CN101021506A - Method for measuring low-speed electric interstifial flow in chip capillary cataphoresis - Google Patents
Method for measuring low-speed electric interstifial flow in chip capillary cataphoresis Download PDFInfo
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- CN101021506A CN101021506A CN200710021018.XA CN200710021018A CN101021506A CN 101021506 A CN101021506 A CN 101021506A CN 200710021018 A CN200710021018 A CN 200710021018A CN 101021506 A CN101021506 A CN 101021506A
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- electroosmotic flow
- probe
- microchip
- mobility
- chip
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Links
- 238000000034 method Methods 0.000 title claims abstract description 22
- 238000001962 electrophoresis Methods 0.000 title claims description 10
- 238000005370 electroosmosis Methods 0.000 claims abstract description 43
- 239000000523 sample Substances 0.000 claims abstract description 33
- 239000000126 substance Substances 0.000 claims abstract description 16
- 239000003792 electrolyte Substances 0.000 abstract description 8
- 238000005251 capillar electrophoresis Methods 0.000 abstract description 6
- 239000008151 electrolyte solution Substances 0.000 abstract description 2
- 239000007853 buffer solution Substances 0.000 abstract 1
- 229940088597 hormone Drugs 0.000 abstract 1
- 239000005556 hormone Substances 0.000 abstract 1
- 239000000463 material Substances 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 239000004372 Polyvinyl alcohol Substances 0.000 description 4
- 239000012530 fluid Substances 0.000 description 4
- 230000007935 neutral effect Effects 0.000 description 4
- 229920002451 polyvinyl alcohol Polymers 0.000 description 4
- 238000013016 damping Methods 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012544 monitoring process Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- GLDQAMYCGOIJDV-UHFFFAOYSA-N 2,3-dihydroxybenzoic acid Chemical compound OC(=O)C1=CC=CC(O)=C1O GLDQAMYCGOIJDV-UHFFFAOYSA-N 0.000 description 2
- YFMPSMITLLBENU-UHFFFAOYSA-N 3,4-dihydroxybenzylamine Chemical compound NCC1=CC=C(O)C(O)=C1 YFMPSMITLLBENU-UHFFFAOYSA-N 0.000 description 2
- 101100148256 Actinobacillus pleuropneumoniae apxIB gene Proteins 0.000 description 2
- 101100148259 Actinobacillus pleuropneumoniae apxIIA gene Proteins 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 229910021607 Silver chloride Inorganic materials 0.000 description 2
- 101150050411 appA gene Proteins 0.000 description 2
- 101150008617 appB gene Proteins 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 230000005684 electric field Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
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Abstract
A method can measure low speed electro-osmotic flow in chip capillary electrophoresis. First of all, measure effective mobility of probe substance by sample zonal method on quick electro-osmotic flow microchip A which being as a reference. The method is simplified as: add a quickly-migrating substance (probe) which can be tested into background electrolyte. Detect electrophoretic pattern when taking hormone background electrolyte with different density from background electrolyte as sample. Based on the peaking time in pattern, effective mobility (Mu eff) of probe substance on reference chip can be calculated. Then, taking same background electrolyte solution without probe as buffer solution and probe substance as sample, detect routine capillary electrophoresis on microchip B with unknown electro-osmotic flow. Apparent mobility (Mu npp) of probe on B can be calculated and curious effective mobility (Mu EOF) can be achieved by the following formula: Mu EOF=Mu npp-Mu eff.
Description
Technical field
The present invention relates to the measurement of low speed electroosmotic flow.Specifically, be based on an electromigratory material and measure the low speed electroosmotic flow indirectly in the constant principle of the ducted effective mobility of microchip.
Background technology
The micro-fluid control chip electrophoretic technology began to develop rapidly [referring to Manz from nineteen nineties, A., Graber, N., Widmer, H.M., Sens.Actuators B 1990, B1 (1-6), 244-248.], in order to satisfy the application of various aspects, often microchannel is carried out chemistry or physical modification, reduce the interaction of solute and tube wall and suppress electroosmotic flow.Therefore the electroosmotic flow property representation to microchannel just seems extremely important.
Commonly used electroosmotic flow assay method has neutral label method [referring to Jorgenson, J.W., Lukacs, K.D., Anal.Chem.1981,53,1298-1302.] and the current monitoring method [referring to: Huang, X., Gordon, M.J., Zare, R.N., Anal.Chem.1988,60,1837-1838.].The most commonly used is neutral label method, utilizes exactly to move under the effect of a kind of neutral compound in electric field and measures electroosmotic flow; The current monitoring method also is the very high method of a kind of utility ratio, the effect that drives by electric osmose in pipeline is with the solution in the alternative kapillary of a same electrolyte solution (comparing with the solution in the kapillary) that slightly dilutes, by electric current time history plot in the observation circuit, calculate electroosmotic flow according to whole times when replaced of pipeline middle and high concentration solution.
When needs are measured low electroosmotic flow or zero electroosmotic flow, difficulty has just appearred, and in neutral label method and current monitoring method, after through very long transit time, signal can become not obvious or not have signal.Up to the present, also not about hanging down the report of electroosmotic flow or zero electroosmotic flow assay method in the microchip capillary electrophoresis.
Summary of the invention
The purpose of this invention is to provide a kind of method of measuring low electroosmotic flow.
Technical scheme of the present invention is as follows:
The measuring method of low speed electroosmotic flow in a kind of chip capillary cataphoresis, it be at first one as the quick electroosmotic flow microchip A of reference on sample area band method [referring to Wang, W., Zhao, L., Jiang, L.P., Zhang, J.R., Zhu, J.J., Chen, H.Y., Electrophoresis 2006,27,5132-5137.] finish the mensuration of the effective mobility of probe substance, be summarized as follows, in back-ground electolyte, add a fast transferring and material can be determined (probe), after the class background electrolyte that is different from background electrolyte concentration is as sample feeding, obtain electrophoretogram, utilized the appearance time among the figure can calculate the effective mobility (μ of probe substance on the reference chip
Eff), then, utilize the identical background electrolyte that does not contain probe as damping fluid, on the microchip B of unknown electroosmotic flow, finish conventional Capillary Electrophoresis with probe substance as sample and detect, can calculate the apparent mobility (μ of probe on microchip B
App), can calculate the electroendosmotic mobility (μ that seeks knowledge according to following formula
EOF), μ
EOF=μ
App-μ
Eff
The effective mobility of a specific probe substance in the damping fluid of determining is invariable, and just the effective mobility of probe substance on reference chip and electroosmotic flow chip to be measured equates,
Therefore, μ
EffA1=μ
EffB1
μ wherein
EffA1And μ
EffB1Be respectively the effective mobility among microchip A and the microchip B.The effective mobility of this probe substance can calculate from its apparent mobility and electroosmotic flow,
μ in the above formula
AppA1, μ
EOFAAnd V
ABe respectively apparent mobility, electroendosmotic mobility and the separation voltage of the probe substance among the microchip A, t
A1And t
A2Be respectively the appearance time of microchip A middle probe material and electroosmotic flow.μ
AppB1, μ
EOFBAnd V
BRepresent the respective amount among the microchip B respectively.L
A, L
Aeff, L
BAnd L
BeffBe respectively length and the effective length of microchip A and microchip B.Therefore, μ
EOFBCan represent with following formula,
If L
A=L
B=L, L
Aeff=L
Beff=L
EffAnd V
A=V
B=V, μ
EOFBCan be calculated as follows and get.
Four, description of drawings
Fig. 1 is an applied microchip synoptic diagram when measuring electroosmotic flow among the present invention, and wherein: A is a buffer pool, and B is the separating pipe outlet, C is a buffer pool, and D is the sample waste liquid pool, and E is sample channel and separating pipe crossing, F is the damping fluid waste liquid pool, and WE is an electrochemical working electrode.。
The electrophoresis synoptic diagram that Fig. 2 records with indirect method on reference chip A for the present invention, t
A1And t
A2Be respectively the appearance time of microchip A middle probe material and electroosmotic flow.
The electrophoresis synoptic diagram that Fig. 3 records with direct method on electroosmotic flow chip B to be measured for the present invention, t
B1And t
B2Be respectively the transit time of microchip B middle probe material and electroosmotic flow.Wherein: the dotted line peak is the signal peak of imaginary electroosmotic flow.
Embodiment
The measurement of the electroosmotic flow of poly dimethyl silica silicon (PDMS) microchip of embodiment 1.Brij 56 modifieds
Electroosmotic flow when being full of phosphate buffer (20mM PBS pH7.00) in the PDMS microchip pipeline of measurement Brij 56 modifieds, with 0.1mM 3,4-dihydroxy benzylamine (DHBA) is as probe substance, 20mM PBS is electrolyte as a setting, not make the PDMS microchip of modification as reference chip A, wherein: AE=0.3cm; EB=3.6cm; CE=ED=0.5cm.In the Capillary Electrophoresis process, separation voltage is set at 800V, detects current potential+1.2V (vs.Ag/AgCl electrode), according to the transit time of measuring gained, t
B1=59.2s, t
A1=41.2s, t
A1=65.0s, the size that calculates electroosmotic flow is (1.40 ± 0.04) * 10
-4Cm
2/ (Vs).Consistent with the document guess value [referring to: Dou, Y.H., Bao, N., Xu, J.J., Meng, F., Chen, H.Y., Electrophoresis 2004,25,3024-3031.].
The measurement of the electroosmotic flow of the PDMS microchip pipeline of embodiment 2. polyvinyl alcohol (PVA) (PVA) modified
Electroosmotic flow when being full of phosphate buffer (20mM PBS pH7.00) in the PDMS microchip pipeline of measurement PVA modified, with 0.1mM 3,4-dihydroxy benzylamine (DHBA) is as probe substance, 20mM PBS is electrolyte as a setting, not make the PDMS microchip of modification as reference chip A, wherein: AE=0.3cm; EB=3.6cm; CE=ED=0.5cm.In the Capillary Electrophoresis process, separation voltage is set at 800V, detects current potential+1.2V (vs.Ag/AgCl electrode), according to the transit time of measuring gained, t
B1=98.7s, t
A1=41.2s, t
A1=65.0s, the size that calculates electroosmotic flow is (0.22 ± 0.02) * 10
-4Cm
2/ (Vs).Consistent with the document guess value [referring to: Wu, D., Luo, Y., Zhou, X., Dai, Z., Lin, B., Electrophoresis 2005,26,211-218.].
Claims (2)
1. the measuring method of low speed electroosmotic flow in the chip capillary cataphoresis is characterized in that: the effective mobility μ that at first records probe substance on a quick electroosmotic flow microchip A as reference
Eff, record the apparent mobility μ of probe substance on the microchip B of electroosmotic flow to be measured then
App, calculate unknown electroendosmotic mobility μ according to following formula
EOF,
μ
EOF=μ
app-μ
eff?。
2. the method for the low electroosmotic flow of fast measuring micro-fluidic chip according to claim 1 is characterized in that: with sample area band method on electroosmotic flow reference chip fast once the property transit time of finishing probe substance and electroosmotic flow measure, directly calculate effective mobility.
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104122316A (en) * | 2014-06-30 | 2014-10-29 | 中国科学院化学研究所 | Method for simultaneously measuring mobility and dielectric mobility of particles by use of distributed direct-current electric field |
CN108007994A (en) * | 2017-06-27 | 2018-05-08 | 北京理工大学 | A kind of method for measuring Capillary Electrophoresis electroosmotic flow |
CN114660283A (en) * | 2022-05-24 | 2022-06-24 | 佛山微奥云生物技术有限公司 | Immunoassay plate type chip based on electrical acceleration and preparation method thereof |
-
2007
- 2007-03-22 CN CNB200710021018XA patent/CN100473986C/en not_active Expired - Fee Related
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104122316A (en) * | 2014-06-30 | 2014-10-29 | 中国科学院化学研究所 | Method for simultaneously measuring mobility and dielectric mobility of particles by use of distributed direct-current electric field |
CN108007994A (en) * | 2017-06-27 | 2018-05-08 | 北京理工大学 | A kind of method for measuring Capillary Electrophoresis electroosmotic flow |
CN108007994B (en) * | 2017-06-27 | 2019-12-03 | 北京理工大学 | A method of measurement Capillary Electrophoresis electroosmotic flow |
CN114660283A (en) * | 2022-05-24 | 2022-06-24 | 佛山微奥云生物技术有限公司 | Immunoassay plate type chip based on electrical acceleration and preparation method thereof |
CN114660283B (en) * | 2022-05-24 | 2022-09-13 | 佛山微奥云生物技术有限公司 | Immunoassay plate type chip based on electrical acceleration and preparation method thereof |
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