CN106198473B - Ratio-type 3-dimensional metal enhances fluorescence Pb2+The structure of biosensor - Google Patents
Ratio-type 3-dimensional metal enhances fluorescence Pb2+The structure of biosensor Download PDFInfo
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- CN106198473B CN106198473B CN201610567187.2A CN201610567187A CN106198473B CN 106198473 B CN106198473 B CN 106198473B CN 201610567187 A CN201610567187 A CN 201610567187A CN 106198473 B CN106198473 B CN 106198473B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
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Abstract
The invention discloses a kind of preparations of Ratio-type 3-dimensional metal enhancing fluorescence paper chip and its paper chip equipment in Pb2+Application in detection.Hydrophilic workspace and the hydrophobic region of paper chip are prepared by wax printing technique, and then measure the fluorescence intensity of two kinds of fluorescence signals by modifying layer by layer in working region growth decahedron silver, the variation by two kinds of fluorescence intensity ratios and Pb2+Concentration relationship curve realizes Pb2+Portable, reliable, highly sensitive detection.
Description
Technical field
The present invention relates to portable, easy to operate, the low, high sensitivity of cost and measurement result Pb with a high credibility2+Analysis
In particular detection technique field is a kind of Ratio-type 3-dimensional metal enhancing fluorescence Pb2+The preparation of biosensor.
Background technology
Pb2+There are in various resources（Such as food, blood, drinking water, industrial wastewater etc.）, it is referred to as a most harmful huge sum of money
A series of the problem of one of category ion, it not only causes environmental pollutions, but also human health is also constituted serious
It threatens, especially children.For example, being chronically exposed to rich in Pb2+In environment, the nervous centralis of the mankind and urinary system can be made to cause
Serious damage, and the damage of intellectual development can be also caused by food chain and drinking water.More seriously Pb2+It can not
By biological decomposition, micro Pb2+Fearful harm may be caused to human health.Therefore, a highly sensitive detection huge sum of money
It is particularly important to belong to ion.In order to reach this purpose, many detection of heavy metal ion methods are established, such as Atomic absorption
Spectrum, chromatography, Ratio-type fluorescence detection method, electrochemical luminescence method, molecular imprinting method, photoelectrochemical method and fluorescent method
Deng.In the above-mentioned methods, fluorescence detection because its is easy to operate, high sensitivity, it is single-minded the advantages that widely paid close attention to.But
In the fluorescence detection method of single fluorogen, because meeting is by the shadows such as prepare liquid environmental instrument factor and fluorogen itself concentration
It rings, will result directly in and be easy to make testing result to generate false positive, the accuracy of testing result needs to be further increased.
Ratio-type fluorescence detection method refers to the fluorogen containing there are two, passes through the fluorescence intensity for measuring two fluorogens, profit
Use the ratio of the two as signal parameter, to measure the analysis method of detectable substance.The ratio signal of fluorescence not light source strength
The influence of degree and instrumental sensitivity.Therefore, Ratiometric fluorescent probe can be with the selectivity of improvement method, sensitivity and dynamic response
Range, to effectively increase the accuracy of testing result.
Micro-fluidic paper substrate analytical equipment, because of its low cost, portable, preparation process is simple, and is easily achieved miniaturization
Detection, is widely used in the fields such as medicine, biological sample, environment diagnosis.The knot of Ratio-type fluorescence detection method and paper chip
It closes, it is perfectly that the low cost of paper chip, the advantage that portable, preparation process is simple is sensitive with Ratio-type fluorescence detection method
The advantages that degree is high, single-minded, measurement result is with a high credibility combines, and is expected to solve heavy metal ion in water resource from far-off regions
Detection.
Invention content
The object of the present invention is to provide a kind of portable, easy to operate, low, high sensitivity of cost and measurement result confidence levels
High Ratio-type 3-dimensional metal enhances fluorescence paper chip, to realize Pb2+Quick, highly sensitive detection.
In order to solve the above-mentioned technical problem, the present invention is three-dimensional by a kind of paper chip with decahedron silver and Ratio-type
Metal-enhanced fluorescence method is realized, it is characterized in that include the following steps：
（1）Design the hydrophobic wax print area of Ratio-type 3-dimensional metal as shown in Fig. 1 enhancing fluorescence paper chip and hydrophilic
Working region；
（2）By wax printer by step（1）The upper hydrophobic pattern of paper chip printing of middle design, the refill that will then obtain
Piece heats under 60-150 oC, until wax melts and be impregnated with entire paper, formation hydrophobic layer；
（3）In step（2）In obtain paper chip hydrophilic working region growth decahedron silver；
（4）By the catalysis chain of graphene quantum dot functionalization, it is defined as DNA1, it is fixed on step（3）Gained paper chip
Hydrophilic region is then rinsed with buffer solution；
（5）By the Pb of dyestuff and ceria functionalization2+It identifies substrate strand, is defined as 17DS, is added drop-wise to step（4）Gained
The hydrophilic region of paper chip, is finally rinsed with buffer solution；
（6）By the Pb of various concentration2+It is added to step（5）The hydrophilic region of gained paper chip, is subsequently placed into fluorescence equipment
In, the Pb in various concentration is measured respectively2+The fluorescence intensity of lower graphene quantum dot and dyestuff；
（7）The variation of two kinds of fluorescence intensity ratios of drafting and Pb2+Concentration relationship curve completes Pb2+Measurement.
It is of the present invention in step（2）In obtain paper chip hydrophilic working region growth decahedron silver step be：
By 1.5 mL, 50 mM sodium citrates, 1.5 mL, 6% polyvinylpyrrolidones, 0.15 mL, 10 mM arginine
And 0.6 mL, 10 mM silver nitrates are added in 6 mL secondary waters, and the fresh preparations of 0.24 mL, 0.1 M are then added under stiring
Sodium borohydride, be made silver seed；It takes the silver-colored seed of 20 μ L to be added drop-wise to working region, places 45 min at room temperature, repeatedly
It is added dropwise three times；The paper chip for dripping silver-colored seed is placed under the blue lamp of 3 W and exposes 10 h.
Step of the present invention（4）Described in DNA1Specific base sequence and its 5 ' terminal modified upper sulfydryl, 3 ' ends are repaiied
Graphene quantum dot on decorations.
Step of the present invention（5）Described in the specific base sequences of 17DS and its 5 ' terminal modified upper ceria, 3 ' end
Dyestuff in modification.
Dyestuff of the present invention is：Hydroxycoumarin, phycoerythrin, phthalocyanine fluorescence probe Cy3, phthalocyanine fluorescence probe
Cy5, phthalocyanine fluorescence probe Cy7, rhodamine, isothiocyanates, roxithromycin, Fluoresceincarboxylic acid.
Ratio-type Pb of the present invention2+Fluoremetry step, paper chip is put into fluorescence ware, is measured respectively different dense
The Pb of degree2+The fluorescence intensity of lower graphene quantum dot and dyestuff, the variation of two kinds of fluorescence intensity ratios of drafting and Pb2+Concentration is closed
It is curve, realizes Pb2+Accurate measurement.
Beneficial effects of the present invention
（1）Ratio-type metal-enhanced fluorescence method, measurement result are with a high credibility.
（2）Ratio-type metal-enhanced fluorescence method and paper chip combination, system low with portable, highly sensitive, required expense
Make the features such as simple, is expected to realize Pb in the limited remote districts of resource2+Portable inspectiont.
（3）Decahedron silver is grown on paper, is increased specific surface area and is effectively reduced the background fluorescence of paper, it is most important that makes
Fluorescence signal is exaggerated several times, substantially increases Pb2+The sensitivity of detection.
Description of the drawings
Fig. 1：The hydrophobic wax bulk print pattern of paper chip.
Specific implementation mode
For a better understanding of the present invention, below by the embodiment content that the present invention is furture elucidated, but the present invention is not
It is limited only to the following examples.
Embodiment 1
Ratio-type 3-dimensional metal enhances biological sensor in Pb2+Application in detection：
（1）Designed ratios type 3-dimensional metal enhances the hydrophobic wax print area of fluorescence paper chip and hydrophilic working region；
（2）The paper chip designed in step 1.1 is printed into upper hydrophobic pattern, the refill that will then obtain by wax printer
Piece heats under 60-150 oC, until wax melts and be impregnated with entire paper, formation hydrophobic layer；
（3）In step（2）In obtain paper chip hydrophilic working region growth decahedron silver：By 1.5 mL, 50 mM
Sodium citrate, 1.5 mL, 6% polyvinylpyrrolidones, 0.15 mL, 10 mM arginine and 0.6 mL, 10 mM silver nitrates are added
Into 6 mL secondary waters, the freshly prepared sodium borohydrides of 0.24 mL, 0.1 M are then added under stiring, silver seed is made；It takes
The silver-colored seed of 20 μ L is added drop-wise to working region, places 45 min at room temperature, is added dropwise repeatedly three times；Silver-colored seed will be dripped
Paper chip, which is placed under the blue lamp of 3 W, exposes 10 h；
（4）By the catalysis chain of graphene quantum dot functionalization, it is defined as DNA1, it is fixed on step（3）Gained paper chip
Hydrophilic region is then rinsed with buffer solution；
（5）By the Pb of dyestuff and ceria functionalization2+It identifies substrate strand, is defined as 17DS, is added drop-wise to step（4）Gained
The hydrophilic region of paper chip, is finally rinsed with buffer solution；
（6）By the Pb of various concentration2+It is added to step（5）The hydrophilic region of gained paper chip, is subsequently placed into fluorescence equipment
In, the Pb in various concentration is measured respectively2+The fluorescence intensity of lower graphene quantum dot and dyestuff；
（7）The variation of two kinds of fluorescence intensity ratios of drafting and Pb2+Concentration relationship curve completes Pb2+Measurement.
SEQUENCE LISTING
<110>University Of Ji'nan
<120>Ratio-type 3-dimensional metal enhances fluorescence Pb2+The structure of biosensor
<130> 2016
<160> 2
<170> PatentIn version 3.3
<210> 1
<211> 51
<212> DNA
<213>It is artificial synthesized
<400> 1
tttttttttt ttttttttca tctcttctcc gagccggtcg aaatagtgag t 51
<210> 2
<211> 21
<212> DNA
<213>It is artificial synthesized
<400> 2
actcactatr aggaagagat g 21
Claims (6)
1. Ratio-type 3-dimensional metal enhances fluorescence Pb2+The construction method of biosensor, it is characterized in that including the following steps：
1.1 designed ratios type 3-dimensional metals enhance the hydrophobic wax print area of fluorescence paper chip and hydrophilic working region；
The paper chip designed in step 1.1 is printed upper hydrophobic pattern by 1.2 by wax printer, then by obtained paper chip in
It is heated under 60-150 oC, until wax melts and be impregnated with entire paper, formation hydrophobic layer；
The hydrophilic working region growth decahedron silver of 1.3 paper chips obtained in step 1.2；
1.4, by the catalysis chain of graphene quantum dot functionalization, are defined as DNA1, it is fixed on the hydrophilic of step 1.3 gained paper chip
Region is then rinsed with buffer solution；
1.5 by the Pb of dyestuff and ceria functionalization2+It identifies substrate strand, is defined as 17DS, is added drop-wise to step 1.4 gained paper
The hydrophilic region of chip, is finally rinsed with buffer solution；
1.6 by the Pb of various concentration2+It is added to the hydrophilic region of step 1.5 gained paper chip, is subsequently placed into fluorescence equipment,
The Pb in various concentration is measured respectively2+The fluorescence intensity of lower graphene quantum dot and dyestuff；
The variation of 1.7 two kinds of fluorescence intensity ratios of drafting and Pb2+Concentration relationship curve completes Pb2+Measurement.
2. Ratio-type 3-dimensional metal enhancing fluorescence Pb according to claim 12+The construction method of biosensor, feature
It is, the hydrophilic working region growth decahedron silver of the paper chip obtained in step 1.2 is as follows：
By 1.5 mL, 50 mM sodium citrates, 1.5 mL, 6% polyvinylpyrrolidones, 0.15 mL, 10 mM arginine and 0.6
10 mM silver nitrates of mL are added in 6 mL secondary waters, and the freshly prepared boron hydrogen of 0.24 mL, 0.1 M is then added under stiring
Change sodium, silver seed is made；It takes the silver-colored seed of 20 μ L to be added drop-wise to working region, places 45 min at room temperature, three are added dropwise repeatedly
It is secondary；The paper chip for dripping silver-colored seed is placed under the blue lamp of 3 W and exposes 10 h.
3. Ratio-type 3-dimensional metal enhancing fluorescence Pb according to claim 12+The construction method of biosensor, feature
It is, DNA described in step 1.41Base sequence and its 5 ' terminal modified upper sulfydryl, 3 ' terminal modified upper graphene quantums
Point.
4. according to the Ratio-type 3-dimensional metal enhancing fluorescence Pb described in claim 12+The construction method of biosensor, it is special
Sign is, the base sequence of 17DS described in step 1.5 and its 5 ' terminal modified upper ceria, 3 ' terminal modified upper dyestuffs.
5. according to the Ratio-type 3-dimensional metal enhancing fluorescence Pb described in claim 12+The construction method of biosensor, it is special
Sign is that the dyestuff described in step 1.5 is：Hydroxycoumarin or phycoerythrin or phthalocyanine fluorescence probe Cy3 or phthalocyanine fluorescence
Probe Cy5 or phthalocyanine fluorescence probe Cy7 or rhodamine or isothiocyanates or roxithromycin or Fluoresceincarboxylic acid.
6. Ratio-type 3-dimensional metal enhancing fluorescence Pb according to claim 12+The construction method of biosensor, feature exist
In Ratio-type Pb2+Fluoremetry step, paper chip is put into fluorescence ware, measures the Pb in various concentration respectively2+Lower graphite
The fluorescence intensity of alkene quantum dot and dyestuff, the variation of two kinds of fluorescence intensity ratios of drafting and Pb2+Concentration relationship curve realizes Pb2+
Accurate measurement.
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| CN106198473B true CN106198473B (en) | 2018-09-14 |
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107643286B (en) * | 2017-09-06 | 2020-04-21 | 济南大学 | Porous CeO2Preparation of nano material and application of nano material in paper-based sensor |
| CN107589113B (en) * | 2017-09-21 | 2019-10-25 | 济南大学 | A kind of method of paper base double mode detection lead ion |
| CN109870437A (en) * | 2019-03-12 | 2019-06-11 | 中山大学 | A kind of multivariate detection chip and its application in alimentary anemia detection |
| CN110487871A (en) * | 2019-08-28 | 2019-11-22 | 济南大学 | A kind of preparation of electrochemistry Ratio-type paper chip and its application in detection of nucleic acids |
| CN113637673B (en) * | 2021-08-12 | 2023-10-13 | 华中科技大学 | DNAzyme combination, long-chain DNAzyme for cleavage of DNA/RNA chimeras and application |
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