CN100410658C - Single cell gel electrophoresis apparatus - Google Patents
Single cell gel electrophoresis apparatus Download PDFInfo
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- CN100410658C CN100410658C CNB2005100960115A CN200510096011A CN100410658C CN 100410658 C CN100410658 C CN 100410658C CN B2005100960115 A CNB2005100960115 A CN B2005100960115A CN 200510096011 A CN200510096011 A CN 200510096011A CN 100410658 C CN100410658 C CN 100410658C
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- electroprobe
- single cell
- gel electrophoresis
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Abstract
The present invention relates to a single cell gel electrophoresis device which comprises a stabilized voltage supply, a voltmeter, a horizontal electrophoresis tank, an electroprobe and a voltage measuring bracket, wherein the stabilized voltage supply is connected with the horizontal electrophoresis tank through a guide wire, and the electroprobe which is connected with the voltmeter through a guide wire is arranged on the voltage measuring bracket. The present invention has the horizontal electrophoresis tank that side plates are arranged on four sides of a bottom plate, two vertical plates which are connected with a transverse plate are arranged in an upper plate of the bottom plate, a leveling mechanism is arranged on the lower surface of the bottom plate, and a level gauge is arranged on the lower surface of the transverse plate. Compared with the existing horizontal electrophoresis tank, the present invention has the advantages of reasonable design, simple structure, convenient operation, rapid adjusting speed, etc. The present invention can be popularized and applied to teaching scientific research, medical clinical examinations, safety surveys of epidemic diseases and occupational diseases as well as biologic experiments.
Description
Technical field
The invention belongs to teaching demonstration experimental technique field, be specifically related to single cell gel electrophoresis apparatus.
Background technology
Single cell gel electrophoresis also claims the comet electrophoresis, is to carry out DNA (deoxyribonucleic acid) (DNA) damage and repair the fluorescence detection method that detects in unicellular level, have simple, fast, characteristics such as sensitivity, original position.During electrophoresis, the dna chain of fracture can shift out in examining under the effect of electric field force, through the size redistribution of agarose molecular sieve effect according to the fracture fragment.Behind fluorescent dyeing, under fluorescent microscope, can be observed the tail of a comet with a bright comet head and a disperse of damaged cell nuclear.Because the distance that the fragment of the dna chain of different breaking degree is moved under same voltage is different, so by this comet image is measured the degree of injury that can judge DNA (deoxyribonucleic acid) on the unicellular level with the computer graphic conformal analysis.Single cell gel electrophoresis has wide practical use in all many-sides such as genetic toxicology, environmental monitoring, epidemiology, food industry, occupational illness protection and clinical diagnosises.Because the detection of single cell gel electrophoresis is very sensitive, so the requirement to electrophoretic apparatus in experimentation is very high, requiring power supply is direct current output, voltage is in 15～30v scope, and the AC ripple of power supply should be as much as possible little, with stablizing of electric field property in the assurance experiment, the necessary low temperature of electrophoresis environment, generally about 4 ℃, the horizontal adjustment of electrophoresis tank should be carried out fast, and the mensuration of unit length voltage should be finished in the shortest time in the electrophoresis tank.The horizontal adjustment of present used electrophoretic apparatus and voltage-regulation need 30 minutes at least, and the adjusting time that the comet electrophoresis allows can not can obtain reliable result to guarantee experiment greater than 5 minutes.
In the electrophoresis step of single cell gel electrophoresis, the glass slide that holds cell sample normally is positioned in the horizontal strip electrophoresis groove of filling electrophoretic buffer, and the general distribution arrangement that requires to be parallel to line of electric force in the electric field of the placement of microslide, so that the migratory direction of cell DNA (deoxyribonucleic acid) part is consistent on each microslide in electrophoresis process.The electric field force that requires to load in electrophoresis process on each microslide cell sample must be consistent and fixing, so all should guarantee to hold in the electrophoresis tank electrophoresis platform of microslide before each experiment is horizontal plane, and the voltage that loads on the microslide can be measured fast and regulate.
When the usage level electrophoresis tank carries out the single cell gel electrophoresis detection at present, usually will be by constantly regulating the height that electrophoresis tank carries, regulate fulcrum and guarantee that by the voltage table measurement plane that the horizontal strip electrophoresis groove holds microslide is a level, the voltage of unit distance is experiment required voltage value in the electrophoresis tank.The whole operation process wastes time and energy, and electrophoretic buffer is contacted with carbon dioxide in air for a long time react, and the pH value changes thereupon.And regulate the level and the long false positive results that will cause experiment of voltage time of electrophoretic apparatus, the reliability of final influence experiment.
After above-mentioned horizontal strip electrophoresis groove is switched on power supply opening, use voltage table earlier, adjust the horizontality that fulcrum is adjusted electrophoresis tank then lentamente, waste time and energy during adjustment, and be difficult to electrophoresis tank is adjusted to horizontal level at each position measurement voltage of voltage groove.
Above-mentioned horizontal strip electrophoresis groove energising back is put into an experiment glass slides earlier in electrophoresis tank, voltage is adjusted at the two ends that then electroprobe are inserted in this microslide again.It is long to expend time in, and vertical with electrophoresis tank of the level between two electroprobes when being not easy to keep measuring, owing to put into an experiment slide glass in electrophoresis buffer solution, can pollute solution in addition.
Summary of the invention
Technical matters to be solved by this invention is to overcome the shortcoming of above-mentioned horizontal strip electrophoresis groove, and a kind of reasonable in design, simple in structure, easy to use, single cell gel electrophoresis apparatus that governing speed is fast is provided.
Solving the problems of the technologies described above the technical scheme that is adopted is: it comprises stabilized voltage supply, voltage table, horizontal strip electrophoresis groove, electroprobe, stabilized voltage supply links to each other with the horizontal strip electrophoresis groove by lead, it also comprises the voltage measurement support, and the electroprobe that is connected with voltage table by lead is arranged on the voltage measurement support.Horizontal strip electrophoresis groove of the present invention is: the side all around at base plate is provided with side plate, in the vertical direction epipleural of plate upper surface, be provided with two risers that connect with transverse slat, be provided with level(l)ing mechanism at the base plate lower surface, be provided with spirit-leveling instrument at the lower surface of transverse slat.
Voltage measurement support of the present invention is: scale is provided with two electroprobe framves that can move on scale, scale is provided with handle, and the electroprobe that links to each other with voltage table by lead is arranged on the electroprobe frame.
Electroprobe frame of the present invention is a grooved.
Handle of the present invention is an insulated handle, and this insulated handle is wooden handle or bakelite handle or plastic handle.
Level(l)ing mechanism of the present invention is the scalable pillar of at least three scalable base plates and reference field relative height.Spirit-leveling instrument of the present invention is spirit level or digital level.
Scalable pillar of the present invention is the threaded adjusting pillar.
The present invention compares with existing horizontal strip electrophoresis groove, has reasonable in design, simple in structure, easy to use, the advantage such as fast of regulating the speed, and can promote the use of in teaching and scientific research, clinical medicine check, epidemic disease and occupational illness safety survey and Bioexperiment.
Description of drawings.
Fig. 1 is a structural representation of the present invention.
Fig. 2 is the front view of voltage measurement support 3 among Fig. 1.
Fig. 3 is the vertical view of Fig. 2.
Fig. 4 is the A-A sectional view of Fig. 2.
Fig. 5 is the front view of horizontal strip electrophoresis groove 4 among Fig. 1.
Fig. 6 is the left view of Fig. 5.
Fig. 7 is the vertical view of Fig. 5.
Fig. 8 is the B-B sectional view of Fig. 5.
Embodiment
The present invention is described in more detail below in conjunction with drawings and Examples, but the invention is not restricted to these embodiment.
In Fig. 1, the single cell gel electrophoresis apparatus of present embodiment is made of stabilized voltage supply 1, voltage table 2, voltage measurement support 3, horizontal strip electrophoresis groove 4, electroprobe 5 connections.
Stabilized voltage supply 1 links to each other with horizontal strip electrophoresis groove 4 by lead, electroprobe 5 is installed on the voltage measurement support 3, present embodiment is equipped with two electroprobes 5 on voltage measurement support 3, electroprobe 5 links to each other with voltage table 2 by lead, during experiment voltage measurement support 3 is placed on the horizontal strip electrophoresis groove 4 stabilized voltage supply 1 and the commodity of voltage table 2 for selling on the market, the voltage adjusting range of stabilized voltage supply 1 is 15～30v, voltage table 2 is a needle dc voltmeter, also can adopt digital voltmeter.
In Fig. 2,3,4, the voltage measurement support 3 of present embodiment is made of scale 3-1, electroprobe frame 3-2, handle 3-3 connection.Be carved with scale on the scale 3-1, under scale, indicate from 1～17 17 numerals by the natural order arrangement, two electroprobe frame 3-2 are installed on the scale 3-1, electroprobe frame 3-2 is a trench structure, electroprobe frame 3-2 can be on scale 3-1 move left and right, be processed with screw hole on the electroprobe frame 3-2, be used to install electroprobe 5.The right-hand member of scale 3-1 is installed with handle 3-3 with the screw threads for fastening connector, and the handle 3-3 of present embodiment is wooden handle, also available bakelite handle or plastic handle, and handle material requires to possess electrical insulating property.
In Fig. 5,6,7,8, the horizontal strip electrophoresis groove 4 of present embodiment is made of riser 4-1, transverse slat 4-2, back scalable pillar 4-3, spirit-leveling instrument 4-4, base plate 4-5, preceding scalable pillar 4-6, side plate 4-7, dividing plate 4-8 connection.Scalable pillar 4-3 base plate 4-5 lower surface is equipped with 2 preceding scalable pillar 4-6 and 1 by thread connection after, scalable pillar 4-3 behind 1 preceding scalable pillar 4-6 and 2 also can be installed, preceding scalable pillar 4-6 and back scalable pillar 4-3 are the threaded adjusting pillar, the level(l)ing mechanism that also can adopt sliding sleeve and restraint screw to constitute, scalable pillar 4-6 and back scalable pillar 4-3 before the rotation, adjustable chassis assembly 4-5 is positioned at horizontal level, side all around at base plate 4-5 is installed with side plate 4-7 with the screw threads for fastening connector, in the vertical direction epipleural 4-7 of base plate 4-5 upper surface, be installed with two riser 4-1 with the screw threads for fastening connector, riser 4-1 goes up and is installed with transverse slat 4-2 with the screw threads for fastening connector, riser 4-1 connects formation electrophoresis tank inner platform with transverse slat 4-2, lower surface at transverse slat 4-2 is installed with spirit-leveling instrument 4-4 with the screw threads for fastening connector, spirit-leveling instrument 4-4 is used to indicate horizontal strip electrophoresis groove 4 to be positioned at horizontal level, the spirit-leveling instrument 4-4 of present embodiment is a spirit level, also can adopt digital spirit-leveling instrument.
Principle of work of the present invention is as follows:
With lead stabilized voltage supply 1 is connected with horizontal strip electrophoresis groove 4 during experiment, turns on the power switch, the electrophoresis buffer solution in the horizontal strip electrophoresis groove 4 is just logical has gone up electricity, has formed a uniform electric field.Pass through the spirit-leveling instrument 4-4 of eye-level electrophoresis tank 4 bottoms earlier, scalable pillar 4-6 and back scalable pillar 4-3 before regulating, with horizontal strip electrophoresis groove 4 to level, then two electroprobes 5 are inserted into respectively in two circular holes of electroprobe frame 3-2, whole electroprobe frame 3-2 is vertical, hand-held wooden handle 3-3, two electroprobe frame 3-2 are adjusted to the length of microslide according to the shown scale of the scale 3-1 on the electroprobe frame 3-2, it is motionless vertically to put into electrophoresis buffer solution, read the reading on the voltage table 2, if reading and desiredly be not inconsistent, then can adjust voltage to requiring, finish electrophoresis preceding horizontal adjustment and voltage adjustment by the output voltage adjustment button of regulating on the stabilized voltage supply 1.Subsequently the glass slide that contains the cell glue-line to be detected is placed on the horizontal strip electrophoresis groove 4 by the direction that is parallel to electric field force, open the power switch of stabilized voltage supply 1, under 4 ℃ of conditions, carry out electrophoresis.In electrophoresis process, the DNA (deoxyribonucleic acid) strand that ruptures in the cell because have negative electricity, can anode move under the effect of electric field force in the environment of electrophoresis liquid.And because the electric field migration force that the different strand of fracture is subjected to is different with the resistance of low melting-point agarose molecular sieve, its distance of moving out of nuclear is also different.After electrophoresis is finished, the glass slide that will contain the cell glue-line places neutralizer to neutralize, so that the pH value is adjusted near about 7.0 from＞13, carry out afterwards from 50～100% gradient Ethanol Treatment, make the low melting-point agarose dehydration, cell can be positioned on the same plane of glass slide because of the attenuation of agarose glue-line subsequently, carries out microscopic examination and computer graphic conformal analysis subsequently.Behind fluorescent dyeing, under fluorescent microscope, can be observed the single-cell gel shape image of damaged cell nuclear, promptly have the tail of a comet of a bright comet head and a disperse.Because the distance that the different dna chain of breaking degree moves under same voltage is different, so can judge dna lesion degree on the unicellular level by this image being measured with the computer graphic conformal analysis.
Claims (6)
1. single cell gel electrophoresis apparatus, comprise stabilized voltage supply (1), voltage table (2), horizontal strip electrophoresis groove (4), electroprobe (5), stabilized voltage supply (1) links to each other with horizontal strip electrophoresis groove (4) by lead, it is characterized in that: it also comprises voltage measurement support (3), and the electroprobe (5) that is connected with voltage table (2) by lead is arranged on the voltage measurement support (3); Said horizontal strip electrophoresis groove (4) is: be provided with side plate (4-7) in the side all around of base plate (4-5), in the vertical direction epipleural (4-7) of base plate (4-5) upper surface, be provided with two risers (4-1) that connect with transverse slat (4-2), be provided with level(l)ing mechanism at base plate (4-5) lower surface, be provided with spirit-leveling instrument at the lower surface of transverse slat (4-2).
2. according to the described single cell gel electrophoresis apparatus of claim 1, it is characterized in that said voltage measurement support (3) is: scale (3-1) is provided with two can go up the electroprobe frame (3-2) that moves at scale (3-1), scale (3-1) is provided with handle (3-3), and the electroprobe (5) that links to each other with voltage table (2) by lead is arranged on the electroprobe frame (3-2).
3. according to the described single cell gel electrophoresis apparatus of claim 2, it is characterized in that: said electroprobe frame (3-2) is a grooved.
4. according to the described single cell gel electrophoresis apparatus of claim 2, it is characterized in that: said handle (3-3) is an insulated handle, and this insulated handle is wooden handle or bakelite handle or plastic handle.
5. according to the described single cell gel electrophoresis apparatus of claim 1, it is characterized in that: said level(l)ing mechanism is the scalable pillar (4-3,4-6) of at least three scalable base plates (4-5) with the reference field relative height; Said spirit-leveling instrument (4-4) is spirit level or digital level.
6. according to the described single cell gel electrophoresis apparatus of claim 5, it is characterized in that: said scalable pillar (4-3,4-6) is the threaded adjusting pillar.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100960115A CN100410658C (en) | 2005-09-12 | 2005-09-12 | Single cell gel electrophoresis apparatus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100960115A CN100410658C (en) | 2005-09-12 | 2005-09-12 | Single cell gel electrophoresis apparatus |
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| Publication Number | Publication Date |
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| CN1737564A CN1737564A (en) | 2006-02-22 |
| CN100410658C true CN100410658C (en) | 2008-08-13 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100960115A Expired - Fee Related CN100410658C (en) | 2005-09-12 | 2005-09-12 | Single cell gel electrophoresis apparatus |
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Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| PL3265796T3 (en) * | 2015-03-06 | 2023-03-20 | 4D Lifetec Ag | Gel electrophoresis system for single cell gel electrophoresis |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2045854U (en) * | 1987-04-28 | 1989-10-11 | 西安医科大学 | Electrophoresis device |
| CN2395274Y (en) * | 1999-06-12 | 2000-09-06 | 辛毅 | Horizontal electrophoretic bath |
| US20030175821A1 (en) * | 2002-03-14 | 2003-09-18 | Hoover Rex A. | Apparatus and method for microscopic comet assay |
| CN1548546A (en) * | 2003-05-09 | 2004-11-24 | 中国科学院大连化学物理研究所 | Continuous unicellular inclusion analyzing method based on microflow control chip platform |
-
2005
- 2005-09-12 CN CNB2005100960115A patent/CN100410658C/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2045854U (en) * | 1987-04-28 | 1989-10-11 | 西安医科大学 | Electrophoresis device |
| CN2395274Y (en) * | 1999-06-12 | 2000-09-06 | 辛毅 | Horizontal electrophoretic bath |
| US20030175821A1 (en) * | 2002-03-14 | 2003-09-18 | Hoover Rex A. | Apparatus and method for microscopic comet assay |
| CN1548546A (en) * | 2003-05-09 | 2004-11-24 | 中国科学院大连化学物理研究所 | Continuous unicellular inclusion analyzing method based on microflow control chip platform |
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| CN1737564A (en) | 2006-02-22 |
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Granted publication date: 20080813 Termination date: 20110912 |