CN110308080A - Utilize the method for functionalized capillary measurement carboxyl polystyrene microsphere concentration - Google Patents
Utilize the method for functionalized capillary measurement carboxyl polystyrene microsphere concentration Download PDFInfo
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- CN110308080A CN110308080A CN201910667462.1A CN201910667462A CN110308080A CN 110308080 A CN110308080 A CN 110308080A CN 201910667462 A CN201910667462 A CN 201910667462A CN 110308080 A CN110308080 A CN 110308080A
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- 239000004793 Polystyrene Substances 0.000 title claims abstract description 33
- 239000004005 microsphere Substances 0.000 title claims abstract description 33
- 229920002223 polystyrene Polymers 0.000 title claims abstract description 33
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 title claims abstract description 32
- 238000000034 method Methods 0.000 title claims abstract description 29
- 238000005259 measurement Methods 0.000 title claims abstract description 14
- 238000005086 pumping Methods 0.000 claims abstract description 16
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 claims abstract description 10
- 229940043267 rhodamine b Drugs 0.000 claims abstract description 10
- 238000004458 analytical method Methods 0.000 claims abstract description 3
- 230000035772 mutation Effects 0.000 claims abstract description 3
- 239000000243 solution Substances 0.000 claims description 70
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 34
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 21
- 239000011780 sodium chloride Substances 0.000 claims description 17
- 238000001228 spectrum Methods 0.000 claims description 11
- 239000011259 mixed solution Substances 0.000 claims description 10
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 8
- 238000012545 processing Methods 0.000 claims description 7
- 238000003384 imaging method Methods 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 4
- 229910052757 nitrogen Inorganic materials 0.000 claims description 4
- 239000004642 Polyimide Substances 0.000 claims description 3
- 229920002521 macromolecule Polymers 0.000 claims description 3
- 239000012528 membrane Substances 0.000 claims description 3
- 229920001721 polyimide Polymers 0.000 claims description 3
- 238000001514 detection method Methods 0.000 abstract description 12
- 239000002245 particle Substances 0.000 abstract description 9
- 239000011521 glass Substances 0.000 abstract description 7
- 238000006073 displacement reaction Methods 0.000 abstract description 5
- 230000035945 sensitivity Effects 0.000 abstract description 5
- 230000002349 favourable effect Effects 0.000 abstract description 3
- 238000000691 measurement method Methods 0.000 abstract description 3
- 229920006254 polymer film Polymers 0.000 abstract 1
- 239000010408 film Substances 0.000 description 13
- 239000010410 layer Substances 0.000 description 13
- 230000003287 optical effect Effects 0.000 description 8
- 230000005284 excitation Effects 0.000 description 6
- 239000000975 dye Substances 0.000 description 5
- 239000002346 layers by function Substances 0.000 description 4
- 230000008569 process Effects 0.000 description 3
- 230000008859 change Effects 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 230000008021 deposition Effects 0.000 description 2
- 238000000695 excitation spectrum Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 229920006389 polyphenyl polymer Polymers 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 1
- 239000005977 Ethylene Substances 0.000 description 1
- 150000001336 alkenes Chemical class 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000004061 bleaching Methods 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000002189 fluorescence spectrum Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920000867 polyelectrolyte Polymers 0.000 description 1
- 239000002096 quantum dot Substances 0.000 description 1
- 238000004557 single molecule detection Methods 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000010409 thin film Substances 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/06—Investigating concentration of particle suspensions
-
- G01N15/075—
Abstract
The invention discloses a kind of methods using functionalized capillary measurement carboxyl polystyrene microsphere concentration, first with pumping rhodamine B in thick-walled capillary chamber, optics geometric resonance mode is formed about in glass wall, then the particle that high molecular polymer film is used to be bonded special function keys is done in capillary tube inner wall, finally includes the displacement of triangle model and star mode using discrete fourier mutation analysis geometric mode.Method provided by the invention, orientation survey special function keys particle, high sensitivity is at low cost, and measurement method is simple, stablizes, and the zlasing mode of favorable repeatability, resonance is higher than the intensity of Whispering-gallery-mode, and detection limits low, no photobleaching.
Description
Technical field
The invention belongs to optical sensing fields, and in particular to a kind of to measure carboxyl polystyrene using functionalized capillary
The method of microballoon concentration.
Background technique
Optical resonator based on Whispering-gallery-mode, comprising microballoon, micro-loop, capillary is widely used in terms of sensing.
Sensing based on Whispering-gallery-mode is based on two methods measurement pattern structure.One is utilize waveguide, prism or other special dresses
It sets and evanescent wave is coupled nearly resonant cavity.It can cause turning for resonance mode when particle or biomolecule to be detected are attached to cavity wall
Become.Evanscent field coupling needs to consider the several factors such as phase matched.The quality factor of the resonance mode of this method be up to 109 or
It is higher, it can be used for Single Molecule Detection.But this method also have disadvantages that limit value its development, need the tunable laser of narrowband at
This height, frangible difficult encapsulation of device etc..Another kind forms stable Whispering-gallery-mode using fluorescence spectrum.This optical resonator needs
Wanting internal has such as luminescent quantum dot or other polymeric materials.These fluorescent materials can by continuous wavelength laser or
Person's LED diode laser etc. excites under free space.This optical resonator does not need narrow-band tunable laser, and accurate
Optical element couple evanescent wave guide cavity mode.
The active optical resonator quality factor with higher of zlasing mode based on Whispering-gallery-mode, stronger hair
Spectrum and relatively narrow line width are penetrated, can be improved the speed and accuracy of sensor, and be applied to the detection of biomolecule.Stable swashs
The optical resonator of optical mode has important application in the context of detection of biomolecule.These devices all use different inorganic
Or gain media of the organic dyestuff as laser.The capillary of zlasing mode has relevant report to the detection of nonspecific molecule,
In addition it has proven to can be used for refractive index sensing application using the thin-walled capillary pipe resonant cavity of the zlasing mode of dyestuff excitation.But
The capillary of zlasing mode is less to the detection the relevant technologies of specific molecular.
Summary of the invention
It is an object of the invention to: it provides and a kind of utilizes functionalized capillary measurement carboxyl polystyrene microsphere concentration
Method, orientation survey special function keys particle, high sensitivity is at low cost, and measurement method is simple, stablizes, favorable repeatability, detection
Limit low, no photobleaching.
The technical solution adopted by the invention is as follows:
A method of carboxyl polystyrene microsphere concentration is measured using functionalized capillary, comprising the following steps:
S1. the polyimides macromolecule membrane for the capillary surface that length is 4-6cm is removed, obtains processing capillary；
S2. it uses NaOH solution with 0.1mL/min pumping 1-3 minutes in processing capillary obtained by S1 step, then uses PAH
Solution and the mixed solution of NaCl solution were with 0.02mL/min pumping 4-6 minute, then mixing using PSS solution and NaCl solution
Solution is closed with 0.02mL/min pumping 4-6 minutes, finally uses PAH solution and the mixed solution of NaCl solution with 0.02mL/min
Pumping 4-6 minutes, obtains functionalized capillary；
Preferably, it uses NaOH solution with 0.1mL/min pumping 2 minutes in processing capillary obtained by S1 step, makes capillary
The interior surfaces of glass band slightly negative electrical charge of pipe；Use PAH solution and the mixed solution of NaCl solution with 0.02mL/min pumping 5 again
Minute, then use PSS solution and NaCl solution to pump 5 minutes with 0.02mL/min, the surface PSS band slightly negative electrical charge can be with
PAH positive surface charge attracts each other to form PAH-PSS film above；Finally use PAH solution and NaCl solution with 0.02mL/
Min is pumped 5 minutes, and band slightly positive charge in the surface PAH can attract each other with PSS negative electrical charge above, and to form PAH-PSS-PAH thin
Film obtains functionalized capillary；
S3. to pumping NHS solution, EDC solution and carboxyl polyphenyl to be measured in functionalized capillary cavity obtained by S2 step
The mixed solution of ethylene microspheres solution, carboxyl polystyrene microsphere is under the catalysis of NHS and EDC and the-NHization on the surface PAH
Reaction is learned, carboxyl polystyrene microsphere is captured by PAH film layer orientation；Rhodamine B solution is pumped after 25-35min；
S4. dye laser is excited using nitrogen lasers, excites the rhodamine B inside the laser excitation capillary of 500nm molten
Liquid, then using the spectrum of imaging spectrometer record geometric resonance mode, spectrum is analyzed using discrete fourier variation, is obtained
To carboxyl polystyrene microsphere concentration.
By reasonably selecting the phase of the component of Fourier transformation and combining their corresponding intensity weighteds, to calculate mode
Displacement.
The present invention is as follows according to measuring principle:
In the interface of thick-walled capillary reflection and refraction occur for light, form stable triangle resonance mode and star resonance
Mode.Triangle model and the free spectral range of star mode are respectively:
Wherein n1,n2,n3Respectively indicate the film layer that particle to be detected in cavity is bonded to, functional layer and capillary
The refractive index of glass wall, r1,r2,r3Respectively indicate the film layer that particle is bonded to, the half of the glass wall of functional layer and capillary
Diameter.Ψ indicates that the half-angular breadth of light incidence functional layer, φ indicate that the half-angular breadth of light incident glass layer, α indicate that light exists
The half-angular breadth of glassy layer refraction.Here N indicates the points of star mode, m intermediate scheme number.Corresponding resonant wavelength are as follows:
When the refractive index and thickness of intracavitary functional layer are certain, when the concentration of the such as particle of intracavitary test substance changes
When, corresponding variation all occurs very much for corresponding refractive index and thickness, and such resonant wavelength will change, and therefore, passes through inspection
Survey the concentration of specific molecular or particle in the displacement energy test chamber of resonant wavelength.
Further, the internal diameter of capillary is 125 μm in S1 step, and outer diameter is 160 μm.
Further, the concentration of NaOH solution is 8-12mol/L in S2 step；Preferably, NaOH solution in S2 step
Concentration is 10mol/L.
Further, the concentration of PAH solution is 1-3mg/mL in S2 step, and the concentration of NaCl solution is 1-4mol/L；It is excellent
Selection of land, the concentration of PAH solution is 2mg/mL in S2 step, and the concentration of NaCl solution is 2.5mol/L.
Further, the concentration of PSS solution is 1-3mg/mL in S2 step, and the concentration of NaCl solution is 1-4mol/L；It is excellent
Selection of land, the concentration of PSS solution is 2mg/mL in S2 step, and the concentration of NaCl solution is 2.5mol/L.
Further, the concentration of NHS solution is 250mmol/L in S3 step, and the concentration of EDC solution is 250mmol/L, sieve
Dan Ming-B solution concentration is 1mmol/L.
Further, the polystyrene microsphere for the surface carboxyl groups that carboxyl polystyrene microsphere is 50nm in S3 step.
Further, the discrete fourier mutation analysis geometric mode used in S4 step includes triangle model and star mode
Displacement.
In conclusion by adopting the above-described technical solution, the beneficial effects of the present invention are:
1, method of the invention, orientation survey special function keys particle, high sensitivity is at low cost, and measurement method is simple, surely
It is fixed, favorable repeatability；
2, the present invention is using the zlasing mode of resonance, and for polystyrene microsphere, the detection of zlasing mode offset is limited to
9.75nmol/L, this is advantageous compared with fluorescence-based device, higher than the intensity of Whispering-gallery-mode, and detection limit is low, unglazed
Bleaching；
3, the present invention utilizes " star and triangle " mode of zlasing mode microcapillary, and at three layers of polyelectrolyte of deposition
Mode position is tracked after combining with the amide of subsequent carboxy-functionalized polystyrene microsphere, is enabled people in mechanical and optics
Monitor that layer-by-layer surface combines in the device of robust；
4, method of the invention can extend in the scheme of other surfaces combination, can according to need using different laser
Wavelength, therefore method of the invention has potential application in detection method of many based on laser.
Detailed description of the invention
In order to illustrate the technical solution of the embodiments of the present invention more clearly, below will be to needed in the embodiment attached
Figure is briefly described, it should be understood that the following drawings illustrates only certain embodiments of the present invention, therefore is not construed as pair
The restriction of range for those of ordinary skill in the art without creative efforts, can also be according to this
A little attached drawings obtain other relevant attached drawings.
Fig. 1 is the rhodamine B exciting light spectrogram of the capillary of naked capillary, the capillary with PE film layer and embodiment；
Fig. 2 (a) is the discrete Fourier transform of the excitation spectrum of the capillary with PE film layer；It (b) is naked capillary, band PE
The sensing figure of the capillary of the capillary and embodiment of film layer.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to the accompanying drawings and embodiments, right
The present invention is further elaborated.It should be appreciated that described herein, specific examples are only used to explain the present invention, not
For limiting the present invention, i.e., described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is logical
The component for the embodiment of the present invention being often described and illustrated herein in the accompanying drawings can be arranged and be designed with a variety of different configurations.
Therefore, the detailed description of the embodiment of the present invention provided in the accompanying drawings is not intended to limit below claimed
The scope of the present invention, but be merely representative of selected embodiment of the invention.Based on the embodiment of the present invention, those skilled in the art
Member's every other embodiment obtained without making creative work, shall fall within the protection scope of the present invention.
It should be noted that the relational terms of term " first " and " second " or the like be used merely to an entity or
Operation is distinguished with another entity or operation, and without necessarily requiring or implying between these entities or operation, there are any
This actual relationship or sequence.Moreover, the terms "include", "comprise" or its any other variant be intended to it is non-exclusive
Property include so that include a series of elements process, method, article or equipment not only include those elements, but also
Further include other elements that are not explicitly listed, or further include for this process, method, article or equipment it is intrinsic
Element.In the absence of more restrictions, the element limited by sentence "including a ...", it is not excluded that including described
There is also other identical elements in the process, method, article or equipment of element.
Feature and performance of the invention are described in further detail with reference to embodiments.
Embodiment
The a kind of of present pre-ferred embodiments offer measures carboxyl polystyrene microsphere concentration using functionalized capillary
Method, the specific steps are as follows:
S1. by internal-and external diameter be respectively 125 μm and 160 μm length be 5cm capillary be placed on logical oxygen 923K furnace
In remove outside polyimides macromolecule membrane, obtain processing capillary, both ends are connected to the plastic tube for pumping liquid
On, it is placed on microscope carrier；
S2. it uses the NaOH solution of 10mol/L with 0.1mL/min pumping 2 minutes in processing capillary obtained by S1 step, makes
The interior surfaces of glass band slightly negative electrical charge of capillary；Again using the PAH solution of 2mg/mL and 2.5mol/L NaCl solution it is mixed
Solution is closed with 0.02mL/min pumping 5 minutes, the surface PAH band slightly positive charge can be mutual with the negative electrical charge of glass surface above
Attraction forms PAH-PSS film；Then use the PSS solution of 2mg/mL and the mixed solution of the NaCl solution of 2.5mol/L with
0.02mL/min is pumped 5 minutes, and band slightly positive charge in the surface PSS can attract each other to form PAH- with PAH positive charge above
PSS-PAH film obtains functionalized capillary；
S3. to pumping the NHS solution of 250mmol/L, 250mmol/L in functionalized capillary cavity obtained by S2 step
The mixed solution of EDC solution and carboxyl polystyrene microsphere solution to be measured, carboxyl polystyrene microsphere are urged NHS's and EDC
Change lower and the surface PAH-NH to chemically react, carboxyl polystyrene microsphere is captured by PAH film layer orientation；It is pumped after 30min
The rhodamine B solution of Pu 1mmol/L；
S4. dye laser is excited using nitrogen lasers, excites the rhodamine B inside the laser excitation capillary of 500nm molten
Liquid, then using the spectrum of imaging spectrometer record geometric resonance mode, spectrum is analyzed using discrete fourier variation, by
Increase in the resonant wavelength of excitation mode with the increase of carboxyl polystyrene microsphere solution concentration, therefore obtains carboxyl polyphenyl second
Alkene microballoon concentration.
For carboxyl polystyrene microsphere, triangle model resonant wavelength is measured with the solution of 50nm carboxyl polystyrene microsphere
The increase of concentration 0.05mg/mL, 0.5mg/mL, 1mg/mL and 2mg/mL and increased separately 0.8nm, 1.15nm, 1.3nm and
1.5nm, so that the sensitivity for obtaining the solution concentration of triangle model measurement polystyrene microsphere is 15pm/nM.Finally use three
The three times of the standard deviation of angle mould formula displacement obtain sensor detection divided by sensitivity and are limited to 9.75nmol/L.With Whispering-gallery-mode
The detection limit (20nmol/L) of strength of fluid sensing compare, detection limit of the invention is lower.
Comparative example
The capillary to naked capillary and with PE film layer pumps rhodamine B solution respectively, then using nitrogen lasers excitation dye
Expect laser, excite the rhodamine B solution inside the laser excitation capillary of 500nm, then is total using imaging spectrometer record geometry
The spectrum of vibration mode analyzes spectrum using discrete fourier variation.Blue shift has occurred in triangle model, and opposite star mode goes out
The red shift of existing opposite direction.
Comparative example obtains the capillary of naked capillary, the capillary with PE film layer and embodiment that imaging spectrometer records
Rhodamine B exciting light spectrogram is as shown in Figure 1.It include being total to different directions are mobile for complicated overlapping in these three spectrum
Vibration mode, tracking motion of defect modes is highly difficult on surface, we form envelope using the peak position of Gaussian function fitting star mode
The resonance shift of line acquisition triangle model.To obtain the red shift of star mode and the blue shift of triangle model.
Spectrum is analyzed using discrete fourier variation, obtains the discrete Fourier transform of the excitation spectrum of capillary
And the misalignment figure of triangle model and star mode, as shown in Figure 2.Because of PE film deposition, star motion of defect modes 34pm, because
The capillary of embodiment, star mode move 81pm again；Under same case, triangle model is displaced respectively in the opposite direction
0.47nm and 0.68nm.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (8)
1. it is a kind of using functionalized capillary measurement carboxyl polystyrene microsphere concentration method, which is characterized in that including with
Lower step:
S1. the polyimides macromolecule membrane for the capillary surface that length is 4-6cm is removed, obtains processing capillary；
S2. it uses NaOH solution with 0.1mL/min pumping 1-3 minutes in processing capillary obtained by S1 step, then uses PAH solution
Mixed solution with NaCl solution was with 0.02mL/min pumping 4-6 minutes, and then the mixing using PSS solution and NaCl solution is molten
Liquid finally used the mixed solution of PAH solution and NaCl solution to pump with 0.02mL/min with 0.02mL/min pumping 4-6 minutes
4-6 minutes, obtain functionalized capillary；
S3. to pumping NHS solution, EDC solution and carboxyl polystyrene to be measured in functionalized capillary cavity obtained by S2 step
The mixed solution of microspheres solution pumps rhodamine B solution after 25-35min；
S4. using nitrogen lasers excite dye laser, then using imaging spectrometer record geometric resonance mode spectrum, using from
It dissipates Fourier's variation to analyze spectrum, obtains carboxyl polystyrene microsphere solution concentration.
2. the method according to claim 1 using functionalized capillary measurement carboxyl polystyrene microsphere concentration,
Be characterized in that: the internal diameter of capillary is 125 μm in the S1 step, and outer diameter is 160 μm.
3. the method according to claim 1 using functionalized capillary measurement carboxyl polystyrene microsphere concentration,
Be characterized in that: the concentration of NaOH solution is 8-12mol/L in the S2 step.
4. the method according to claim 1 using functionalized capillary measurement carboxyl polystyrene microsphere concentration,
Be characterized in that: the concentration of PAH solution is 1-3mg/mL in the S2 step, and the concentration of NaCl solution is 1-4mol/L.
5. the method according to claim 1 using functionalized capillary measurement carboxyl polystyrene microsphere concentration,
Be characterized in that: the concentration of PSS solution is 1-3mg/mL in the S2 step, and the concentration of NaCl solution is 1-4mol/L.
6. the method according to claim 1 using functionalized capillary measurement carboxyl polystyrene microsphere concentration,
Be characterized in that: the concentration of NHS solution is 250mmol/L in the S3 step, and the concentration of EDC solution is 250mmol/L, Luo Dan
The concentration of bright-B solution is 1mmol/L.
7. the method according to claim 1 using functionalized capillary measurement carboxyl polystyrene microsphere concentration,
Be characterized in that: carboxyl polystyrene microsphere is the polystyrene microsphere of the surface carboxyl groups of 50nm in the S3 step.
8. the method according to claim 1 using functionalized capillary measurement carboxyl polystyrene microsphere concentration,
Be characterized in that: the discrete fourier mutation analysis geometric mode used in the S4 step includes the position of triangle model and star mode
It moves.
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