CN108722506A - A method of control micro-fluidic chip inner hydrophilic modification effect - Google Patents
A method of control micro-fluidic chip inner hydrophilic modification effect Download PDFInfo
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Abstract
A method of control micro-fluidic chip inner hydrophilic modification effect, belong to micro-fluidic chip hydrophiling control technology field, the specificity of the aggregation-induced emission molecule modified the present invention is based on two boric acid base groups and the polyvinyl alcohol (PVA) containing hydroxyl, Selective recognition effect, to carrying out fluorescent visual imaging label inside the micro-fluidic chip Jing Guo hydrophilicity-imparting treatment, and analysis and Control condition is measured to its intrinsic fluorescence intensity and distribution situation by fluorescence microscope or laser confocal microscope.This method realizes the visualized in situ control to microchannel in micro-fluidic chip, it can solve the problems, such as that in situ detection can not be implemented to chip interior in traditional measurement, be it is a kind of it is safe, lossless, quickly and effectively visualize recognition methods, can be widely applied in material to the rapid screening of hydrophilicity-imparting treatment effect.
Description
Technical field
The invention belongs to micro-fluidic chip hydrophiling technical fields, and in particular to one kind is to hydrophiling in elastic PDMS chips
The method of the dyeing of carry out specific identification and fluorescence imaging control after modification.
Background technology
Micro-fluidic chip is closed because it can realize sample preparation, separation, detection simultaneously in micro-meter scale environment
Note.It is widely used in chemistry, biology and environment neck due to the advantages that its accuracy is high, sensitivity is good, reactant dosage is few
Domain, wherein dimethyl silicone polymer (PDMS) become miniflow due to the features such as its optical clarity is good, it is simple to prepare, excellent spring
Control the main making material of chip.In order to play important function of the PDMS micro-fluidic chips in example reaction system, to its table
Face hydrophilicity-imparting treatment has great importance.
Common method of hydrophilizing includes Polymer adsorption, acid processing, plasma or ultraviolet lighting etc..Due to micro-
The duct of fluidic chip is other in the micron-scale, is mostly based at the simulation to extraneous material surface to the evaluation of hydrophilic effect at present
Reason is difficult to realize to the direct characterization inside micro-fluidic chip duct.And have with stress, the structure of outer surface inside duct
Institute's difference, external simulation test are difficult to represent the true horizon of duct inner hydrophilic modification effect.Therefore, there is an urgent need to one
Kind original position, lossless specific recognition method, realize true, effective control to micro-fluidic duct inner hydrophilic effect.
The method of the present invention is that there are two the aggregation-induced emission molecules (DB-TPE) of boric acid base group to polymer based on tool
The selectivity bonding identification of hydroxyl, handles the micro-fluidic chip inner hydrophilicization after PVA is modified on polyvinyl alcohol (PVA)
Effect carries out Visual evaluation, and can expand in the monitoring of other hydrophilicity-imparting treatment rear surface hydroxy radical contents.This method is safe, fast
It is fast, effective, it can be used for the in situ detection and visual control method to microchannel hydrophiling in micro-fluidic chip, be micro-fluidic core
Effect of the piece in initial adsorbent material reaction lays the foundation.
Invention content
Based on hydrophilic important function in above-mentioned micro-fluidic chip, in order to overcome prior art problem, the present invention to provide
A kind of hydrophilic effect visual control method in situ.
The technical scheme is that：DB-TPE solution is prepared, it is its ionization of~10 guarantees to adjust pH value of solution.By quality point
Number for 3% polyvinyl alcohol (PVA) take not same amount be added in DB-TPE solution, obtained after fully shaking fluorescence property enhancing
Composite material.PVA and DB-TPE is verified by check experiment and IR Characterization, and specificity, stable knot are formed in the form of covalent bond
It closes.Based on this, is modified inside the micro-fluidic chip duct that PDMS is constructed, dyed with DB-TPE after washing, after dyeing with PVA
Extra DB-TPE is cleaned with deionized water again.By laser scanning co-focusing microscope to micro-fluidic chip duct inside
Fluorescence distribution situation carries out qualitative, quantitative analysis, realizes that the visualization to duct inner hydrophilic degree is screened.In addition, this
Method is equally applicable in the Visual evaluation of other hydrophilicity-imparting treatment modes, has the convenience and reliability of operation.
This method can in situ visualize the hydrophily in chip interior duct, efficiently solve conventional method and exist
The limitation of microcosmos area, quick, lossless imaging method are of great significance to chip modification and subsequent applications.
PDMS micro-fluidic chip internal gutter hydrophiling control methods, which is characterized in that include the following steps：
(1) alkaline solution of the fluorescent material with boric acid modified is prepared；
(2) preparation of PDMS micro-fluidic chips
It is preferred that：Design simultaneously photolithographic basic mode plate, the mold as micro-fluidic chip；With volume ratio 1:10 by curing agent and
PDMS is mixed, and is poured the mixture into mold slots, and 30min discharge bubbles are vacuumized；This mold is positioned over to 80 DEG C of baking
Cured in case；Finally the PDMS after solidification is removed from mold spare；
(3) hydrophilicity-imparting treatment of PDMS micro-fluidic chips
The PDMS chip deionized waters prepared are cleaned by ultrasonic 5-20min；The hydrophilicity-imparting treatment reagent that will have been configured
Solution injects in the micro-fluidic ducts PDMS, and solution is sucked out after retaining different time sections；With behind deionized water rinse duct 3-4 times
It is spare；
(4) fluorescent marker of PDMS micro-fluidic chips
What injection step (1) had configured in the PDMS chips after step (3) hydrophilicity-imparting treatment is glimmering with boric acid modified
The aqueous solution of stimulative substance retains 2-10min and solution is sucked out, spare afterwards with deionized water rinse duct 3-4 time；
(5) co-focusing imaging is handled
The chip handled well in step (4) is purged with nitrogen and is done, is placed directly on glass slide, is inverted in laser scanning
Laser Scanning Confocal Microscope carries out imaging；Fluorescence Intensity Assays are carried out to obtained images, interception specific region is to fluorescence
Distribution situation measures；
Hydrophilicity-imparting treatment reagent solution is the inorganic matter of material containing hydroxy groups or the solution of organic matter, be such as different alcoholysis degrees,
The polyvinyl alcohol of different polymerization degree, different viscosities, different molecular weight, other material containing hydroxy groups include the tools such as polyethylene glycol, glycerine
Standby different structure and small molecule or polymer with hydroxyl group, soda acid processing includes with dense H2SO4-H2O2、HCl-H2O2、
The strong acid and strong bases solution such as NaOH.
When according to different hydrophilic reagent treatment solution type, concentration in the further set-up procedure of result (3) and retaining
Between further adjust hydrophilic effect.
Fluorescent material is the gathering induced luminescence material of one or more boric acid modifieds, the fluorescence with boric acid base group
Matter, the tetraphenylethylene molecule (DB-TPE) of preferably double boric acid modifieds；More preferably by the tetraphenylethylene of double boric acid modifieds point
Sub (DB-TPE) is dissolved in dimethyl sulfoxide (DMSO), obtains the storing solution of a concentration of 15-25mM, and K is added2CO3And KHCO3The pH of composition
The buffer solution that value is 10；The volume fraction of final dimethyl sulfoxide (DMSO) is 1-5%, a concentration of 50-1000 μm of ol/L of DB-TPE.
The micro-fluidic chip duct marked by fluorescent material is observed with fluorescence microscope or laser scanning co-focusing microscope
Its fluorescence intensity change situation and mean pixel distribution.
The present invention is based on rich hydroxylcontaining materials and boric acid base group can form covalent B-O keys, to lure the aggregation of boric acid modified
Light emitting molecule is led for the specific recognition to hydroxylcontaining materials, and is thus applied to hydrophilic visualization in microchannel and comments
Valence.This method is not necessarily to carry out the destructive processing such as cutting to micro-fluidic chip, and original position, safety, nothing are carried out to micron level duct
The fluorescent marker of damage.The boronate fluorescent molecular amount very little used, the label time is short, response is rapid, result is stable, identification information
It is reliable accurate.It is lossless to the hydrophilic visualization Study of recognition of micro-fluidic chip that the present invention realizes a kind of original position, for into one
The outstanding micro-fluidic chip of step screening effect provides foundation.
Description of the drawings
Fig. 1 is that boric acid base group modifies fluorescence emission spectrum of the aggregation-induced emission molecule (DB-TPE) in alkaline solution
Figure, excitation wavelength 330nm, illustration are the fluorescence photo of the solution in the UV lamp.
Fig. 2 is polyvinyl alcohol (PVA) and DB-TPE combination figures, A be 3% by mass fraction polyvinyl alcohol
(PVA) it is that composite material after not same amount PVA is added that solution, which takes the fluorescence spectra obtained in not same amount addition DB-TPE solution, B,
The figure of fluorescence intensity changes at 465nm, C are the quantum yield situation of change of composite material after not same amount PVA is added.
Fig. 3 is that different polymer are added in DB-TPE solution：Polyvinylidene fluoride (PVDF), poly- (diallyl dimethyl
Ammonium chloride) (PDDA), kayexalate (PSS) fluorescence intensity change value of the compound at 465nm afterwards；
Fig. 4 is the infrared transmission spectra figure of composite material after not same amount PVA is added.
Fig. 5 is Laser Scanning Confocal Microscope image；Upper surface is bright after A and B is respectively PDMS chips injection TPEDB and cleans
Laser Scanning Confocal Microscope image under field and details in a play not acted out on stage, but told through dialogues；Lower surface is bright after C and D is respectively PDMS chips injection TPEDB and cleans
Laser Scanning Confocal Microscope image under field and details in a play not acted out on stage, but told through dialogues.
Fig. 6 is the light field Laser Scanning Confocal Microscope after injecting TPEDB and cleaning after PDMS chips handle different time with PVA
Image, picture size are 1200 × 1200 μm2；
Fig. 7 is the details in a play not acted out on stage, but told through dialogues Laser Scanning Confocal Microscope after injecting TPEDB and cleaning after PDMS chips handle different time with PVA
Image, picture size are 1200 × 1200 μm2。
Fig. 8 is PDMS chips PVA processing different time, injects details in a play not acted out on stage, but told through dialogues Laser Scanning Confocal Microscope imaging after TPEDB and cleaning
Mean pixel point analysis.
Fig. 9 is after PDMS modules handle different time with PVA, in the contact that extraneous environmental simulation hydrophily is tested
Angle.
Figure 10 be distinct methods hydrophilicity-imparting treatment PDMS after, inject TPEDB and clean after details in a play not acted out on stage, but told through dialogues Laser Scanning Confocal Microscope at
As figure：Wherein A is acid solution (VH2O:VHCl:VH2O2=5:1:1) polyethylene glycol that, B is 3%, the glycerine that C is 3%, D 3%
PVA solution, illustration be arrow at intercept fluorescence intensity profile, picture size be 800 × 800 μm2。
Specific implementation mode
With reference to embodiment, the present invention will be further described, but the present invention is not limited to following embodiments.
Embodiment 1：
(1) configuration of DB-TPE solution
Tetraphenylethylene molecule (DB-TPE) 0.042g for weighing double boric acid modifieds is dissolved in 5.0mL dimethyl sulfoxide (DMSO)s, is surpassed
Sound dissolves 15min, obtains the storing solution of a concentration of 20.0mM.Configure the buffer solution of pH=10：Weigh the K of 1.12g2CO3、
The KHCO of 0.19g3It is dissolved in 96.0mL deionized waters.3.5mL dimethyl sulfoxide (DMSO)s are added in the buffer solution, and continuous
The TPEDB storing solution 0.5mL of configured 10.0mM are added with stirring, a concentration of 100 μM of solution is obtained.The solution is carried out
Fluorescence spectrum is tested, as shown in Figure 1, exciting to obtain the transmitting spectrogram of DB-TPE using 330nm, there is fluorescent emission at 415nm
Peak, intensity are relatively low.
(2) PVA and DB-TPE composite materials are constructed
The PVA solids for weighing 3.0g are dissolved in 90 DEG C of hot water of 100mL, and stirring 2h obtains the solution of clear homogeneous.It should
It is spare that solution is cooled to room temperature.Not same amount is taken to be added dropwise to the DB-TPE that is equipped in step (1) PVA solution prepared molten
Liquid, while constantly oscillation, obtain final PVA contents be respectively 0.00%, 0.15%, 0.30%, 0.60%, 0.90%,
1.35% and 1.80%, and by the suspension after persistent oscillation 5 minutes.It is obtained into transparent membrane in 60 DEG C of baking oven drying 2h.It is right
Obtained composite material carries out fluorescence spectrum test, obtains fluorescence spectra as shown in Figure 2 A, compound as PVA contents increase
The fluorescence of material first increases to be reduced afterwards, and the fluorescence intensity change at 460nm is as shown in Figure 2 B.By to composite material quantum
The measurement of yield, as Fig. 2 C show that the trend of first increases and then decreases is equally presented in its quantum yield.
(3) repercussion study of PVA and DB-TPE composite materials
Other not hydroxyl polymer-containing such as PVDF, PSS and PDDA that equally applicable mass fraction is 3%, by itself and DB-
TPE is mixed, and the ultimate density of polymer is 0.6% in compound.(Fig. 3) is characterized in the fluorescence intensity of 460nm to it,
It was found that these three polymer cannot effectively be combined with DB-TPE, the fluorescence intensity of compound has no bright compared with original DB-TPE
Aobvious variation.Obtained laminated film in step (2) is subjected to IR Characterization, as shown in figure 4, its 1310-1430cm-1Place is shape
At B-O keys vibration peak, it was demonstrated that effective combination of DB-TPE and PVA.
(4) preparation of PDMS micro-fluidic chips
It designs and photoetching has 500 μm wide, 500 μm of depths Metal Substrate template, the mold as micro-fluidic chip.Take 20mL
PDMS and 2mL curing agent uniformly mix, vacuumized exclude bubble after pour into mold, vacuumize again discharge generate
Bubble.This mold is positioned over 2h in 80 DEG C of baking oven to cure.Finally the PDMS after solidification is removed from mold spare.
(5) hydrophilicity-imparting treatment of PDMS micro-fluidic chips
The PDMS chip deionized waters prepared are cleaned by ultrasonic 10min, are purged with nitrogen dry.The quality that will have been configured
The PVA solution that score is 3% is injected in the micro-fluidic ducts PDMS, respectively by PVA after 0,1,2,3,5,10,15 and 20min of reservation
Solution is sucked out.Duct is cleaned with deionized water 3 times, is removed spare after extra PVA.
(6) fluorescent marker of PDMS micro-fluidic chips
The DB-TPE solution that the continuous injection of PDMS chip relays after above-mentioned PVA modifieds has configured, retains 5min.Later
DB-TPE solution is sucked out, duct is cleaned 3 times with deionized water, removes extra DB-TPE.PDMS chips after cleaning are used
Nitrogen purging is dry, is placed on glass slide, is covered and sealed with coverslip, ensures the smooth of surface, smooth and transparent.
(7) co-focusing imaging is handled
The laser scanning co-focusing microscope with 20 times of enlargement ratios is selected, light source is made with the ultraviolet light of 405nm, and right
The transmitting light of 430-520nm carries out Image Acquisition.For the PDMS chips modified without PVA, after DB-TPE dyeing at
As scheming as shown, smooth surface is neat, upper and lower surface does not have observable fluorescence color lump.To by PVA modifications
PDMS chips, surface condition do not cause its pattern big influence as shown in fig. 6, smooth under light field.And with
PVA modify the time growth, from can obviously be observed in Fig. 7 the chip DB-TPE dyeing after fluorescence distribution it is more uniform,
Fluorescence intensity gradually increases.The average pixel value of fluorescence in duct is analyzed, its mean pixel is obtained and increases with the modification time
It grows and increases (such as Fig. 8), it was demonstrated that its hydrophily constantly enhances.And hydrophilic enhancing is tested with conventional method in outer surface simulation
Contact angle result it is consistent (such as Fig. 9).
In order to prove the versatility of this method, hydrophiling is carried out to PDMS micro-fluidic chips using other hydrophilic treatment methods
Modification.And the modification result of four kinds of methods is compareed：A is acid solution (VH2O:VHCl:VH2O2=5:1:1), B be 3% it is poly-
Ethylene glycol, the glycerine that C is 3%, the PVA solution that D is 3%.From in Figure 10, four kinds of methods generate it after surface modification
Hydroxyl shows fluorescence after DB-TPE dyeing.And the fluorescence intensity profile intercepted from arrow is found out, first three methods
Handle that obtained iridescent bulk strength is weak, dispersion is uneven.Show that the hydrophilicity-imparting treatment effect of these three methods is uneven, hydrophilic
Property it is weak, and PVA processing after have preferable hydrophilicity.
Claims (6)
1.PDMS micro-fluidic chip internal gutter hydrophiling control methods, which is characterized in that include the following steps：
(1) alkaline solution of the fluorescent material with boric acid modified is prepared；Fluorescent material is the poly- of one or more boric acid modifieds
Collect induced luminescence material, the fluorescent material with boric acid base group；
(2) preparation of PDMS micro-fluidic chips
(3) hydrophilicity-imparting treatment of PDMS micro-fluidic chips
The PDMS chip deionized waters prepared are cleaned by ultrasonic 5-20min；The hydrophilicity-imparting treatment reagent solution that will have been configured
It injects in the micro-fluidic ducts PDMS, solution is sucked out after retaining different time sections；With the 3-4 standby in deionized water rinse duct
With；Hydrophilicity-imparting treatment reagent solution is the inorganic matter of material containing hydroxy groups or the solution of organic matter；
(4) fluorescent marker of PDMS micro-fluidic chips
The fluorescence with boric acid modified that injection step (1) has configured in PDMS chips after step (3) hydrophilicity-imparting treatment
The aqueous solution of matter retains 2-10min and solution is sucked out, spare afterwards with deionized water rinse duct 3-4 time；
(5) co-focusing imaging is handled
The chip handled well in step (4) is purged with nitrogen and is done, is placed directly on glass slide, laser scanning copolymerization is inverted in
Focusing microscope carries out imaging；Fluorescence Intensity Assays are carried out to obtained images, interception specific region is to fluorescence distribution
Situation measures；
According to different hydrophilic reagent treatment solution type, concentration and retention time in the further set-up procedure of result (3) come
Further adjust hydrophilic effect.
2. PDMS micro-fluidic chips internal gutter hydrophiling control method described in accordance with the claim 1, which is characterized in that step
(2) preparation of PDMS micro-fluidic chips：Design simultaneously photolithographic basic mode plate, the mold as micro-fluidic chip；With volume ratio 1:
10 mix curing agent and PDMS, pour the mixture into mold slots, vacuumize 30min discharge bubbles；This mold is put
It is placed in 80 DEG C of baking oven and is cured；Finally the PDMS after solidification is removed from mold spare.
3. PDMS micro-fluidic chips internal gutter hydrophiling control method described in accordance with the claim 1, which is characterized in that fluorescence
Substance is the tetraphenylethylene molecule (DB-TPE) of double boric acid modifieds.
4. PDMS micro-fluidic chips internal gutter hydrophiling control method described in accordance with the claim 3, which is characterized in that step
(1) it is：The tetraphenylethylene molecule (DB-TPE) of double boric acid modifieds is dissolved in dimethyl sulfoxide (DMSO), a concentration of 15-25mM is obtained
Storing solution, be added K2CO3And KHCO3The buffer solution that the pH value of composition is 10；The volume fraction of final dimethyl sulfoxide (DMSO) is 1-
A concentration of 50-1000 μm of ol/L of 5%, DB-TPE.
5. PDMS micro-fluidic chips internal gutter hydrophiling control method described in accordance with the claim 1, which is characterized in that glimmering
The micro-fluidic chip duct of stimulative substance label is observed its fluorescence intensity with fluorescence microscope or laser scanning co-focusing microscope and is become
Change situation and mean pixel distribution.
6. PDMS micro-fluidic chips internal gutter hydrophiling control method described in accordance with the claim 1, which is characterized in that hydrophilic
Change reagent is the polyvinyl alcohol of different alcoholysis degrees, different polymerization degree, different viscosities, different molecular weight, polyethylene glycol, glycerine, acid
Alkali process solution, soda acid processing solution are selected from dense H2SO4-H2O2、HCl-H2O2, NaOH strong acid and strong base solution.
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| CN114588956A (en) * | 2020-12-03 | 2022-06-07 | 中国科学院大连化学物理研究所 | Microfluidic chip glutathione local hydrophilic modification method based on laminar flow effect |
| CN114088697A (en) * | 2021-11-12 | 2022-02-25 | 南通大学 | Microfluidic chip device for biogenic amine detection and application |
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