CN101000290A - Sample enrichment chip, manufacturing method and enrichment method and on micronano structure - Google Patents
Sample enrichment chip, manufacturing method and enrichment method and on micronano structure Download PDFInfo
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- CN101000290A CN101000290A CN 200710036415 CN200710036415A CN101000290A CN 101000290 A CN101000290 A CN 101000290A CN 200710036415 CN200710036415 CN 200710036415 CN 200710036415 A CN200710036415 A CN 200710036415A CN 101000290 A CN101000290 A CN 101000290A
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Abstract
The invention relates to sample enrichment chip, manufacturing method and enrichment method based on micro nanostructure. Its feature is that the enrichment chip is used quartz glass as backing material, formed by enrichment nanometer channel and micro sample transmitting pipeline. It applies MEMS technique to process the channel and pipeline one the quartz glass, strictly control channel depth to make it satisfy ion falling demand, utilizes low temperature bonding method to process for the substrate and covering piece, fills sample into chip pipeline, adds direct voltage in the sample pool to form electric field in nanometer channel. Because of Debye layers in the channel are superposed, while ion falling zoon is formed beside the channel. The sample moved under the action of the electric field is enriched beside the channel to form sample enrichment zoon. The chip has small volume, will not destroy enrichment component while enriching.
Description
Technical field
The present invention relates to a kind ofly relate to a kind of example enrichment chip, method for making or rather and subside the example enrichment method of principle based on example enrichment chip, the method for making of micro nano structure efficiently, belong to the analytical chemistry field based on ion based on micro nano structure.
Background technology
Micro-total analysis system (claiming chip lab again) is the notion that is proposed by people such as Manz in the nineties in last century, be that micro-electromechanical technology (MEMS) and analytical chemistry detection are combined, specimen preparation related in the fields such as biological and chemical, biological with chemical reaction, separate, basic operation unit such as detection is all integrated or be partially integrated on more than one square centimeters the chip, in order to finishing different biological or chemical courses of reaction, and the technology that its product is analyzed.Through ten years development, very fast to development aspect nucleic acid, protein, organic compound and separate inorganic ions, analysis and the detection system on microchip, there has been integrated commercialization micro-total analysis system to occur, as the MCE-2010 type microchip electrophoresis system of Agilent company 2100 micro-analysis systems and the release of Tianjin, island company.Yet, the enrichment of biological sample with separate micro-system development and relatively lag behind, this is because the diversity (as blood, urine, ight soil, contaminant water etc.) of biological sample on the one hand, form have nothing in common with each other (gas, liquid, solid etc.); On the other hand, biological sample also has multiple complicated ingredient, is detecting phase mutual interference when separating.And become the bottleneck that restricts whole micro-total analysis system integrated development, also be one of emphasis of research at present.The chip that is used for biological sample separation and enrichment is made of some microchannels and microcavity body usually, according to its performed specific function and task, may also can comprise some other assembly, as electrode, heating/cooling device, filter membrane, piezoelectric ceramic piece etc.What the chip preparation was used is the micro fabrication of comparative maturity in microelectronics industry and the MEMS (micro electro mechanical system) industry, as photoetching, reactive ion etching, little injection casting and polymeric membrane placing, on base materials such as glass, plastics, silicon chip, process the microstructure of the micron-scale that is used for biological sample separation or reaction.The physics, chemistry and the biological property that on microchip, mostly the separation and the enrichment of composition to be measured in the biological sample are to utilize composition to be measured itself, conventional enrichment method and microchip enrichment method are as broad as long on the enrichment principle, but after system's microminiaturization, can combine the principle of multiple separation and enrichment easily, improve and separate and bioaccumulation efficiency, formed the new feature of biological sample separation and enrichment.
Miniature organism example enrichment and separating chips should have the characteristics of the following aspects preferably: 1) higher enrichment ratio is arranged; 2) nonspecific residual component is less; 3) enrichment speed is very fast; 4) process of enrichment does not influence the character of enrichment composition.Still the report that rarely has at present this respect.According to these four characteristics.The present invention proposes a kind of example enrichment chip, method for making and enrichment method based on micro nano structure.
Summary of the invention
The object of the present invention is to provide a kind of example enrichment chip, method for making and enrichment method based on micro nano structure.Specifically on silica glass material, adopt photoetching, nanoscale wet etching and the low-temperature bonding fabrication techniques nanometric sample enrichment chip of standard, and realize a kind of example enrichment method of high power example enrichment therein.
The gordian technique that the present invention need solve is:
1, the control of nano-channel corrosion depth
In the nanoscale wet etching method, corrosion depth is relevant with a plurality of factors, comprising: the environment temperature of material property, corrosive liquid proportioning, corrosion.Under the prerequisite that material property is determined, the present invention adopts under the water-bath corrodes operation, and the temperature of control corrosion operating environment is at 40 ℃; The proportioning of strict control corrosive liquid.Control the nanoscale Corrosion of Pipeline degree of depth thus.
2, the configuration of enrichment damping fluid
The ion concentration of damping fluid directly affects the thickness of pipeline debye layer, just affects concentration effect.
3, the control of extra electric field
In order to realize better sample enrichment, must rationally control extra electric field, thereby control the distribution of electric field under the micro-nano-scale and the distribution of buffering system intermediate ion.
The technical solution used in the present invention is: at first use MEMS technology and go out nano-channel and sample transport channel in the quartz glass surface working, strict control nanometer trench depth makes it meet ion and subsides requirement; Utilize low-temperature bonding method, will accomplish fluently the substrate and the cover plate low-temperature bonding of sample well.In the chip pipeline, pour into the sample that needs enrichment then, between sample cell, add DC voltage, in the nanometer ditch, form electric field; Since the stack of debye layer in the nano-channel, and subside band at the other ion that forms of nanometer ditch; The sample that moves under electric field action owing to can't subside band and in the other enrichment of nanometer ditch, form the example enrichment band by ion.Last switched voltage is shifted the example enrichment band onto surveyed area and is detected.(concrete processing step and enrichment method see embodiment 1,2 for details)
Advantage then of the present invention and beneficial effect are:
1, the processing base material of the present invention's employing is a quartz glass, and its source is abundant, and the processing technology maturation can be produced in batches, and can reuse.
2, miniature organism example enrichment chip volume of the present invention is little, is easy to carry, and does not rely on main equipments such as hydro-extractor, operates comparatively simple.
What 3, adopt in the enrichment process is the physics enrichment method that ion is subside band, does not therefore destroy the structure of enrichment composition in enrichment process.
4, adopting quartz glass enrichment chip, this chip are convenient to clean, and can repeat example enrichment.
According to the example enrichment chip that above-mentioned preparation method and enrichment method provide based on micro nano structure, be base material with the quartz glass, to form by enrichment nano-channel and micron order sample transmission pipeline, enrichment nano-channel frame is at two micron order pipeline enclosures.The described nano-channel degree of depth is 50nm.Micron order sample transmission pipeline is wide 50 microns, dark 10 microns.
Description of drawings
Fig. 1 enrichment chip structural representation: 1 is the enrichment nano-channel, and 2 is micron order sample transmission pipeline
Fig. 2 nanotubes manufacture craft process flow diagram: (a) chromium plating on quartz substrate (b) steaming HMDS (c) resist coating 6809 (d) photoetching (e) corrosion chromium layer (f) glass corrosion (g) removes photoresist and chromium layer (h) bonding
Fig. 3 nano-channel structural drawing: (a) nano-channel AFM design sketch (b) nano-channel structure photo is the nano-channel of frame at two microns pipeline enclosures in the circle
Fig. 4 enrichment process synoptic diagram: (a) sample feeding (b) example enrichment (c) band detects
Embodiment
Embodiment 1 is based on the manufacture craft process of the example enrichment chip of micro nano structure
One, at first, use MEMS technology and go out nano-channel and nanoscale transmission pipeline in the quartz glass surface working
A. the micron order transmission pipeline is made
1) photoetching for the first time
1. clean: use acetone, ethanol, deionized water ultrasonic cleaning quartz glass substrate 5min respectively; Nitrogen dries up; Oven dry,
2. evaporate HMDO, resist coating 6809 whirl coatings, speed is 4000rpm, the whirl coating time is 30s,
3. baking before: 100 ℃, 6min
4. photoetching: with 12mW/cm
2The intensity photoetching, the machine photoetching, 12s,
5. develop: 18s,
6. the film 10s that feels secure,
7. 120 ℃ of baking oven 30min of post bake;
2) micron order corrosive pipeline
1. dechromise: use by ammonium ceric nitrate 400g+ perchloric acid 110mL, deionized water is settled to the chromium corrosive liquid of 1.76L preparation and removes visuals chromium layer,
2. washed with de-ionized water is 10 times,
3. glass corrosion: the volume ratio of putting into 40-50 ℃ of constant temperature is HF: H
2O
2: HAC=2: corrode 40-50min in the quartz glass corrosive liquid of forming at 1: 1, obtain widely 50 microns, dark 10 microns micron order transfer tubes arrive,
4. washed with de-ionized water is 10 times;
B. the making of nano-groove
1) photoetching for the second time
1. clean: use acetone, ethanol, deionized water ultrasonic cleaning quartz glass substrate 5min respectively; Nitrogen dries up; Oven dry 20min,
2. whirl coating: get rid of photoresist 6809 with 4000rpm speed, thickness is 0.9 μ m,
3. baking before: 100 ℃, 6min,
4. aim at photoetching: working strength is 12mW/cm
2Litho machine photoetching 12s,
5. develop: 18s,
6. the film 10s that feels secure,
7. 120 ℃ of baking oven 30min of post bake;
2) making of nano-channel
1. dechromise: use by ammonium ceric nitrate 400g+ perchloric acid 110mL, deionized water is settled to the chromium corrosive liquid of 1.76L preparation and removes visuals chromium layer
2. nanometer corrosion: what put into 40-50 ℃ of constant temperature is HF: H by volume ratio
2O
2: HAC=2: the quartz glass corrosive liquid of forming at 5: 5 stirs, and 60s is dark about 50nm;
3) punching: diamond hole diameter on the substrate relevant position is the hole of 2mmol/L, as liquid storage tank.
Two, low-temperature bonding
1. acetone removes the remained on surface photoresist, ethanol and washed with de-ionized water,
2. dechromise in the surface: with substrate put into prepare with step 2) described chromium corrosive liquid, the 3min layer that dechromises in ultrasonic pond,
3. acetone, ethanol, deionized water ultrasonic cleaning substrate, cover plate 5min
4. the Piranha washing lotion is cleaned. substrate and cover plate are put into by volume ratio H
2SO
4: H
2O
2=4: the 1 Piranha washing lotion of forming, in hot plate; Increase surface affinity; Washed with de-ionized water,
5. again by volume ratio NH
3H
2O: H
2O: H
2O
2=6: hot plate 30min in the affine washing lotion that form at 3: 1; Washed with de-ionized water 10 times,
6. the pre-bonding of applying chip in the water: directly substrate and cover plate are fitted in deionized water, transfer in the vacuum drying chamber then. set the vacuum drying chamber temperature and be 100 ℃ and open vacuum pump, close vacuum pump after continuing to vacuumize 30min, keep 100 ℃,
7. bonding: slowly rise to powered-down natural cooling behind 200 ℃ of 4～6h, realize that low-temperature bonding makes the nanometer enrichment chip.
The enrichment method of embodiment 2 enrichment chips
On the chip basis behind embodiment 1 low-temperature bonding, adopt following two processing steps to realize enrichment
1, the surface treatment of chip: use the NaOH solution of 1mol/L to charge in the good chip of firm bonding, wash 3 times after, leave standstill 30min, the use deionized water rinsing.
2, adopting the albumen of FITC fluorochrome label is sample, and 0.75mmol/L boric acid is damping fluid; Bring the sample accent into sample to the nanoparticle downcast area with the 30V DC voltage, shown in Fig. 4 (a); Sample strip is because the ion that nano-area causes is subside, and constantly accumulation, enrichment is shown in Fig. 4 (b); Working voltage will compress good sample and transfer band, introduce surveyed area, carry out sample detection, shown in Fig. 4 (c).
Claims (10)
1, a kind of example enrichment chip based on micro nano structure is characterized in that described enrichment chip is to be base material with the quartz glass, is made up of enrichment nano-channel and micron order sample transmission pipeline, and enrichment nano-channel frame is at two micron order pipeline enclosures.
2, by the described example enrichment chip of claim 1, it is characterized in that the described nano-channel degree of depth is 50nm based on micro nano structure.
3, by the described example enrichment chip of claim 1, it is characterized in that described micron order sample transmission pipeline is wide 50 microns, dark 10 microns based on micro nano structure.
4, preparation is as the method for claim 1,2 or 3 described example enrichment chips based on micro nano structure, it is characterized in that at first using MEMS technology and go out nano-channel and micron order sample transmission raceway groove in the quartz glass surface working, the control nano-channel degree of depth makes it meet ion and subsides requirement; Then, utilize low-temperature bonding method, will accomplish fluently the substrate and the cover plate low-temperature bonding of sample well.
5, the example enrichment chip production method based on micro nano structure as claimed in claim 4 is characterized in that preparing concrete steps and is:
1) photoetching for the first time
1. clean: use acetone, ethanol, deionized water ultrasonic cleaning substrate 5min respectively; Nitrogen dries up; Oven dry,
2. evaporate HMDO, resist coating 6809 whirl coatings, speed is 4000rpm, the whirl coating time is 30s,
3. baking before: 100 ℃, 6min
4. photoetching: with 12mW/cm
2The intensity photoetching, the machine photoetching, 12s,
5. develop: 18s,
6. the film 10s that feels secure,
7. 120 ℃ of baking oven 30min of post bake;
2) micron order corrosive pipeline
1. dechromise: use by ammonium ceric nitrate 400g+ perchloric acid 110mL, deionized water is settled to the chromium corrosive liquid of 1.76L preparation and removes visuals chromium layer,
2. washed with de-ionized water is 10 times,
3. glass corrosion: the volume ratio of putting into 40-50 ℃ of constant temperature is HF: H
2O
2: HAC=2: corrode 40-50min in the quartz glass corrosive liquid of forming at 1: 1, obtain widely 50 microns, dark 10 microns micron order transfer tubes arrive,
4. washed with de-ionized water is 10 times;
3) photoetching for the second time
1. clean: use acetone, ethanol, deionized water ultrasonic cleaning substrate 5min respectively; Nitrogen dries up; Oven dry 20min,
2. whirl coating: get rid of photoresist 6809 with 4000rpm speed, thickness is 0.9 μ m,
3. baking before: 100 ℃, 6min,
4. aim at photoetching: working strength is 12mW/cm
2Litho machine photoetching 12s,
5. develop: 18s,
6. the film 10s that feels secure,
7. 120 ℃ of baking oven 30min of post bake;
4) making of nano-channel
1. dechromise: use by ammonium ceric nitrate 400g+ perchloric acid 110mL, deionized water is settled to the chromium corrosive liquid of 1.76L preparation and removes visuals chromium layer
2. nanometer corrosion: what put into 40-50 ℃ of constant temperature is HE: H by volume ratio
2O
2: HAC=2: the quartz glass corrosive liquid of forming at 5: 5 stirs, and 60s is dark about 50nm;
5) punching: diamond hole diameter on the substrate relevant position is the hole of 2mmol/L, as liquid storage tank.
6) low-temperature bonding
1. acetone removes the remained on surface photoresist, ethanol and washed with de-ionized water,
2. dechromise in the surface: with substrate put into prepare with step 2) described chromium corrosive liquid, the 3min layer that dechromises in ultrasonic pond,
3. acetone, ethanol, deionized water ultrasonic cleaning substrate, cover plate 5min
4. the Piranha washing lotion is cleaned. substrate and cover plate are put into by volume ratio H
2SO
4: H
2O
2=4: the 1 Piranha washing lotion of forming, in hot plate; Increase surface affinity; Washed with de-ionized water,
5. again by volume ratio NH
3H
2O: H
2O: H
2O
2=6: hot plate 30min in the affine washing lotion that form at 3: 1; Washed with de-ionized water 10 times,
6. the pre-bonding of applying chip in the water: directly substrate and cover plate are fitted in deionized water, transfer in the vacuum drying chamber then. set the vacuum drying chamber temperature and be 100 ℃ and open vacuum pump, close vacuum pump after continuing to vacuumize 30min, keep 100 ℃,
7. bonding: slowly rise to powered-down natural cooling behind 200 ℃ of 4～6h, realize that low-temperature bonding makes the nanometer enrichment chip.
6, by the described example enrichment chip production method based on micro nano structure of claim 5, cleaning the back bake out temperature when it is characterized in that photoetching for the first time is 120 ℃, and the time is 20min.
7, by the described example enrichment chip production method based on micro nano structure of claim 5, the whirl coating time is 30s when it is characterized in that photoetching for the second time.
8, by the described example enrichment chip production method of claim 5 based on micro nano structure, it is characterized in that in the low-temperature bonding technology, substrate and cover plate are put into the Piranha washing lotion hot plate 10-15min.
9, the enrichment method of the example enrichment chip based on micro nano structure as claimed in claim 1, after it is characterized in that substrate and cover plate low-temperature bonding, adopt ion to subside the physics enrichment method of band, in the chip pipeline, pour into earlier the sample that needs enrichment, between sample cell, add DC voltage, in the nanometer ditch, form electric field; The stack of debye layer in nano-channel makes the other ion that forms of nanometer ditch subside band; The sample that moves under electric field action be owing to can't subside band by ion, and can only be in the other enrichment of nanometer ditch, thereby forms the example enrichment band.
10, by the enrichment method of the described example enrichment chip based on micro nano structure of claim 9, it is characterized in that the concrete steps of enrichment are:
1. the surface treatment of chip: use the NaOH solution of 1mol/L to charge in the good chip of firm bonding, wash 3 times after, leave standstill 30min, use deionized water rinsing again;
2. the albumen that adopts the FITC fluorochrome label is sample, and 0.75mmol/L boric acid is damping fluid; Bring the sample accent into sample to the nanoparticle downcast area with the 30V DC voltage, sample strip is subside at the ion that nano-area causes, and constantly piles up and enrichment.
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| CN101859074A (en) * | 2010-07-15 | 2010-10-13 | 深圳市路维电子有限公司 | Cleaning solution for dry plate developing tank and cleaning method thereof |
| CN101796412B (en) * | 2007-07-23 | 2013-03-27 | 纳诺恩科技有限公司 | Chip for analyzing fluids |
| CN103105415A (en) * | 2013-01-14 | 2013-05-15 | 江苏大学 | Experimental chip for inspecting characteristics of hot air bubbles in nanometer channel and manufacturing method of experimental chip |
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| CN104513984B (en) * | 2013-09-30 | 2017-02-15 | 苏州博洋化学股份有限公司 | Method for preparing silver-chromium stripping liquid |
| WO2017045264A1 (en) * | 2015-09-18 | 2017-03-23 | 北京工业大学 | Plasma-assisted microstructure alignment and pre-bonding method of glass or quartz chip |
| CN109060739A (en) * | 2018-07-13 | 2018-12-21 | 中国科学院上海微系统与信息技术研究所 | A method of detection hydrogen peroxide |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101796412B (en) * | 2007-07-23 | 2013-03-27 | 纳诺恩科技有限公司 | Chip for analyzing fluids |
| CN101859074A (en) * | 2010-07-15 | 2010-10-13 | 深圳市路维电子有限公司 | Cleaning solution for dry plate developing tank and cleaning method thereof |
| CN101859074B (en) * | 2010-07-15 | 2012-05-23 | 深圳市路维电子有限公司 | Cleaning method for dry plate developing tank |
| CN103105415A (en) * | 2013-01-14 | 2013-05-15 | 江苏大学 | Experimental chip for inspecting characteristics of hot air bubbles in nanometer channel and manufacturing method of experimental chip |
| CN104513984B (en) * | 2013-09-30 | 2017-02-15 | 苏州博洋化学股份有限公司 | Method for preparing silver-chromium stripping liquid |
| CN104868054A (en) * | 2015-04-28 | 2015-08-26 | 吉林建筑大学 | Method for fixing flexible material substrate by utilizing photoresist |
| WO2017045264A1 (en) * | 2015-09-18 | 2017-03-23 | 北京工业大学 | Plasma-assisted microstructure alignment and pre-bonding method of glass or quartz chip |
| US9842749B2 (en) | 2015-09-18 | 2017-12-12 | Beijing University Of Technology | Plasma assisted method of accurate alignment and pre-bonding for microstructure including glass or quartz chip |
| CN105912766A (en) * | 2016-04-07 | 2016-08-31 | 东北电力大学 | Electric nanofluid enrichment loss mechanism simulation method |
| CN109060739A (en) * | 2018-07-13 | 2018-12-21 | 中国科学院上海微系统与信息技术研究所 | A method of detection hydrogen peroxide |
| CN109991346A (en) * | 2019-04-18 | 2019-07-09 | 南京大学 | A kind of micro-fluidic Ultraviolet Oxidation device for organic nitrogen analysis |
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