CN101283241A - System and method for the identification and quantification of a biological sample suspended in a liquid - Google Patents
System and method for the identification and quantification of a biological sample suspended in a liquid Download PDFInfo
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- CN101283241A CN101283241A CNA2006800370712A CN200680037071A CN101283241A CN 101283241 A CN101283241 A CN 101283241A CN A2006800370712 A CNA2006800370712 A CN A2006800370712A CN 200680037071 A CN200680037071 A CN 200680037071A CN 101283241 A CN101283241 A CN 101283241A
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Abstract
A system for the identification and quantification of a biological sample suspended in a liquid includes a fluorescence excitation module with at least one excitation light source; a sample interface module optically coupled to the fluorescence excitation module for positioning a biological sample to receive excitation light from the at least one excitation light source; a fluorescence emission module optically coupled to the sample interface module and comprising at least one detection device for detecting fluorescence excitation-emission matrices of the biological sample; and a computer module operatively coupled to the fluorescence emission module. The computer module performs multivariate analysis on the fluorescence excitation-emission matrices of the biological sample to identify and quantify the biological sample. The multivariate analysis may comprise extended partial least squared analysis for identification and quantification of the biological sample. A method for the identification and quantification of a biological sample suspended in a liquid is also provided.
Description
The cross reference of related application
It is on August 8th, 2005 that the application requires the applying date, and application number is No.60/706, the right of priority of 489 U.S. Provisional Patent Application " identification and the system that quantitatively is suspended in the biological sample in the liquid ", and introduce its full content for your guidance.
Background technology
Technical field
Present invention relates in general to the system and method that a kind of identification and quantification are suspended in biological sample in the liquid.More precisely, the present invention relates to a kind of fluorescence excitation emission matrix utilization multivariate analysis and discern the system and method that quantizes biological sample biological sample.
The explanation of association area
In the bacteriology, decoration method is used to discern Gram-positive and these two kinds general bacterium groups of Gram-negative, and can not discern bacterial species.Use chromogenic culture medium can separate and discern some microorganisms of human pathology, might kind but can not discern institute.Use chemical dyeing method can discern about 20000 kinds of different bacterium kinds at present.Yet these methods are still existing huge problem aspect the time of bacterium identification, for example, and the organism (approximately ancillary cost is 24 hours) that uses automation equipment application standard chemistry method to need ability acquisition in 18-24 hour to separate.
In order to obtain the response time faster, various spectral techniques are developed.For example, FFIR (FTIR) just is used in the analysis of biological sample to obtain the response time faster.The step of FITR spectroscopic methodology is as follows.At first, collect isolated bacterium group from urethral infection (UTI) patient urine sample.50 ℃ of samples of drying 30 minutes are down analyzed, at 4000cm
-1To 600cm
-1Wavelength coverage in collected specimens spectrum, spectra collection will carry out under the frequency of 20Hz.In order to improve signal to noise ratio (S/N ratio), 256 spectrum are superposeed with average.The sample identification of the analysis showed that to information is feasible.
Raman spectrum also may can be used for discerning and the quantitative method of bacteria samples after deliberation.The step of Raman spectrum is as follows.At first, utilize the dispersive Raman spectrum instrument (Ramascope) of the semiconductor laser with low-power (30mW) near infrared 780nm to collect spectrum under sampled point, the sampled point representative value is 3mW.Sample is a bacterial suspension (3 * 10
9Every milliliter in individual cell).Spectrum sample 60 seconds.Information analysis shows that the identification of biological sample is feasible.Yet the degree of confidence of identification is not high.
Further research and propose the use fluorescence spectrum and carried out quick bacteria identification.For example, use many fluorescence excitation spectrums, then again set of biomolecules is carried out selective excitation to carry out best bacterial species identification to select best excitation wavelength.
The U.S. Patent No. 6834237 of people such as Noergaard invention discloses a kind of categorizing system characterizes the biological sample that separates under the condition of the environment that comprises an animal organism sample that separates method of following the trail of.The biological sample that separates is selected from body fluid or tissue sample.Tissue sample and conditional independence.An example of this method is the urine sample of getting smoker and non-smoker, if detect emission light in the urine sample, then is the smoker.
The U.S. Patent No. 6773922,6426045,6365109 and 6087182 of people such as Jeng invention discloses a kind of instrument and method that is used for the concentration of location parameter such as at least a biological sample analysis thing.This instrument and method use visible absorption spectrum to obtain the concentration value of some analyte, use infrared absorption spectrum to obtain the concentration value of other analytes.
The U.S. Patent No. 5938617 of Vo-Dinh invention discloses a kind of optical excitation sample that utilizes different wave length, and the synchronized sampling emissive porwer is with the system of identification sample biopathogen.The mechanism that this system adopts is for to be exposed to sample under the exciting radiation, to obtain emitted radiation.Biopathogen can be virus and bacterium.
Yet above-mentioned each method and/or system are not to use reagent to need the complicated sample preparation exactly, and this makes method and/or system more be difficult to operation, and is easier to misoperation.The required time of specimen preparation also makes said method and/or system not be suitable for diagnosis fast.
Summary of the invention
Correspondingly, the system that the purpose of this invention is to provide biological sample in a kind of identification and the quantification liquid.The present invention further purpose provides a kind of quick analysis system.Another object of the present invention provide a kind of do not need to use reagent and discern and the system that quantizes biological sample in the liquid to reduce Master Cost.
The present invention be directed to the system that the biological sample that is suspended in the liquid is discerned and quantized.This system comprises the fluorescence excitation module that has an excitation source at least; Sample interface module with fluorescence excitation module optical coupled, this module is used to locate biological sample to accept the exciting light of at least one excitation source; Fluorescent emission module with sample interface module optical coupled, this module comprise that at least one is used for the detection of biological fluorescent and excites-pick-up unit of emission matrix; A computer module that effectively is coupled with the fluorescent emission module.This computer module carries out multivariate analysis with identification and quantification biological sample to the fluorescence excitation-emission matrix of biological sample.Multivariate analysis can comprise that the analysis of expansion partial least square method is with identification and quantification biological sample.
System can comprise further that one absorbs module and a diffuse reflection module.Absorb module use from least one excitation source or separately the light that sends of modulated light source realize the absorptiometry of biological sample.Absorptiometry can combine with the fluorescence excitation-emission matrix of biological sample and discern and quantize biological sample.Absorbing module both can be that monochromator also can be the filter wheel with photomultiplier.The diffuse reflection module use from least one excitation source or separately the light that sends of modulated light source realize the diffuse reflectance measurement of biological sample.Diffuse reflectance measurement can combine with the fluorescence-excitation-emission matrix of biological sample and discern and quantize biological sample.The diffuse reflection module can be the monochromator with semiconductor detector or photomultiplier.
At least one excitation source can be the combination in any of continuous light source, pulse flashing light, semiconductor laser, tunable laser or above-mentioned light source.At least one excitation source can be selected wavelength by using grating monochromator, the filter wheel that has narrowband light filter, acousto-optic turnable filter, liquid crystal tunable optical filter, circular variable filter, linear variable filter or above-mentioned combination in any.
At least one pick-up unit of fluorescent emission module both can be that the raster monochromator with solid-state detector also can be the non-raster monochromator with multichannel array detector.The fluorescent emission module can further include the gate electronics that is used for the optical sampling degree of depth in the controlling liquid and optimizes the signal to noise ratio (S/N ratio) feature.
Affiliated system can further include display device, and this display device is used for showing the identification and the quantification of biological sample.
The present invention also further relates to a kind of method that is used for detecting and quantizing to be suspended in the liquid biological sample.Described method comprises step: an excitation source a) is provided; B) use the excitation light source excites biological sample; C) spectral information of detection of biological sample, the form of spectral information are to excite-emission matrix, absorptiometry, diffuse reflectance measurement or above-mentioned combination in any; D) spectral information is carried out multivariate analysis with identification and quantification biological sample, described multivariate analysis can comprise that the analysis of expansion partial least square method is with identification and quantification biological sample.
Excitation source can be the combination in any of continuous light source, pulse flashing light, semiconductor laser, tunable laser or above-mentioned light source.At least one excitation source can be selected wavelength by using grating monochromator, the filter wheel that has narrowband light filter, acousto-optic turnable filter, liquid crystal tunable optical filter, circular variable filter, linear variable filter or above-mentioned combination in any.
Described method can further include step: e) show the identification and the quantification of biological sample.Data formatting and data pre-service can be carried out before step d), and described multivariate analysis can comprise that the analysis of expansion partial least square method quantizes biological sample with identification.
With reference to following with reference to the description of accompanying drawing and additional claim, the function of these and other features of the present invention and method of operating and associated cell structures becomes more clear.The supplemental instruction of following description and accompanying drawing also is the part of this instructions, and wherein Reference numeral is represented the appropriate section in each accompanying drawing.Odd number in instructions and claims " one " also comprises its plural form, unless its content has clearly illustrated reverse situation.
Description of drawings
Fig. 1 is according to a kind of general illustration of discerning and quantizing the biological sample system in the liquid that is suspended in of the present invention
Fig. 2 is according to a kind of quadrature (right-angle) configuration schematic diagram of discerning and quantizing the biological sample system in the liquid that is suspended in of the present invention
Fig. 3 is according to a kind of front (front-face) configuration schematic diagram of discerning and quantizing the biological sample system in the liquid that is suspended in of the present invention
Fig. 4 is the detail drawing of the sample interface module of positive configuration among Fig. 3
Fig. 5 is the chart that concerns of subduing Klebsiella pneumoniae concentration in positive fluorescence intensity and the phosphate buffer
Fig. 6 be subdue Klebsiella pneumoniae in fluorescent emission intensity and the phosphate buffer excitation wavelength concern chart
Fig. 7 is a chart of subduing Escherichia coli concentration relationship in quadrature fluorescence intensity and the water
Fig. 8 is the chart that concerns of subduing Escherichia coli concentration in positive fluorescence intensity and the phosphate buffer
Fig. 9 is the chart that concerns of subduing Escherichia coli concentration in quadrature fluorescence intensity and the human urine
Embodiment
For purposes of illustration, term hereinafter " on " D score " right side " " left side " " level " " vertically " " top " " bottom " " side " " vertically " and above-mentioned derivatives be all relevant with invention, refers to position in the accompanying drawing.Yet, should be understood to, the present invention also can take the different alternative changing value except that the phase counterstatement.Also can be regarded as in the accompanying drawing and the device specifies described in the following explanation only for demonstration example of the present invention.Therefore, the concrete size that relates among the embodiment disclosed herein and other physical features should not be construed as limitation of the present invention.
System and method of the present invention can quick identification and is quantized in the liquid biological sample and do not need to add reagent in liquid.The present invention can be used for following environment: to microorganism in the timely point-of-care bio-medical analysis of bacterium in the human body fluid and virus, the identification seawater with the content monitoring of the navigation that is controlled at the coastal marine site of the U.S., detection and Identification biological warfare agent, Food ﹠ Drink industry and potable water and waste water.
Fig. 1 is the system 1 of biological sample in identification and the quantification liquid, comprises fluorescence excitation module 3, sample interface module 5, fluorescent emission module 7, computer module 9, display device 11, absorbs module 13 and diffuse reflection module 15.
Referring to Fig. 2,3 and Fig. 1, the system of the present invention 1 configurable optical arrangement that various fluorescence excitations and collection are arranged.For example, system 1 can be configured to orthogonal configuration 1 ' (see figure 2) or positive configuration 1 " (see figure 3).
Optical interface comprises mirror 25 and lens 37, is used for as much as possible directed and focuses on the light that excitation source 19 produces.A selectable scheme of the present invention is, uses single or branch optical fiber as optical interface.The appropriate location that sample color comparison tube 23 is used in system holds the biological sample that is suspended in the liquid.Fig. 4 is more detailed positive configuration 1 " the synoptic diagram of sample interface module 5.Sample interface module 5 comprises that one is used for focusing on the lens 40 of the light that fluorescent emission module 3 sends.Lens 40 can be the coupling of CVI PXF-50.8-90.8-UV lens and CVI BXF-50.8-90.8-UV lens, and these lens are by CVI Laser LLC, 200 Dorado SE, and Albuquerque, NM 87123 companies make.The light that focused on of lens 40 is reflexed to sample color comparison tube 23 by mirror 41 and 42 then.Mirror 41 can be Newport20D10.AL2, and mirror 42 can be for Newport 10D10.AL2, by NewportCorporation, and 1791Deere Avenue, Irvine, CA92606 company makes.
The light that sample color comparison tube 23 is reflected points to fluorescent emission module 7 by focusing, hole 45 and the lens 46 of hole 43 and process lens 44.Lens 44 can be CVI PXF-50.8-77.3-UV lens, and lens 46 can be the CVIPXF-50.8-40.7-UV lens, all are that CVI Laser LLC company makes.
The light that sees through sample color comparison tube 23 is reflected by mirror 47 and 48.Mirror 47 can be Newport 10D10.AL2, and mirror 48 is Newport 20D10.AL2, is made by Newport Corporation company.Light is then focused on by lens 49, and by iris device 50, the final sensing absorbs module 13.Lens 49 can be the coupling by CVIPXF-50.8-90.8-UV lens and CVI BXF-50.8-312.0-UV lens, and these lens are made by CVI Laser LLC company.
All optical modules of sample interface 5 are located by using suitable clamper, for example by Thorlabs, and Inc., 435 Route, 206 North, Newton, clamper and locating device that NJ07860 company produces.Optical interface can comprise that also beam dump 51 and 52 is used for the reflect stray light of lowering apparatus inside, and its position as shown in Figure 4.
The polarized light (not shown) is a polarizer, is used for from exciting of coming out of the liquid medium that is suspended with biological sample and/or launches light beam and elastic scattering light.
After computer module 9 was finished polynary Spectrum Analysis, display device 11 showed relevant information to the user.These information can include but not limited to identification and quantification, identification probability and the quantitative statistics data of biological sample.Display device 11 can be any suitable display device, includes but not limited to CRT monitor, plasma display, rear projection display, LCD display or similar.
During operation, system 1,1 ' and 1 " carry out following steps.At first, excitation source 19 sends exciting light, and 21 pairs of wavelength selection systems excite light wavelength to select, the biological sample that exciting light points in the sample color comparison tube 23 by mirror 25 again.Thereby exciting light excites biological sample.The spectral information of biological sample detects by detecting device 29 and detecting device 31, and the form of spectral information is to excite-emission matrix, the absorptiometry that absorbs module 13 and the diffuse reflectance measurement of diffuse reflection module. Signal Processing electronics 33 and 35 pairs of spectral informations are handled, and then carry out data formatting and pre-service by 9 pairs of spectral informations of computer module.Computer module comprises the analysis of expansion partial least square method in interior multivariate analysis, biological sample is discerned and quantized to the data and the pretreated spectral information of layout then.At last, be the user interpretation purpose, show the identification and the quantification of biological sample.
Described below is each embodiment of the present invention.Embodiment only is used for explanation and is not used in the restriction scope of invention.
Following form is 1.6 * 10 for not containing the phosphate buffer (table 1) of Klebsiella pneumoniae and containing concentration
7Exciting-emission matrix of the phosphate buffer (table 2) of the Klebsiella pneumoniae of CFU/mL.These excite-and emission matrix is by using front configuration-system 1 as shown in Figure 3 to produce.
Table 1 (not containing Klebsiella pneumoniae)
| 420 | 8.95E+02 | 8.54E+02 | 6.10E+02 | 4.83E+02 | 4.06E+02 | 1.47E+03 |
| 424 | 9.88E+02 | 8.98E+02 | 6.03E+02 | 4.77E+02 | 4.27E+02 | 1.62E+03 |
| 428 | 1.05E+03 | 8.44E+02 | 5.95E+02 | 4.81E+02 | 4.35E+02 | 1.49E+03 |
| 432 | 1.02E+03 | 8.50E+02 | 5.99E+02 | 4.55E+02 | 3.88E+02 | 1.38E+03 |
| 436 | 1.08E+03 | 9.45E+02 | 5.67E+02 | 4.72E+02 | 3.82E+02 | 1.62E+03 |
| 440 | 1.08E+03 | 8.54E+02 | 5.54E+02 | 4.72E+02 | 3.86E+02 | 1.50E+03 |
Table 2 (containing Klebsiella pneumoniae)
Fig. 4 and Fig. 5 are seen in gathering of above table.Fig. 4 has illustrated the graph of a relation of subduing Klebsiella pneumoniae concentration in positive fluorescence intensity and the phosphate buffer, and Fig. 5 has illustrated the graph of a relation of subduing the excitation wavelength of Klebsiella pneumoniae in emissive porwer and the phosphate buffer.
Following form is a water (table 3) and to contain concentration be 3.9 * 10
7Exciting-emission matrix of the water (table 4) of the Escherichia coli of CFU/mL.These excite-and emission matrix is by using orthogonal configuration system 1 shown in Figure 2 to produce.
Table 3 (water)
Table 4 (water that contains Escherichia coli)
| 332 | 3.42E+03 | 6.07E+03 | 1.09E+04 | 1.52E+04 | 1.29E+04 | 1.14E+04 |
| 336 | 3.29E+03 | 5.72E+03 | 1.02E+04 | 1.45E+04 | 1.23E+04 | 1.19E+04 |
| 340 | 3.01E+03 | 5.30E+03 | 9.48E+03 | 1.34E+04 | 1.13E+04 | 5.33E+03 |
| 344 | 2.68E+03 | 4.96E+03 | 8.81E+03 | 1.28E+04 | 1.11E+04 | 3.74E+03 |
| 348 | 2.74E+03 | 4.77E+03 | 8.24E+03 | 1.24E+04 | 1.01E+04 | 3.63E+03 |
| 352 | 2.57E+03 | 4.62E+03 | 8.06E+03 | 1.14E+04 | 9.78E+03 | 3.55E+03 |
| 356 | 2.48E+03 | 4.27E+03 | 7.55E+03 | 1.05E+04 | 9.24E+03 | 3.32E+03 |
| 360 | 2.41E+03 | 4.03E+03 | 6.93E+03 | 9.87E+03 | 8.86E+03 | 3.13E+03 |
| 364 | 2.31E+03 | 3.78E+03 | 6.38E+03 | 8.98E+03 | 7.99E+03 | 2.76E+03 |
| 368 | 2.16E+03 | 3.50E+03 | 5.63E+03 | 7.91E+03 | 7.02E+03 | 2.61E+03 |
| 372 | 2.14E+03 | 3.21E+03 | 5.24E+03 | 7.37E+03 | 6.42E+03 | 2.76E+03 |
| 376 | 2.02E+03 | 3.13E+03 | 4.61E+03 | 6.57E+03 | 6.01E+03 | 2.81E+03 |
| 380 | 2.06E+03 | 2.73E+03 | 4.35E+03 | 5.86E+03 | 5.31E+03 | 2.19E+03 |
| 384 | 1.91E+03 | 2.55E+03 | 3.86E+03 | 5.31E+03 | 4.77E+03 | 2.10E+03 |
| 388 | 1.75E+03 | 2.41E+03 | 3.50E+03 | 5.01E+03 | 4.36E+03 | 1.96E+03 |
| 392 | 1.60E+03 | 2.08E+03 | 3.00E+03 | 4.35E+03 | 3.66E+03 | 1.99E+03 |
| 396 | 1.44E+03 | 1.99E+03 | 2.73E+03 | 3.77E+03 | 3.25E+03 | 1.54E+03 |
| 400 | 1.26E+03 | 1.64E+03 | 2.15E+03 | 3.40E+03 | 3.04E+03 | 1.42E+03 |
Fig. 6 is seen in gathering of above table.Fig. 6 has illustrated the graph of a relation of subduing the concentration of Escherichia coli in quadrature fluorescence intensity and the water.
Following form is 5.7 * 10 for not containing the phosphate buffered solution (table 5) of Escherichia coli and containing concentration
7Exciting-emission matrix of the phosphate buffered solution (table 6) of the Escherichia coli of CFU/mL.These excite-and emission matrix produces by using front configuration-system shown in Figure 31.
Table 5 (not containing Escherichia coli)
| 348 | 5.65E+02 | 5.49E+02 | 4.56E+02 | 8.77E+02 |
| 352 | 5.82E+02 | 5.21E+02 | 4.37E+02 | 1.14E+03 |
| 356 | 5.56E+02 | 5.10E+02 | 4.35E+02 | 1.07E+03 |
| 360 | 5.80E+02 | 4.80E+02 | 4.31E+02 | 7.29E+02 |
| 364 | 5.54E+02 | 4.71E+02 | 4.35E+02 | 7.62E+02 |
| 368 | 5.29E+02 | 4.33E+02 | 3.99E+02 | 7.47E+02 |
| 372 | 5.03E+02 | 4.25E+02 | 3.81E+02 | 9.70E+02 |
| 376 | 4.75E+02 | 4.08E+02 | 3.55E+02 | 1.53E+03 |
| 380 | 4.96E+02 | 4.19E+02 | 3.65E+02 | 1.11E+03 |
| 384 | 4.80E+02 | 4.25E+02 | 3.67E+02 | 9.47E+02 |
| 388 | 4.85E+02 | 4.03E+02 | 3.61E+02 | 8.83E+02 |
| 392 | 4.71E+02 | 4.00E+02 | 3.51E+02 | 9.18E+02 |
| 396 | 4.56E+02 | 3.89E+02 | 3.57E+02 | 1.29E+03 |
| 400 | 4.47E+02 | 3.74E+02 | 3.46E+02 | 7.78E+02 |
Table 6 (containing Escherichia coli)
Fig. 7 is seen in gathering of above table.Fig. 7 illustrated subdue positive fluorescence intensity with
The graph of a relation of the concentration of Escherichia coli in the phosphate buffer.
Following form is 8.9 * 10 for not containing the human urine (table 7) of Escherichia coli and containing concentration
7Exciting-emission matrix of the human urine (table 8) of the Escherichia coli of CFU/mL.These excite-and the emission matrix formula produces by using orthogonal configuration system shown in Figure 21.
Table 7 (human urine)
Table 8 (human urine that contains Escherichia coli)
Fig. 8 gathers above table.Fig. 8 has illustrated the relation of subduing Escherichia coli concentration in quadrature fluorescence intensity and the human urine.
Although the present invention for illustrative purposes, to thinking that at present the most practical preferred embodiment describes in detail, can be understood as these detailed descriptions only is for illustrative purposes.The present invention is not limited in disclosed embodiment, and is opposite but for the modification in the spirit and scope that cover additional requirement be equal to replacement.For example, the combination to one or more technical characterictics of different embodiment within the possible range can be understood as in desired extent of the present invention.
Claims (20)
1. an identification and quantize to be suspended in the system of biological sample in the liquid comprises:
The fluorescence excitation module that comprises at least one excitation source;
With the sample interface module of fluorescence excitation module optical coupled, be used to locate the exciting light of biological sample, and transmission light is to fluorescent absorption and diffuse reflection module to accept to send from least one excitation source;
With the fluorescent emission module of sample interface module optical coupled, comprise at least one detection of biological fluorescent to excite-pick-up unit of emission matrix; And
With the computer module of the effective coupling of fluorescent emission module,
Described computer module carries out multivariate analysis with identification and quantification biological sample to the fluorescence excitation-emission matrix of biological sample.
2. the system as claimed in claim 1 comprises that also one absorbs a module and a diffuse reflection module.
3. system as claimed in claim 2, described absorption module are used to the absorptiometry that one light in an at least one excitation source and an independent modulated light source carries out biological sample.
4. system as claimed in claim 3, described absorptiometry is discerned in conjunction with the fluorescence excitation-emission matrix of biological sample and is quantized biological sample.
5. system as claimed in claim 2, described absorption module is the filter wheel that a monochromator or has photomultiplier.
6. system as claimed in claim 2, described diffuse reflection module are used to the diffuse reflectance measurement that one light in an at least one excitation source and an independent modulated light source carries out biological sample.
7. system as claimed in claim 6, described diffuse reflectance measurement is discerned in conjunction with the fluorescence excitation-emission matrix of biological sample and is quantized biological sample.
8. system as claimed in claim 7, described diffuse reflection module are in monochromator, semiconductor detector and the photomultiplier.
9. the system as claimed in claim 1, described at least one excitation source is a continuous light source, a light-pulse generator, semiconductor laser instrument, a tunable laser or above-mentioned combination in any.
10. the system as claimed in claim 1, the wavelength of described at least one excitation source uses grating monochromator, has filter wheel, acousto-optic turnable filter, liquid crystal tunable optical filter, circular variable filter, linear variable filter or the above-mentioned combination in any of narrowband light filter by selectivity.
11. the system as claimed in claim 1, at least one pick-up unit of described fluorescent emission module are to have the raster monochromator of solid-state detector and have in the non-raster monochromator of multichannel array detecting device one.
12. the system as claimed in claim 1, described fluorescent emission module comprise that further the gate electronics is with the optical sampling degree of depth in the controlling liquid and optimization signal to noise ratio (S/N ratio).
13. the system as claimed in claim 1, described multivariate analysis comprise that expansion partial least square method analysis discerns and quantize biological sample.
14. the system as claimed in claim 1 also further comprises the display device of the identification and the quantification that are used to show biological sample.
15. the method discerning and quantize to be suspended in biological sample in the liquid comprises following
Step:
A) provide an excitation source;
B) excitation light source excites biological sample;
C) spectral information of detection of biological sample, spectral information is following form: excite-emission matrix, absorptiometry, diffuse reflectance measurement or above-mentioned combination in any; And
D) spectral information is carried out multivariate analysis with identification and quantification biological sample.
16. method as claimed in claim 15, described excitation source are continuous light source, light-pulse generator, semiconductor laser, tunable laser or above-mentioned combination in any.
17. method as claimed in claim 15, the wavelength of described excitation source is selected by using grating monochromator, the filter wheel that has narrowband light filter, acousto-optic turnable filter, liquid crystal tunable optical filter, circular variable filter, linear variable filter or above-mentioned combination in any.
18. method as claimed in claim 15, described multivariate analysis comprise that expansion partial least square method analysis discerns and quantize biological sample.
19. method as claimed in claim 15 further comprises step: e) identification and the quantification of demonstration biological sample
20. method as claimed in claim 15, advanced line data formatting and data processing before step d).
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