CN102331415A - Method for positioning capillary tube array by using raman spectral imaging - Google Patents
Method for positioning capillary tube array by using raman spectral imaging Download PDFInfo
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- CN102331415A CN102331415A CN201110160548A CN201110160548A CN102331415A CN 102331415 A CN102331415 A CN 102331415A CN 201110160548 A CN201110160548 A CN 201110160548A CN 201110160548 A CN201110160548 A CN 201110160548A CN 102331415 A CN102331415 A CN 102331415A
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Abstract
The invention provides a method for positioning a capillary tube array on a charge coupled device by using a raman spectrum. The method comprises the following steps of: (1) irradiating a capillary tube bundle by using laser to emit raman light; (2) imaging on the charge coupled device (CCD) through a raman imager; and (3) analyzing a spectrogram on the CCD to position each capillary tube in the capillary tube bundle. In the method, the raman spectrum of quartz or water is excited by using double-beam laser to realize spectral imaging; and thus, positioning and parallelism adjustment of the capillary tube on the CCD are realized, error during trace reagent analysis of the capillary tube array can be eliminated, and the analysis accuracy is improved.
Description
Technical field
The present invention relates to a kind of method of locating capillary array image space on charge coupled cell, belong to high flux trace analysis field.
Background technology
Because kapillary has good heat dissipation, and bear the voltage height, sample size is little, it is desired efficient to have an instrumental analysis, fast, amount of samples waits the excellent characteristics of fundamental sum less.Kapillary two ends institute making alive may be up to 30kV; The longitudinal electric field intensity of separation capillary can reach more than the 400V/cm; Thereby lock out operation can reach very high separation efficiency (theoretical cam curve reaches more than the 400000/m, is up to the 107/m order of magnitude) in a short period of time.Because internal diameter capillaceous very little (general＜100 μ m), to internal diameter 50 μ m, length is the kapillary of 50cm, its volume less than 1 μ L, and sampling volume is in the nL level, and sample concentration can be lower than 10～4mol/L.
Advantages such as in addition, capillary electrophoresis technique also has easy robotization, and is easy and simple to handle, and solvent consumption is few, and environmental pollution is little.The application of capillary electrophoresis technique at first concentrates on amino acid, carbohydrate, on the compartment analysis of biomolecule such as nucleic acid and protein, along with the continuous development of this technology and perfect, its use gradually to medical and health, food chemical industry, the infiltration of fields such as environment.Capillary electrophoresis technique also is applied to the high speed order-checking of DNA, the high efficiency separation of protein, carbohydrate analysis, cell analysis, chiral resolution, the mensuration of physicochemical constant, production engineering control etc.
In order to improve the flux of capillary detection, capillary array is widely used in the biochemical analysis as high flux electrophoresis analysis means.Carry out the qualitative and quantitative analysis of trace reagent at field widespread use capillary arrays such as medical treatment, chemical, bio-pharmaceuticals, forensic analysises.Capillary array can be realized the analysis of reagent in batches simultaneously.But, make capillary array have error when carrying out the qualitative and quantitative analysis of trace reagent because that intercapillary position relation exists is uncertain.Still do not eliminate at present the mature technology of this error.
Summary of the invention
In order simultaneously many capillaries to be analyzed, need position the error when carrying out the qualitative or quantitative test of trace reagent to the image space of kapillary on charge coupled cell (CCD) to eliminate capillary array.
The present invention provides a kind of method of utilizing Raman spectrum imaging location capillary array; Said method comprises: Raman light is sent with the laser beam irradiation capillary bundle in (1); (2) be imaged on the charge coupled cell (CCD) through the Raman imager, the spectrogram that (3) are analyzed on the charge coupled cell CCD positions each kapillary in the capillary bundle.
Further, said method step (1) with the laser beam irradiation capillary bundle send Raman light be meant laser beam by quartz capillaceous or the pipe in the solution scattering send Raman light.
Further, said Raman imager is imaged on the CCD Raman light through getting optical slits, grating beam splitting and condenser lens.
Further, said method step (1) sends Raman light with the water scattering in the laser beam irradiation capillary bundle, and used laser beam is a double laser beam, and the wavelength of said double laser beam is respectively 488nm and 514.5nm.
As optimal way, said odd number kapillary is gathered the Raman light that the 488nm laser instrument excites, and the even numbers kapillary is gathered the Raman light that the 514.5nm laser instrument excites.
As another optimal way, said even numbers kapillary is gathered the Raman light that the 488nm laser instrument excites, and the odd number kapillary is gathered the Raman light that the 514.5nm laser instrument excites.
Further, be transverse axis with the pixel count in the said method step (3), spectral intensity is that the longitudinal axis obtains the free-air correction spectrogram, obtains the effective width between every capillary center and the every capillary through peak identification.
Location positioning technology of the present invention adopts double laser beam, is respectively 488nm and 514.5nm.The characteristic Raman peaks of water is positioned at 3400cm
-1The place.For 488nm (20491.8cm
-1), the raman characteristic peak of water scattering is positioned at 17091.8cm
-1, corresponding wavelength is 585nm.For 514.5nm (19436.3cm
-1), the raman characteristic peak of water scattering is positioned at 16036.3cm
-1, corresponding wavelength is 623.6nm.Fig. 2 is the characteristic Raman peaks of water scattering.As can be seen from the figure, its spectrum mainly is distributed in 2800cm
-1To 3800cm
-1Between the wave number.
488nm and 514.5nm laser are projected on the data acquisition CCD by the characteristic Raman peaks of water scattering can obtain Fig. 3.Blue light (left side) is to gather 488nm laser by the Raman light intensity of water scattering; What ruddiness (right side) was corresponding is to gather 514.5nm laser by the Raman fluorescence intensity of water scattering; The corresponding capillary in each peak adopts adjacent capillary space to gather the characteristics of the Raman light that different wave length laser is scattered, and promptly the odd number kapillary is gathered the Raman light that the 488nm laser instrument excites; The even numbers kapillary is gathered the Raman light that the 514.5nm laser instrument excites, and vice versa.We can see from figure, and 488nm and 514.5nm laser can be separated by the Raman peaks of water scattering just, and just in time are distributed within the CCD sensing range: 500～660nm.This has met just gathers two groups of demands that data are proofreaied and correct in the free-air correction.Fig. 4 has provided the frame spectrogram on the free-air correction CCD, from Fig. 4, can obviously see corresponding two bright spots (raman peak values) on every capillary.
Fig. 4 is carried out transverse axis data summations as spectral intensity, and it is as shown in Figure 5 to be with longitudinal axis pixel that the horizontal ordinate mapping provides the free-air correction spectrogram.Can provide every capillary center (as shown in Figure 6) and effective width (getting one or two pixel about general) through the identification of simple peak.
Beneficial effect of the present invention:
(1) adopt the Raman spectrum of laser excitation quartz or water to realize light spectrum image-forming;
(2) can adopt single or multi-laser excites;
(3) realize location positioning and the depth of parallelism adjustment of kapillary on CCD;
(4) the method implement device is simple in structure, and is easy to operate.
(5) can eliminate capillary array and carry out the error of trace reagent when analyzing, improve accuracy of analysis.
Description of drawings
Fig. 1 is a localization method operation chart of the present invention;
Fig. 2 is the Raman spectrogram of water: the characteristic Raman peaks of water scattering;
Fig. 3 is the Raman spectrogram of water: 488nm and 514.5nm laser are projected on the data acquisition CCD by the characteristic Raman peaks of water scattering
Fig. 4 is the image of capillary array Raman light on CCD;
Fig. 5 is a locus positioning analysis synoptic diagram.Transverse axis is a number of picture elements, and the longitudinal axis is the spectral intensity mapping
Fig. 6 analyzes data for space orientation
Wherein: the 1-laser beam, the 2-capillary array, the 3-single capillary, the 4-Raman light,
5-Raman spectrum imaging CCD
Embodiment
Combine accompanying drawing and embodiment that the inventive method is done further explain at present.
Fig. 1 is the operation chart of the inventive method.Impinge upon the detection window place of capillary bundle 2 with laser beam 1; Laser is by the water scattering in the solution in the quartzy or pipe of single capillary 3; Send Raman light 4, through getting optical slits, grating beam splitting; Be imaged on after the lens focus on the CCD 5, the spectrogram of analyzing on the CCD 5 carries out location positioning to capillary bundle.
The location positioning technology adopts double laser beam, is respectively 488nm and 514.5nm.488nm and 514.5nm laser are projected on the data acquisition CCD by the characteristic Raman peaks of water scattering can obtain Fig. 3.Blue light (left side) is to gather 488nm laser by the Raman light intensity of water scattering; What ruddiness (right side) was corresponding is to gather 514.5nm laser by the Raman fluorescence intensity of water scattering; The corresponding capillary in each peak adopts adjacent capillary space to gather the characteristics of the Raman light that different wave length laser is scattered, and promptly the odd number kapillary is gathered the Raman light that the 488nm laser instrument excites; The even numbers kapillary is gathered the Raman light that the 514.5nm laser instrument excites, and vice versa.488nm and 514.5nm laser can be separated by the Raman peaks of water scattering just, and just in time are distributed within the CCD sensing range: 500～660nm.This has met just gathers two groups of demands that data are proofreaied and correct in the free-air correction.Fig. 4 has provided the frame spectrogram on the free-air correction CCD, from Fig. 4, can obviously see corresponding two bright spots (raman peak values) on every capillary.
Fig. 4 is carried out transverse axis data summations as spectral intensity, and it is as shown in Figure 5 to be with longitudinal axis pixel that the horizontal ordinate mapping provides the free-air correction spectrogram.Can provide every capillary center (as shown in Figure 6) and effective width (getting one or two pixel about general) through the identification of simple peak.
Although through some preferred embodiment with reference to invention; Invention is described; But those of ordinary skill in the art should be appreciated that and can make various changes to it in form with on the details, and the spirit and scope of the present invention that do not depart from appended claims and limited.
Claims (7)
1. method of utilizing Raman spectrum imaging location capillary array; It is characterized in that; Said method comprises: Raman light is sent with the laser beam irradiation capillary bundle in (1); (2) be imaged on the charge coupled cell CCD through the Raman imager, the spectrogram that (3) are analyzed on the charge coupled cell CCD positions each kapillary in the capillary bundle.
2. according to the described method of utilizing Raman spectrum imaging location capillary array of claim 1; It is characterized in that, said method step (1) with the laser beam irradiation capillary bundle send Raman light be meant laser beam by quartz capillaceous or the pipe in the solution scattering send Raman light.
3. according to the described method of utilizing Raman spectrum imaging location capillary array of claim 1, it is characterized in that said Raman imager is imaged on the CCD Raman light through getting optical slits, grating beam splitting and condenser lens.
4. according to the described method of utilizing Raman spectrum imaging location capillary array of claim 2; It is characterized in that; Raman light is sent in aqueous solvent scattering in the said laser beam irradiation capillary bundle, and used laser beam is a double laser beam, and the wavelength of said double laser beam is respectively 488nm and 514.5nm.
5. according to the described method of utilizing Raman spectrum imaging location capillary array of claim 4, it is characterized in that said odd number kapillary is gathered the Raman light that the 488nm laser instrument excites, and the even numbers kapillary is gathered the Raman light that the 514.5nm laser instrument excites.
6. according to the described method of utilizing Raman spectrum imaging location capillary array of claim 4, it is characterized in that said even numbers kapillary is gathered the Raman light that the 488nm laser instrument excites, and the odd number kapillary is gathered the Raman light that the 514.5nm laser instrument excites.
7. according to the described method of utilizing Raman spectrum location capillary array image space on charge coupled cell of claim 1; It is characterized in that; Be transverse axis with the pixel count in the said method step (3); Spectral intensity is that the longitudinal axis obtains the free-air correction spectrogram, obtains the effective width between every capillary center and the every capillary through peak identification.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102411024A (en) * | 2011-06-15 | 2012-04-11 | 公安部第一研究所 | Capillary array electrophoresis detection method based on spatial correction and spectral correction |
| CN108760646A (en) * | 2018-06-22 | 2018-11-06 | 西安科锐盛创新科技有限公司 | Chiral sensor part and fluid chiral detection system |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102411024A (en) * | 2011-06-15 | 2012-04-11 | 公安部第一研究所 | Capillary array electrophoresis detection method based on spatial correction and spectral correction |
| CN102411024B (en) * | 2011-06-15 | 2014-07-09 | 公安部第一研究所 | Capillary array electrophoresis detection method based on spatial correction and spectral correction |
| CN108760646A (en) * | 2018-06-22 | 2018-11-06 | 西安科锐盛创新科技有限公司 | Chiral sensor part and fluid chiral detection system |
| CN108760646B (en) * | 2018-06-22 | 2020-12-22 | 浙江清华柔性电子技术研究院 | Chiral sensing device and fluid chiral detection system |
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