CN101806740B - Detection method of human plasma surface enhanced raman spectroscopy by integrating main component analysis - Google Patents
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Abstract
The invention provides a detection method of human plasma surface enhanced raman spectroscopy by integrating main component analysis; the method comprises the following steps that: a human plasma sample in physiological conditions is obtained, and is reduced by hydroxylamine hydrochloride to prepare silver sol; the silver sol and the human plasma sample are uniformly mixed by equal volume and incubated for 2h at 4DEG C, to measure the plasma surface enhanced raman spectroscopy; the measurement to the plasma surface enhanced raman spectroscopy can also adopt laser in different polarization states to excite the human plasma sample; and a plasma surface enhanced raman spectroscopy database is built, and the scattergraph distribution corresponding to the surface enhanced raman spectroscopy of different human plasma is obtained by integrating main component analysis. The detection method ensures the activity of biomolecules in the plasma, and has the advantages of simplicity, fastness and strong reliability. In addition, an SERS technology enables the sample to obtain very strong raman spectroscopy signals at very low laser power, so that the repeatability of the spectrum signals is good, thereby preventing carbonation and damage of high-power laser to the biological sample.
Description
Technical field
The present invention relates to a kind of human plasma surface enhanced raman spectroscopy in conjunction with the multivariable analysis detection method, more specifically say so blood of human body is carried out blood plasma centrifugal treating under the aseptic condition, obtain being in the human plasma solution under the physiological status, detect the SERS signal of blood plasma then by Surface enhanced raman spectroscopy, and utilize the multivariable analysis technology to carry out the method for statistical study.Belong to biomedical sector.
Background technology
Raman spectroscopy can provide the vibrational spectrum of molecule, has the fine structure information and the characteristic fingerprint of molecule in the spectrum, and Raman spectrum has become one of important technology of article evaluation and Molecular Detection.And Raman spectroscopy is applied to the research of life science, by human body tissue or intracellular biomacromolecule, as protein, nucleic acid, the Raman spectrum of materials such as lipid, come the variation of analyst soma or eucaryotic cell structure or component, become the hot subject of present Related Research Domain.But conventional Raman spectrum exists a little less than the signal, and the shortcoming that disturbed by autofluorescence, therefore, need strengthen Raman signal, processing and amplifying.Surface enhanced raman spectroscopy (Surface-enhanced Raman spectroscopy, be called for short SERS) is exactly one of means of the enhancing Raman signal used always.This technology is utilized the effect of molecule and metal (as Au, Ag, Cu and Pt etc.) the surface generation absorption that some is coarse, with the Raman scattering intensity increase by 10 of these molecules
4~ 10
14Times, and suppressed the autofluorescence signal effectively.Characteristics such as the SERS technology has the spatial resolution height, and is highly sensitive, and the information content is abundant have been widely used in the fields such as material evaluation and molecular structure detection.
Chirality is natural base attribute, have many molecules often to have to be mirror image mutually but two kinds of versions that can not be overlapping fully at occurring in nature, the molecule of these two kinds of forms is as people's right-hand man, and this have the compound molecule of chirality factor to be called mapping or optical isomer.The chirality of vital movement and biomolecule is closely related, and for example, to have only glycocoll be not chirality to needed 20 several amino acids in the human body, and other is chiral molecules; Again such as: the selectivity of enzymic catalytic reaction has reflected the requirement that molecule is three-dimensional, and other many processes are also all being permeated the influence of the three-dimensional factor of molecule.Pathology when taking place in biomolecule, and subtle change might take place the space multistory structure of biomolecule, causes that further the biomolecule chirality changes.And the minor alteration of biomolecule space multistory structure is difficult to detect with common non-polarized Raman laser.The supercoil electromagnetic field that circularly polarized laser produced can be sensitive detect the small variation of biomolecule chirality.Therefore, utilize the variation of circularly polarized laser detectable biomolecule chirality, the information that discloses secrets of life, pathology is had great importance.
It mainly is that blood and the plasma sample that will obtain carries out direct Raman detection that present Chinese scholars utilizes Raman spectroscopy to carry out blood testing, but does not all obtain desirable effect.The weak point of these researchs is a little less than the conventional raman spectral signal that autofluorescence is disturbed strong, and bigger, the consuming time length of the required excitation light power of conventional laser Raman detection, easily sample is caused damage.And utilizing non-polarized Raman laser, linearly polarized laser or circularly polarized laser as exciting light, elargol carries out check and analysis in conjunction with principal component analysis (PCA) to human plasma for the Surface enhanced raman spectroscopy technology that strengthens matrix, does not see relevant report yet.
Summary of the invention
The objective of the invention is to deficiency and problem at existence in the present blood Raman spectrum detection, provide a kind of plasma surface enhanced raman spectroscopy technology of utilizing in conjunction with principal component analytical method, it carried out blood plasma centrifugal treating under the aseptic condition to blood of human body before this, obtain being in the human plasma solution under the physiological status, and utilize the SERS detection means to realize the detection of human plasma Surface enhanced raman spectroscopy.Plasma sample preprocessing process required time of the present invention is 2h, and only be 10 seconds detection time, therefore guarantees the activity of biomolecule in the blood plasma in measuring process, has simply fast the advantage of good reliability.And the SERS technology makes sample just can obtain very strong raman spectral signal under very low laser power, spectral signal good reproducibility, carbonization, the damage phenomenon of having avoided high power laser light that biological sample is caused.Based on the plasma surface enhanced raman spectroscopy of different people, set up the plasma surface enhanced raman spectroscopy database, utilize the multivariate statistical analysis method, for analyst's plasma surface enhanced raman spectroscopy provides a kind of new method.
For realizing that purpose of the present invention adopts technical scheme as follows:
(1) extract blood of human body and add anti-coagulants and carry out centrifugal treating aseptic condition under, acquisition is in the human plasma sample under the physiological status, utilizes oxammonium hydrochloride reduction preparation of silver colloidal sol;
(2) silver sol and human plasma sample mix by equal-volume and 4
0Hatching two hours laggard promoting circulation of blood slurry Surface enhanced raman spectroscopy under the C condition measures;
(3) plasma surface enhanced raman spectroscopy is measured and can be adopted laser in different polarization states to excite the human plasma sample;
(4) set up the plasma surface enhanced raman spectroscopy database, utilize principal component analysis (PCA) and T check to obtain the corresponding scatter plot distributions of Surface enhanced raman spectroscopy of different human body blood plasma.
Wherein the described plasma surface enhanced raman spectroscopy measurement of step (2) is the mixed solution of human plasma and elargol to be dripped carry out surface-enhanced Raman measure on purity is 99.99% aluminium flake, and emphasis detects 450-4000cm
-1Wave-number range.
The described laser in different polarization states of step (3) can be used for human plasma sample, purifying protein, DNA, RNA sample and carry out conventional Raman spectrum check and analysis.
The described laser in different polarization states of step (3) can be non-polarized Raman laser, linearly polarized laser, Left-hand circular polarization laser or right-hand circular polarization laser.Utilize circularly polarized laser to excite and to obtain the information that relevant biomolecule chirality changes.
The described plasma surface enhanced raman spectroscopy database of step (4) is made up of the plasma surface enhanced raman spectroscopy detection data of different human body; Before described plasma surface enhanced raman spectroscopy database is set up, earlier different people blood plasma SERS spectrum is utilized fitting of a polynomial elimination fluorescence background and carries out the area normalization processing, to remove the inconsistent influences that cause of experiment condition such as excitation light power fluctuation, gathering difference.
It can be urine, serum, lymph liquid, celiolymph, urine, saliva, tear, sweat, cell extract, tissue homogenate, vaginal secretion or seminal fluid that the present invention adopts the substitute of the described blood plasma of technical scheme, also can be purifying protein, DNA or RNA sample.
Advantage of the present invention is to utilize non-polarized Raman laser, linearly polarized laser and circularly polarized laser to carry out the method that Surface enhanced raman spectroscopy is measured as exciting light, can obtain high-quality plasma surface and strengthen Raman signal, can obtain the information that relevant biomolecule chirality changes in the different people blood plasma molecule.Obtain the scatter plot distributions of the Surface enhanced raman spectroscopy correspondence of different human body blood plasma in conjunction with principal component analysis (PCA).For realizing providing important references to quick, the harmless detection of different people plasma surface enhanced raman spectroscopy.
Description of drawings
Fig. 1 is that the average surface of the A group blood plasma that records of the present invention strengthens Raman spectrum;
Fig. 2 is that the average surface of the B group blood plasma that records of the present invention strengthens Raman spectrum;
Fig. 3 is that the present invention is used for calculating parameter a
1, a
4, a
8Needed the first, the 4th and the 8th major component spectrum
Fig. 4 is A group blood plasma and the B group plasma surface enhanced raman spectroscopy PCA score scatter plot distributions that the present invention utilizes PC1 and PC4 to draw next, selecting of circular black represented A group human plasma Surface enhanced raman spectroscopy among the figure, and red triangled tip is represented B group human plasma Surface enhanced raman spectroscopy;
Fig. 5 is A group blood plasma and the B group plasma surface enhanced raman spectroscopy PCA score scatter plot distributions that the present invention utilizes PC1 and PC8 to draw next, selecting of circular black represented A group human plasma Surface enhanced raman spectroscopy among the figure, and red triangled tip is represented B group human plasma Surface enhanced raman spectroscopy;
Fig. 6. be first human plasma average SERS spectrum comparison diagram under non-polarized Raman laser, Left-hand circular polarization laser and right-hand circular polarization laser excitation respectively;
Fig. 7. be second human plasma average SERS spectrum comparison diagram under non-polarized Raman laser, Left-hand circular polarization laser and right-hand circular polarization laser excitation respectively;
Embodiment
The present invention is described below according to concrete implementation detail:
(1) pre-service of silver sol pre-preparation and plasma sample
The sodium hydroxide solution (0.1mol) of 4.5 ml is joined in the 5ml oxammonium hydrochloride solution (0.06mol), then potpourri is added to fast in the 90ml liquor argenti nitratis ophthalmicus (0.0011mol), evenly stir until obtaining uniform newborn grey solution.With 10000 rev/mins in hydro-extractor, centrifugal 10 minutes, make the elargol layering, supernatant is abandoned, take off silver sol that layer concentrates at room temperature lucifuge seal up for safekeeping standby.
Aseptic condition extracts down the fasting blood overnight of different people between point-8 in mornings 7, adds EDTA and prevents blood clotting and centrifugal (2000 rev/mins) 15 minutes.Upper serum is abandoned, take off layer blood plasma as sample.Utilize this method to obtain two groups of different people plasma samples (compile and be A group and B group) respectively.Utilizing liquid-transfering gun to take out each sample blood plasma 200 μ l from the A group adds through aseptic disinfecting in vitro.And add each 200 μ l of silver sol behind previous preparation centrifugal in the test tube with liquid-transfering gun, mix with silver sol volume ratio 1:1 according to blood plasma.Mixed solution is fully stirred, and it is even as far as possible that blood plasma is mixed with silver sol, makes A group blood plasma-silver sol mixed solution.Utilizing pipettor to take out each 200 μ l of each sample blood plasma from the B group adds through aseptic disinfecting in vitro.And add each 200 μ l of silver sol behind previous preparation centrifugal in the test tube with pipettor, mix with silver sol volume ratio 1:1 according to blood plasma.Mixed solution is fully stirred, and it is even as far as possible that blood plasma is mixed with silver sol, makes B group blood plasma-silver sol mixed solution.All mixed solutions that make are put into the refrigerator that is set at 4 ℃ hatched two hours.
(2) Surface enhanced raman spectroscopy test sample process
With liquid-transfering gun the blood plasma-silver sol mixed liquor that mixes being moved to purity is on the 99.99% aluminium flake sample stage, naturally dry, utilize the Raman spectrometer test sample, used exciting light is non-polarized Raman laser, linearly polarized laser, Left-hand circular polarization laser or right-hand circular polarization laser, emphasis detects 450-4000cm
-1Wave-number range is to obtain the Surface enhanced raman spectroscopy of blood plasma.The setting measurement parameter: integral time 10s, excitation wavelength 785nm, excitation light power 5mw.
(3) Surface enhanced raman spectroscopy of different people blood plasma is carried out principal component analysis (PCA)
Different blood plasma are carried out principal component analysis (PCA), need set up the plasma surface enhanced raman spectroscopy database.Before setting up the spectra database model, earlier the Surface enhanced raman spectroscopy of blood plasma to be carried out area normalization and handle to remove the inconsistent influences that cause of experiment condition such as excitation light power fluctuation, gathering difference.When setting up database, consider some individual differences that exist between the different people, for guaranteeing statistical, must gather the Surface enhanced raman spectroscopy data of the different people blood plasma of abundant quantity.The plasma surface enhanced raman spectroscopy database is made up of the plasma surface enhanced raman spectroscopy detection data of different human body.Setting up on the basis of database, utilize PCA to analyze and obtain the pairing score of each major component (PC score), then utilize the Independent-Sample T test among the SPSS, select to have most three PCA of significant difference must assign to further to draw scatter plot distributions of A group and B group different people plasma surface enhanced raman spectroscopy.
The first step of setting up blood plasma SERS database is that spectrum is carried out normalization.Because the information spinner in the spectrum will be composed on the peak relative intensity from each, and the absolute strength at spectrum peak and laser power fluctuation, gathering situation are relevant, and normalization can be eliminated this influence.The present invention adopts spectral line is carried out normalized by the normalized method of integral area.Second step of setting up model is to draw the averaged spectrum that averaged spectrum and the B of the A group in composition data storehouse organize.The averaged spectrum computing formula is:
, wherein
Be averaging spectrum.Set forth for convenient, we represent wave number with the different spectrum of subscript m mark with subscript i and j.
Next utilize the canonical algorithm of principal component analysis (PCA), establish the principal element spectrum of describing the spectrum change feature.At first, the covariance matrix of each wave number after the calculating normalization, that is:
Compute matrix
Eigenvalue and eigenvector because a few eigenvalue that only has by size order to stand out has the numerical value that is higher than noise, the back owing to be weaker than noise and can be left in the basket, so we only get preceding 20 eigenvalues
,
And its corresponding eigenvector
,
.
Composition principal element spectrum.Because the orthonomality of eigenvector can carry out decomposing by the linearity of principal element spectrum, that is: to calibration spectrum
Wherein
Be the spectrum after the normalization,
Be expansion coefficient (score).{
Formed orthonormal base mistake collection, and Zhang Chengyi new space is called the principal element space, and this space can be by matrix before
Doing spatial variations obtains.Through top variation, every spectrum is mapped as a point in principal element space.
Be to use the concrete computation process that PCA analyzes below:
(1) at first utilize different people blood plasma SERS spectrum fitting of a polynomial to eliminate fluorescence background;
(2) the different people blood plasma SERS spectrum of eliminating fluorescence background being carried out area normalization handles;
(3) utilizing SPSS software that the different people blood plasma SERS spectrum of handling through (1), (2) is carried out PCA analyzes;
(4) utilize the T check to obtain three the PCA score of significant difference and the scatter plot distributions of drawing are arranged most;
Through repetition test, excite down in non-polarized Raman laser, the present invention confirms that by the T check PCA analyzes back major component 1, major component 4 and major component 8 these three major components and has significant difference.Our scatter diagram of major component 1 and major component 4, major component 1 and major component 8 that further draws.
Above-mentioned non-polarized Raman laser can adopt linearly polarized laser, Left-hand circular polarization laser or right-hand circular polarization laser to substitute.
(4) blood plasma SERS database carries out PCA to analyze concrete steps as follows:
1. different people blood plasma is prepared silver sol-blood plasma mixed solution according to step ();
2. measure plasma surface enhanced raman spectroscopy according to step (two); A group and B group are measured the blood plasma spectrum that is not less than 30 examples respectively.
3. every routine blood plasma SERS spectrum is all carried out area normalization.
4. to the blood plasma SERS spectrum in the spectra database, utilize the PCA canonical algorithm to calculate each principal component scores..
5. utilize the T check to calculate the major component that significant difference is arranged most and be PC1, PC4, PC8.
6. the Surface enhanced raman spectroscopy PCA score scatter plot distributions of different people blood plasma promptly is a measurement result of the present invention.
All data of the present invention and numerical value are all recorded out by actual measurement, and the identification conclusion of blood plasma is provided by the Surface enhanced raman spectroscopy data are objective, do not rely on observer's subjective judgement.The plasma sample preparation can be finished in 2 hours, and the spectral measurement time can be controlled in two minutes.The principal component analysis (PCA) of spectrum is calculated and can be obtained the result in 10 minutes.Therefore human plasma surface enhanced raman spectroscopy by integrating main component analysis has boundless application prospect at biomedical sector.
Below be several specific embodiment of the present invention, further describe the present invention, but the present invention be not limited only to this.
Embodiment 1
Extract the fasting blood overnight of two groups of different people between point-8 in mornings 7, add EDTA and prevented blood clotting and centrifugal (2000 rev/mins) 15 minutes.Upper serum is abandoned, take off layer blood plasma as sample.Totally 33 parts of first group of human plasmas are compiled the group into A.Totally 43 parts of second group of human plasmas are compiled the group into B; Utilizing liquid-transfering gun to take out each sample blood plasma 200 μ l from the A group adds through aseptic disinfecting in vitro.And add each 200 μ l of silver sol behind previous preparation centrifugal in the test tube with liquid-transfering gun, mix with silver sol volume ratio 1:1 according to blood plasma.Mixed solution is fully stirred, and it is even as far as possible that blood plasma is mixed with silver sol, makes A group blood plasma-silver sol mixed solution.Utilizing pipettor to take out each 200 μ l of each sample blood plasma from the B group adds through aseptic disinfecting in vitro.And add each 200 μ l of silver sol behind previous preparation centrifugal in the test tube with pipettor, mix with silver sol volume ratio 1:1 according to blood plasma.Mixed solution is fully stirred, and it is even as far as possible that blood plasma is mixed with silver sol, makes B group blood plasma-silver sol mixed solution.All mixed solutions that make are put into the refrigerator that is set at 4 ℃ hatched two hours.
With liquid-transfering gun the blood plasma-silver sol mixed liquor that mixes being moved to purity is on the 99.99% aluminium flake sample stage, dries naturally, utilizes confocal Raman spectra instrument test sample, and emphasis detects 450-4000cm
-1Wave-number range is to obtain the Surface enhanced raman spectroscopy of blood plasma.The setting measurement parameter: integral time 10s, excitation wavelength 785nm, excitation light power 5mw.Used exciting light is a non-polarized Raman laser when measuring SERS spectrum.
Before setting up the plasma surface enhanced raman spectroscopy database, earlier the Surface enhanced raman spectroscopy of blood plasma to be carried out area normalization and handle to remove the inconsistent influences that cause of experiment condition such as excitation light power fluctuation, gathering difference.Setting up on the basis of database, drawing the averaged spectrum (as shown in Figure 1) of A group in the composition data storehouse and the averaged spectrum (as shown in Figure 2) of B group.Utilize principal component analysis (PCA) to obtain the pairing score of each major component (PC score), then utilize the Independent-Sample T test among the SPSS, select to have most three PCA scores of significant difference, be PC1, PC4 and PC8, the scatter plot distributions of the A that further draws group and B group plasma surface enhanced raman spectroscopy.Accompanying drawing 3 is the first, the 4th and the 8th major component spectrum, and accompanying drawing 4 is scatter plot distributions of PC1 and PC4, and accompanying drawing 5 is scatter diagrams of PC1 and PC8.
Extract the fasting blood overnight of two different people between point-8 in mornings 7, add EDTA and prevented blood clotting and centrifugal (2000 rev/mins) 15 minutes.Upper serum is abandoned, take off layer blood plasma as sample.Utilizing liquid-transfering gun to take out sample blood plasma 200 μ l from first plasma sample adds through aseptic disinfecting in vitro.And add each 200 μ l of silver sol behind previous preparation centrifugal in the test tube with liquid-transfering gun, mix with silver sol volume ratio 1:1 according to blood plasma.Mixed solution is fully stirred, and it is even as far as possible that blood plasma is mixed with silver sol, makes first blood plasma-silver sol mixed solution.Utilizing pipettor to take out each 200 μ l of sample blood plasma from second human plasma adds through aseptic disinfecting in vitro.And add each 200 μ l of silver sol behind previous preparation centrifugal in the test tube with pipettor, mix with silver sol volume ratio 1:1 according to blood plasma.Mixed solution is fully stirred, and it is even as far as possible that blood plasma is mixed with silver sol, makes control group silver sol-blood plasma mixed solution.All mixed solutions that make are put into the refrigerator that is set at 4 ℃ hatched two hours.
Utilizing liquid-transfering gun that the silver sol-blood plasma mixed liquor that mixes is moved to purity is on the 99.99% aluminium flake sample stage, naturally dry, utilize confocal Raman spectra instrument test sample, used exciting light is respectively to use non-polarized Raman laser when measuring SERS spectrum, Left-hand circular polarization laser and right-hand circular polarization laser.Emphasis detects 450-1730cm
-1Wave-number range is to obtain the Surface enhanced raman spectroscopy of blood plasma.The setting measurement parameter: integral time 10s, excitation wavelength 785nm, excitation light power 5mw.Test obtains two groups of data shown in accompanying drawing 6, accompanying drawing 7., especially utilize circularly polarized light to excite as can be seen and can obtain to excite the information that more manys about the molecular chiral variation at the average SERS comparison diagram of the exciting light of different polarization states from these two human plasmas than nonpolarized light in the contrast of shadow region bands of a spectrum scope.Show that the variation that utilizes circularly polarized laser detectable biomolecule chirality can have great importance to the information that discloses secrets of life, pathology.
Claims (8)
1. the detection method of a human plasma surface enhanced raman spectroscopy by integrating main component analysis is characterized in that this method is made up of following three steps:
(1) extract blood of human body and add anti-coagulants and carry out centrifugal treating aseptic condition under, acquisition is in the human plasma sample under the physiological status, utilizes oxammonium hydrochloride reduction preparation of silver colloidal sol;
(2) silver sol and human plasma sample mix by equal-volume and 4
0Hatching two hours laggard promoting circulation of blood slurry Surface enhanced raman spectroscopy under the C condition measures;
(3) set up the plasma surface enhanced raman spectroscopy database, utilize principal component analysis (PCA) and T check to obtain the corresponding scatter plot distributions of Surface enhanced raman spectroscopy of different human body blood plasma.
2. the detection method of a kind of human plasma surface enhanced raman spectroscopy by integrating main component analysis according to claim 1, it is characterized in that: the described plasma surface enhanced raman spectroscopy measurement of step (2) is the mixed solution of human plasma sample and silver sol to be dripped carry out Surface enhanced raman spectroscopy measure on purity is 99.99% aluminium flake, and emphasis detects 450-4000cm
-1Wave-number range.
3. the detection method of a kind of human plasma surface enhanced raman spectroscopy by integrating main component analysis according to claim 1 is characterized in that: the described plasma surface enhanced raman spectroscopy database of step (3) detects data by the plasma surface enhanced raman spectroscopy of different human body and forms.
4. according to the detection method of claim 1 or 3 described a kind of human plasma surface enhanced raman spectroscopy by integrating main component analysis, it is characterized in that: before described plasma surface enhanced raman spectroscopy database is set up, earlier different people blood plasma SERS spectrum is utilized fitting of a polynomial elimination fluorescence background and carries out the area normalization processing.
5. a human plasma surface enhanced raman spectroscopy is in conjunction with the detection method of different polarization states laser excitation, and it is characterized in that: concrete steps are as follows:
(1) extract blood of human body and add anti-coagulants and carry out centrifugal treating aseptic condition under, acquisition is in the human plasma sample under the physiological status, utilizes oxammonium hydrochloride reduction preparation of silver colloidal sol;
(2) silver sol and human plasma sample mix by equal-volume and 4
0Hatching two hours laggard promoting circulation of blood slurry Surface enhanced raman spectroscopy under the C condition measures;
(3) plasma surface enhanced raman spectroscopy is measured and is adopted laser in different polarization states to excite the human plasma sample.
(4) set up the plasma surface enhanced raman spectroscopy database, utilize principal component analysis (PCA) to obtain the scatter plot distributions of the Surface enhanced raman spectroscopy correspondence of different human body blood plasma.
6. human plasma surface enhanced raman spectroscopy according to claim 5 is characterized in that in conjunction with the detection method of different polarization states laser excitation: described laser in different polarization states is non-polarized Raman laser, linearly polarized laser, Left-hand circular polarization laser or right-hand circular polarization laser.
7. human plasma surface enhanced raman spectroscopy according to claim 5 is characterized in that in conjunction with the detection method of different polarization states laser excitation: described different polarization states laser is used for human plasma sample, purifying protein, DNA or RNA sample are carried out the check and analysis of Surface enhanced raman spectroscopy.
8. human plasma surface enhanced raman spectroscopy according to claim 5 is in conjunction with the detection method of different polarization states laser excitation, and the substitute that it is characterized in that described blood plasma is urine, serum, lymph liquid, celiolymph, saliva, tear, sweat, cell extract, tissue homogenate, vaginal secretion, seminal fluid, purifying protein, DNA or RNA sample.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102095718A (en) * | 2010-12-25 | 2011-06-15 | 福建师范大学 | Raman spectrum detecting device based on different polarization and excitation light sources |
| CN102175664A (en) * | 2011-02-17 | 2011-09-07 | 福建师范大学 | Method for detecting surface enhanced Raman spectra of blood RNA |
| EP2700933A1 (en) * | 2012-08-20 | 2014-02-26 | Consejo Superior De Investigaciones Científicas (CSIC) | Raman, infrared, or Raman-Infrared analysis of peripheral blood plasma protein structure and its relation to cognitive development in Alzheimer's disease |
| US10067060B2 (en) | 2013-01-30 | 2018-09-04 | Hewlett-Packard Development Company, L.P. | Polarization selective surface enhanced raman spectroscopy |
| EP2951562A4 (en) * | 2013-01-30 | 2015-12-09 | Hewlett Packard Development Co | Polarization selective surface enhanced raman spectroscopy |
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| CN103968946B (en) * | 2014-05-19 | 2016-01-13 | 中国人民解放军第二军医大学 | A kind of acquisition method of surface-enhanced Raman two-dimensional correlation spectra |
| CN104914089B (en) * | 2015-06-18 | 2017-10-27 | 清华大学 | The method for carrying out semi-quantitative analysis to trace mixture with SERS |
| CN106546572B (en) * | 2015-12-13 | 2018-06-19 | 中国科学院大连化学物理研究所 | A kind of short wavelength laser chirality Raman spectrometer |
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| CN111751349A (en) * | 2020-06-29 | 2020-10-09 | 陕西未来健康科技有限公司 | Method and system for label-free analyte detection |
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|---|---|---|---|---|
| US20060135861A1 (en) * | 2003-02-06 | 2006-06-22 | Koninklijke Philips Electronics N.V. | Apparatus and method for blood analysis |
| CN1444045A (en) * | 2003-04-15 | 2003-09-24 | 吉林大学 | Surface enhancement Raman scattering labelling immunodetection method |
| US7524671B2 (en) * | 2005-01-27 | 2009-04-28 | Prescient Medical, Inc. | Handheld raman blood analyzer |
| US7651851B2 (en) * | 2005-01-27 | 2010-01-26 | Prescient Medical, Inc. | Handheld Raman body fluid analyzer |
| CN2777536Y (en) * | 2005-03-14 | 2006-05-03 | 河南大学 | Adsorption substrate for Raman scattering analyzing tester |
| CN1837791A (en) * | 2006-04-26 | 2006-09-27 | 大连理工大学 | Near field enhanced Raman molecular fingerprint spectrum analysis method |
| US20100105020A1 (en) * | 2006-10-24 | 2010-04-29 | Koninklijke Philips Electronics N.V. | Quantitative measurement of glycated hemoglobin |
| CN101482509A (en) * | 2009-03-03 | 2009-07-15 | 福建师范大学 | Method for detecting animal active unicellular sample by surface reinforced Raman spectrum |
-
2010
- 2010-04-19 CN CN2010101492679A patent/CN101806740B/en not_active Expired - Fee Related
- 2010-06-21 WO PCT/CN2010/074142 patent/WO2011130938A1/en active Application Filing
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| Publication number | Publication date |
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| WO2011130938A1 (en) | 2011-10-27 |
| CN101806740A (en) | 2010-08-18 |
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