CN102879361B - Capillary electrophoresis image-forming detecting system - Google Patents
Capillary electrophoresis image-forming detecting system Download PDFInfo
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- CN102879361B CN102879361B CN201110194139.0A CN201110194139A CN102879361B CN 102879361 B CN102879361 B CN 102879361B CN 201110194139 A CN201110194139 A CN 201110194139A CN 102879361 B CN102879361 B CN 102879361B
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Abstract
The present invention relates to technical field of optical detection, particularly relate to a kind of detection system. Capillary electrophoresis image-forming detecting system, including capillary electrophoresis chip, light source, photoelectric detector, the signal output part of photoelectric detector connects microprocessor system, and photoelectric detector includes ccd image sensor, and the signal output part of ccd image sensor connects microprocessor system. DNA sample enters from the starting point of capillary electrophoresis chip, containing fluorescent material in DNA sample. Owing to adopting technique scheme, the present invention significantly reduces the volume and weight that detecting of capillary electrophoresis is broadening, decrease whole system, also has the remarkable advantages such as electrophoresis spectral peak with low cost, that detect ideal.
Description
Technical field
The present invention relates to technical field of optical detection, particularly relate to a kind of detection system.
Background technology
Capillary electrophoresis is a kind of important separation detection technique. Capillary electrophoresis is in conjunction with sieving media for biomacromolecule, and as DNA and protein have good separating power, this is one of the important foundation of modern life science research. Along with the development in the fields such as life sciences, the application demand for capillary electrophoresis is also continuously increased. Its range of application is just from simple academic research field towards more practical field extension.
In the various detection techniques of capillary electrophoresis, induced fluorescence detection is one of the highest mode of sensitivity, is particularly suitable for the detection of biomacromolecule. In detection process, the detection width of detection device can make the electrophoretogram peak width of sample more than actual zone width, is called that detection is broadening. Detect and broadening reduce the peak-to-peak resolution capability of adjacent electrophoretogram. Induced fluorescence detection generally adopts photomultiplier tube (PMT) that fluorescence is carried out light-intensity test, and the detection width of photomultiplier tube depends on photosurface size. Owing to the photosurface of photomultiplier tube is relatively big, limit the reduction of detection width, thus limiting the raising of electrophoresis resolution capability. Photomultiplier tube there is also the defects such as price is partially expensive, can only differentiate light intensity, volume is bigger.
Summary of the invention
It is an object of the invention to provide a kind of capillary electrophoresis image-forming detecting system, solve above technical problem.
Technical problem solved by the invention can realize by the following technical solutions:
Capillary electrophoresis image-forming detecting system, including a capillary electrophoresis chip, light source, photoelectric detector, the signal output part of described photoelectric detector connects a microprocessor system being used for data receiver, process and control, it is characterized in that, described photoelectric detector includes a ccd image sensor (charge-coupled image sensor), and the signal output part of described ccd image sensor connects described microprocessor system;
DNA sample enters from the starting point of described capillary electrophoresis chip, containing fluorescent material in described DNA sample.
DNA sample containing fluorescent material is excited fluorescence under the effect of light source, and ccd image sensor receives optical signal, and optical signal is changed into the signal of telecommunication, sends microprocessor system to and processes. The detection width of ccd image sensor depends on the size of the single pixel in array. The present invention adopts ccd image sensor as photoelectric detector, compare with the detection mode of traditional employing photomultiplier tube (PMT), substantially reduce detection width, and decrease the volume and weight of whole system, also there is the remarkable advantages such as electrophoretogram with low cost, that detect ideal. It addition, adopt ccd image sensor can not only differentiate light intensity, moreover it is possible to differentiate the situation of DNA fragmentation.
Described capillary electrophoresis chip is contained within sieving media, and sieving media can be gel.
Described ccd image sensor can adopt one-dimensional line array CCD, it would however also be possible to employ the area array CCD of two dimension.
Described ccd image sensor contains at least three pixel, and the signal corresponding to pixel described at least three received is overlapped or is averaged by described microprocessor system, in order to eliminate clutter or interference.
Described microprocessor system takes three pixels P1, P2, P3, and P1 is nearest apart from described capillary electrophoresis chip starting point, and P3 is farthest; It is L1, L2, L3 that three pixels P1, P2, P3 separate effective length accordingly, then obtains: L1 < L2 < L3;
In the capillary electrophoresis running DNA sample and detection process, described microprocessor system receives the signal of each pixel, it is the electrophoretogram of light intensity that the signal processing of each pixel is become transverse axis to be time shaft, the longitudinal axis by described microprocessor system, it is F1, F2, F3 respectively, wherein, the electrophoresis spectral peak of same DNA fragmentation occurs the earliest in F1, occurs the latest in F3;
It is modified with pixel P2 for benchmark, the transverse axis of F1 is multiplied by correction factor (F2/F1) and obtains F1 ', the transverse axis of F2 is multiplied by correction factor (F2/F2) and obtains F2 ', the transverse axis of F3 is multiplied by correction factor (F2/F3) and obtains F3 ', in F1 ', F2 ', F3 ', the position that the electrophoresis spectral peak of same DNA fragmentation occurs is identical;
For F1 ', it is interpolated with each time point of F2 ' time shaft, namely obtains the F1 ' numerical value at each time point of F2 ' time shaft, be designated as F1 ". F1 " time shaft and F2 ' identical, the shape of electrophoretogram is then identical with F1 ';
In like manner, F2 ', F3 ' are interpolated process, obtain F2 ", F3 ";
After interpolation processing, F1 ", F2 ", F3 " time shaft unified, then microprocessor system is to F1 ", F2 ", F3 " be overlapped or be averaging, obtain the detection signal that signal to noise ratio improves.
Described microprocessor system connects a display device, and shows in the way of electrophoretogram chart. So that tester more intuitively checks.
When described ccd image sensor adopts the area array CCD of two dimension, being perpendicular to each row's pixel of described capillary electrophoresis chip as a big pixel, the signal of pixel contained by a big pixel adopts aforesaid way to be overlapped or be averaged. Above-mentioned design, the often one-dimensional of area array CCD of two dimension has been carried out a signal processing by microprocessor system, and the detection signal so obtained is more accurate, and electrophoretic image is even more ideal.
Beneficial effect: owing to adopting technique scheme, the present invention significantly reduces the volume and weight that detecting of capillary electrophoresis is broadening, decrease whole system, also has the remarkable advantages such as electrophoresis spectral peak with low cost, that detect ideal. It addition, adopt ccd image sensor can not only differentiate light intensity, moreover it is possible to differentiate the situation of DNA fragmentation.
Accompanying drawing explanation
Fig. 1 is the part-structure schematic diagram of the present invention;
Fig. 2 is the electrophoretic image before the pixel of the present invention is not revised;
Fig. 3 is the revised electrophoretic image of pixel of the present invention;
Fig. 4 is the pixel superposition of the present invention or the electrophoretic image after being averaging.
Detailed description of the invention
For the technological means making the present invention realize, creation characteristic, reach purpose and effect and be easy to understand, the present invention is expanded on further below in conjunction with being specifically illustrating.
With reference to Fig. 1, capillary electrophoresis image-forming detecting system, including a capillary electrophoresis chip 1, light source 2, photoelectric detector, the signal output part of photoelectric detector connects a microprocessor system being used for data receiver, process and control, photoelectric detector includes a ccd image sensor 3 (charge-coupled image sensor), and the signal output part of ccd image sensor 3 connects microprocessor system. DNA sample enters from the starting point of capillary electrophoresis chip 1, containing fluorescent material in DNA sample. DNA sample containing fluorescent material is excited fluorescence under the effect of light source 2, and ccd image sensor 3 receives optical signal, and optical signal is changed into the signal of telecommunication, sends microprocessor system to and processes. The detection width of ccd image sensor 3 depends on the size of the single pixel in array. The present invention adopts ccd image sensor 3 as photoelectric detector, compare with the detection mode of traditional employing photomultiplier tube (PMT), substantially reduce detection width, and decrease the volume and weight of whole system, also there is the remarkable advantages such as electrophoretogram with low cost, that detect ideal. It addition, adopt ccd image sensor 3 can not only differentiate light intensity, moreover it is possible to differentiate the situation of DNA fragmentation.
Capillary electrophoresis chip 1 is contained within sieving media, and sieving media can be gel.
Ccd image sensor 3 can adopt one-dimensional line array CCD, it would however also be possible to employ the area array CCD of two dimension. Ccd image sensor 3 is containing at least three pixel, and the signal corresponding at least three pixel received is overlapped or is averaged by microprocessor system, in order to eliminate clutter or interference.
With reference to Fig. 2, Fig. 3, Fig. 4, microprocessor system takes three pixels P1, P2, P3, and P1 distance capillary electrophoresis chip 1 starting point is nearest, and P3 is farthest. It is L1, L2, L3 that three pixels P1, P2, P3 separate effective length accordingly, then obtains: L1 < L2 < L3. In the capillary electrophoresis running DNA sample and detection process, microprocessor system receives the signal of each pixel, it is the electrophoretogram of light intensity that the signal processing of each pixel is become transverse axis to be time shaft, the longitudinal axis by microprocessor system, it is F1, F2, F3 respectively, wherein, the electrophoresis spectral peak of same DNA fragmentation occurs the earliest in F1, occurs the latest in F3. It is modified with pixel P2 for benchmark, the transverse axis of F1 is multiplied by correction factor (F2/F1) and obtains F1 ', the transverse axis of F2 is multiplied by correction factor (F2/F2) and obtains F2 ', the transverse axis of F3 is multiplied by correction factor (F2/F3) and obtains F3 ', in F1 ', F2 ', F3 ', the position that the electrophoresis spectral peak of same DNA fragmentation occurs is identical.
In principle, F1 ', F2 ', F3 ' are added or are averaging, just completes data handling procedure, obtain the detection signal that signal to noise ratio improves. But, when actual data process, F1 ', F2 ', F3 ' cannot directly be added, because their time shaft is discrete, and are different.All pixels of array detector are synchronous workings, thus set revise before the time shaft of F1, F2, F3 be all (1,2,3,4,5 ...). If the correction factor of P1 is 0.9, P2's is 1, P3's is 1.1, then revise after obtain F1 ', F2 ', F3 ' time shaft be respectively (0.9,1.8,2.7,3.6,4.5 ...) (1,2,3,4,5 ...) (1.1,2.2,3.3,4.4,5.5 ...), it is clear that cannot directly be added or be averaging. Therefore, F1 ', F2 ', F3 ' are also needed to process as follows step:
For F1 ', it is interpolated with each time point of F2 ' time shaft, namely obtains the F1 ' numerical value at each time point of F2 ' time shaft, be designated as F1 ". F1 " time shaft and F2 ' identical, the shape of electrophoretogram is then identical with F1 '; In like manner, F2 ', F3 ' are interpolated process, obtain F2 ", F3 ". After interpolation processing, F1 ", F2 ", F3 " time shaft unified, then microprocessor system is to F1 ", F2 ", F3 " be overlapped or be averaging, obtain the detection signal that signal to noise ratio improves.
Microprocessor system connects a display device, with reference to Fig. 2, Fig. 3, Fig. 4, shows in the way of electrophoretogram chart. So that tester more intuitively checks.
When ccd image sensor 3 adopts the area array CCD of two dimension, being perpendicular to each row's pixel of capillary electrophoresis chip 1 as a big pixel, the signal of pixel contained by a big pixel adopts aforesaid way to be overlapped or be averaged. Above-mentioned design, the often one-dimensional of area array CCD of two dimension has been carried out a signal processing by microprocessor system, and the detection signal so obtained is more accurate, and electrophoretic image is even more ideal.
Embodiment one:
Ccd image sensor 3 is containing 50 pixels, and the starting point of the 1st pixel distance capillary electrophoresis chip 1 is nearest. Microprocessor system receives the signal corresponding to 50 pixels, and microprocessor system takes the testing result of the 1st, 25,50 pixels and is modified processing. With reference to Fig. 1, electrophoretic image F1, F2, the F3 before the 1st, 25,50 pixel correcting process, with reference to Fig. 2, experimental data is as follows:
| Time | 1st pixel light intensity | 2nd pixel light intensity | 3rd pixel light intensity |
| 9.2000000e+002 | -1.3407025e-002 | -3.6926299e-003 | 1.0683385e-002 |
| 1.0000000e+003 | 3.6260740e-003 | 9.7773260e-001 | -1.6528910e-003 3 --> |
| 1.0500000e+003 | 3.9882975e-003 | 4.9744645e-001 | 1.1123748e-002 |
| 1.1000000e+003 | 4.2007982e-003 | 9.4923964e-003 | 1.1443517e-002 |
| 1.1200000e+003 | -9.6449040e-004 | -2.2012810e-002 | 8.4521139e-003 |
F1, F2, F3 are modified by microprocessor system, being modified for benchmark with the 25th pixel, obtain F1 ', F2 ', F3 ', F1 ', F2 ', F3 ' are interpolated process and obtain F1 by microprocessor system ", F2 ", F3 "; with reference to Fig. 3, experimental data is as follows:
| Time | 1st pixel light intensity | 2nd pixel light intensity | 3rd pixel light intensity |
| 9.2000000e+002 | -2.8618540e-003 | -3.6926299e-003 | 5.3875831e-003 |
| 1.0000000e+003 | 1.0071722e+000 | 9.7773260e-001 | 1.0145737e+000 |
| 1.0500000e+003 | 4.9906783e-001 | 4.9744645e-001 | 4.7715608e-001 |
| 1.1000000e+003 | -2.6204747e-003 | 9.4923964e-003 | 1.4033733e-003 |
| 1.1200000e+003 | 9.3533850e-003 | -2.2012810e-002 | -9.7249412e-003 |
Microprocessor system is to F1 ", F2 ", F3 " be averaging, obtain the detection signal F that signal to noise ratio improves, with reference to Fig. 4, experimental data is as follows:
| Time | 3 pixels be averaged after light intensity |
| 9.2000000e+002 | -1.7618987e-003 |
| 1.0000000e+003 | 9.9985629e-001 |
| 1.0500000e+003 | 4.9171983e-001 |
| 1.1000000e+003 | 1.1816860e-006 |
| 1.1200000e+003 | 1.7002843e-003 |
The ultimate principle of the present invention and principal character and advantages of the present invention have more than been shown and described. Skilled person will appreciate that of the industry; the present invention is not restricted to the described embodiments; described in above-described embodiment and description is that principles of the invention is described; 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 both fall within the claimed scope of the invention. Claimed scope is defined by appending claims and equivalent thereof.
Claims (4)
1. capillary electrophoresis image-forming detecting system, including a capillary electrophoresis chip, light source, photoelectric detector, the signal output part of described photoelectric detector connects a microprocessor system being used for data receiver, process and control, it is characterized in that, described photoelectric detector includes a ccd image sensor, and the signal output part of described ccd image sensor connects described microprocessor system;
DNA sample enters from the starting point of described capillary electrophoresis chip, containing fluorescent material in described DNA sample;
Described ccd image sensor contains at least three pixel, and described microprocessor system takes three pixels P1, P2, P3, and P1 is nearest apart from described capillary electrophoresis chip starting point, and P3 is farthest; It is L1, L2, L3 that three pixels P1, P2, P3 separate effective length accordingly, and described microprocessor system obtains then: L1 < L2 < L3;
In the capillary electrophoresis running DNA sample and detection process, described microprocessor system receives the signal of each pixel, it is the electrophoretogram of light intensity that the signal processing of each pixel is become transverse axis to be time shaft, the longitudinal axis by described microprocessor system, it is F1, F2, F3 respectively, wherein, the electrophoresis spectral peak of same DNA fragmentation occurs the earliest in F1, occurs the latest in F3;
It is modified with pixel P2 for benchmark, the transverse axis of F1 is multiplied by correction factor and obtains the transverse axis of F1 ', F2 and be multiplied by correction factor and obtain the transverse axis of F2 ', F3 and be multiplied by correction factor and obtain F3 ', in F1 ', F2 ', F3 ', the position that the electrophoresis spectral peak of same DNA fragmentation occurs is identical;
For F1 ', it is interpolated with each time point of F2 ' time shaft, namely obtains the F1 ' numerical value at each time point of F2 ' time shaft, be designated as F1 "; F1 " time shaft and F2 ' identical, the shape of electrophoretogram is then identical with F1 ';
In like manner, F2 ', F3 ' are interpolated process, obtain F2 ", F3 ";
After interpolation processing, F1 ", F2 ", F3 " time shaft unified, then microprocessor system is to F1 ", F2 ", F3 " be overlapped or be averaging, obtain the detection signal that signal to noise ratio improves;
Described microprocessor system connects a display device, and described display device is used for showing electrophoretogram chart.
2. capillary electrophoresis image-forming detecting system according to claim 1, it is characterised in that described capillary electrophoresis chip is contained within sieving media.
3. capillary electrophoresis image-forming detecting system according to claim 1, it is characterised in that described ccd image sensor adopts one-dimensional line array CCD.
4. capillary electrophoresis image-forming detecting system according to claim 1, it is characterised in that described ccd image sensor adopts the area array CCD of two dimension.
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| US6613212B1 (en) * | 1998-01-30 | 2003-09-02 | Centre National De La Recherche Scientifique | Multiple capillary electrophoresis systems |
| CN1560627A (en) * | 2004-02-26 | 2005-01-05 | 复旦大学 | Multi-chromaticity-electrophoresis separating and detecting system based on array capillary electrofocasing |
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| CN1700016A (en) * | 2005-05-27 | 2005-11-23 | 中国科学院大连化学物理研究所 | Capillary electrophoresis fluorescence - non-contact conductance combination detector |
| CN101865876A (en) * | 2009-04-15 | 2010-10-20 | 上海培清科技有限公司 | Fully automatic molecular electrophoresis gel imaging analyser |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE19803753C1 (en) * | 1998-01-30 | 1999-12-02 | Max Planck Gesellschaft | Device and method for capillary electrophoresis |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6613212B1 (en) * | 1998-01-30 | 2003-09-02 | Centre National De La Recherche Scientifique | Multiple capillary electrophoresis systems |
| CN1560627A (en) * | 2004-02-26 | 2005-01-05 | 复旦大学 | Multi-chromaticity-electrophoresis separating and detecting system based on array capillary electrofocasing |
| CN1595117A (en) * | 2004-06-18 | 2005-03-16 | 中国科学院上海有机化学研究所 | Online self-calibration laser induced fluorescence detection method based on electric charge coupling apparatus |
| CN1700016A (en) * | 2005-05-27 | 2005-11-23 | 中国科学院大连化学物理研究所 | Capillary electrophoresis fluorescence - non-contact conductance combination detector |
| CN101865876A (en) * | 2009-04-15 | 2010-10-20 | 上海培清科技有限公司 | Fully automatic molecular electrophoresis gel imaging analyser |
Non-Patent Citations (2)
| Title |
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