CN103079032A - High-definition capillary electrophoresis gel imaging system - Google Patents
High-definition capillary electrophoresis gel imaging system Download PDFInfo
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Abstract
The invention relates to the biochemical technical field, and relates to a high-definition capillary electrophoresis gel imaging system, which comprises a lens and a focusing control system, wherein the focusing control system comprises an image acquisition module, a data processing and analyzing module, a serial port communication module and a hardware control drive circuit, wherein the image acquisition module is connected with the data processing and analyzing module; the data processing and analyzing module is connected with the serial port communication module; the serial port communication module is connected with a system controller; the system controller is connected with a drive motor through the hardware control drive circuit; the drive motor is connected with the lens; the lens comprises an aperture ring and a focusing ring; an aperture ring drive circuit is connected with an aperture drive motor; the aperture drive motor is connected with the aperture ring; the focusing drive circuit is connected with the focusing drive motor; and the focusing drive motor is connected with the focusing ring.
Description
Technical field
The present invention relates to the bioscience technical field, relate in particular to a kind of gel imaging system.
Background technology
Gel imaging equipment is a kind of dark box type imaging device of taking pictures.Domestic gel imaging equipment still adopts the quality of traditional human eye subjective visual evaluation image at present, and finish the Obtaining Accurate of image by manual adjustments aperture ring, focusing ring and zoom ring, the picture quality of obtaining when the hand labor amount is large is unsatisfactory.
Summary of the invention
The object of the present invention is to provide a kind of Capillary Electrophoresis gel imaging system of high definition, to solve the problems of the technologies described above.
Technical problem solved by the invention can realize by the following technical solutions:
A kind of Capillary Electrophoresis gel imaging system of high definition, comprise that a camera lens, is used for controlling the focus control system of described camera lens, it is characterized in that, described focus control system comprises an image capture module, a Data Management Analysis module, a serial communication module, a hardware controls drive circuit; The signal output part of described image capture module connects the signal input part of described Data Management Analysis module, the signal output part of described Data Management Analysis module connects the signal input part of described serial communication modular, and the signal output part of described serial communication modular connects a system controller;
Described system controller connects a drive motors by described hardware controls drive circuit, and described drive motors connects described camera lens;
Described camera lens comprises an aperture ring, a focusing ring, and described drive motors comprises an aperture drive motors, a focusing drive motors, and described hardware controls drive circuit comprises an aperture drive circuit, a focusing drive circuit; Described system controller connects respectively described aperture drive circuit and described focusing drive circuit; Described aperture drive circuit connects described aperture drive motors, and described aperture drive motors connects described aperture ring; Described focusing drive circuit connects described focusing drive motors, and described focusing drive motors connects described focusing ring.So that the present invention according to different image informations, controls different drive motors, finish forward and reverse motion control of aperture, focusing and zoom.
When the present invention uses, image capture module is used for Real-time Collection present image information, the Data Management Analysis module is used for the preliminary treatment of current image information and the evaluation of picture quality, and provides control signal, and serial communication modular is used for control signal by sending to system controller.The control signal control drive motors adjustable lens that system controller is read according to solution is realized the automatic focus of gel imaging system.
The present invention has abandoned the quality of traditional human eye subjective visual evaluation image, and the mode by manual adjustments, adds the automatic focus mode but adopt on the basis of former Manual focusing control system, has realized self-focusing purpose.
Image capture module, also comprise a capillary electrophoresis chip, light source, photoelectric detector, the signal output part of described photoelectric detector connects a Data Management Analysis module for data receiver, processing and control, described photoelectric detector comprises a ccd image sensor (charge coupled device), and the signal output part of described ccd image sensor connects described Data Management Analysis module;
The DNA sample enters from the starting point of described capillary electrophoresis chip, contains fluorescent material in the described DNA sample.
The DNA sample that the contains fluorescent material fluorescence that under the effect of light source, is excited, the ccd image sensor receiving optical signals, and light signal changed into the signal of telecommunication, send the Data Management Analysis module to and process.The detection width of ccd image sensor depends on the size of the single pixel in the array.The present invention adopts ccd image sensor as photoelectric detector, with the detection mode of traditional employing photomultiplier (PMT) relatively, greatly reduce detection width, and reduced the volume and weight of whole system, also had the remarkable advantages such as electrophoretogram ideal with low cost, detected.In addition, adopt ccd image sensor can not only differentiate light intensity, can also differentiate the situation of dna fragmentation.
Described ccd image sensor contains at least three pixels, and at least three corresponding signals of described pixel that described Data Management Analysis module will be received superpose or are averaged, in order to eliminate clutter or interference.
Also comprise a slave computer, described slave computer is as described system controller, and the signal input part of described slave computer connects described serial communication modular by the RS232 serial ports, and the signal output part of slave computer connects described drive motors by relay.Serial communication modular is used for control signal is sent to slave computer (single-chip microcomputer) by the RS232 serial ports.The control signal control relay that slave computer is read according to solution drives the drive motors adjustable lens, realizes the automatic focus of gel imaging system.
Described image capture module comprises an optical-electrical converter, an AD converter, described camera lens rear is provided with a ccd image sensor, described ccd image sensor connects described optical-electrical converter, described photovoltaic converter connects described AD converter, and described AD converter connects described Data Management Analysis module.
Also comprise a camera bellows, establishing one in the described camera bellows has article carrying platform, a light source, and described article carrying platform is arranged on described light source rear, and described camera lens is arranged on described article carrying platform rear.The present invention collects the image information of article on the article carrying platform by ccd image sensor and image capture module.
Described image capture module also connects a display unit, and described camera lens, described ccd image sensor form the used industrial camera of a gel imaging equipment.Described image capture module is to form according to writing with the supporting kit of the used industrial camera of gel imaging equipment, and its function is accurately to read in real time every frame image information, and extracts useful view data part.Image capture module also with image information display in the preview window at program control master interface, be convenient to eye-observation automatic focus state.
Described Data Management Analysis module comprises an image pretreatment module, an image quality evaluation module one focused search algorithm control module.The image preliminary treatment comprises that image denoising is processed and image brightness strengthens processing, and image denoising is processed and adopted median filtering method; Described image brightness strengthens processes the Retinex algorithm that adopts based on the human eye vision color constancy, and with the reference exposure amount of the image brightness average after this algorithm process as image.
The image quality evaluation module comprises image brightness evaluation and image definition evaluation, with the brightness average of image in the focus window as the brightness evaluation function, case verification and comparison through multiple evaluation function, proposition with based on the gray scale square difference evaluation function of spatial domain as Image Definition, its expression formula is:
Focused search algorithm control module refers to calculate the control output variable according to the control model of setting up in exposure and the focusing, namely corresponding aperture ring and the change amount of focusing ring are controlled model and are mainly contained variable step search by hill climbing algorithm model, function-fitting method model and fuzzy control model.The control model all can adopt the control model based on the VC++ language development.
Drive motors preferably adopts stepping motor, and described slave computer is decoded to the control signal that receives, and drives each road stepping motor of Control and moves respectively, finishes forward and reverse motion control of aperture, focusing, zoom.
Beneficial effect: owing to adopting technique scheme, compared with prior art, the present invention is incorporated into Techniques of Automatic Focusing in the gel imaging system, automatically finishes exposure, focusing operation; The searching algorithm that the Function Fitting that proposes combines with climbing method can be skipped the Local Extremum of Jiao Qu far away effectively, approaches fast and find global optimum's point; The fuzzy control theory is incorporated in the search control of automatic exposure and focusing, efficiently solves system's dynamic response slowly and the large problem of steady-state error.Therefore, the native system design has improved high efficiency, accuracy and the stability of gel imaging system to a great extent.
Description of drawings
Fig. 1 is a kind of system schematic of the present invention;
Fig. 2 is another kind of system schematic of the present invention;
Fig. 3 is variable step search by hill climbing algorithm model figure of the present invention;
Fig. 4 is function-fitting method illustraton of model of the present invention;
Fig. 5 is Fuzzy PID illustraton of model of the present invention.
Embodiment
For technological means, creation characteristic that the present invention is realized, reach purpose and effect is easy to understand, further set forth the present invention below in conjunction with concrete diagram.
With reference to Fig. 1, a kind of Capillary Electrophoresis gel imaging system of high definition, comprise that a camera lens 2, is used for the focus control system of control camera lens 2, camera lens 2 rears are provided with a ccd image sensor 9, and the focus control system comprises an image capture module 3, a Data Management Analysis module 4, a serial communication module 5, a hardware controls drive circuit 6.The signal input part of the signal output part connection data Treatment Analysis module 4 of image capture module 3, the signal output part of Data Management Analysis module 4 connects the signal input part of serial communication modular 5, and the signal output part of serial communication modular 5 connects a system controller 7.System controller 7 connects a drive motors 8 by hardware controls drive circuit 6, and drive motors 8 comes connection lens 2 by system actuator, in order to change the position of camera lens 2.
Image capture module, also comprise a capillary electrophoresis chip, light source, photoelectric detector, the signal output part of described photoelectric detector connects a Data Management Analysis module for data receiver, processing and control, described photoelectric detector comprises a ccd image sensor (charge coupled device), and the signal output part of described ccd image sensor connects described Data Management Analysis module;
The DNA sample enters from the starting point of described capillary electrophoresis chip, contains fluorescent material in the described DNA sample.
The DNA sample that the contains fluorescent material fluorescence that under the effect of light source, is excited, the ccd image sensor receiving optical signals, and light signal changed into the signal of telecommunication, send the Data Management Analysis module to and process.The detection width of ccd image sensor depends on the size of the single pixel in the array.The present invention adopts ccd image sensor as photoelectric detector, with the detection mode of traditional employing photomultiplier (PMT) relatively, greatly reduce detection width, and reduced the volume and weight of whole system, also had the remarkable advantages such as electrophoretogram ideal with low cost, detected.In addition, adopt ccd image sensor can not only differentiate light intensity, can also differentiate the situation of dna fragmentation.
Described ccd image sensor can adopt the line array CCD of one dimension, also can adopt the area array CCD of two dimension.
Described ccd image sensor contains at least three pixels, and at least three corresponding signals of described pixel that described Data Management Analysis module will be received superpose or are averaged, in order to eliminate clutter or interference.
Described Data Management Analysis module is got three pixel P1, P2, P3, and the described capillary electrophoresis chip starting point of P1 distance is nearest, and P3 farthest; It is L1, L2, L3 that three pixel P1, P2, P3 separate effective length accordingly, then obtains: L1＜L2＜L3;
In the Capillary Electrophoresis and testing process of operation DNA sample, described Data Management Analysis module is received the signal of each pixel, it is that time shaft, the longitudinal axis are the electrophoretogram of light intensity that described Data Management Analysis module is processed into a transverse axis with the signal of each pixel, respectively F1, F2, F3, wherein, the electrophoretogram peak of same dna fragmentation occurs the earliest in F1, occurs the latest in F3;
Revise take pixel P2 as benchmark, the transverse axis of F1 multiply by correction factor (F2/F1) and obtains F1 ', the transverse axis of F2 multiply by correction factor (F2/F2) and obtains F2 ', the transverse axis of F3 multiply by correction factor (F2/F3) and obtains F3 ', in F1 ', F2 ', F3 ', the position that the electrophoretogram peak of same dna fragmentation occurs is identical;
For F1 ', carry out interpolation with each time point of F2 ' time shaft, namely obtain F1 ' at the numerical value of each time point of F2 ' time shaft, be designated as F1 ".F1 " time shaft identical with F2 ', the shape of electrophoretogram is then identical with F1 ';
In like manner, F2 ', F3 ' are carried out interpolation processing, obtain F2 ", F3 ";
After the interpolation processing, F1 ", F2 ", F3 " time shaft unified, so the Data Management Analysis module is to F1 ", F2 ", F3 " superpose or be averaging, obtain the detection signal that signal to noise ratio improves.
Described Data Management Analysis module connects a display unit, and shows in the mode of electrophoretogram chart.So that the tester more intuitively checks.
When described ccd image sensor adopted the area array CCD of two dimension, as a large pixel, the signal of a contained pixel of large pixel adopted aforesaid way to superpose or is averaged perpendicular to each row's pixel of described capillary electrophoresis chip.Above-mentioned design, the Data Management Analysis module has been carried out a signal processing to every one dimension of the area array CCD of two dimension, and the detection signal that obtains like this is more accurate, and electrophoretic image is even more ideal.
With reference to Fig. 2, comprise that also establishing one in a camera bellows 1, the camera bellows 1 has article carrying platform 11, a light source 12, article carrying platform 11 is arranged on light source 12 rears, and camera lens 2 is arranged on article carrying platform 11 rears, and ccd image sensor 9 is arranged on camera lens 2 rears.
Data Management Analysis module 4 comprises an image pretreatment module 41, an image quality evaluation module 42 1 focused search algorithm control modules 43.Image pretreatment module 41 comprises that image denoising is processed and image brightness strengthens processing, and image denoising is processed and adopted median filtering method; Image brightness strengthens processes the Retinex algorithm that adopts based on the human eye vision color constancy, and with the reference exposure amount of the image brightness average after this algorithm process as image.
Image quality evaluation module 42 comprises image brightness evaluation and image definition evaluation, with the brightness average of image in the focus window as the brightness evaluation function, case verification and comparison through multiple evaluation function, proposition with based on the gray scale square difference evaluation function of spatial domain as Image Definition, its expression formula is:
Focused search algorithm control module 43 refers to calculate the control output variable according to the control model of setting up in exposure and the focusing, namely corresponding aperture ring and the change amount of focusing ring are controlled model and are mainly contained variable step search by hill climbing algorithm model, function-fitting method model and fuzzy control model.The control model all can adopt the control model based on the VC++ language development.
Also comprise a slave computer, slave computer of the present invention can be one-chip computer module, and slave computer is as system controller 7, and the display interface of slave computer is as display unit.Display interface comprises image preview window and Image Saving window etc.The signal input part of slave computer connects serial communication modular 5 by the RS232 serial ports, and the signal output part of slave computer connects drive motors 8 by relay.Serial communication modular 5 is used for control signal is sent to slave computer (single-chip microcomputer) by the RS232 serial ports.The control signal control relay that slave computer is read according to solution drives drive motors 8 adjustable lens 2, realizes the automatic focus of gel imaging system.
The drive motors 8 preferred stepping motors that adopt, slave computer is decoded to the control signal that receives, and drives each road stepping motor of Control and moves respectively, finishes forward and reverse motion control of aperture, focusing, zoom.
When the present invention used, the photosurface by ccd image sensor 9 carried out imaging to article on the article carrying platform 11.Image capture module 3 is used for Real-time Collection present image information, Data Management Analysis module 4 is used for the preliminary treatment of current image information and the evaluation of picture quality, and providing control signal, serial communication modular 5 is used for control signal by sending to system controller 7.Control signal control drive motors 8 adjustable lens 2 that system controller 7 is read according to solution are realized the automatic focus of gel imaging system.
The present invention has abandoned the quality of traditional human eye subjective visual evaluation image, and the mode by manual adjustments, adds the automatic focus mode but adopt on the basis of former Manual focusing control system, has realized self-focusing purpose.
During the invention process, the gel biological molecular preparation is placed on the article carrying platform 11 of 1 li in camera bellows, open light source 12, start the focus control system, open the image preview window of display unit, can see the image formation state of the photosurface of current C CD transducer 9, then calling graph image brightness evaluation module and image pretreatment module 41, present image is carried out analyzing and processing, obtain image brightness value and reference exposure value, these two values are compared, its difference is as the decision signal of aperture regulated quantity, call the control of diaphragm module (fuzzy control), calculate corresponding drive motors 8 output step-lengths according to difference signal, and this control signal is sent to system controller by the RS232 serial ports, drive system actuator finishes aperture and regulates, repeat said process until reach the target exposure, namely realize the automatic exposure of gel imaging system.
Automatic exposure enters auto-focus process after finishing, calling graph image sharpness evaluation module and focusing control module.The image definition evaluation curve as shown in Figure 3, abscissa represents camera lens 2 positions, ordinate represents the evaluation function value, when evaluation function was obtained maximum, imaging was the most clear on the photosurface of corresponding place, place, camera lens 2 position ccd sensor 9.With the difference of the sharpness evaluation function value of the front and back two two field pictures decision signal as the focusing control procedure.For how quick and precisely finding the global extremum point, the present invention has provided three kinds of focusing control models: variable step search by hill climbing algorithm model, Function Fitting model and fuzzy control model.
Variable step search by hill climbing algorithm model as shown in Figure 3, camera lens 2 is nearest with the photosurface of ccd sensor 9 during initial condition, the focusing beginning, drive motors moves with fixed step size by positive direction and gathers image, when the evaluation function difference is negative, change the drive motors direction of motion, step-length reduces by half and moves by opposite direction, gather simultaneously image, when difference again for negative, step-length reduces by half and by positive direction motion, simultaneously image data again, repeat this process until satisfy the search end condition, focusing is finished.
The Function Fitting model as shown in Figure 4, initial condition is constant, drive motors moves along positive direction, gather three frame image datas (camera lens 2 positions that this three two field picture is corresponding are not arbitrarily fixed, but are chosen by experiment test), take camera lens 2 position coordinateses as independent variable x, the evaluation function value is dependent variable y, simulate a secondary parabolic curve that Open Side Down, calculate the maximum y1 of curve, and drive motors moves to corresponding x1 position.Can adopt little step length searching very near positive burnt position this moment, finds global extremum point position, finishes focusing.
Fuzzy control model as shown in Figure 5, this control model is different from the above two parts and is, it has not only utilized the polarity information of evaluation function difference, simultaneously also according to difference size adaptation regulating electric machine step-length, determine rational fuzzy control parameter with reference to Heuristics and a large amount of experiment tests, the dynamic responding speed of Effective Raise system and stable state accuracy (are annotated: σ, T0 and the T1 condition decision threshold for setting in the model).
The focus control system also can not adopt above-mentioned three kinds of models, but utilize the aperture drive motors that aperture ring is adjusted to suitable brightness by the aperture drive circuit first, then regulate focusing ring by focusing drive circuit utilization focusing drive motors, make image be put into maximum, then regulate the focal length of focusing ring by focusing drive circuit utilization focusing drive motors, obtain the most clearly image, at last image scaling imaging after suitable size.This control method is the simplest.
Above demonstration and described basic principle of the present invention and principal character and advantage of the present invention.The technical staff of the industry should understand; the present invention is not restricted to the described embodiments; that describes in above-described embodiment and the specification just illustrates principle of the present invention; without departing from the spirit and scope of the present invention; the present invention also has various changes and modifications, and these changes and improvements all fall in the claimed scope of the invention.The claimed scope of the present invention is defined by appending claims and equivalent thereof.
Claims (7)
1. the Capillary Electrophoresis gel imaging system of a high definition, comprise that a camera lens, is used for controlling the focus control system of described camera lens, it is characterized in that, described focus control system comprises an image capture module, a Data Management Analysis module, a serial communication module, a hardware controls drive circuit; The signal output part of described image capture module connects the signal input part of described Data Management Analysis module, the signal output part of described Data Management Analysis module connects the signal input part of described serial communication modular, and the signal output part of described serial communication modular connects a system controller;
Described system controller connects a drive motors by described hardware controls drive circuit, and described drive motors connects described camera lens;
Described camera lens comprises an aperture ring, a focusing ring, and described drive motors comprises an aperture drive motors, a focusing drive motors, and described hardware controls drive circuit comprises an aperture drive circuit, a focusing drive circuit; Described system controller connects respectively described aperture drive circuit and described focusing drive circuit; Described aperture drive circuit connects described aperture drive motors, and described aperture drive motors connects described aperture ring; Described focusing drive circuit connects described focusing drive motors, and described focusing drive motors connects described focusing ring.
2. the Capillary Electrophoresis gel imaging system of a kind of high definition according to claim 1, it is characterized in that, image capture module, also comprise a capillary electrophoresis chip, light source, photoelectric detector, the signal output part of described photoelectric detector connects a Data Management Analysis module for data receiver, processing and control, described photoelectric detector comprises a ccd image sensor, and the signal output part of described ccd image sensor connects described Data Management Analysis module;
The DNA sample enters from the starting point of described capillary electrophoresis chip, contains fluorescent material in the described DNA sample.
3. the Capillary Electrophoresis gel imaging system of a kind of high definition according to claim 2, it is characterized in that, described ccd image sensor contains at least three pixels, and at least three corresponding signals of described pixel that described Data Management Analysis module will be received superpose or are averaged.
4. according to claim 1, the Capillary Electrophoresis gel imaging system of 2 or 3 described a kind of high definition, it is characterized in that, also comprise a slave computer, described slave computer is as described system controller, the signal input part of described slave computer connects described serial communication modular by the RS232 serial ports, and the signal output part of slave computer connects described drive motors by relay.
5. the Capillary Electrophoresis gel imaging system of a kind of high definition according to claim 1, it is characterized in that, described image capture module also comprises an optical-electrical converter, an AD converter, described camera lens rear is provided with a ccd image sensor, described ccd image sensor connects described optical-electrical converter, described photovoltaic converter connects described AD converter, and described AD converter connects described Data Management Analysis module.
6. according to claim 1, the Capillary Electrophoresis gel imaging system of 2 or 3 described a kind of high definition, it is characterized in that, described Data Management Analysis module comprises an image pretreatment module, an image quality evaluation module, a focused search algorithm control module;
The image preliminary treatment comprises that image denoising is processed and image brightness strengthens processing, and image denoising is processed and adopted median filtering method; Described image brightness strengthens processes the Retinex algorithm that adopts based on the human eye vision color constancy, and with the reference exposure amount of the image brightness average after this algorithm process as image;
The image quality evaluation module comprises image brightness evaluation and image definition evaluation, with the brightness average of image in the focus window as the brightness evaluation function, with based on the gray scale square difference evaluation function of spatial domain as Image Definition, its expression formula is:
, F(I wherein) and expression evaluation function value, I(x, y) the presentation video data, x, y represent the pixel position;
Focused search algorithm control module refers to calculate the control output variable according to the control model of setting up in exposure and the focusing, namely corresponding aperture ring and the change amount of focusing ring are controlled model and are mainly contained variable step search by hill climbing algorithm model, function-fitting method model and fuzzy control model.
7. the Capillary Electrophoresis gel imaging system of a kind of high definition according to claim 6, it is characterized in that, also comprise a camera bellows, establishing one in the described camera bellows has article carrying platform, a light source, described article carrying platform is arranged on described light source rear, and described camera lens is arranged on described article carrying platform rear;
Described image capture module also connects a display unit;
During use, the gel biological molecular preparation is placed on the article carrying platform in the camera bellows, open light source, start the focus control system, open the image preview window of display unit, can see the image formation state of the photosurface of current C CD transducer, then calling graph image brightness evaluation module and image pretreatment module, present image is carried out analyzing and processing, obtain image brightness value and reference exposure value, these two values are compared, its difference is as the decision signal of aperture regulated quantity, call the control of diaphragm module, calculate corresponding drive motors output step-length according to difference signal, and this control signal is sent to system controller by the RS232 serial ports, drive system actuator finishes aperture and regulates, repeat said process until reach the target exposure, namely realize the automatic exposure of gel imaging system;
Enter auto-focus process after automatic exposure finishes, calling graph image sharpness evaluation module and focusing control module, according to the image definition evaluation curve with the difference of the sharpness evaluation function value of the front and back two two field pictures decision signal as the focusing control procedure.
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| CN109733329A (en) * | 2019-01-22 | 2019-05-10 | 南京信息职业技术学院 | Automobile defogging method and device based on machine vision |
| CN111510631A (en) * | 2020-04-27 | 2020-08-07 | 成都国铁电气设备有限公司 | Rapid automatic focusing method and focusing device |
| CN113810616A (en) * | 2021-09-27 | 2021-12-17 | 季华实验室 | Aperture focal length adjusting method, system, electronic device and storage medium |
| WO2024092985A1 (en) * | 2022-10-31 | 2024-05-10 | 上海交通大学 | Intrinsic fluorescence imaging detection system for protein gel electrophoresis and method |
| CN118150531A (en) * | 2023-04-21 | 2024-06-07 | 深圳市真迈生物科技有限公司 | Imaging system, nucleic acid fragment analysis apparatus, analysis method, and storage medium |
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