CN110227563A - The encapsulating method and PDMS micro-fluidic chip of PDMS micro-fluidic chip vaporization prevention - Google Patents
The encapsulating method and PDMS micro-fluidic chip of PDMS micro-fluidic chip vaporization prevention Download PDFInfo
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- CN110227563A CN110227563A CN201910393044.8A CN201910393044A CN110227563A CN 110227563 A CN110227563 A CN 110227563A CN 201910393044 A CN201910393044 A CN 201910393044A CN 110227563 A CN110227563 A CN 110227563A
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502707—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the manufacture of the container or its components
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0887—Laminated structure
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/12—Specific details about materials
Abstract
The present invention provides the encapsulating method and PDMS micro-fluidic chip of a kind of PDMS micro-fluidic chip vaporization prevention.The encapsulating method is included in cleaned glass on piece and forms photoresist layer；Exposure mask with micro-fluidic chip patterning is fitted in into photoresist layer surface, exposure, development obtain template；Template is laid in glass baseplate surface, and it tapes along template edge, the thickness of adhesive tape is greater than the thickness of template, suffocating treatment is carried out to glass substrate and the paraffin oil dispersion liquid of dimethyl silicone polymer will be poured, vacuumize process, the first transparent glass slide is sticked, the first transparent glass slide is solidified, separating treatment, obtains chip；Vacuum plasma handles chip and the second transparent glass slide with holes, and the two is aligned and is bonded；Connect the female Luer of with closure.The micro-fluidic chip that the present invention obtains can effectively prevent evaporating, thus avoid the problems such as the diminution that micro- reaction member generates in temperature changing process, volatilizing, merging, deform.
Description
Technical field
The invention belongs to micro fluidic chip technical fields, and in particular to a kind of sealing side of PDMS micro-fluidic chip vaporization prevention
Method and PDMS micro-fluidic chip.
Background technique
Polymerase chain reaction (Polymerase chain reaction, abbreviation PCR) has in molecular biology field
Extremely important status has developed into the experimental technique of a routine.From initial regular-PCR by continuously improving, by
Gradually derivative reverse transcription PCR, multiplex PCR, nest-type PRC, immuno-PCR etc..It is also gradually developed from initial qualitative analysis simultaneously
For quantitative analysis.By the continuous development of more than ten years, real-time quantitative PCR (real time PCR, abbreviation qPCR) is because of its sensitivity
High, high specificity and the quantitatively advantages such as accurate are increasingly becoming the most common technological means of genetic analysis and obtain in medical diagnosis on disease
To being widely applied.But since many factors can only also carry out " relative quantification " qPCR.Therefore need one kind can be with
The new technology of " absolute quantitation " is all limited to solve the problems, such as nucleic acid quantification medium sensitivity and accuracy.Based on such demand,
Digital pcr (digital PCR, abbreviation dPCR) technology is come into being.The technology need not rely on any reference substance or external standard, only
Only it will achieve the purpose that absolute quantitation by directly counting the positive signal of single lattice nucleic acid molecules, therefore, sensitivity and essence
Exactness all very advantageous, it is only necessary to which considerably less sample size can be used for quantitative analysis, these features make digital pcr technology fast
Speed is used widely in every field, and is developed and formed micro- reaction chamber/orifice plate (Micro-chamber), micro-fluidic core
Piece (Microfluidic chip) and droplet PCR system (droplet digital PCR system, ddPCR).
Micro-fluidic chip is generally by two layers of dimethyl silicone polymer (Polydimethylsiloxane, PDMS) film and bottom
The coverslip composition of layer, to constitute corresponding fluidized bed and control layer.PDMS film is processed by soft lithography, from
And the gas passage and fluid passage of right-angled intersection are formed, fluid can be rapidly divided by the control of corresponding valve flat
Capable reaction member.PDMS material is a kind of the macromolecule organic silicon compound, has optical clear, hydrophobicity and waterproofness etc. special
Point.Because its is at low cost, the strong, transparent air-permeable using simple, bio-compatibility, and becomes and be widely used in the poly- of micro-fluidic equal fields
Close object material.However, just because of the gas permeability of PDMS material, frequently result in its manufactured digital pcr chip leakproofness at
There is phenomena such as reducing, volatilizing, merge, deforming in temperature changing process in problem, micro- reaction member in digital pcr chip,
The inhomogenous even reaction failure of micro- reaction member is caused, this makes PDMS material in terms of digital pcr micro-fluidic chip
Using being also extremely restricted.
Summary of the invention
It existing in temperature change reduce, volatilize, deform for micro- reaction member of current digital pcr micro-fluidic chip
Phenomena such as lead to react inhomogenous or even failure, the present invention provide a kind of sealing side of PDMS micro-fluidic chip vaporization prevention
Method.
Further, the present invention also provides the PDMS micro-fluidic chips obtained by above-mentioned encapsulating method.
For achieving the above object, technical scheme is as follows:
A kind of encapsulating method of PDMS micro-fluidic chip vaporization prevention, comprising the following steps:
Photoresist is coated on clean glass sheet surface by step S01., then carries out the processing that is heating and curing, and forms stacking
In the photoresist layer of the glass sheet surface；
Exposure mask with micro-fluidic chip patterning is fitted in the photoresist layer surface by step S02., and is exposed
Light, development treatment obtain the template with micro-fluidic chip patterning；
The template is laid in glass baseplate surface by step S03., and in the glass baseplate surface along the mould
Edges of boards paste one layer tape along surrounding, so that the thickness of the adhesive tape is greater than the thickness of the template, using insulating liquid described in attaching
The glass substrate of adhesive tape carries out suffocating treatment, one layer of separation layer is formed in template surface, then by the stone of dimethyl silicone polymer
Wax oil dispersion liquid, which pours, divides the paraffin oil of the dimethyl silicone polymer uniformly
It is distributed in the template surface, and sticks the first transparent glass slide, curing process is carried out to the described first transparent glass slide, obtains band
The chip of structure；
The chip of the band structure and the template are carried out separating treatment by step S04.；
The chip of the band structure and the with holes second transparent glass slide are placed under vacuum condition by step S05. to carry out etc.
Ion processing, then by the two alignment fitting, heat treatment；
Step S06. coats one layer of glue in the with holes described second transparent slide surface, and by Rule of with closure
Connector is adhered to the described second transparent slide surface, is aligned the female Luer and the hole of the described second transparent glass slide,
The inlet and outlet of micro-fluidic chip is formed, heat treatment obtains PDMS micro-fluidic chip.
Correspondingly, a kind of PDMS micro-fluidic chip, including the first transparent glass slide, it is laminated in the described first transparent glass slide
The PDMS film layer on surface, the second transparent glass slide for being laminated in the PDMS film surface；
The second transparent slide surface has several through-holes, and the female Luer of with closure is connected on the through-hole.
Technical effect of the invention are as follows:
Compared with the existing technology, the encapsulating method of the PDMS micro-fluidic chip vaporization prevention of the above-mentioned offer of the present invention, by PDMS
Progress plasma treatment is bonded with the second transparent glass slide again after film layer is attached to transparent slide surface, thus in PDMS
Film surface forms air-locked transparent structural layers, reduces the area that PDMS film layer is contacted with air, and in inlet and outlet
Connection solves micro- in digital pcr chip with the female Luer of sealing cover so that playing has the effect of vaporization prevention performance
Diminution that reaction member generates in temperature changing process, the problems such as volatilizing, merge, deforming.
PDMS micro-fluidic chip provided by the invention is air-locked transparent due to foring in two apparent surface of PDMS film layer
Structure sheaf, and the female Luer of sealing cover is had in import and export connection, the effect for preventing evaporation is effectively acted as, is avoided
Diminution that micro- reaction member generates in temperature changing process, the problems such as volatilizing, merge, deforming.
Detailed description of the invention
It to describe the technical solutions in the embodiments of the present invention more clearly, below will be to needed in the embodiment
Attached drawing is briefly described, it should be apparent that, drawings in the following description are only some embodiments of the invention, for ability
For the those of ordinary skill of domain, without creative efforts, it can also be obtained according to these attached drawings other attached
Figure.
Fig. 1 is the clean sheet glass signal that the encapsulating method of PDMS micro-fluidic chip vaporization prevention provided by the invention provides
Figure；
Fig. 2 is that the encapsulating method of PDMS micro-fluidic chip vaporization prevention provided by the invention is coated in clean glass sheet surface
Form the schematic diagram of photoresist layer；
Fig. 3, which is bonded for the encapsulating method of PDMS micro-fluidic chip vaporization prevention provided by the invention on photoresist layer surface, to be had
The schematic diagram of the mask process of micro-fluidic chip patterning；
Fig. 4 is that obtained template is laid in glass by the encapsulating method of PDMS micro-fluidic chip vaporization prevention provided by the invention
Substrate surface and the structural schematic diagram formed in glass baseplate surface along template edge surrounding patch one layer tape；
Fig. 5 is that the encapsulating method of PDMS micro-fluidic chip vaporization prevention provided by the invention pours PDMS paraffin in the template
The cured structural schematic diagram for forming PDMS film layer of oil solution；
Fig. 6 is that the encapsulating method of PDMS micro-fluidic chip vaporization prevention provided by the invention is bonded first in PDMS film surface
The structural schematic diagram that transparent glass slide is formed；
Fig. 7 is the encapsulating method of PDMS micro-fluidic chip vaporization prevention provided by the invention by the first transparent glass slide and second
Transparent glass slide carries out plasma treatment and is bonded the structural schematic diagram of formation；
Fig. 8 is that the encapsulating method of PDMS micro-fluidic chip vaporization prevention provided by the invention connects in the second transparent slide surface
The PDMS microfluidic chip structure schematic diagram obtained after the female Luer of tape splicing sealing cover；
Wherein, 1- sheet glass；2- photoresist (template)；3- exposure mask；4- glass substrate；5- adhesive tape；6-PDMS film layer；7-
One transparent glass slide；The transparent glass slide of 8- second, 81- through-hole；The female Luer of 9- with closure.
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.
Fig. 1~7 are please referred to, the present invention provides a kind of encapsulating method of PDMS micro-fluidic chip vaporization prevention, including following step
It is rapid:
The sheet glass 1 of clean dried is provided, Fig. 1 is specifically detailed in.The sheet glass 1 should clean drying, avoid remained on surface
Grease stain, stain and influence processing effect.
A layer photoresist layer 2 is coated on the surface of sheet glass 1 by the way of spin coating, is specifically detailed in Fig. 2.Form photoetching
The photoresist of glue-line 2 can be positive photoresist or negative photoresist.Specifically, positive photoresist is selected from polymethylacrylic acid
Any one of methyl esters (PMMA), adjacent diazo naphthoquinone-phenolic resin (DQN)；The negative photoresist SU8 photoresist.
Specifically, photoresist is dripped and carries out spin-coat process on 1 surface of sheet glass, stood about 5 minutes, obtain a layer photoresist
Layer 2 then carries out the processing that is heating and curing to photoresist layer 2, and heating temperature is 50~70 DEG C, can such as use 55 DEG C, 60 DEG C, 65 DEG C
Solidified, the photoresist layer 2 being cured.By the exposure mask with micro-fluidic chip patterning after cooled to room temperature
3 are fitted in 2 surface of photoresist layer, and are exposed, development treatment, obtain template 2 (number and the light formed by photoresist layer 2
The number of photoresist layer is identical), specifically it is detailed in Fig. 3.
In exposure process, processing is exposed using ultraviolet light, and use ethyl lactate or tetramethylammonium hydroxide
(TMAH) develop, development terminates, and judges whether to develop completely using isopropanol.The template 2 formed by photoresist layer 2
It is upper that there is micro-fluidic chip patterning.
The glass substrate 4 for taking cleaning dry, it is ensured that the size of glass substrate 4 is greater than the size of template 2, then by template 2
It is laid in 4 surface of glass substrate, sticks one layer tape 5 along 2 surrounding edge of template on 4 surface of glass substrate, it is ensured that adhesive tape 5
Thickness be greater than template 2 thickness, in favor of formed PDMS film layer 6.It is carried out with glass substrate 4 of the insulating liquid to joint adhesive band 5
Suffocating treatment, fumigation time are 5~10min, form one layer of separation layer on 2 surface of template by fumigating, subsequent to prevent
PDMS film layer 6 and template 2 bond, and are specifically detailed in Fig. 4.
The insulating liquid be dichlorodimethylsilane, the insulating liquid formed separation layer it is subsequent can be by 6 He of PDMS film layer
Template 2 is spaced.The separation layer of formation is dichlorodimethylsilane film layer.
The paraffin oil dispersion liquid of dimethyl silicone polymer (Polydimethythylsiloxane, referred to as: PDMS) is poured
2 surface of template, then vacuumize process, so that PDMS is filled up completely in the micro-fluidic chip patterning of template 2.It vacuumizes
Time generally can be 20~40min, such as can form PDMS film layer 6 with vacuumize process 25min, 30min, 35min.Then
6 surface of PDMS film layer is bonded the first transparent glass slide 7, and is heated, so that the first transparent glass slide 7 and PDMS film layer
6 bondings are integral, and heating temperature is 50~70 DEG C, and heating time is 1~2h, are specifically detailed in Fig. 6.
Preferably, the PDMS film layer 6 of formation has micro-fluidic chip pattern, and the thickness of PDMS film layer 6 is not more than
0.25mm.If the thickness of PDMS film layer 6 is blocked up, the area contacted with air increases, and finished product is easy evaporation during the reaction,
Lead to deformation.
The PDMS film layer 6 for being bonded the first transparent glass slide 7 is separated with template 2, is obtained with structured chip,
This includes the first transparent glass slide 7 and PDMS film layer 6 with structured chip.
The chip of band structure and the with holes second transparent glass slide 8 are respectively placed under vacuum condition and carried out at plasma
Reason, so that the two, convenient for fitting, the second transparent glass slide 8 has several through-holes 81, and through-hole 81 is mainly used for being subsequently formed miniflow
The inlet and outlet of chip is controlled, two entrances and one outlet are such as formed.After plasma treatment, by the second transparent glass slide
8 and the chip of band structure carry out alignment fitting, and heat, so that the two closely bonds.During fitting, the second transparent load
Slide 8 is Nian Jie with PDMS film layer 6, to form " Sanming City of the first transparent glass slide 8 of transparent glass slide 7-PDMS film layer 6- second
Control " structure, is specifically detailed in Fig. 7.
It is after the second transparent 8 surface of glass slide coats one layer of glue, the female Luer 9 and through-hole 81 that have sealing cover is right
It is bonded, and is heated, and heating temperature is 50~70 DEG C, so that the female Luer 9 of with closure and the second transparent load
Slide 8 forms good bonding effect.
Encapsulating method provided by the invention carries out plasma treatment again after PDMS film layer is attached to transparent slide surface
It is bonded with the second transparent glass slide, to form air-locked transparent structural layers in PDMS film surface, is subtracted as much as possible
The area that few PDMS film layer is contacted with air, and the female Luer of sealing cover is had in inlet and outlet connection, to play
Have the effect of vaporization prevention performance, solves the contracting that micro- reaction member in digital pcr chip generates in temperature changing process
Small, the problems such as volatilizing, merge, deforming.
The present invention provides the PDMS micro-fluidic chip obtained using above-mentioned encapsulating method as a result,.It specifically is detailed in Fig. 8, it should
PDMS micro-fluidic chip includes the first transparent glass slide 7, the PDMS film layer 6 for being laminated in the first transparent 7 surface of glass slide, is laminated in
The transparent glass slide 8 of the second of 6 surface of PDMS film layer；
There are several through-holes 81 on second transparent 8 surface of glass slide, and the female Luer 9 of with closure is connected on through-hole 81.By
The female Luer 9 of with closure is connected to PDMS film layer 6, and forms the inlet and outlet of PDMS micro-fluidic chip.
The PDMS micro-fluidic chip, due to foring air-locked transparent structural layers in two apparent surface of PDMS film layer, and
And the female Luer of sealing cover is had in import and export bonding, the effect for preventing evaporation is effectively acted as, thus avoid micro- anti-
The diminution of answering unit to generate in temperature changing process, the problems such as volatilizing, merge, deforming.
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. within mind and principle should all include within protection scope of the present invention.
Claims (10)
1. a kind of encapsulating method of PDMS micro-fluidic chip vaporization prevention, which comprises the following steps:
Photoresist is coated on clean glass sheet surface by step S01., then carries out the processing that is heating and curing, and formation is layered in institute
State the photoresist layer of glass sheet surface；
Exposure mask with micro-fluidic chip patterning is fitted in the photoresist layer surface by step S02., and be exposed,
Development treatment obtains the template with micro-fluidic chip patterning；
The template is laid in glass baseplate surface by step S03., and in the glass baseplate surface along the template side
One layer tape is pasted along surrounding, the thickness of the adhesive tape is made to be greater than the thickness of the template, using insulating liquid to the attaching adhesive tape
Glass substrate carry out suffocating treatment, template surface formed one layer of separation layer, then by the paraffin oil of dimethyl silicone polymer
Dispersion liquid, which is poured, is distributed the paraffin oil homogeneous dispersion of the dimethyl silicone polymer to the template surface, vacuumize process
In the template surface, and the first transparent glass slide is sticked, curing process is carried out to the described first transparent glass slide, obtains band knot
The chip of structure；
The chip of the band structure and the template are carried out separating treatment by step S04.；
The chip of the band structure and the with holes second transparent glass slide are placed under vacuum condition and carry out plasma by step S05.
Processing, then by the two alignment fitting, heat treatment；
Step S06. coats one layer of glue in the with holes described second transparent slide surface, and by the female Luer of with closure
It is pasted on the described second transparent slide surface, is aligned the female Luer and the hole of the described second transparent glass slide, is formed
The inlet and outlet of micro-fluidic chip, heat treatment, obtains PDMS micro-fluidic chip.
2. the encapsulating method of PDMS micro-fluidic chip vaporization prevention as described in claim 1, which is characterized in that the poly dimethyl
The PDMS layer thickness that the paraffin oil dispersion liquid of siloxanes is formed by curing is not more than 0.25mm.
3. the encapsulating method of PDMS micro-fluidic chip vaporization prevention as described in claim 1, which is characterized in that the photoresist is
Positive photoresist or negative photoresist.
4. the encapsulating method of PDMS micro-fluidic chip vaporization prevention as claimed in claim 3, which is characterized in that the positive-tone photo
Glue is selected from any one of polymethyl methacrylate, adjacent diazo naphthoquinone-phenolic resin；The negative photoresist is selected from SU8 light
Photoresist.
5. the encapsulating method of PDMS micro-fluidic chip vaporization prevention as claimed in claim 1, which is characterized in that the plasma treatment
Time is 2~5min.
6. the encapsulating method of PDMS micro-fluidic chip vaporization prevention as described in claim 1, which is characterized in that add in step S05
The temperature of heat treatment is 50~70 DEG C.
7. the encapsulating method of PDMS micro-fluidic chip vaporization prevention as described in claim 1, which is characterized in that step S03's consolidates
Changing processing is that 1~3h is kept the temperature at 50~70 DEG C.
8. the encapsulating method of PDMS micro-fluidic chip vaporization prevention as described in claim 1, which is characterized in that developer solution is lactic acid
Any one of ethyl ester, tetramethylammonium hydroxide.
9. a kind of PDMS micro-fluidic chip, which is characterized in that including the first transparent glass slide, be laminated in the described first transparent load glass
The PDMS film layer on piece surface, the second transparent glass slide for being laminated in the PDMS film surface；
The second transparent slide surface has several through-holes, and the female Luer of with closure is connected on the through-hole.
10. PDMS micro-fluidic chip as claimed in claim 9, which is characterized in that the thickness of the PDMS film layer is not more than
0.25mm。
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| CN112403542A (en) * | 2020-11-05 | 2021-02-26 | 清华大学 | Micro-fluidic chip for quantitatively researching biofilm and manufacturing method thereof |
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