CN101957337B - Method and system for analyzing electrophoresis components - Google Patents
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Abstract
The invention relates to a method and a system for analyzing electrophoresis components, wherein the method comprises the following steps: prestoring a corresponding relation of the contents of the electrophoresis components and the electric fields of the electrophoresis components when electrophoresis is carried out on all the electrophoresis components under the action of different electric fields for electrophoresis and in electrophoresis liquid with different pH values; and detecting the electric fields of the components to be detected when the electrophoresis is carried out on the electrophoresis components under the action of the set electric fields for electrophoresis and in the electrophoresis liquid with a set pH value, and obtaining the contents of the electrophoresis components to be detected according to the corresponding relation of the contents of the electrophoresis components and the electric fields of the electrophoresis components when electrophoresis is carried out on all the electrophoresis components under the action of the set electric fields for electrophoresis and in the electrophoresis liquid with the set pH value. By adopting the technical scheme of the invention, the contents of the electrophoresis components can be analyzed quantitatively in real time.
Description
Technical field
The present invention relates to biochemical field, particularly relate to a kind of electrophoretic fractians analytical approach and system.
Background technology
Electrophoresis refers to charged particle or molecule mobile phenomenon in direct current electrophoresis electric field, at biochemical field, can be protein, nucleic acid molecules, gel molecular etc. as charged particle or the molecule of electrophoretic fractians.Electrophoresis is under the effect of electrophoresis electric field, carry out in the electrophoresis liquid of certain pH value, the migration velocity of electrophoretic fractians by its with the quantity of electric charge and distribute to determine, after the pH value of electrophoresis electric field, electrophoresis liquid and electrophoretic fractians were determined, the migration velocity of electrophoretic fractians had also just been determined.Take protein as example, as a kind of ampholyte, protein will be with electric charge under certain PH condition, the kind of protein is different, with charge species, the quantity of electric charge and distribution thereof also different, therefore, the potpourri of different proteins is placed on the starting point of electrophoresis tank, behind the beginning electrophoresis, because the migration velocity of different proteins under same electrophoresis electric field action is different, thereby after after a while, homologous protein forms the electrophoresis band of this protein because migration velocity is identical, and different types of protein is arranged in different electrophoresis bands and by separately.Therefore, electrophoresis can be used as a kind of means that different types of Separation of Proteins is come.
Electrophoresis just with the means of the qualitative separation of different types of electrophoretic fractians, wants further to analyze various electrophoretic fractianses, also needs to carry out quantitative detection.The technology of existing quantitative detection electrophoretic fractians utilizes optics, spectroscopy, electrochemical method to carry out.The technology that quantitatively detects electrophoretic fractians take gel electrophoresis is example, prior art is at first carried out the electrophoresis of gel, different types of gel is separated, then all gels of separating are peeled off respectively gel slab, dry fixing, dyeing and decolouring, optical scanning obtains spectrum, draw the operations such as spectrum correction curve, each step operation described here is all very time-consuming, take drying fixedly operation as example, need to carry out respectively drying to all gels of separating with the gel drying instrument processes, and the baking temperature in this operation, dried gel humidity and drying time length all can affect the quality that electrophoretic fractians is analyzed, other operations also are like this.
This shows, utilize prior art quantitatively to detect electrophoretic fractians, the operation that need to carry out is a lot, and each step operation is all very consuming time, and therefore, analyzing each time all will be through just obtaining testing result for a long time.
Summary of the invention
Technical matters to be solved by this invention provides a kind of electrophoretic fractians analytical approach and system, content that can real-time quantitative ground analytical electrophoresis composition.
The technical scheme that the present invention solves the problems of the technologies described above is as follows: a kind of electrophoretic fractians analytical approach, and the method comprises:
The various electrophoretic fractianses of pre-save are at the electrophoretic fractians content during electrophoresis and the corresponding relation of composition electric field in the electrophoresis liquid under the different electrophoresis electric field actions, in different PH;
Electrophoretic fractians to be detected set under the electrophoresis electric field action, when setting in the electrophoresis liquid of pH value electrophoresis, detect the composition electric field of described electrophoretic fractians to be detected, according to described electrophoretic fractians to be detected set under the electrophoresis electric field action, electrophoretic fractians content when setting electrophoresis in the electrophoresis liquid of pH value and the corresponding relation of composition electric field, obtain the content of described electrophoretic fractians to be detected.
In addition, the present invention also provides a kind of electrophoretic fractians analytic system, and this system comprises memory storage and detection and analytical equipment, wherein:
Described memory storage is used for, and preserves various electrophoretic fractianses at the electrophoretic fractians content during electrophoresis and the corresponding relation of composition electric field in the electrophoresis liquid under the different electrophoresis electric field actions, in different PH;
Described detection and analytical equipment are used for, when electrophoretic fractians to be detected is being set under the electrophoresis electric field action, when setting in the electrophoresis liquid of pH value electrophoresis, detected the composition electric field of described electrophoretic fractians to be detected; From described memory storage obtain described electrophoretic fractians to be detected set under the electrophoresis electric field action, electrophoretic fractians content when setting electrophoresis the electrophoresis liquid of pH value and the corresponding relation of composition electric field; According to described memory storage obtain described electrophoretic fractians to be detected set under the electrophoresis electric field action, electrophoretic fractians content when setting electrophoresis in the electrophoresis liquid of pH value and the corresponding relation of composition electric field, obtain the content of described electrophoretic fractians to be detected.
The invention has the beneficial effects as follows: among the present invention, since pre-save various electrophoretic fractianses under different electrophoresis electric field actions, electrophoretic fractians content in the electrophoresis liquid of different PH during electrophoresis and the corresponding relation of composition electric field, when electrophoretic fractians to be detected is being set under the electrophoresis electric field action, in setting the electrophoresis liquid of pH value during electrophoresis, only need to detect the composition electric field of electrophoretic fractians to be detected, can set under the electrophoresis electric field action at this according to electrophoretic fractians to be detected, electrophoretic fractians content when this sets electrophoresis in the electrophoresis liquid of pH value and the corresponding relation of composition electric field, obtain the content of electrophoretic fractians to be detected, therefore, utilize technical scheme of the present invention, in electrophoresis, get final product the content that real-time quantitative ground obtains electrophoretic fractians to be detected.
Description of drawings
Fig. 1 is electrophoretic fractians analytical approach process flow diagram provided by the invention;
Fig. 2 is the cross-sectional configuration figure of thin film transistor (TFT);
Fig. 3 is the schematic diagram that thin film transistor (TFT) detects the composition electric field;
Fig. 4 is electrophoretic fractians analytic system structural drawing provided by the invention;
Fig. 5 is the vertical view of electrophoretic fractians analytic system the first embodiment provided by the invention;
Fig. 6 is the vertical view of three kinds of protein embodiment of electrophoresis in electrophoretic fractians analytic system provided by the invention;
Fig. 7 is the situation of change figure of the detecting device output current that causes respectively of three kinds of protein;
Fig. 8 is the vertical view of electrophoretic fractians analytic system the second embodiment provided by the invention;
Fig. 9 is detection in the two dimensional electrophoresis elemental analysis system provided by the invention and the example structure figure of analytical equipment.
Embodiment
Below in conjunction with accompanying drawing principle of the present invention and feature are described, institute gives an actual example and only is used for explaining the present invention, is not be used to limiting scope of the present invention.
Fig. 1 is electrophoretic fractians analytical approach process flow diagram provided by the invention.As shown in Figure 1, the method comprises:
Step 101: the various electrophoretic fractianses of pre-save are at the electrophoretic fractians content during electrophoresis and the corresponding relation of composition electric field in the electrophoresis liquid under the different electrophoresis electric field actions, in different PH;
Here, the electrophoresis electric field is the power that makes electrophoretic fractians generation electrophoresis, is DC electric field, and among the present invention, the direction of an electric field of electrophoresis electric field is horizontal direction.The intensity of electrophoresis electric field varies in size, and the migration velocity of electrophoretic fractians is also different, and electrophoresis electric field intensity is larger, and then the migration velocity of electrophoretic fractians is also just larger.Therefore, among the present invention, need the various electrophoretic fractianses of pre-save the electrophoretic fractians content during electrophoresis and corresponding relation of composition electric field under different electrophoresis electric field actions.
Electrophoretic fractians comprises the biomolecule such as range protein, nucleic acid molecules, gel molecular.When in electrophoresis liquid, carrying out electrophoresis, all carry a certain amount of electric charge above the various electrophoretic fractianses, the electric field of these charge generation is called the composition electric field of electrophoretic fractians, because the pH value of electrophoresis liquid is different, various electrophoretic fractianses with the quantity of electric charge and distribute just different, the composition electric field that these electric charges produce and the migration velocity of electrophoretic fractians are also all different, therefore, among the present invention, need the various electrophoretic fractianses of pre-save the electrophoretic fractians content during electrophoresis and corresponding relation of composition electric field in the electrophoresis liquid of different PH.
Step 102: electrophoretic fractians to be detected is being set under the electrophoresis electric field action, when setting in the electrophoresis liquid of pH value electrophoresis, is being detected the composition electric field of electrophoretic fractians to be detected.
Here, when electrophoretic fractians to be detected is detected, the PH of electrophoresis liquid is for setting pH value, the electrophoresis electric field is also for setting the electrophoresis electric field, the electrophoresis electric field of the pH value of this setting and setting all is included in the scope of the different PH described in the step 101 and different electrophoresis electric fields, like this, the present invention just can pass through query steps 101 described corresponding relations, comes real-time quantitative ground analytical electrophoresis composition.
Electric charge on the electrophoretic fractians to be detected can produce the composition electric field, the composition electric field can obtain its intensity by detection means, thereby according to the conversion method of electric field and the quantity of electric charge, obtain the quantity of electric charge on the electrophoretic fractians to be detected, and then obtain the content of this electrophoretic fractians to be detected.If multiple electrophoretic fractians to be detected is arranged, then at first need to determine the kind of the different electrophoretic fractianses to be detected separated in the electrophoresis process, and then determine the content of each electrophoretic fractians to be detected.Because the specific charge of every kind of electrophoretic fractians in the certain electrophoresis liquid of pH value is given data, when the electrophoresis electric field intensity of electrophoresis liquid is determined, the migration velocity of every kind of electrophoretic fractians has also just been determined, therefore, the mix ingredients of multiple electrophoretic fractians to be detected is through after the electrophoresis of certain hour, can determine by the position of isolated different electrophoresis bands the kind of each electrophoretic fractians to be detected, recycle the quantity of electric charge on the formed electrophoresis band of each electrophoretic fractians to be detected, determine the content of this electrophoretic fractians.
Step 103: according to electrophoretic fractians to be detected set under the electrophoresis electric field action, electrophoretic fractians content when setting electrophoresis in the electrophoresis liquid of pH value and the corresponding relation of composition electric field, obtain the content of electrophoretic fractians to be detected.
Here, obtain the composition electric field of electrophoretic fractians to be detected in step 102 after, just can according to this electrophoretic fractians to be detected of pre-save described in the step 101 set under the electrophoresis electric field action, electrophoretic fractians content when setting electrophoresis in the electrophoresis liquid of pH value and the corresponding relation of composition electric field, obtain the content of electrophoretic fractians to be detected.
The content of electrophoretic fractians to be detected can be the quality of electrophoretic fractians to be detected, or be the volume of electrophoretic fractians to be detected, or be the quantity of the molecule of electrophoretic fractians to be detected, or be the mass percent of electrophoretic fractians to be detected in all electrophoretic fractianses to be detected, or be the percent by volume of electrophoretic fractians to be detected in all electrophoretic fractianses to be detected, or be the ratio of molecular amounts in the molecular amounts of all electrophoretic fractianses to be detected of electrophoretic fractians to be detected.
In the step 102, the method that detects the composition electric field of electrophoretic fractians to be detected is: the variable quantity that detects the detecting device output electrical signals that the composition electric field of electrophoretic fractians to be detected causes; According to the variable quantity of detecting device output electrical signals, determine the composition electric field of electrophoretic fractians to be detected.
If electrophoretic fractians to be detected further is two or more electrophoretic fractians to be detected; The method that then detects the composition electric field of electrophoretic fractians to be detected in the step 102 is:
The various electrophoretic fractianses of pre-save are at the electrophoretic fractians kind during electrophoresis and the corresponding relation of its translational speed in the electrophoresis liquid under the different electrophoresis electric field actions, in different PH;
Each electrophoretic fractians to be detected set under the electrophoresis electric field action, when setting in the electrophoresis liquid of pH value electrophoresis, detect the translational speed of each electrophoretic fractians to be detected, according to each electrophoretic fractians to be detected set under the electrophoresis electric field action, electrophoretic fractians kind when setting electrophoresis in the electrophoresis liquid of pH value and the corresponding relation of its translational speed, determine the kind of each electrophoretic fractians to be detected;
Detect the variable quantity of the caused detecting device output electrical signals of composition electric field of every kind of electrophoretic fractians to be detected; According to the variable quantity of the caused detecting device output electrical signals of composition electric field of every kind of electrophoretic fractians to be detected, determine the composition electric field of this electrophoretic fractians to be detected.
Here, detecting device does not have output electrical signals in the time spent of doing that is not subject to adding the composition electric field, perhaps export a constant electric signal, perhaps export an electric signal that changes, the electric signal of this variation can be the electric signal of sine gauge rule variation, or step signal, or the electric signal of other variations.When the composition electric field action of electrophoretic fractians to be detected during in detecting device, can make the output electrical signals of detecting device that certain variation occurs, for example, detecting device output electrical signals not when not adding the composition electric field is when composition electric field action output electrical signals during in detecting device; Detecting device is not exported a constant electric signal when perhaps adding the composition electric field, when the composition electric field action is exported another electric signal during in detecting device, this electric signal is along with the composition electric field strengthens gradually near detecting device gradually, when the composition electric field is nearest from detecting device, this output electrical signals reaches peak value, then along with the composition electric field gradually away from detecting device, the electric signal of output weakens gradually, the effect of ultimate constituent electric field disappears, and output electrical signals reverts to original constant electric signal; The electric signal that detecting device output one changes when perhaps not adding the composition electric field for example, is exported the signal that sinusoidal rule changes, when composition electric field action during in detecting device, the output electrical signals of detecting device is amplitude-modulated signal, and its amplitude is the direct ratio function of composition electric field intensity, etc.
Utilize the variable quantity of detecting device output electrical signals, can according to the response pattern of the known output signal of detecting device to input signal, determine the composition electric field of electrophoretic fractians to be detected.For example, if detecting device is exported a constant electric signal when not adding the composition electric field, when the composition electric field action is exported another electric signal during in detecting device, this electric signal is along with the composition electric field strengthens gradually near detecting device gradually, when the composition electric field is nearest from detecting device, this output electrical signals reaches peak value, then along with the composition electric field gradually away from detecting device, the electric signal of output weakens gradually, the effect of ultimate constituent electric field disappears, output electrical signals reverts to original constant electric signal, so, and can be according to the response pattern of following the intrinsic output signal of detecting device to input signal, the peak value of output electrical signals is deducted the constant electric signal of original output, as the variable quantity of output electrical signals, according to the size of this variable quantity, determine the composition electric field of electrophoretic fractians to be detected.In addition, in the situation of carrying out the position of the starting point of electrophoresis and detecting device all fixing at electrophoretic fractians, when the pH value of electrophoresis liquid and electrophoresis electric field have all been set, the time that can occur according to the electrical signal peak of electrophoresis start time and detecting device output, calculate the migration velocity of electrophoretic fractians to be detected, and then utilize this migration velocity, and the various electrophoretic fractianses of pre-save are determined the kind of electrophoretic fractians to be detected at the electrophoretic fractians kind during electrophoresis and the corresponding relation of its translational speed in the electrophoresis liquid under the different electrophoresis electric field actions, in different PH.
In the electrophoretic fractians analytical approach provided by the invention, detecting device can have multiple choices, for example, can be thin film transistor (TFT), detecting device that also can output electrical signals changes when utilizing the composition electric field influence of other charge generation on being subject to electrophoretic fractians to be detected, thin film transistor (TFT) is specifically as follows amorphous silicon film transistor (TFT), or is polycrystalline SiTFT, or other thin film transistor (TFT)s.
Here, because the detecting device among the present invention is the detecting device of output electrical signals, when being subject to extra electric field and affecting, its output electrical signals can change in real time, therefore, this detecting device can be connected with the real-time analysis device, intensity that can this composition electric field of Analysis deterrmination when detecting device is subject to adding the impact of composition electric field, and then kind, migration velocity and the content etc. of definite electrophoretic fractians to be detected, and error is very little, with respect to prior art, electrophoretic fractians analytical approach provided by the invention is simple to operate, effective, real-time is good and accuracy is high.
Fig. 2 is the cross-sectional configuration figure of thin film transistor (TFT).As shown in Figure 2, thin film transistor (TFT) is made on the substrate 201, and this substrate can be glass, also can be monocrystalline silicon, or is other materials.Thin film transistor (TFT) has two grids, be that bottom grid 202 and electric charge bring out grid 203, the material of bottom grid 202 doped crystalline silicon of can attaching most importance to, also can be other materials, bottom grid 202 can be used as the break-make that control electrode is controlled the thin film transistor (TFT) output current, also can control with drain electrode 205 break-make of thin film transistor (TFT) output current.In addition, the source electrode of thin film transistor (TFT) is 204.There is insulation course 207 outside of bringing out grid 203 at bottom grid 202 and electric charge, be thin film semiconductive layer 206 between this dielectric layers 207, the material of thin film semiconductive layer 206 can be amorphous silicon, the material that also can have for other characteristic of semiconductor, thin film semiconductive layer 206 is used for receiving the effect of extra electric field, output current size between source electrode 204 and the drain electrode 205 is changed, when DC Electric Field during in thin film semiconductive layer 206, output current between the source electrode 204 of thin film transistor (TFT) and the drain electrode 205 can change, and present this Changing Pattern, be the transfer characteristics of thin film transistor (TFT), be known rules.
Fig. 3 is the schematic diagram that thin film transistor (TFT) detects the composition electric field.As shown in Figure 3, after offset side 301 places of thin film transistor (TFT) add bias voltage, can between drain electrode and source electrode, produce constant output current, be in conducting state when reading control electrode 302 control bottom grids 202, and the electrophoretic fractians to be detected with electric charge, such as protein electrophorese band, nucleic acid molecules etc., when bringing out grid 203 near electric charge, the formed composition electric field action of electric charge in the electrophoretic fractians to be detected is in the thin film semiconductive layer of thin film transistor (TFT), thus the size of change output current.The output current of thin film transistor (TFT) enters analytical equipment 4022, and analytical equipment 4022 is by analyzing the variable quantity of this output current, the content of Analysis deterrmination electrophoretic fractians to be detected.Because the quantity of electric charge and distribution in the electrophoretic fractians to be detected have determined the composition electric field, and the output current variable quantity of composition electric field and thin film transistor (TFT) has specific proportionate relationship, therefore, analytical equipment 4022 is analyzed the output current variable quantity that obtains thin film transistor (TFT), just can determine electrophoretic fractians to be detected with total charge dosage, for each electrophoretic fractians to be detected, because the molecule of the single electrophoretic fractians to be detected electrically charged amount of institute and be distributed as known, therefore, obtain electrophoretic fractians to be detected with total charge dosage after, just can determine the molecular amounts of electrophoretic fractians to be detected, and then determine the quality of electrophoretic fractians to be detected, volume and this tittle be shared ratio in all electrophoretic fractianses to be detected.
In the step 101, preserving various electrophoretic fractianses under different electrophoresis electric field actions, before electrophoretic fractians content in the electrophoresis liquid of different PH during electrophoresis and the corresponding relation of composition electric field, the method further comprises: determine that various electrophoretic fractianses are under different electrophoresis electric field actions, electrophoretic fractians content in the electrophoresis liquid of different PH during electrophoresis and the corresponding relation of composition electric field, the method of determining is: according to every kind of electrophoretic fractians of unit content under different electrophoresis electric field actions, CHARGE DISTRIBUTION in the electrophoresis liquid of different PH during electrophoresis, calculate the composition electric field of this electrophoretic fractians, obtain every kind of electrophoretic fractians of unit content under different electrophoresis electric field actions, electrophoretic fractians content in the electrophoresis liquid of different PH during electrophoresis and the corresponding relation of composition electric field;
Then according to electrophoretic fractians to be detected set under the electrophoresis electric field action, electrophoretic fractians content when setting electrophoresis in the electrophoresis liquid of pH value and the corresponding relation of composition electric field, the method that obtains the content of electrophoretic fractians to be detected is:
Calculate electrophoretic fractians to be detected set under the electrophoresis electric field action, when setting in the electrophoresis liquid of pH value electrophoresis, proportionate relationship between the composition electric field of the composition electric field of electrophoretic fractians to be detected and the electrophoretic fractians to be detected of unit content obtains the proportionate relationship between the content of electrophoretic fractians to be detected of the content of electrophoretic fractians to be detected and unit content;
According to the proportionate relationship between the content of the electrophoretic fractians to be detected of the content of electrophoretic fractians to be detected and unit content, obtain the content of electrophoretic fractians to be detected.
Here, the method for utilizing CHARGE DISTRIBUTION to calculate electric field is existing mature technology.At the every kind of electrophoretic fractians that utilizes unit content under different electrophoresis electric field actions, CHARGE DISTRIBUTION in the electrophoresis liquid of different PH during electrophoresis, calculate after the composition electric field of this electrophoretic fractians, because the content of this electrophoretic fractians is unit content, therefore, just can obtain the content of this electrophoretic fractians, it is the corresponding relation between the composition electric field of unit content and this electrophoretic fractians, and then, after detection obtains the composition electric field of electrophoretic fractians to be detected in step 102, can utilize the proportionate relationship between the composition electric field of the composition electric field of electrophoretic fractians to be detected and unit content composition to be detected, determine the content of this electrophoretic fractians to be detected.
Fig. 4 is electrophoretic fractians analytic system structural drawing provided by the invention.As shown in Figure 4, this system comprises memory storage 401 and detection and analytical equipment 402, wherein:
Detect with analytical equipment 402 and be used for, when electrophoretic fractians to be detected is being set under the electrophoresis electric field action, when setting in the electrophoresis liquid of pH value electrophoresis, detected the composition electric field of electrophoretic fractians to be detected; From memory storage 401 obtain electrophoretic fractians to be detected set under the electrophoresis electric field action, electrophoretic fractians content when setting electrophoresis the electrophoresis liquid of pH value and the corresponding relation of composition electric field; According to electrophoretic fractians to be detected set under the electrophoresis electric field action, electrophoretic fractians content when setting electrophoresis in the electrophoresis liquid of pH value and the corresponding relation of composition electric field, obtain the content of electrophoretic fractians to be detected.
Detection comprises detecting device 4021 and analytical equipment 4022 with analytical equipment 402, wherein:
Detecting device 4021 is used for, and the composition electric field of the electrophoretic fractians to be detected that self is received is made response, responds as output electrical signals changes, and the variable quantity of output electrical signals is sent to analytical equipment 4022;
Further, electrophoretic fractians to be detected can be two or more electrophoretic fractianses to be detected; At this moment, detect the kind that needs at first to determine each electrophoretic fractians to be detected with analytical equipment 402, then detect the variable quantity of the caused output electrical signals of composition electric field of each electrophoretic fractians to be detected, and then determine the content of each electrophoretic fractians to be detected;
Detection comprises detecting device 4021 and analytical equipment 4022 with analytical equipment 402, wherein:
Detecting device 4021 is used for, when each electrophoretic fractians to be detected setting under the electrophoresis electric field action, when setting in the electrophoresis liquid of pH value electrophoresis, detect the translational speed of each electrophoretic fractians to be detected, and the translational speed of each electrophoretic fractians to be detected is sent to analytical equipment 4022; The composition electric field of every kind of electrophoretic fractians to be detected that self is received is made response, responds as output electrical signals changes, and the variable quantity of the caused output electrical signals of composition electric field of every kind of electrophoretic fractians to be detected is sent to analytical equipment 4022;
In the electrophoretic fractians analytic system provided by the invention, detecting device has a lot of middle selections, for example, can be TFT, or be polycrystalline SiTFT, or be other thin film transistor (TFT)s, certainly, detecting device that also can output electrical signals changes when utilizing the composition electric field influence of other charge generation on being subject to electrophoretic fractians to be detected.
In addition, among the present invention, detecting device also can be more than one detecting device, for example, a detecting device can be, also the linear array of plural detecting device composition can be, the orientation of each detecting device is parallel with the direction of electrophoresis electric field, for example, the orientation of each detecting device is identical with the direction of electrophoresis electric field, or opposite; Because the orientation of each detecting device is parallel with the direction of electrophoresis electric field, and electrophoretic fractians to be detected is under the effect of electrophoresis electric field, carry out electrophoresis along the direction parallel with the direction of electrophoresis electric field, therefore, each detecting device can be successively detects the composition electric field of electrophoretic fractians to be detected, thereby determine its content, simultaneously, because the position of each detecting device and the position that electrophoretic fractians to be detected carries out the initiating terminal of electrophoresis are all fixed, the present invention can also determine the migration velocity of this electrophoretic fractians to be detected.Detecting device can also be the two-dimensional surface array that a plurality of detecting devices form, and this two-dimensional surface array is comprised of a plurality of linear arrays that are parallel to each other, and the detecting device on every linear array simultaneously or do not detect simultaneously.
Among the present invention, electrophoretic fractians to be detected is to carry out electrophoresis in electrophoresis tank, and detecting device is positioned at the below of electrophoresis tank.
Here, detecting device is positioned at the below of electrophoresis tank, when then electrophoretic fractians to be detected carries out electrophoresis in electrophoresis tank, the composition electric field of electrophoretic fractians to be detected just can closely act on detecting device, like this, it is very high that the precision of detecting device and sensitivity just can reach, thereby make the relative prior art of analysis result of the present invention more accurate.
Fig. 5 is the vertical view of electrophoretic fractians analytic system the first embodiment provided by the invention.As shown in Figure 5, electrophoretic fractians to be detected carries out electrophoresis in this system in electrophoresis tank 501, and the reference position of electrophoresis is the straight line place at 502 places, and under the effect of electrophoresis electric field, electrophoretic fractians to be detected carries out electrophoresis along electrophoresis direction 503.This system only uses a detecting device, is positioned at the below of electrophoresis tank, and is positioned on the electrophoresis path of electrophoretic fractians to be detected, and its drawing reference numeral is 4021, and the output current of detecting device 4021 enters analytical equipment 4022.
Fig. 6 is the vertical view of three kinds of protein embodiment of electrophoresis in electrophoretic fractians analytic system provided by the invention.As shown in Figure 6, three kinds of protein carry out electrophoresis in electrophoresis tank 501, because migration velocity is different, three kinds of protein form three independently electrophoresis bands along electrophoresis direction 503, the drawing reference numeral of these three electrophoresis bands is respectively 601,602 and 603, as seen from Figure 6, the migration velocity of electrophoresis band 601 is the fastest, and the migration velocity of electrophoresis band 603 is the slowest, like this, electrophoresis band 601,602 and 603 successively through being positioned at the detecting device 4021 of electrophoresis tank below, its composition electric field also just acts on detecting device 4021 successively, and the output current of detecting device 4021 is changed, and the output current of the 4022 pairs of detecting devices 4021 of analytical equipment that are connected with detecting device 4021 detects, thereby calculate the variable quantity of output current, and then the content of these three kinds of protein of Analysis deterrmination.The situation of change of the detecting device output current that these three kinds of protein cause respectively as shown in Figure 7.
Fig. 7 is the situation of change figure of the detecting device output current that causes respectively of three kinds of protein.Among Fig. 7, transverse axis represents the time, and the longitudinal axis represents output current or the voltage of detecting device.As shown in Figure 7, when not having DC Electric Field, the output current of detecting device is steady current, electrophoresis band 601 when three kinds of each self-formings of protein shown in Fig. 6,602 and 603 pass through detecting device 4021 successively, when its composition electric field also just acts on detecting device 4021 successively, the output current of detecting device 4021 changes, form drawing reference numeral shown in Figure 7 and be respectively 701,702 and 703 current peak, can be calculated the migration velocity of three kinds of protein by the time of occurrence of three peak values, and then obtain these three kinds of protein kind separately, can be determined the composition electric field of three kinds of protein by the size of three peak values, and then determine these three kinds of protein content separately.
Fig. 8 is the vertical view of electrophoretic fractians analytic system the second embodiment provided by the invention.As shown in Figure 8, detecting device in this system is the linear array that a plurality of detecting devices form, this linear array is positioned at the below of electrophoresis tank 501, the orientation of each detecting device is parallel with electrophoresis direction 503, namely parallel with the direction of electrophoresis electric field, the output signal of each detecting device all enters analytical equipment 4022.Three kinds of protein carry out electrophoresis in electrophoresis tank 501, form three electrophoresis bands that are separated from each other 601,602 and 603.Like this, the control electrode of reading of each detecting device 4021 makes corresponding detecting device be in conducting state successively, thereby each detecting device 4021 outputs signal to analytical equipment 4022 successively, analytical equipment 4022 is analyzed by the output signal to each detecting device, can determine the composition electric field of these three kinds of protein, and then determine these three kinds of protein content separately.
Fig. 9 is detection in the two dimensional electrophoresis elemental analysis system provided by the invention and the example structure figure of analytical equipment.As shown in Figure 9, detecting device in this detection and the analytical equipment is the two-dimensional surface array that a plurality of detecting devices form, all detecting devices all are positioned at the below of electrophoresis tank 501, each detecting device obtains bias voltage by offset line 902, the output electrical signals of each row detecting device enters same amplifier 904, electric signal after each amplifier amplifies enters analog to digital converter 906 by analog switch 905, thereby carries out next step analysis.Circled among Fig. 9 is that the electric charge of detecting device brings out grid 203.This detection and analytical equipment have many and read control line 901, read the break-make that control line 901 can be controlled the bottom grid 202 of delegation's detecting device for every, can be to amplifier 904 output electrical signals thereby control this row detecting device, perhaps can not be to amplifier 904 output electrical signals.Gate drivers 903 can be controlled the level height that each bar is read control line 901, and then the bottom grid 202 that control is respectively read in a plurality of detecting devices that control line 901 connects is in conducting state or not on-state, and gate drivers 903 can adopt TFT-liquid crystal (LCD) gate drivers.Among the embodiment shown in Figure 9, any time only has one to read the bottom grid 202 that control line 901 connects and be in conducting state, and when bottom grid 202 is in conducting state, amplifier 904 output electrical signals that these bottom grid 202 place detecting devices can connect to this row detecting device.
This shows, the present invention has the following advantages:
(1) among the present invention, since pre-save various electrophoretic fractianses under different electrophoresis electric field actions, electrophoretic fractians content in the electrophoresis liquid of different PH during electrophoresis and the corresponding relation of composition electric field, when electrophoretic fractians to be detected is being set under the electrophoresis electric field action, in setting the electrophoresis liquid of pH value during electrophoresis, only need to detect the composition electric field of electrophoretic fractians to be detected, can set under the electrophoresis electric field action at this according to electrophoretic fractians to be detected, electrophoretic fractians content when this sets electrophoresis in the electrophoresis liquid of pH value and the corresponding relation of composition electric field, obtain the content of electrophoretic fractians to be detected, therefore, utilize technical scheme of the present invention, in electrophoresis, get final product the content that real-time quantitative ground obtains electrophoretic fractians to be detected.
(2) among the present invention, because the detecting device that adopts is the detecting device of output electrical signals, when being subject to extra electric field and affecting, its output electrical signals can change in real time, therefore, this detecting device can be connected with the real-time analysis device, intensity that can this composition electric field of Analysis deterrmination when detecting device is subject to adding the impact of composition electric field, and then the kind of definite electrophoretic fractians to be detected, migration velocity and content etc., and error is very little, with respect to prior art, electrophoretic fractians analytical approach provided by the invention is simple to operate, effectively, real-time is good and accuracy is high.
(3) among the present invention, detecting device both can be a detecting device, can be linear array or the two-dimensional surface array of a plurality of detecting devices compositions again, different detector number and arrangement mode can have different purposes, for example, linear array can be used for the composition that real-time quantitative detects protein, gel etc., and the two-dimensional surface array can also further be applied to detect nucleic acid molecules, such as the order-checking of DNA, therefore, of the present inventionly be of wide application generally, and have a extensive future.
(4) among the present invention, since pre-save various electrophoretic fractianses under different electrophoresis electric field actions, electrophoretic fractians content in the electrophoresis liquid of different PH during electrophoresis and the corresponding relation of composition electric field, therefore, utilize technical scheme of the present invention to come real-time quantitative to detect the content of electrophoretic fractians to be detected, only need to utilize detecting device automatically to detect the composition electric field of this electrophoretic fractians to be detected, can utilize analytical equipment automatically to analyze in real time, whole process does not need manually substantially, therefore, technical scheme of the present invention is simple to operate, do not need a large amount of study to grasp, with respect to prior art, can reduce corresponding operating personnel's training cost, use cost is very low.
The above only is preferred embodiment of the present invention, and is in order to limit the present invention, within the spirit and principles in the present invention not all, any modification of doing, is equal to replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (8)
1. an electrophoretic fractians analytical approach is characterized in that, the method comprises:
The various electrophoretic fractianses of pre-save are at the electrophoretic fractians content during electrophoresis and the corresponding relation of composition electric field in the electrophoresis liquid under the different electrophoresis electric field actions, in different PH;
The various electrophoretic fractianses of pre-save are at the electrophoretic fractians kind during electrophoresis and the corresponding relation of its translational speed in the electrophoresis liquid under the different electrophoresis electric field actions, in different PH;
Two or more each electrophoretic fractians to be detected set under the electrophoresis electric field action, when setting in the electrophoresis liquid of pH value electrophoresis, detect the translational speed of each electrophoretic fractians to be detected, according to each electrophoretic fractians to be detected set under the electrophoresis electric field action, electrophoretic fractians kind when setting electrophoresis in the electrophoresis liquid of pH value and the corresponding relation of its translational speed, determine the kind of each electrophoretic fractians to be detected; Electrophoretic fractians to be detected set under the electrophoresis electric field action, when setting in the electrophoresis liquid of pH value electrophoresis, detect the described composition electric field of determining each electrophoretic fractians to be detected of kind, according to described electrophoretic fractians to be detected set under the electrophoresis electric field action, electrophoretic fractians content when setting electrophoresis in the electrophoresis liquid of pH value and the corresponding relation of composition electric field, obtain the content of described electrophoretic fractians to be detected.
2. method according to claim 1 is characterized in that, the method that detects the composition electric field of described electrophoretic fractians to be detected is: the variable quantity that detects the detecting device output electrical signals that the composition electric field of described electrophoretic fractians to be detected causes; According to the variable quantity of described detecting device output electrical signals, determine the composition electric field of described electrophoretic fractians to be detected.
3. method according to claim 2 is characterized in that, described detecting device is amorphous silicon film transistor.
4. method according to claim 1 and 2, it is characterized in that, preserving various electrophoretic fractianses under different electrophoresis electric field actions, before electrophoretic fractians content in the electrophoresis liquid of different PH during electrophoresis and the corresponding relation of composition electric field, the method further comprises: determine that various electrophoretic fractianses are under different electrophoresis electric field actions, electrophoretic fractians content in the electrophoresis liquid of different PH during electrophoresis and the corresponding relation of composition electric field, the method of determining is: according to every kind of electrophoretic fractians of unit content under different electrophoresis electric field actions, CHARGE DISTRIBUTION in the electrophoresis liquid of different PH during electrophoresis, calculate the composition electric field of this electrophoretic fractians, obtain every kind of electrophoretic fractians of unit content under different electrophoresis electric field actions, electrophoretic fractians content in the electrophoresis liquid of different PH during electrophoresis and the corresponding relation of composition electric field;
Then described according to described electrophoretic fractians to be detected set under the electrophoresis electric field action, electrophoretic fractians content when setting electrophoresis in the electrophoresis liquid of pH value and the corresponding relation of composition electric field, the method that obtains the content of described electrophoretic fractians to be detected is:
Calculate described electrophoretic fractians to be detected set under the electrophoresis electric field action, when setting in the electrophoresis liquid of pH value electrophoresis, proportionate relationship between the composition electric field of the composition electric field of described electrophoretic fractians to be detected and the electrophoretic fractians to be detected of described unit content obtains the proportionate relationship between the content of electrophoretic fractians to be detected of the content of described electrophoretic fractians to be detected and described unit content;
According to the proportionate relationship between the content of the electrophoretic fractians to be detected of the content of described electrophoretic fractians to be detected and described unit content, obtain the content of described electrophoretic fractians to be detected.
5. an electrophoretic fractians analytic system is characterized in that, this system comprises memory storage and detection and analytical equipment, wherein:
Described memory storage is used for, and preserves various electrophoretic fractianses at the electrophoretic fractians content during electrophoresis and the corresponding relation of composition electric field in the electrophoresis liquid under the different electrophoresis electric field actions, in different PH; And
Preserve various electrophoretic fractianses at the electrophoretic fractians kind during electrophoresis and the corresponding relation of its translational speed in the electrophoresis liquid under the different electrophoresis electric field actions, in different PH;
Described detection and analytical equipment are used for, when two or more each electrophoretic fractians to be detected are being set under the electrophoresis electric field action, when setting in the electrophoresis liquid of pH value electrophoresis, detected the translational speed of each electrophoretic fractians to be detected; Obtain each electrophoretic fractians to be detected at the electrophoretic fractians kind during electrophoresis and the corresponding relation of its translational speed under the described setting electrophoresis electric field action, the electrophoresis liquid of described setting pH value from described memory storage, and the translational speed of each electrophoretic fractians to be detected, determine the kind of each electrophoretic fractians to be detected; When two or more electrophoretic fractianses to be detected are being set under the electrophoresis electric field action, when setting in the electrophoresis liquid of pH value electrophoresis, detected the composition electric field of the electrophoretic fractians to be detected of described definite kind; From described memory storage obtain described electrophoretic fractians to be detected set under the electrophoresis electric field action, electrophoretic fractians content when setting electrophoresis the electrophoresis liquid of pH value and the corresponding relation of composition electric field; According to described electrophoretic fractians to be detected set under the electrophoresis electric field action, electrophoretic fractians content when setting electrophoresis in the electrophoresis liquid of pH value and the corresponding relation of composition electric field, obtain the content of described electrophoretic fractians to be detected.
6. system according to claim 5 is characterized in that, described detection and analytical equipment comprise detecting device and analytical equipment, wherein:
Described detecting device is used for, when each electrophoretic fractians to be detected setting under the electrophoresis electric field action, when setting in the electrophoresis liquid of pH value electrophoresis, detect the translational speed of each electrophoretic fractians to be detected, and the translational speed of each electrophoretic fractians to be detected is sent to described analytical equipment; The composition electric field of every kind of electrophoretic fractians to be detected that self is received is made response, and described response is that output electrical signals changes, and the variable quantity of the caused output electrical signals of composition electric field of every kind of electrophoretic fractians to be detected is sent to described analytical equipment;
Described analytical equipment is used for, obtain each electrophoretic fractians to be detected at the electrophoretic fractians kind during electrophoresis and the corresponding relation of its translational speed under the described setting electrophoresis electric field action, the electrophoresis liquid of described setting pH value from described memory storage, and the translational speed of each electrophoretic fractians to be detected, determine the kind of each electrophoretic fractians to be detected; According to the variable quantity of the caused described detecting device output electrical signals of composition electric field of every kind of electrophoretic fractians to be detected, determine the composition electric field of this electrophoretic fractians to be detected.
7. system according to claim 6 is characterized in that, described detecting device is thin film transistor (TFT).
8. system according to claim 6 is characterized in that,
Described detecting device is more than one detecting device; And if detecting device is plural detecting device, then the orientation of each detecting device is parallel with the direction of described electrophoresis electric field;
And/or,
Described electrophoretic fractians to be detected carries out electrophoresis in electrophoresis tank, described detecting device is positioned at the below of described electrophoresis tank.
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