CN108387422A - A kind of large biological molecule transferring film, coloring system and the equipment of efficient high sensitivity - Google Patents
A kind of large biological molecule transferring film, coloring system and the equipment of efficient high sensitivity Download PDFInfo
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- CN108387422A CN108387422A CN201810153291.6A CN201810153291A CN108387422A CN 108387422 A CN108387422 A CN 108387422A CN 201810153291 A CN201810153291 A CN 201810153291A CN 108387422 A CN108387422 A CN 108387422A
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Abstract
The present invention discloses a kind of large biological molecule transferring film of efficient high sensitivity, coloring system, including working cell, is equipped at least one channel for testing and analyzing sample；The associated operation part with working cell creates at least one dyeing or transferring film channel parameters, and each dyeing or transferring film channel parameters include electric field cycle-index at least once；Invention additionally discloses a kind of large biological molecule transferring film, the equipment of coloring system based on efficient high sensitivity, including pedestal and shell, hull outside is equipped with operation panel, the system power supply and operating circuit plate that pedestal and enclosure interior are equipped with workspace and are connected with workspace, a drawing and pulling type autonomous working room is at least arranged in the workspace, and the positive-negative electrode plate for being provided with electric field is installed in the operating room.The system or equipment of the rapid dyeing and transferring film of the motion control large biological molecule disclosed in this invention that can utilize electric field driven biology charged particle, while also ensuring the highly sensitive requirement of biological detection.
Description
The application is application number 201310025508.2, applying date 2013.01.22, a kind of entitled " the efficient Gao Ling of application
The divisional application of large biological molecule transferring film, coloring system and the equipment of sensitivity ".
Technical field
The present invention relates to a kind of biological detection equipment technical field, more particularly to it is a kind of using electric field driven biology or
Efficiently highly sensitive large biological molecule transferring film, coloring system and equipment are realized in the movement of other charged particles.
Background technology
In DC electric field, charged particle is to referred to as electrophoresis (electropho- the phenomenon that tape symbol opposite electrode movement
resis).Russia physicist Pe н ce are found that electrophoresis first within 1809, but until the Tiselius of nineteen thirty-seven Sweden
After the moving boundary electrophoresis (boundary electrophoresis) for establishing protein isolate matter, electrophoretic techniques just starts to apply.
In the age in this century 60-70, since the media such as filter paper, polyacrylamide gel are introduced sequentially into electrophoresis, electrophoretic techniques is able to rapidly
Development.It is very extensive that colourful electrophoretic type makes it apply, main other than being analyzed for the separation of small-molecule substance
It is easy to operate since electrophoresis equipment is simple for protein, nucleic acid, enzyme, or even the research of virus and cell, there is high-resolution
Rate and selective feature, it has also become common technology in medical test.
Biological ployose such as peptide, protein, nucleic acid, oligosaccharides etc. or its mixture pass through gel electrophoresis analysis, usual mesh
Sample be loaded in a medium, such as polyacrylamide gel, according to the difference of molecular size, charge and other physical properties
Different, migration in the electric field obtains different bands, it is detected after band is combined by large biological molecule with staining reagent after electrophoresis or
These biological ployoses are transferred to the analysis experiment that next step is carried out on NC, PVDF or nylon membrane by person.Wherein, more in biology
In terms of polymers dyeing, the biology that a large amount of colouring methods and reagent are developed for use in dyeing medium as gel divides greatly
Son, these reagents can be divided into four kinds：The first staining reagent includes the organic dyestuff being attached on polymer, as coomassie is bright
Blue staining reagent makes protein band become au bleu so as to by being visually observed；Second of staining reagent is poly- including being attached to
The fluorescent dye on object is closed, as EB combination DNA or RNA make DNA or RNA take on a red color when ultraviolet light irradiates；The third dyeing
Reagent includes silver staining reagent；4th kind be dyeing background reagent, that is, be referred to as negative staining.These dyestuffs also carry charge mostly,
It can be moved in DC electric field.Factor due to influencing electrophoretic mobility mainly has electric field strength, the pH value of solution and solution
Ionic strength etc. is related, therefore mobility in the medium for applying electric field of large biological molecule and dyestuff and differs, and biology is big
Molecular is larger, mobile slow in the electric field, will not be removed from medium within the time of restriction, organic dye molecule is opposite
Large biological molecule molecule comes much smaller, and the dye molecule not combined with large biological molecule molecule can be removed from medium rapidly,
It therefore can large biological molecule (combining dyestuff) be retained in medium by so that dyestuff is removed from medium in same electric field
In mode realize the colour developing of large biological molecule, so as to obtain in a short time, high-resolution, background influence be small or even nothing
The analysis image of the large biological molecule of background, if but dyestuff and large biological molecule merely through primary combinations of meeting, it is clever to some
Sensitivity requires relatively high experiment, then the case where being difficult to meet the requirements, or even be susceptible to missing inspection in the detection.
With the fast development of the subjects such as biomedicine, also promoted rapidly in biomacromolecule detection technical aspect, but
There has been no the motion control large biological molecules that set of system or equipment can utilize electric field driven biology charged particle currently on the market
Rapid dyeing and transferring film, and realize highly sensitive requirement simultaneously.
Invention content
The technical problem to be solved by the present invention is in view of the deficiencies of the prior art, provide one kind to utilize electric field driven
The system of the rapid dyeing and transferring film of the motion control large biological molecule of biological charged particle, while also ensuring biological detection
Highly sensitive requirement.
The present invention also provides a kind of large biological molecule transferring film of the efficient high sensitivity using the system, dyeing installations.
The purpose of the present invention can be realized by following technical proposal：
A kind of large biological molecule transferring film of efficient high sensitivity, coloring system, including working cell are equipped at least one
Channel is for testing and analyzing sample；The associated operation part with working cell, creates at least one dyeing or transferring film
Channel parameters, preferably 1-10, more preferable 1-4, each dyeing or transferring film channel parameters include electric field cycle at least once
Number, preferably 1-10 times, it is 2-5 times more preferable.
It is arranged while multi-channel parameter, dyeing or the transferring film of the large biological molecule of different samples may be implemented, electric field follows
The selection of ring number can adjust time that dyestuff reciprocating passes through large biological molecule to be dyed according to the requirement of different experiments
Number also ensures the sensitivity of dyeing while ensureing that experiment is efficient.
The operation part, the following relevant parameters of at least one of setting：Channel number, dyeing/transferring film, Gel numbers,
Working time, operating voltage, operating current, validation of information, startup/pause, analysis/processing of input data.
The setting of above-mentioned parameter, especially working time, the setting of operating voltage, operating current, can be according to different realities
The speed for the requirement adjustment dyestuff flowing tested, improves conventional efficient；The parameters such as the joint passage label, dyeing/transferring film, Gel numbers
Setting can simultaneously will dyeing and transferring film function be integrated into set of system, realize set of system be carried out at the same time dyeing transferring film
Process.
Further, the acceleration detection large biological molecule system further includes for recording operation and testing result data
Storage device, carry out storage/export of data and state, most convenient solves the problems, such as experimentally to client.
Further, the acceleration detection large biological molecule system further include for policer operation normal operation and
Judge the monitoring system whether work terminates.
It is may determine that by monitoring system either with or without sample to be analysed is put into the working cell of machine, if be not put into
Or it is put into mistake, machine does not work, and damage to avoid machine no-load running to machine or is put into wrong machine operation and causes
Sample destruction, cause the waste of expensive biological sample；It can also judge whether machine operation is completed, include mainly：Machine
Device time to rear power cut-off and prompts that work terminates or machine passes through and detects machine itself according to the time service of setting
Operating current changes, and judges whether work terminates, is prompted after power cut-off.
A kind of large biological molecule transferring film, the equipment of coloring system based on efficient high sensitivity, including pedestal and shell are used
In providing the inside and outside support of machine, hull outside is equipped with operation panel, and pedestal and enclosure interior are equipped with workspace and and work
Make the connected system power supply in area and a drawing and pulling type autonomous working room, preferably 1- is at least arranged in operating circuit plate, the workspace
10, more preferable 1-4, the positive-negative electrode plate for being provided with electric field is installed in the operating room.
The setting of multiple autonomous working rooms with multiple running parameter independent operatings and can be carried out at the same time, and realize same instrument
It carries out, and does not interfere with each other while the dyeing of the multiple samples of device or transferring film.
The chamber bottom is equipped with saddle, and the positive electrode is fixedly mounted on saddle；Installation can be beaten above operating room
The lid opened, lid are equipped with negative electrode and the gland that can uniformly press to electrode plate and sandwich structure to be analyzed locking dress
It sets.
The pressure locking device by positioned at lid surface runner, the eccentric wheel that is fixedly connected with runner and with bias
Opposite clamp composition is taken turns, the eccentric wheel bottom is wedge type slide unit structure, and the clamp is fixedly connected with a pressing plate, the pressing plate
Lower section is equipped with negative electrode or is directly fixedly connected with negative electrode.
When runner of the rotation positioned at lid surface, the eccentric wheel of driven by rotating wheel lower part, by operating room's cover lock, after
Continuous rotating wheel, the wedge-shaped slide unit in driven by rotating wheel lower part coordinate with clamp, pressing plate are made to move up and down, so as to according to dyeing or
The thickness of transferring film sandwich structure adjusts pressure size, and electrode plate is enable to be put down with dyeing therein or transferring film sandwich structure
The contact of whole uniform close further increases dyeing or the sensitivity of transferring film.
The contour bolt that pressing plate is cased with spring by four is spirally connected with lid, keeps pressing plate steady and can perceive down
The resistance of square sandwich structure carrys out automatic distribution pressure, so that it is guaranteed that electric current is uniformly distributed.
The large biological molecule transferring film based on efficient high sensitivity, the equipment of coloring system further include display panel,
Working gas outlet and purifier, wherein the display panel is TFT liquid crystal or touch LCD screen, the purification
Device, which is located in working gas outlet, to be surveyed, and the inside is equipped with activated carbon,
The display screen of TFT liquid crystal or touch LCD screen can clearly indicate parameter setting interface and experiment carry out state, only
Makeup, which is set, to be purified it before exhaust gas discharge, reduced environmental pollution and the harm to operating personnel.
The operation panel is any one in microswitch, touch key-press, remote control, touch screen, film key.
The equipment further includes dyeing consumptive material and transferring film consumptive material, the dyeing consumptive material include dyeing carrier, dyestuff
And dye solution, or invaded the dyeing carrier containing dyestuff and dye solution；The transferring film consumptive material include transferring film carrier,
Film and equilibration buffer, or soaked transferring film carrier and film containing equilibration buffer.
The dyeing carrier and transferring film carrier is filter paper, gel or pure liquid phase；The dye solution
It is one group, wherein：The part A adjacent with negative electrode mainly includes the coomassie brilliant blue R250 (CBB- of (mass ratio) 0.1%-3%
R250), the acetic acid of the isopropanol of (volume ratio) 5%-50% and (volume ratio) 1%-30%, remaining is water；Preferably：0.1%
Coomassie brilliant blue R250 (CBB-R250), 20% isopropanol, 10% acetic acid and 69.9% water；It is adjacent with positive electrode
Part B include mainly the EDTA (ethylenediamine tetra-acetic acid) of 5-500mmol/L, 5-500mmol/L Na2HPO4,5-
The NaH2PO4 of 500mmol/L and the acetic acid for accounting for overall solution volume 1%-20%, preferably：EDTA, 30mmol/L of 80mmol/L
Na2HPO4,45mmol/L NaH2PO4 and account for 10% acetic acid of overall solution volume；The equilibration buffer includes mainly：1-
The N of trishydroxymethylaminomethane (Tris) and 1-200mmol/L of 200mmol/L, N- bicine N-s (Bicine) are excellent
Select the Bicine of the Tris and 100mmol/L of 100mmol/L.
Wherein, part A mainly makes one ionization dyestuff of the proteopexy in gel while offer in dye solution
Liquid environment, part B mainly provide suitable acidic environment；Equilibration buffer in protein During migration mainly for providing
One constant electric field.
The present invention provides a kind of large biological molecule transferring film, coloring system and the equipment of efficient high sensitivity, furtherly
Bright, this invention provides a kind of large biological molecule transferring film of efficient high sensitivity, coloring system and may be implemented PAGE
In large biological molecule dyeing or transfer equipment, make large biological molecule dyeing and During migration positive and negative two electrode plate it
Between highly sensitive completion in a short period of time, such as more peptide or proteins in medium, RNA, DNA, oligosaccharide or one are contained
One or several kinds of complex mixture in polypeptide, protein, RNA, DNA or oligosaccharide, is entered by electric field driven dye molecule
Medium carrys out stained biological macromolecular, or the analysis detection that next step is carried out on other carriers is transferred by electric field.
In dyeing course, electric field power drive dye molecule is by gel and interaction of biomacromolecules, to make the biology in gel
Macromolecular is colored, and an electric field cycle-index means that dyestuff is combined and detached with the primary of large biological molecule, passes through change
Electric field when dynamic dyeing can adjust the speed that dyestuff flows and reciprocating passes through the number of large biological molecule to be dyed, can
Different sensitivity is realized for different testing goals, is capable of the completion protein staining of high quality and high speed, effect
More than tradition and current product and method on the market；During migration is equally completed between positive and negative two electrode plate, electric field
Driving protein passes through gel to reach on film and is adsorbed on film, the movement velocity of protein can by positive and negative two electrode it
Between size of current be adjusted, the time of transferring film can complete within ten minutes；The present invention will dye and transferring film function is whole
It closes into an equipment, realizes that equipment completes dyeing During migration, it can be achieved that multichannel is carried out at the same time, can dye simultaneously
Or transferring film or the quantity of the glue of dyeing plus transferring film greatly increase, and funds and lab space have been saved while improving efficiency.
In addition, the experiment proved that, it a kind of large biological molecule transferring film of efficient high sensitivity provided by the present invention, coloring system and sets
Standby, reciprocal electric field cycle can realize the separating effect of more preferably large biological molecule on same limited carrier.
In conclusion the invention has the advantages that：
1) efficiently：The time of dyeing and transferring film, while multichannel are greatly shortened using the principle of electric field driven charged ion
It is carried out at the same time dyeing, transferring film or the two to be carried out at the same time, or even can also be carried out at the same time the separation of large biological molecule, substantially increase reality
Test efficiency.
2) sensitive：The speed and reciprocating of adjustment dyestuff flowing pass through the number of large biological molecule to be dyed, are protecting
The good separating effect of the sensitivity for also ensureing to dye while testing efficient or large biological molecule is confirmed, in addition, gland contracts
Tight device enables electrode plate closely to be contacted with dyeing therein or transferring film sandwich structure even uniform, further increases dye
The sensitivity of color or transferring film.
3) economic and environment-friendly：Dyeing and transferring film function are integrated into an equipment, realize that an equipment is completed dyeing and turned
Membrane process has saved funds and lab space, while gas cleaning plant can purify exhaust gas, reduce environmental pollution
With the harm to operating personnel.
Description of the drawings
Fig. 1 is the flow diagram of the system of acceleration large biological molecule dyeing of the present invention and transferring film；
Fig. 2 is the external structure schematic diagram of the equipment of acceleration large biological molecule dyeing of the present invention and transferring film；
Fig. 3 is the locking device schematic diagram of the equipment of acceleration large biological molecule dyeing of the present invention and transferring film；
Fig. 4 is the locking device eccentric wheel bottom knot of the equipment of acceleration large biological molecule dyeing of the present invention and transferring film
Structure schematic diagram；
Fig. 5 is the locking device clamp and pressing plate knot of the equipment of acceleration large biological molecule dyeing of the present invention and transferring film
Structure schematic diagram；
Fig. 6 is the gland structure schematic diagram of the equipment of acceleration large biological molecule dyeing of the present invention and transferring film；
Fig. 7 is the structural schematic diagram of bolt in gland structure of the present invention；
Fig. 8-12 is display schematic diagram when operation panel operates；
Figure 13 is the schematic diagram of the sandwich structure between electrode in dyeing course；
Figure 14 is the alternating electric field distribution map of upper/lower electrode in the direction of time；
Figure 15 is the schematic diagram of the sandwich structure between electrode in During migration；
Figure 16 is to be incubated the result of exposure after conventional method (wet method) transferring film；
Figure 17 is the result that exposure is incubated after system of the present invention and the quick transferring film of instrument；
Figure 18 is the experimental result dyed with conventional coloring method；
Figure 19 is the experimental result of system of the present invention and instrument rapid dyeing；
In attached drawing：1 pedestal, 2 shells, 3 film keys, 4 be TFT liquid crystal, 5 workspaces, 6 operating rooms, 7 grooves, and 8 data are defeated
Outgoing interface, 9 circuit thermovents, 10 way circuits switch, 11 lids, 12 gland locking devices, 13 runners, 14 eccentric wheels, 15 wedge shapes
Slide unit, 16 clamps, 17 pressing plates, 18 springs, 19 bolts.
Specific implementation mode
The invention will be further described for the following example, it must further be noted that, the present invention is not limited only to this：
Embodiment 1
A kind of large biological molecule that can be directed to different testing goals and realize the efficient high sensitivity of different sensitivity
Transferring film, coloring system, including working cell are equipped with 2-6 channel for testing and analyzing sample；It is associated with working cell
Operation part, create dyeing or the transferring film channel parameters of 2-6 accordingly, each dyeing or transferring film channel parameters are according to inspection
The difference of purpose is surveyed, required sensitivity difference is arranged different electric field cycle-indexes.
Embodiment 2
The system of a kind of acceleration detection large biological molecule as shown in Figure 1 comprising working cell is equipped with 4 channels
For testing and analyzing sample；Operation part associated with working cell can create 4 dyeing or transferring film channel ginsengs simultaneously
Number, such as：Channel number, dyeing/transferring film, Gel numbers, working time, operating voltage, operating current, electric field cycle-index, information
Confirmation, startup/pause, analysis/processing of input data etc.；Monitoring device is used for the normal operation of policer operation, when machine
Some specific channel is not put into sample to be analysed in working cell, or is put into mistake, and machine does not work and prompted, also
It may determine that whether the work in each channel is completed, include mainly：Machine operation according to the time service of setting, after tie by the time
Beam work and prompt work terminate or machine by detect machine itself operating current change, judge whether work terminates,
It is prompted after power cut-off；Storage device, record operation and testing result data, the data can be read by external reader, such as：
Time<2012,04.20.,21:39:11>:start main,search Cell.
Time<2012,04.20.,21:39:11>:finded cell A,B
Time<2012,04.20.,21:39:11>:enter main function.
Time<2012,04.20.,21:49:10>:cell A staining,one mini gel,7min；
Time<2012,04.20.,21:56:11>:running,waiting data.
Time<2012,04.20.,21:56:12>:finished
Time<2012,04.20.,21:49:10>:start main,search Cell.
Time<2012,04.20.,21:49:10>:finded cell A,B.
Time<2012,04.20.,21:49:11>:enter main function.
Time<2012,04.20.,21:50:11>:cell B transfer,one mini gel,9min；
Time<2012,04.20.,21:50:12>:running,waiting data.
Time<2012,04.20.,21:51:11>:Error.over current.
Embodiment 3
A kind of large biological molecule transferring film, the equipment of coloring system based on efficient high sensitivity, including pedestal and shell are used
In providing the inside and outside support of machine, case top is equipped with touch key-press, and pedestal and enclosure interior are equipped with workspace and and work
Make the connected system power supply in area and operating circuit plate；2-6 drawing and pulling type independent operating room is set in workspace, is pacified in operating room
Equipped with the positive-negative electrode plate for providing electric field.
Embodiment 4
A kind of large biological molecule transferring film, the equipment of coloring system based on efficient high sensitivity, including pedestal and shell are used
In providing the inside and outside support of machine, case top is equipped with touch key-press, and pedestal and enclosure interior are equipped with workspace and and work
Make the connected system power supply in area and operating circuit plate；The independent operating room of 2-6 drawing and pulling type, chamber bottom are set in workspace
Saddle is installed, positive electrode is fixedly mounted on saddle；Openable lid is installed above operating room, lid be equipped with negative electrode and
The gland locking device that can uniformly press to electrode plate and sandwich structure to be analyzed.
Embodiment 5
A kind of large biological molecule transferring film, the equipment of coloring system based on efficient high sensitivity as shown in Figure 2-5, including
Pedestal 1 and shell 2, for providing machine inside and outside support, 2 upper right of shell is equipped with film key 3 and TFT liquid crystal 4,
Pedestal 1 and shell 2 are internally provided with workspace 5 and the system power supply being connected with workspace 5 and operating circuit plate, the setting of workspace 54
A independent operating room 6 of drawing and pulling type, can dye simultaneously or transferring film or the quantity of glue of dyeing plus transferring film are up to：mini
Gel, 8 or midi Gel 4,6 pull face of operating room is equipped with groove 7, and convenient for hand-held pull, chamber bottom is equipped with support
Platform, positive electrode are fixedly mounted on saddle；Openable lid 11 is installed above operating room, lid 11 be equipped with can to electrode plate and
The gland locking device 12 that sandwich structure to be analyzed uniformly presses, pressure locking device 12 positioned at 11 surface of lid by turning
Wheel 13, the eccentric wheel 14 being fixedly connected with runner 13 and the clamp 16 opposite with eccentric wheel 14 composition, 14 bottom of eccentric wheel are wedge
15 structure of type slide unit, clamp 16 are fixed in a plastic presser plate 17, the contour bolt 19 that pressing plate 17 is cased with spring 18 by four
It is spirally connected with lid 11, keeps pressing plate 17 steady and the resistance that can perceive lower section sandwich structure carrys out automatic distribution pressure,
So that it is guaranteed that electric current is uniformly distributed.Metal negative electrode is fixed by the plastic presser plate of the bayonet and lid 11 of side, plastics pressure
Plate 17 is fixed by contour screw and lid 11；Equipment left back is additionally provided with working gas outlet and purifier, purification dress
On the inside of working gas outlet, the inside is equipped with activated carbon, and data output interface 8 and circuit heat dissipation are additionally provided on the right side of instrument
Mouth 9, lower right are equipped with way circuit switch 10, and bottom is equipped with four limbs rubber feet.
Embodiment 6
Referring to the instrument described in embodiment 5, the instrument described in the present embodiment is also configured with dyeing consumptive material and transferring film consumptive material,
Wherein dyeing consumptive material is to have invaded filter paper or gel containing dyestuff and dye solution, and transferring film consumptive material is to have soaked containing equalizing and buffering
The filter paper or gel and film of liquid；Wherein, dye solution is one group：The part A adjacent with negative electrode includes mainly 0.1%
Coomassie brilliant blue R250 (CBB-R250), 20% isopropanol, 10% acetic acid and 69.9% water；It is adjacent with positive electrode
Part B includes mainly Na2HPO4,45mmol/L NaH2PO4 of EDTA, 30mmol/L of 80mmol/L and accounts for overall solution volume
10% acetic acid；Equilibration buffer includes mainly：Concentration is the Tris and Bicine of 100mmol/L.
Instrument described in the present embodiment, system are additionally provided with a set of operating voltage, operating current and electric field cycle-index
When using mating dyeing consumptive material and transferring film consumptive material this default parameters can be used directly, more in the default setting of equal key parameters
It is convenient and efficient.
Embodiment 7
The display content of operation panel when remaining parameter setting flow of instrument described in embodiment 6 and operation：
1) as shown in fig. 6, when machine operation can automatic detection instrument operating room, have the operating room of task can be aobvious
Show and show Staining/Transfer on panel, operator can select job category, and the interface of task can not shown
None, in the case of such as Fig. 6, the stainging of Cell A is acquiescence focus, "enter" key" is pressed, into next setting window
Mouthful.
2) Cell A show that acquiescence focus is One as shown in fig. 7, operator's selection is needed to any Gel processing
Mini Gel, press "enter" key", into next setting window.
3) Cell A are shown as shown in figure 8, instrument work can be arranged in operator by the key up and down of operation panel
Time, acquiescence focus are 7min, "enter" key" are pressed, into next setting window.
4) prompt message is step 1) -3) the confirmation prompts of all operation settings, as shown in figure 9, the focus of acquiescence is
Start presses "enter" key", which starts to work, while entering next interface.
5) as shown in Figure 10 when working, it is shown as job category+activity bar+count down time+pause symbol, in this shape
Under state, pause symbol is focus, presses Enter, operating room's break-off；It simultaneously can select that other are arranged by upper lower key
Operating room.
Embodiment 8
The dyeing of large biological molecule transferring film, the equipment of coloring system disclosed in this invention based on efficient high sensitivity or
During migration operating process：
1. dyeing course
As shown in figure 11, lower layer's filter paper, gel, upper layer filter paper are sequentially placed between the upper/lower electrode of instrument operating room,
It closes the lid of instrument operating room and is pushed into workspace, the channel parameters of operating room are set and start instrument operation.Wherein：
Lower layer's filter paper：Lower layer's filter paper mainly provides specific buffer system and makes electric field energy in this sandwich structure
The distribution of even stabilization, while the environment so that protein belt positive charge being provided.Alcohol component therein makes gel surface shrink, egg
White matter or nucleic acid cannot run out from gel.
Gel：Gel is the main carriers of large biological molecule, mainly there is polyacrylamide gel and Ago-Gel, biology
Macromolecular is mainly protein and nucleic acid.
Upper layer filter paper：Specific buffer system is mainly provided and so that electric field energy is uniform and stable in this sandwich structure
Distribution, meanwhile, alcohol component therein makes glue surface shrinkage, positively charged protein that cannot run out from gel.Wherein
It also can be with staining for protein or the negative electrical charge dyestuff of nucleic acid.
As shown in figure 12, when instrument works, upper electrode provides the cathode of electric field, and lower electrode provides the anode of electric field,
Electronegative dye molecule passes through gel to reach lower layer's filter paper from upper layer filter paper from top to down；A：Dye molecule reaches lower layer's filter paper
Afterwards, machine can provide a reversed electric field, by controlling electric field duration so that dye molecule reaches gel from lower layer's filter paper
In；B：Subsequent machine provides a positive field again so that dye molecule turns again in lower layer's filter paper.AB steps 3 are repeated to arrive
10 times, last machine provides a lasting positive field and excess dyestuff is fully fallen in lower layer's filter paper, can make in this way
The sensitivity that must be dyed improves, while will not so that the time is too short and dyeing sensitivity is made to decline.
2. During migration
As shown in figure 13, it is sequentially placed into lower layer's filter paper, film, gel, upper layer filter paper between the upper/lower electrode of instrument, closes
It the lid of instrument operating room and is pushed into workspace, this job note is controlled according to the operating process of instrument shown in embodiment 5
Member.Wherein：
Lower layer's filter paper：Lower layer's filter paper mainly provides specific buffer system and makes electric field energy in this sandwich structure
The distribution of even stabilization, while the environment so that protein belt negative electrical charge being provided.
Film：Predominantly pvdf membrane and NC films, film can be that protein is adsorbed on its surface, and the macromoleculars such as protein cannot be worn
Cross film.
Gel：Gel is the main carriers of large biological molecule, mainly there is polyacrylamide gel and Ago-Gel, biology
Macromolecular is mainly protein and nucleic acid.
Upper layer filter paper：Specific buffer system is mainly provided and so that electric field energy is uniform and stable in this sandwich structure
Distribution.
When instrument works, upper electrode persistently provides the cathode of electric field, and lower electrode persistently provides the anode of electric field, larger
Electric current make protein or the particle of the negative point of other band quickly be transferred out from gel, while with the surface of film
In conjunction with.Protein or other electronegative biomolecule can carry out the experiment of next step after being combined with film.
The experimental result that the present invention carries out protein transferring film, dyeing and separating experiment is as follows：
1. transferring film is tested
Laboratory sample is His- albumen, and sample amount is 1 (A):5ul,2(B):4ul,3(C):3ul,4(D):2ul,5(E):
1ul,6(F):0.5ul,7(E):0.1ul after the same terms electrophoresis, uses conventional method (wet method) transferring film 100 minutes respectively, and
With system of the present invention and the quick transferring film of instrument 10 minutes, two panels film is incubated 1,2 with same condition together after transferring film
Antibody, darkroom expose 30 seconds, Figure 14 be incubated after conventional method (wet method) transferring film exposure as a result, Figure 15 is instrument of the present invention
The result of exposure is incubated after the quick transferring film of device.The result shows that with system of the present invention and instrument, turn in convenient and efficient improve
While membrane efficiency, transferring film sensitivity also significantly improves for conventional method.
2. Coloration experiment
Laboratory sample is bovine serum albumin(BSA), and applied sample amount is：1(A):1000ng,2(B):300ng,3(C):100ng,4
(D):30ng,5(E):10ng,6(F):3ng, Figure 16 use the experimental result that conventional coloring method is dyed：1h is dyed, so
Rear decoloring 3h (in three times, first time 0.5h, second of 1h, the 3rd 1.5h), wherein：Dyeing liquor：0.1%R250,50% first
Alcohol, 10% acetic acid, 40% water；Destainer：50% methanol, 10% acetic acid, 40% water.Figure 17 be with system of the present invention and
Instrument dyes the result of 6min.The result shows that substantially reducing dyeing time with system of the present invention and instrument, and dye
Sensitivity is also significantly improved than traditional dyeing method, and same sample conventional method can detect that applied sample amount is 30ng, and use this hair
The bright system and instrument can detect to 10ng, three times up to conventional method of sensitivity.
Claims (10)
1. a kind of large biological molecule transferring film of efficient high sensitivity, coloring system, including working cell are equipped at least one logical
Road is for testing and analyzing sample；The associated operation part with working cell, creates at least one dyeing or transferring film is logical
Road parameter, each dyeing or transferring film channel parameters include electric field cycle-index at least once.
2. the large biological molecule transferring film of efficient high sensitivity according to claim 1, coloring system, it is characterised in that：Institute
The operation part stated, the following relevant parameters of at least one of setting：Channel number, dyeing/transferring film, Gel numbers, the working time,
Operating voltage, operating current, electric field cycle-index, validation of information, startup/pause, analysis/processing of input data.
3. special according to the efficient highly sensitive large biological molecule transferring film of claims 1 or 2 any one of them, coloring system
Sign is：The acceleration detection large biological molecule system further includes being set for recording the storage of operation and testing result data
It is standby.
4. special according to the efficient highly sensitive large biological molecule transferring film of claims 1 or 2 any one of them, coloring system
Sign is：The acceleration detection large biological molecule system further includes the normal operation for policer operation and judges that work is
The monitoring system of no end.
5. a kind of large biological molecule transferring film, the equipment of coloring system based on efficient high sensitivity described in claim 1, including
Pedestal and shell for providing the inside and outside support of machine, hull outside are equipped with operation panel, and pedestal and enclosure interior are set
There are workspace and the system power supply being connected with workspace and operating circuit plate, the workspace that one drawing and pulling type independence is at least set
Operating room, in the operating room installation be provided with the positive-negative electrode plate of electric field.
6. large biological molecule transferring film, the equipment of coloring system according to claim 5 based on efficient high sensitivity, special
Sign is：The chamber bottom is equipped with saddle, and the positive electrode is fixedly mounted on saddle；Installation can open above operating room
Lid, lid be equipped with negative electrode and can uniformly press to electrode plate and sandwich structure to be analyzed gland locking dress
It sets.
7. large biological molecule transferring film, the equipment of coloring system according to claim 6 based on efficient high sensitivity, special
Sign is：The pressure locking device by positioned at lid surface runner, the eccentric wheel that is fixedly connected with runner and with bias
Opposite clamp composition is taken turns, the eccentric wheel bottom is wedge type slide unit structure, and the clamp is fixedly connected with a pressing plate, the pressing plate
Lower section is equipped with negative electrode or is directly fixedly connected with negative electrode.
8. large biological molecule transferring film, the equipment of coloring system according to claim 7 based on efficient high sensitivity, special
Sign is：The contour bolt that the pressing plate is cased with spring by four is spirally connected with lid.
9. according to large biological molecule transferring film of the claim 5-8 any one of them based on efficient high sensitivity, coloring system
Equipment, it is characterised in that：The equipment further includes display panel, working gas outlet and purifier, wherein described
Display panel is TFT liquid crystal or touch LCD screen, and the purifier, which is located in working gas outlet, to be surveyed, and the inside is equipped with work
Property charcoal.
10. according to large biological molecule transferring film of the claim 5-8 any one of them based on efficient high sensitivity, coloring system
Equipment, it is characterised in that：The operation panel is microswitch, touch key-press, remote control, touches in screen, film key
Any one.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012102468958A CN102749369A (en) | 2012-07-17 | 2012-07-17 | High-efficiency and high-sensitivity biomacromolecule film transferring and dyeing system and equipment |
| CN2012102468958 | 2012-07-17 | ||
| CN201310025508.2A CN104075916A (en) | 2012-07-17 | 2013-01-22 | High-efficiency and high-sensitivity biomacromolecule trans-membrane and staining system and device |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310025508.2A Division CN104075916A (en) | 2012-07-17 | 2013-01-22 | High-efficiency and high-sensitivity biomacromolecule trans-membrane and staining system and device |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108387422A true CN108387422A (en) | 2018-08-10 |
Family
ID=47029731
Family Applications (3)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012102468958A Pending CN102749369A (en) | 2012-07-17 | 2012-07-17 | High-efficiency and high-sensitivity biomacromolecule film transferring and dyeing system and equipment |
| CN201810153291.6A Pending CN108387422A (en) | 2012-07-17 | 2013-01-22 | A kind of large biological molecule transferring film, coloring system and the equipment of efficient high sensitivity |
| CN201310025508.2A Pending CN104075916A (en) | 2012-07-17 | 2013-01-22 | High-efficiency and high-sensitivity biomacromolecule trans-membrane and staining system and device |
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| CN2012102468958A Pending CN102749369A (en) | 2012-07-17 | 2012-07-17 | High-efficiency and high-sensitivity biomacromolecule film transferring and dyeing system and equipment |
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| CN201310025508.2A Pending CN104075916A (en) | 2012-07-17 | 2013-01-22 | High-efficiency and high-sensitivity biomacromolecule trans-membrane and staining system and device |
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| CN (3) | CN102749369A (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103852364A (en) * | 2012-11-29 | 2014-06-11 | 南京金斯瑞生物科技有限公司 | Biopolymer automatic staining method, and staining device |
| US20150153307A1 (en) | 2009-06-29 | 2015-06-04 | Nanjingjinsirui Science & Technology Biology Corporation | System for rapid electrophoresis binding method and related kits and compositions |
| CN110249220B (en) * | 2017-04-24 | 2023-01-31 | 赛默飞世尔（上海）仪器有限公司 | Biological transfer apparatus, biological transfer system, and control method |
| CN108425200B (en) * | 2018-05-13 | 2024-03-12 | 新疆天通兴业农牧科技有限公司 | Water-saving mineralized cashmere dyeing device |
| JP7103144B2 (en) * | 2018-10-12 | 2022-07-20 | 株式会社島津製作所 | Automatic analyzer |
| CN111982638B (en) * | 2020-08-20 | 2023-11-10 | 诒福生物科技南通有限公司 | Biological tissue staining method, apparatus, computer device, and storage medium |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101936837A (en) * | 2009-06-26 | 2011-01-05 | 南京金斯瑞生物科技有限公司 | Rapid staining method for biopolymers |
| US20110297544A1 (en) * | 2009-12-10 | 2011-12-08 | Bio-Rad Laboratories, Inc. | Electrotransfer cassette with integrated electrical contacts and locking mechanism |
| US20110297545A1 (en) * | 2009-12-11 | 2011-12-08 | Bio-Rad Laboratories, Inc. | Instrument for independent electrotransfer in multiple cassettes |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AUPR398601A0 (en) * | 2001-03-26 | 2001-04-26 | Proteome Systems Ltd | Blotting system and apparatus |
| CN101539496B (en) * | 2009-04-17 | 2011-05-18 | 首都医科大学 | Method used for selective dyeing after electrophoretic separation of protein |
-
2012
- 2012-07-17 CN CN2012102468958A patent/CN102749369A/en active Pending
-
2013
- 2013-01-22 CN CN201810153291.6A patent/CN108387422A/en active Pending
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101936837A (en) * | 2009-06-26 | 2011-01-05 | 南京金斯瑞生物科技有限公司 | Rapid staining method for biopolymers |
| US20110297544A1 (en) * | 2009-12-10 | 2011-12-08 | Bio-Rad Laboratories, Inc. | Electrotransfer cassette with integrated electrical contacts and locking mechanism |
| US20110297545A1 (en) * | 2009-12-11 | 2011-12-08 | Bio-Rad Laboratories, Inc. | Instrument for independent electrotransfer in multiple cassettes |
Non-Patent Citations (2)
| Title |
|---|
| 胡琳: "《植物脱毒技术》", 31 July 2007 * |
| 陈宏: "《基因工程实验技术》", 31 August 2008 * |
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| Publication number | Publication date |
|---|---|
| CN104075916A (en) | 2014-10-01 |
| CN102749369A (en) | 2012-10-24 |
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