CN100394172C - Micro-capillary electropheresis apparatus - Google Patents
Micro-capillary electropheresis apparatus Download PDFInfo
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- CN100394172C CN100394172C CNB2005100057462A CN200510005746A CN100394172C CN 100394172 C CN100394172 C CN 100394172C CN B2005100057462 A CNB2005100057462 A CN B2005100057462A CN 200510005746 A CN200510005746 A CN 200510005746A CN 100394172 C CN100394172 C CN 100394172C
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Abstract
The present invention relates to a micro-capillary electropheresis apparatus which is mainly composed of a capillary electrophoresis system, an electrophoresis spectrogram collection system and an image collection system, wherein in the capillary electrophoresis system, two liquid bottles are connected by a capillary to form a capillary channel, and the two liquid bottles are respectively connected with both ends of a high-voltage power supply; in the electrophoresis spectrogram collection system, one side edge of a capillary is provided with a light source, the other side edge of the capillary corresponding to one side edge is provided with a photoelectric converter, and the photoelectric converter is electrically connected with an electrophoresis spectrogram collection display; in the image collection system, a light source of a microscope is arranged on one side edge of a capillary, and the other side edge of the capillary corresponding to one side edge is provided with a microscope; the microscope is connected with an image collection apparatus, and the image collection apparatus is connected with an image display processor. The present invention can realize real-time observation of an electrophoresis spectrogram and a detection object, high-speed electrophoresis of micro images, single-cell or single-particle analysis and simultaneous rapid analysis of congener and non-congener multiple particles.
Description
Technical field
The present invention relates to Capillary Electrophoresis, two kinds of technology of micro-imaging, belong to Capillary Electrophoresis and micro-imaging coupling new technology.
Background technology
Capillary Electrophoresis belongs to the new separation technology over nearly 20 years, and micro-imaging then belongs to the observation technology of comparative maturity, but both couplings then belong to the innovative technology invention.Capillary Electrophoresis is one of current research focus, and microscope is one of important means of biology and other scientific research.At present, the detection means of Capillary Electrophoresis, no matter be ultraviolet (UV), laser-induced fluorescence (LIF) (LIF), still diode array (DAD) or other means all can't make the experimenter obtain electrophoresis pattern and measured object pattern proterties simultaneously, to overlap peak, can only ignore measured object, by the various conditions of conversion, make the peak separately, otherwise can't analyze; Traditional microelectrophoresis can be observed the shape characteristic of particulate samples, but can't realize efficient, at a high speed and the high automation operation resemble the Capillary Electrophoresis, does not also possess the spectral analysis function, therefore has significant deficiency on using.So combine as advantage with Capillary Electrophoresis and micro-imaging, can realize micro-image high speed electrophoresis, unicellular or single-particle analysis, the express-analysis simultaneously of similar and non-similar multiparticle, behavior is intuitively observed, and has great value and economic benefit.
When high pressure is added to two ends capillaceous because the localization charge affects of capillary tube inner wall can produce electric osmose, have electrophoresis if measured object has electric charge, its finally show as the two vector and.In most aqueous solution, the inside surface meeting of quartz or glass capillary is because dissociating of silicon hydroxyl, and producing negative localization electric charge, many organic materials such as teflon, polystyrene etc. also can produce negative localization electric charge because of residual carboxyl, consequently produce the electric osmose of pointing to negative pole.Because in Capillary Electrophoresis, the numerical value of electric osmose is usually than big one to two order of magnitude of electrophoresis numerical value, so can realize the motion in the same way of all samples component.Have under negative localization electric charge (can change electrically by the coating sometimes) situation at capillary tube inner wall, the peak order that goes out that separates the back measured object is: positive ion, neutral molecule, negative ion.Different components because of its with the difference of electric charge, size, structure etc., finally realize separating (neutral molecule total with electric osmose with fast, must not separate).Move to the other end by an end capillaceous.In the process of motion, each component of sample is concentrated and separates.When sample process electrophoresis spectrogram acquisition system, note the electrophoresis pattern of sample by it; When sample process image collection processing system, note the image and the processing of sample by it.
Micro-imaging is to utilize microscope to amplify and the electronic image capture technology, measured object is carried out image amplify, collect and handle, and finally realizes the effect of observing samples particle directly perceived.
Summary of the invention
The object of the invention is to provide a kind of micro-capillary electropheresis apparatus, can obtain electrophoresis pattern and the online proterties of measured object simultaneously, particularly utilizes the coupling of the two to obtain more more conclusive observed results directly perceived.The present invention can expand to the monitoring of a plurality of point and mobile monitoring or tracking monitor, so that obtain more multidata.
For achieving the above object, micro-capillary electropheresis apparatus provided by the invention mainly is made up of capillary electrophoresis system, electrophoresis spectrogram acquisition system and image capturing system; Wherein:
Capillary electrophoresis system: connect with kapillary between two liquid bottles, form capillary channel, two liquid bottles connect the two ends of high-voltage power supply respectively;
The electrophoresis pattern acquisition system: kapillary one side is provided with light source, and another side of corresponding kapillary is provided with photoelectric commutator, and photoelectric commutator and electrophoresis pattern are collected display and electrically connected;
Image capturing system: a microscope light source is arranged on a side capillaceous, and another side of corresponding kapillary is provided with microscope, and this microscope connects an image acquisition device, connects image display processor by image acquisition device.
Light source in the electrophoresis pattern acquisition system is laser instrument, light emitting diode or deuterium lamp, tungsten lamp, Halogen lamp LED.
Be furnished with optical filter and slit on the light source in the electrophoresis pattern acquisition system, shape of slit and size are optional.
Microscope in the image capturing system is optical microscope or adds object lens assemblings by light source that wherein object lens are magnifier, microscope camera lens or CCD.
Used image capturing system is a plurality of settings, and multiple spot is monitored simultaneously on a capillary.
Electrophoresis spectrogram acquisition system and image capturing system can move separately and control, also linkage control.
Translational speed, the path of electrophoresis spectrogram acquisition system and image capturing system are adjustable.
Description of drawings
Fig. 1 is a micro-capillary electropheresis apparatus flow process frame diagram.
Fig. 2 is the structural representation of micro-capillary electropheresis apparatus of the present invention, and 1 for high-voltage power supply, 2 is that kapillary, 3 is the liquid bottle of electrode, 4 for sealing among the figure, constitutes electrophoresis system; 5 is that light source, 6 is collected display for photoelectric commutator, 9 for electrophoresis pattern for slit, 8 for optical filter, 7, constitutes the electrophoresis pattern acquisition system; 10 is that microscope light source, 11 is microscope optical filter and focusing system, and 12 is that micro objective, 13 is that image acquisition device, 14 is image display processor, composing images acquisition system.
Fig. 3 be the erythrocyte of rabbit in micro-capillary electropheresis apparatus of the present invention, adopt UV-detector, the electrophoresis spectrogram that obtains at the 214nm wavelength.
Image (A is single cell dispersion image, and B is the cell aggregation volume image) when Fig. 4 measures high-voltage power supply for the erythrocyte of rabbit in micro-capillary electropheresis apparatus of the present invention in the motion.
The erythrocyte that Fig. 5 behaves adopts UV-detector in micro-capillary electropheresis apparatus of the present invention, the electrophoresis spectrogram that obtains at the 214nm wavelength.
Image (A is single cell dispersion image, and B is a cell aggregation autologous mononuclear cell image together) when the erythrocyte that Fig. 6 behaves is measured high-voltage power supply in micro-capillary electropheresis apparatus of the present invention in the motion.
Embodiment
See also Fig. 1 and Fig. 2, the concrete structure of micro-capillary electropheresis apparatus of the present invention is:
Connect with kapillary 2 between two liquid bottles 4, form capillary channel, two liquid bottles also connect high-voltage power supply 1 by electrode 3 respectively, constitute electrophoresis system with this.Making for transparent material of kapillary 2, its diameter, shape are unrestricted, can select and change according to needs.High-voltage power supply is known direct supply, can regulate the voltage and current of the different sizes of output.
Be respectively equipped with electrophoresis pattern acquisition system and image capturing system around kapillary 2.Wherein the formation of electrophoresis pattern acquisition system is: at kapillary 2 one sides light source 5 is set, corresponding is provided with photoelectric commutator 8 at kapillary 2 another sides, photoelectric commutator 8 is collected display 9 with electrophoresis pattern and is connected, this electrophoresis pattern is collected display can adopt the chromatographic work station of Dalian Chemiclophysics Inst., Chinese Academy of Sciences, and other chromatographic work stations also can; In order to adapt to different samples, also be provided with optical filter 6 and slit 7 on the light source 5.The light source 5 that the present invention adopts can be laser instrument, light emitting diode or deuterium lamp, tungsten lamp, Halogen lamp LED, and can dispose suitable optical filter according to detecting needs.Slit 7 comprises different shape and size, and can regulate.
Image capturing system also is to be provided with around kapillary 2: a microscope light source 10, be arranged on a side of kapillary 2, corresponding another side at kapillary 2 is provided with microscope 12, this microscope 12 connects an image acquisition device 13, connect image display processor 14 again by image acquisition device 13, this image acquisition device and image display processor can adopt the CCD of the TK-C1381EG type that Japanese JVC company produces and the OK-C30A image pick-up card of robotization research institute of the Chinese Academy of Sciences; The same with aforementioned electrophoresis pattern acquisition system, microscope light source 10 is provided with optical filter and focusing system 11, and about configuration optical filter and focusing system on microscope light source are known technologies, the present invention is not described in detail.The present invention can select and changes according to the device that different microscopes or different light sources add the object lens assembling, and used microscope can adopt various optical microscopes or add the object lens assembling by light source, and its object lens can be magnifier, microscope camera lens or CCD.Used micro imaging system not only is confined to one, can monitor simultaneously by multiple spot on a capillary, and the microscope of each point or light source add object lens and can be the same or different.
Micro-capillary electropheresis apparatus of the present invention, the moving and control system of its collection point, comprise electrophoresis spectrogram acquisition system and image collection processing system move and control can move separately and control, also linkage and control, its translational speed, the path is adjustable.
Micro-capillary electropheresis apparatus of the present invention, moving and control system of its electrophoresis pattern acquisition system and image capturing system collection point, comprise in moving or the relative position after moving can be various forms, its move mode can be with manually control, or Electric Machine Control.What control system of the present invention adopted is known technology.
When being used by the above-mentioned micro-capillary electropheresis apparatus that constitutes, sample (comprising: electric field, electric osmose, viscous force, gravity, the pressure when sometimes can pressure auxiliary) under the common promotion of various effects, is moved to the other end by an end capillaceous.In the process of motion, each component of sample is concentrated and separates.When sample process electrophoresis pattern acquisition system, note the electrophoresis pattern of sample by it; When sample process image collection processing system, note the image and the processing of sample by it, and in the respective record system, carry out signal demonstration and online or processed offline in real time.
Use the micro-capillary electropheresis apparatus of embodiment 1, quartz capillary (internal diameter 75 μ m) is through CH
3OH, H
2O, NaOH, H
2O pressure washing successively is clean, and pressure injects 0.15mmol/LTris-H
3BO
3, after stablizing, pressure reduction enters the erythrocyte of rabbit.Adopt UV to detect, collect the electrophoresis pattern of 214nm, can obtain the cell ultraviolet absorption peak and see Fig. 3; Add CCD and computer with inverted microscope, collect image and see Fig. 4 (A is single cell dispersion image, and B is the cell aggregation volume image), can see the motion process of cell.
Embodiment 3
Use the micro-capillary electropheresis apparatus of embodiment 1, after the method identical with embodiment 1 usefulness cleaned kapillary, pressure injected 0.15mmol/L Tris-H
3BO
3, after stablizing, pressure reduction enters people's erythrocyte.Adopt UV to detect, collect the electrophoresis pattern of 214nm, can obtain the cell ultraviolet absorption peak and see Fig. 5; Add CCD and computer with inverted microscope, collect image and see Fig. 6 (A is single cell dispersion image, and B is cell aggregation and individual cells image together), can see the motion process of cell.
By the foregoing description as can be seen, the present invention has following characteristics with Capillary Electrophoresis and micro-imaging technique coupling:
Can realize micro-image high speed electrophoresis, unicellular or single-particle analysis, the express-analysis simultaneously of similar and non-similar multiparticle, and behavior is intuitively observed.
1. the electrophoresis peak that large particulate matter such as cell, bacterium are formed can give conclusive qualitative.
2. when cell or other material electrophoresis, different component image difference directly is that sample component is qualitative per sample, removes the hardship of the standard items that must obtain based on very high purity from, and the processing in the acquisition process is to the influence of measured object, and the research that this pair cell is learned is particularly important.
3. can be by the scanning of image being obtained the quantitative information of sample component, and carry out parallel with electrophoresis spectrogram quantitative result.
4. the Combined Treatment of the data by the two acquisition or multiple spot monitoring, can obtain separately utilization separately the information that can't obtain.
5. can be used for the research that the measured object proterties changes under the high pressure.
6. can obtain measured object information at the volley by tracking monitor individually or simultaneously, as the relation of proterties variation with action time to spectrogram and image.
Claims (8)
1. a micro-capillary electropheresis apparatus mainly is made up of capillary electrophoresis system, electrophoresis spectrogram acquisition system and image capturing system; Wherein:
Capillary electrophoresis system: connect with kapillary between two liquid bottles, form capillary channel, two liquid bottles connect the two ends of high-voltage power supply respectively;
The electrophoresis pattern acquisition system: kapillary one side is provided with light source, and another side of corresponding kapillary is provided with photoelectric commutator, and photoelectric commutator and electrophoresis pattern are collected display and electrically connected;
Image capturing system: a microscope light source is arranged on a side capillaceous, and another side of corresponding kapillary is provided with microscope, and this microscope connects an image acquisition device, connects image display processor by image acquisition device;
Used image capturing system is a plurality of settings, and multiple spot is monitored simultaneously on a capillary.
2. the micro-capillary electropheresis apparatus of claim 1 is characterized in that, the light source in the electrophoresis pattern acquisition system is laser instrument, light emitting diode or deuterium lamp, tungsten lamp, Halogen lamp LED.
3. claim 1 or 2 micro-capillary electropheresis apparatus is characterized in that being furnished with optical filter and slit on the light source in the electrophoresis pattern acquisition system.
4. the micro-capillary electropheresis apparatus of claim 1 is characterized in that, the microscope in the image capturing system is optical microscope or adds object lens assemblings by light source.
5. the micro-capillary electropheresis apparatus of claim 4 is characterized in that, object lens are magnifier, microscope camera lens or CCD.
6. the micro-capillary electropheresis apparatus of claim 1 is characterized in that, used shape of slit and size are optional.
7. the micro-capillary electropheresis apparatus of claim 1 is characterized in that, electrophoresis spectrogram acquisition system and image capturing system are to move separately and control, or interlock control.
8. the micro-capillary electropheresis apparatus of claim 1 is characterized in that, translational speed, the path of electrophoresis spectrogram acquisition system and image capturing system are adjustable.
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| CNB2005100057462A CN100394172C (en) | 2005-01-25 | 2005-01-25 | Micro-capillary electropheresis apparatus |
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| CNB2005100057462A CN100394172C (en) | 2005-01-25 | 2005-01-25 | Micro-capillary electropheresis apparatus |
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| CN100394172C true CN100394172C (en) | 2008-06-11 |
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| CN104049025A (en) * | 2014-01-23 | 2014-09-17 | 杭州师范大学 | Capillary electrophoresis analysis system utilizing micro-injection pump to drive liquid flows |
| CN103914695B (en) * | 2014-04-25 | 2017-01-11 | 中山大学 | Device and method for micro-electrophoresis image recognition |
| CN104777606A (en) * | 2015-03-27 | 2015-07-15 | 余启佳 | Magnifying glass device for natural laminar motion of fluids |
| CN104778877A (en) * | 2015-03-27 | 2015-07-15 | 余启佳 | Liquid device for natural laminar motion of fluids |
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| CN104795004A (en) * | 2015-03-27 | 2015-07-22 | 余启佳 | Fluid natural laminar flow motion displayer device |
| CN104766511A (en) * | 2015-03-27 | 2015-07-08 | 余启佳 | Fluid natural laminar motion monitor device |
| CN111693413A (en) * | 2020-04-23 | 2020-09-22 | 杭州兴浩晖生物科技有限公司 | Variable grating micro-signal separation device and manufacturing method thereof |
| CN112924536A (en) * | 2021-03-01 | 2021-06-08 | 无锡职业技术学院 | Quality control method before and after plating diamond-like layer on piston for high-pressure pump |
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