead ions, eight kinds of other metal ions (sodium, calcium, zinc, potassium, copper, iron, ferrous iron and manganese) are respectively tested, and as can be seen from fig. 5, the sensing device has obvious signal response to the lead ions, and has very weak response to the other metal ions. The test result proves that the sensing device has good specificity detection on lead ions.
The above are preferred embodiments of the present invention, and all changes made according to the technical scheme of the present invention that produce functional effects do not exceed the scope of the technical scheme of the present invention belong to the protection scope of the present invention.
Claims (7)
1. The utility model provides an implementation device of plumbous ion sensor in light miniflow microbubble chamber which characterized in that: the micro-bubble type micro-flow sensor comprises a micro-bubble cavity, a micro-flow pump, an analyte container, and a tunable laser controller, a tunable laser, an attenuation sheet, a polaroid, a fused cone optical fiber, a data acquisition card and a computer which are sequentially connected, wherein the computer is in signal connection with the data acquisition card and the tunable laser controller; the two ends of the micro-bubble cavity are respectively connected with the micro-flow pump and the analyte container through the guide pipes, and the micro-flow pump pumps the analyte in the analyte container into the micro-bubble cavity; the fused cone optical fiber is arranged on one side of the micro-bubble cavity, one end of the fused cone optical fiber is connected with the output end of the polaroid, and the other end of the fused cone optical fiber is connected with the data acquisition card;
the computer sends a signal to the tunable laser controller to control the tunable laser to emit laser, the light sequentially passes through the attenuator and the polaroid to enter the fused-cone optical fiber, the fused-cone optical fiber couples the laser into the micro-bubble cavity, the laser which is continuously transmitted is received by the data acquisition card and is sent to the computer for processing, and the computer monitors the resonance wavelength of the received micro-bubble cavity to realize the real-time monitoring of the lead ions.
2. The device for realizing the optical microfluidic microbubble cavity lead ion sensor as claimed in claim 1, wherein: an acrylic box for fixing is arranged on the outer side of the micro-bubble cavity.
3. The device for realizing the optical microfluidic microbubble cavity lead ion sensor as claimed in claim 1, wherein: the tunable laser is 1520 and 1630nm laser.
4. A method for realizing an optical microfluid microbubble cavity lead ion sensor is characterized by comprising the following steps: the method comprises the following steps:
s1: introducing piranha solution into the micro-bubble cavity to attach negative charges to the inner wall of the micro-bubble cavity;
s2: introducing polylysine into the micro-cavity to attach positive charges to the inner wall of the micro-cavity;
s3: introducing DNA polymerase chain into the micro-bubble cavity to attach the DNA polymerase chain to the inner wall of the micro-bubble cavity;
s4: introducing a substrate DNA chain into the microcavity, wherein the DNA chain and the substrate chain form a DNA double-chain structure, so that GR-5 DNA double chains are attached to the inner wall of the microcavity;
s5: introducing an analysis sample to be detected into the microbubble cavity;
s6: other metal ions are introduced into the microbubble cavity to test the specificity of the microbubble cavity.
5. The method for implementing the optical microfluidic microbubble cavity lead ion sensor according to claim 4, wherein the method comprises the following steps: before the step S1, introducing an ethanol water solution with a refractive index gradient of five hundred thousandths into the micro-cavity, and monitoring the change of the resonance wavelength of the micro-cavity to obtain the refractive index sensitivity of the micro-cavity.
6. The method for implementing the optical microfluidic microbubble cavity lead ion sensor according to claim 4, wherein the method comprises the following steps: the analysis sample to be detected in step S5 is a lead ion solution.
7. The method for implementing the optical microfluidic microbubble cavity lead ion sensor according to claim 4, wherein the method comprises the following steps: the other metal ions in step S6 are eight kinds of metal ions, such as sodium, calcium, zinc, potassium, copper, iron, ferrous iron, and manganese, respectively.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112113923A (en) * | 2020-09-24 | 2020-12-22 | 中山大学 | Micro-bubble cavity coupling CO based on whispering gallery mode2Sensor and manufacturing method |
| CN112710630A (en) * | 2020-11-24 | 2021-04-27 | 深圳大学 | Composite micro-cavity and using method and preparation method thereof |
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| CN104096606A (en) * | 2014-06-08 | 2014-10-15 | 青岛博睿林新材料有限公司 | Microfluidic chip able to separate lead and zinc ions |
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| EP3408276A1 (en) * | 2016-01-29 | 2018-12-05 | Uniwersytet Warszawski | 5'-phosphorothiolate mrna 5'-end (cap) analogs, mrna comprising the same, method of obtaining and uses thereof |
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| CN102798624A (en) * | 2012-08-08 | 2012-11-28 | 中国科学院长春光学精密机械与物理研究所 | Near-field Raman biosensor based on echo wall mode |
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| EP3408276A1 (en) * | 2016-01-29 | 2018-12-05 | Uniwersytet Warszawski | 5'-phosphorothiolate mrna 5'-end (cap) analogs, mrna comprising the same, method of obtaining and uses thereof |
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Cited By (2)
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| CN112710630A (en) * | 2020-11-24 | 2021-04-27 | 深圳大学 | Composite micro-cavity and using method and preparation method thereof |
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