CN107421922A - Double light path transmissions and the method for fluorescence spectral measuring free hemoglobin content - Google Patents
Double light path transmissions and the method for fluorescence spectral measuring free hemoglobin content Download PDFInfo
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- CN107421922A CN107421922A CN201710529535.1A CN201710529535A CN107421922A CN 107421922 A CN107421922 A CN 107421922A CN 201710529535 A CN201710529535 A CN 201710529535A CN 107421922 A CN107421922 A CN 107421922A
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Abstract
The invention discloses a kind of double light path transmissions and the method for fluorescence spectral measuring free hemoglobin content, including：Light source includes transmitted light source and fluorescence excitation light source, and transmitted light source transmits to blood sample in blood bag, and fluorescence excitation light source excites to blood sample in blood bag；Displacement platform controls light source movement, spectrum acquisition device acquisition of transmission spectrum and fluorescence spectrum under the premise of ensureing that light source light extraction optical port and spectrum acquisition device entrance slit are coaxial；Ask logarithm to be absorbed spectrum light intensity ratio under each wavelength of transmitted spectrum under two light paths, and combine two fluorescence spectrums normalized together, with reference to the data of chemical analysis, founding mathematical models；The transmitted spectrum and fluorescence spectrum under unknown two light paths of blood sample are gathered, the ratio of two transmitted spectrums is asked and brings mathematical modeling into after the absorption spectrum and two fluorescence spectrums normalization that logarithm obtains and is calculated, obtains the content of free hemoglobin.
Description
Technical field
The present invention relates to spectrum complicated solution concentration analysis stoichiometry field, more particularly to a kind of double light path transmissions and glimmering
The method of light spectral measurement free hemoglobin content.
Background technology
In the prior art, more ripe mode is to detect containing for free hemoglobin in blood bag by chemical analysis
Amount, there are the high outstanding advantages of accuracy, but the mode of chemical analysis can not meet quick, non-contact and free of contamination need
Ask, spectral measurement is also possible to realize the content detection of free hemoglobin in blood bag due to its non-contact, free of contamination characteristic.
In spectral detection, according to Lambert-Beer's law：The incident intensity I of each wavelength is measured respectively0And output intensity
I, the absorbance A of each wavelength is calculated by formula (1).∈ is material absorptivity under a certain wavelength, and c is the dense of material
Degree, b is optical path length.
In fact, fail to measure incident intensity I for various reasons0, such as：Incident intensity I0It is difficult to measure by force very much, but
If in incident intensity I0It is basicly stable it is constant in the case of, only measure an output intensity I can also obtain good result.But
Influence that spectral detection changes due to light source and measure influences of container and be difficult to reach the precision for measuring needs.
The influence of light source is mainly shown as the change of spatial distribution and light intensity.The reason for causing light source to change, has a lot, such as
Light source voltage change, filament aging, variation of ambient temperature etc..In spectrum analysis, rarely have document to introduce light source to measurement accuracy
Influence, and reduce the intensity of light source change on measurement accuracy influence method.In research in the early time, with the mode of calibration come
Some interference are eliminated, are such as calibrated with water, but because light intensity is too strong, are difficult to operate in practice.Also have in many scholars' utilizations
Property attenuator or optical fibre light splitting mode measure incident intensity I0.By taking neutral filter as an example (following discussion unless stated otherwise,
Discussed on some wavelength), the output intensity I that measurement passes through neutral filtern, then the light intensity I of light source0 nAbsorbance can be used
A and output intensity InTo represent：
Then sample is replaced into neutral filter, measures the output intensity I of sampleS,
NoticeSo
Formula does not have in (4)(namely) occur, illustrate that the intensity (and its spectrum) of light source does not interfere with the survey to sample
Amount, as long as all measurements are all calibrated using same neutral attenuator, that is, keep lgIn+AnFor constant constant.
But different occasions are difficult to find duplicate neutral filter, and it is difficult to ensure that the position of sample and neutral filter
Put unanimously, add difficulty to measurement and be difficult to ensure that measurement accuracy.
For the complexity of blood constituent, what simple transmitted spectrum obtained is the information of whole blood, and specific aim is poor, to enter
One step improves the measurement accuracy of free hemoglobin, with reference to fluorescence it is with strong points the characteristics of, but by fluorescence excitation light intensity, light path
Influence that is long and surveying constituent concentration, causes fluorescence to have serious self-absorption problem, so as to cause obtained fluorescence spectrum tool
Have very strong non-linear, it is impossible to which the feature of material is surveyed in reaction well.
The content of the invention
The invention provides a kind of double light path transmissions and the method for fluorescence spectral measuring free hemoglobin content, add
The information content of free hemoglobin, transmitted light source change and the influence of blood bag are not only eliminated, and measure with strong points, greatly suppression
The non-linear effects of spectrum have been made, have improved the precision of free hemoglobin content analysis in blood bag.Realize quick, pollution-free
Blood bag in free hemoglobin measurement, it is workable.It is described below：
A kind of double light path transmissions and the method for fluorescence spectral measuring free hemoglobin content, methods described include following step
Suddenly：
The light extraction optical port of light source is close to blood bag and coaxial with the entrance slit of spectrum acquisition device, and light source includes transmitted light source
And fluorescence excitation light source, transmitted light source transmit to blood sample, fluorescent exciting source excitation free hemoglobin produces glimmering
Light, spectrum acquisition device acquisition of transmission spectrum and fluorescence spectrum；
Displacement platform is ensureing that transmitted light source and fluorescence excitation light source light extraction optical port and spectrum acquisition device entrance slit are same
Under the premise of axle, transmitted light source and fluorescence excitation light source movement are controlled, by spectrum acquisition device acquisition of transmission spectrum and fluorescence spectrum；
Ask logarithm to be absorbed spectrum light intensity ratio under each wavelength of two opening position transmitted spectrums, and combine two
Fluorescence spectrum normalized under position, with reference to the data of chemical analysis, founding mathematical models；
The transmitted spectrum and fluorescence spectrum at unknown blood sample two positions are gathered, the ratio of two transmitted spectrums is sought into logarithm
Obtained absorption spectrum and two fluorescence spectrums are brought mathematical modeling into after normalizing and calculated, and obtain free hemoglobin
Content；
The transmitted spectrum and fluorescence spectrum are to cause the spectrum under the nonlinear factor collective effect of spectrum, and increase is free
Hemoglobin information content；Eliminate transmitted light source change and the influence of blood bag；Suppress the non-linear effects of spectrum；Improve in blood bag and swim
From the precision of content of hemoglobin analysis.
Wherein, displacement platform control transmitted light source and fluorescence excitation light source movement, by spectrum acquisition device acquisition of transmission light
Spectrum and it is specially the step of fluorescence spectrum：
Transmitted light source and fluorescence excitation light source are transmitted and excited respectively to blood sample at a of position, are connect by spectrum
Receiving apparatus acquisition of transmission spectrum and fluorescence spectrum；
Displacement platform controls transmitted light source and fluorescence excitation light source to be moved to position b, by spectrum acquisition device acquisition of transmission
Spectrum and fluorescence spectrum；
Or,
Transmitted light source and fluorescence excitation light source are transmitted and excited respectively to the blood sample in blood bag, by spectrum acquisition
Device acquisition of transmission spectrum and fluorescence spectrum at a of position；
Displacement platform control spectrum acquisition device is moved to position b, gathers transmitted spectrum and fluorescence spectrum at the b of position；
Or,
The blood sample in blood bag is transmitted and swashed respectively by transmitted light source and fluorescence excitation light source at a of position
Hair, at position a ' places by spectrum acquisition device acquisition of transmission spectrum and fluorescence spectrum；
Displacement platform controls transmitted light source and fluorescence excitation light source to be moved to position b, and spectrum acquisition device is moved to position
B ' places, by spectrum acquisition device acquisition of transmission spectrum and fluorescence spectrum.
Wherein, methods described also includes：
One optical fiber is set at light source, as incident optical, and ensures incident optical and spectrum acquisition device entrance slit
It is close to blood bag and coaxial；
Or,
One optical fiber is set at spectrum acquisition device, as the output optical fiber, and ensures the output optical fiber and light source light extraction optical port
It is close to blood bag and coaxial；
Or,
Incident optical and the output optical fiber are set respectively at light source and spectrum acquisition device, and ensure incident optical and outgoing
Optical fiber is close to blood bag and coaxial.
Wherein, a positions are the first position of incident optical, and incident optical and the output optical fiber phase are gathered by spectrum acquisition device
To the transmitted spectrum and fluorescence spectrum under a of position；Displacement platform is controlled on the premise of ensureing that incident optical and the output optical fiber are coaxial
Incident optical processed is moved at the b of position, and incident optical and the transmission under the output optical fiber relative position b are gathered by spectrum acquisition device
Spectrum and fluorescence spectrum.
Wherein, a positions are the first position of the output optical fiber, and incident optical and the output optical fiber phase are gathered by spectrum acquisition device
To the transmitted spectrum and fluorescence spectrum under a of position；Displacement platform is controlled on the premise of ensureing that incident optical and the output optical fiber are coaxial
The output optical fiber processed is moved at the b of position, and incident optical and the transmission under the output optical fiber relative position b are gathered by spectrum acquisition device
Spectrum and fluorescence spectrum.
Wherein, a, a ' be respectively incident optical and the output optical fiber first position, by spectrum acquisition device gather incident light
The fine position a, a relative with the output optical fiber ' under transmitted spectrum and fluorescence spectrum；Displacement platform is ensureing incident optical and outgoing
On the premise of fiber-coaxial, incident optical and the output optical fiber is controlled to be moved respectively to position b, b ' place, adopted by spectrum acquisition device
Collect the transmitted spectrum and fluorescence spectrum under incident optical position b, b ' relative with the output optical fiber.
Wherein, the lit transmissive light source is super continuous wide range laser, the super continuous wide range laser covering visible light wave band,
Or near infrared light wave band, or both combination；Fluorescence excitation light source is ultraviolet lamp, and above-mentioned light source can directly light or through incidence
Fiber optic conduction.
Wherein, institute's translation stage is stepper motor；The spectrum acquisition device is spectrometer.
Further, the transmitted light source is xenon lamp wide spectrum light source or bromine tungsten filament lamp wide spectrum light source, covering visible light wave band or
Near infrared light wave band, or both combination；The fluorescence excitation light source is ultraviolet laser tube or ultra-violet light-emitting pipe, and above-mentioned light source can
Directly light or conducted through incident optical；Institute's translation stage is magnet attraction device.
Further, the mathematical modeling utilizes principal component analysis, artificial neural network, PLS, support
Vector machine, signal analysis or statistical method are established.
The beneficial effect of technical scheme provided by the invention is：
1st, the present invention changes light path by command displacement platform, the blood bag in the case where different light path strong points gather same transmitted light source
Under the transmitted spectrum of interior blood and same fluorescence excitation light source in blood bag blood fluorescence spectrum, realize accordingly in blood bag dissociate
The Non-Destructive Testing of content of hemoglobin；
2nd, the characteristic of the invention for making full use of free hemoglobin to be produced fluorescence by ultraviolet excitation, but due in light
Cheng Fang is different with ultraviolet light incidence depth upwards and produces different fluorescence intensities, and excites fluorescence to produce position and received bit
The distance difference put can cause the self-absorption of fluorescence different, and in blood free hemoglobin simultaneously by other material concentrations
Influence, when ultraviolet light is by the more of other material absorbings, the ultraviolet light that the material that can be excited receives is fewer, therefore causes to obtain
Spectrum have it is very strong non-linear；
3rd, the present invention obtained transmitted spectrum of measurement and fluorescence spectrum are above-mentioned to cause the nonlinear factor of spectrum to be made jointly
Spectrum under, the information content of free hemoglobin is added, not only eliminate transmitted light source change and the influence of blood bag, and survey
Measure with strong points, greatly inhibit the non-linear effects of spectrum；
4th, the present invention improves the precision of free hemoglobin content analysis in blood bag；Realize quick, free of contamination blood
The measurement of free hemoglobin, workable in bag；
Brief description of the drawings
Fig. 1 is to combine double light path transmitted spectrum principle schematics；
Fig. 2 is the method schematic diagram of double light path transmissions and fluorescence spectral measuring free hemoglobin content in embodiment 1；
Fig. 3 is double light path transmissions and another signal of method of fluorescence spectral measuring free hemoglobin content in embodiment 2
Figure；
Fig. 4 is double light path transmissions and another signal of method of fluorescence spectral measuring free hemoglobin content in embodiment 3
Figure；
Fig. 5 is double light path transmissions and another signal of method of fluorescence spectral measuring free hemoglobin content in embodiment 4
Figure；
Fig. 6 is double light path transmissions and another signal of method of fluorescence spectral measuring free hemoglobin content in embodiment 5
Figure；
Fig. 7 is double light path transmissions and another signal of method of fluorescence spectral measuring free hemoglobin content in embodiment 6
Figure；
Fig. 8 is double light path transmissions and another signal of method of fluorescence spectral measuring free hemoglobin content in embodiment 7
Figure；
Fig. 9 is double light path transmissions and another signal of method of fluorescence spectral measuring free hemoglobin content in embodiment 8
Figure.
In accompanying drawing, the list of parts representated by each label is as follows：
1：First light path； 2：Second light path；
3：Light source；Including transmitted light source and fluorescence excitation light source 4：Incident optical；
5：Blood bag； 6：Displacement platform；
7：Spectrum acquisition device； 8：The output optical fiber；
a、a’：First position； b、b’：The second place.
Embodiment
To make the object, technical solutions and advantages of the present invention clearer, embodiment of the present invention is made below further
It is described in detail on ground.
Double light path transmission spectrum methods are according to Lambert-Beer's law, as shown in figure 1, setting the first light path 1 and second respectively
Light path 2.Derivation is as follows：
Wherein, A1It is the absorbance of the first light path 1, A2It is the absorbance of the second light path 2.IOIt is the incident light of the first light path 1
Light intensity, while be also the second light path 2 incident light light intensity, I1It is the output intensity of the first light path 1, I2It is the second light path 2
Output intensity, b1It is the optical length of the first light path 1, b2It is the optical length of the second light path 2, △ b are the difference of two optical lengths, ∈ extinctions
Coefficient is that c surveys material concentration.
The absorbance of double light path spectroscopic methodologies is still linear with optical path difference it can be seen from formula (7), meet lambert-
Beer law, and with light intensity of incident light IOIt is unrelated.Therefore, double light path methods are not influenceed by light source in theory, are deducted simultaneously
The influence of blood bag in itself.
Due to the complexity of blood constituent, what simple transmitted spectrum obtained is the information of whole blood, and specific aim is poor, to enter
One step improves the measurement accuracy of free hemoglobin, the characteristics of with reference to exciting fluorescence with strong points, but due on light path direction
Different fluorescence intensities is produced with ultraviolet light incidence depth difference, and the distance for exciting fluorescence to produce position and receiving position
Difference can cause the self-absorption of fluorescence different, and free hemoglobin is influenceed by other material concentrations simultaneously in blood,
When ultraviolet light is by the more of other material absorbings, the ultraviolet light that the material that can be excited receives is fewer, therefore causes the light obtained
Spectrum is with very strong non-linear.
The transmitted spectrum and fluorescence spectrum that double light path measurements obtain are above-mentioned to cause the nonlinear factor collective effect of spectrum
Under spectrum, add the information content of free hemoglobin, not only eliminate transmitted light source change and the influence of blood bag, and measure
It is with strong points, the non-linear effects of spectrum are greatly inhibited, improve the precision of free hemoglobin content analysis in blood bag.It is real
Show the measurement of free hemoglobin in quick, free of contamination blood bag, it is workable.
Embodiment 1
Double light path transmissions provided in an embodiment of the present invention and the method for fluorescence spectral measuring free hemoglobin content, are made
The device used as shown in Fig. 2 including：Light source 3, blood bag 5, displacement platform 6 and spectrum acquisition device 7.
Wherein, ensure that the light extraction optical port of light source 3 is close to blood bag 5 and coaxial with the entrance slit of spectrum acquisition device 7, light source 3 exists
First position a (corresponding first light path 1) place is transmitted and excited to the blood sample in blood bag 5, adopted by spectrum acquisition device 7
Collect transmitted spectrum and fluorescence spectrum.Then it is emitted by displacement platform 6 in the guarantee light extraction optical port of light source 3 and spectrum acquisition device 7
Control light source to be moved to second place b (corresponding second light path 2) on the premise of slit is coaxial, gathered by spectrum acquisition device 7
Penetrate spectrum and fluorescence spectrum.
Logarithm is asked to be absorbed spectrum light intensity ratio under each wavelength of the transmitted spectrum of two opening positions of a, b collection,
The absorption spectrum is normalized with reference to two fluorescence spectrums, and method for normalizing is：
Ag=A/max (A) (8)
In formula (8), AgTo normalize absorbance, max (A) is the absorbance maximum on different wave length, and A is extinction
Degree.With reference to the data of chemical analysis, principal component analysis (PCA, principal component analysis) or artificial is utilized
Neutral net (ANN, artificial neural network) or PLS (PLSR, particle least
Squares calibration analysis) or SVMs (SVM, support vector machines) signal point
The methods of analysis or statistics, can founding mathematical models.
The embodiment of the present invention is not repeated the process of above-mentioned founding mathematical models, known to those skilled in the art.
The transmitted spectrum and fluorescence spectrum of position at unknown blood sample a, b two are gathered, the ratio of two transmitted spectrums is asked
The absorption spectrum and two fluorescence spectrums that logarithm obtains are brought the above-mentioned mathematical modeling established into after normalizing and calculated,
Obtain the content of free hemoglobin.
To the model of each device in addition to specified otherwise is done, the model of other devices is not limited the embodiment of the present invention,
As long as the device of above-mentioned function can be completed.
In summary, the embodiments of the invention provide a kind of double light path transmissions and fluorescence spectral measuring free hemoglobin to contain
The method of amount, transmitted light source change and the influence of blood bag are not only eliminated, and measurement is with strong points, greatly inhibits the non-of spectrum
Linear effect, improve the precision of free hemoglobin content analysis in blood bag.Realize in quick, free of contamination blood bag and dissociate
The measurement of hemoglobin, it is workable.
Embodiment 2
The embodiment of the present invention differs only in embodiment 1, and the move mode of light source 3 and spectrum acquisition device 7 is not
Together, it is described below：
Referring to Fig. 3, ensure that the light extraction optical port of light source 3 is close to blood bag 5 and coaxial, light source 3 with the entrance slit of spectrum acquisition device 7
Blood sample in blood bag 5 is transmitted and excited, by spectrum acquisition device 7 in first position a acquisition of transmission spectrum and
Fluorescence spectrum.Then by displacement platform 6 before ensureing that the light extraction optical port of light source 3 and the entrance slit of spectrum acquisition device 7 are coaxial
Put control spectrum acquisition device 7 and be moved to second place b, gather transmitted spectrum and fluorescence spectrum at the b of the second place.
Wherein, the step of content of follow-up normalization, founding mathematical models and calculating free hemoglobin is with implementing
Example 1 is identical, and the embodiment of the present invention is not repeated this.
To the model of each device in addition to specified otherwise is done, the model of other devices is not limited the embodiment of the present invention,
As long as the device of above-mentioned function can be completed.
In summary, the embodiments of the invention provide a kind of double light path transmissions and fluorescence spectral measuring free hemoglobin to contain
The method of amount, transmitted light source change and the influence of blood bag are not only eliminated, and measurement is with strong points, greatly inhibits the non-of spectrum
Linear effect, improve the precision of free hemoglobin content analysis in blood bag.Realize in quick, free of contamination blood bag and dissociate
The measurement of hemoglobin, it is workable.
Embodiment 3
The embodiment of the present invention differs only in embodiment 1, and the moving direction of light source 3 and spectrum acquisition device 7 is not
Together, it is described below：
Referring to Fig. 4, ensure that light source 3 is close to blood bag 5 with spectrum acquisition device 7 and ensures that the light extraction optical port of light source 3 and spectrum connect
The entrance slit of receiving apparatus 7 is coaxial, and the blood sample in blood bag 5 is transmitted and excited by light source 3 at a of first position,
First position a ' places are then ensureing light source by the acquisition of transmission spectrum of spectrum acquisition device 7 and fluorescence spectrum by displacement platform 6
On the premise of 3 light extraction optical ports and the entrance slit of spectrum acquisition device 7 are coaxial, light source 3 and spectrum acquisition device 7 is controlled to move respectively
To the second place b, b ' place, gather the transmitted spectrum and fluorescence spectrum of the opening position.
Wherein, the step of content of follow-up normalization, founding mathematical models and calculating free hemoglobin is with implementing
Example 1 is identical, and the embodiment of the present invention is not repeated this.
To the model of each device in addition to specified otherwise is done, the model of other devices is not limited the embodiment of the present invention,
As long as the device of above-mentioned function can be completed.
In summary, the embodiments of the invention provide a kind of double light path transmissions and fluorescence spectral measuring free hemoglobin to contain
The method of amount, transmitted light source change and the influence of blood bag are not only eliminated, and measurement is with strong points, greatly inhibits the non-of spectrum
Linear effect, improve the precision of free hemoglobin content analysis in blood bag.Realize in quick, free of contamination blood bag and dissociate
The measurement of hemoglobin, it is workable.
Embodiment 4
During specific implementation, due to the limitation of space structure, it is possible that light source 3 can not be close to spectrum acquisition device 7
The situation of blood bag 5, at this moment can be by setting an optical fiber, as incident optical 4 respectively at light source 3 and spectrum acquisition device 7
With the output optical fiber 8.
Referring to Fig. 5, light source 3 is transmitted and excited to the blood sample in blood bag 5 by incident optical 4, is connect by spectrum
Receiving apparatus 7 passes through the acquisition of transmission spectrum of the output optical fiber 8 and fluorescence spectrum, and incident optical 4 is close to blood bag 5 respectively with the output optical fiber 8
And ensure coaxially, a positions are the first position of incident optical 4, and spectrum acquisition device 7 gathers incident optical 4 and the phase of the output optical fiber 8
To the transmitted spectrum and fluorescence spectrum under a of the first position；Incident optical 4 and emergent light are then being ensured by displacement platform 6
On the premise of fine 8 positions are still coaxial, control incident optical 4 is moved at the b of the second place, and spectrum acquisition device 5 gathers incidence
Transmitted spectrum and fluorescence spectrum under the optical fiber 4 and relative second position b of the output optical fiber 8.
Wherein, the step of content of follow-up normalization, founding mathematical models and calculating free hemoglobin is with implementing
Example 1 is identical, and the embodiment of the present invention is not repeated this.
To the model of each device in addition to specified otherwise is done, the model of other devices is not limited the embodiment of the present invention,
As long as the device of above-mentioned function can be completed.
In summary, the embodiments of the invention provide a kind of double light path transmissions and fluorescence spectral measuring free hemoglobin to contain
The method of amount, transmitted light source change and the influence of blood bag are not only eliminated, and measurement is with strong points, greatly inhibits the non-of spectrum
Linear effect, improve the precision of free hemoglobin content analysis in blood bag.Realize in quick, free of contamination blood bag and dissociate
The measurement of hemoglobin, it is workable.
Embodiment 5
The embodiment of the present invention is different only in that with embodiment 4, the output optical fiber 8, sets with first position a, second place b
Difference is put, it is described below：
Referring to Fig. 6, light source 3 is transmitted and excited to the blood sample in blood bag 5 by incident optical 4, is connect by spectrum
Receiving apparatus 7 passes through the acquisition of transmission spectrum of the output optical fiber 8 and fluorescence spectrum, and incident optical 4 is close to blood bag 5 respectively with the output optical fiber 8
And ensure coaxially, a positions are the first position of the output optical fiber 8, and spectrum acquisition device 7 gathers incident optical 4 and the phase of the output optical fiber 8
To the transmitted spectrum and fluorescence spectrum under a of the first position；Incident optical 4 and emergent light are then being ensured by displacement platform 6
On the premise of fine 8 positions are still coaxial, control the output optical fiber 8 is moved at the b of the second place, and spectrum acquisition device 5 gathers incidence
Transmitted spectrum and fluorescence spectrum under the optical fiber 4 and relative second position b of the output optical fiber 8.
Wherein, the step of content of follow-up normalization, founding mathematical models and calculating free hemoglobin is with implementing
Example 1 is identical, and the embodiment of the present invention is not repeated this.
To the model of each device in addition to specified otherwise is done, the model of other devices is not limited the embodiment of the present invention,
As long as the device of above-mentioned function can be completed.
In summary, the embodiments of the invention provide a kind of double light path transmissions and fluorescence spectral measuring free hemoglobin to contain
The method of amount, transmitted light source change and the influence of blood bag are not only eliminated, and measurement is with strong points, greatly inhibits the non-of spectrum
Linear effect, improve the precision of free hemoglobin content analysis in blood bag.Realize in quick, free of contamination blood bag and dissociate
The measurement of hemoglobin, it is workable.
Embodiment 6
The embodiment of the present invention is different only in that with embodiment 4, incident optical 4, the output optical fiber 8, with first position a, the
Two position b setting is different, described below：
Referring to Fig. 7, light source 3 is transmitted and excited to the blood sample in blood bag 5 by incident optical 4, is connect by spectrum
Receiving apparatus 7 passes through the acquisition of transmission spectrum of the output optical fiber 8 and fluorescence spectrum, and incident optical 4 is close to blood bag 5 respectively with the output optical fiber 8
And ensure coaxial, a, a ' be respectively incident optical 4 and the output optical fiber 8 first position, spectrum acquisition device 7 gathers incident optical
4 transmitted spectrum and fluorescence spectrum with the output optical fiber 8 under relative first position a, a '；Then ensured by displacement platform 6
Incident optical 4 and the position of the output optical fiber 8 remain unchanged it is coaxial on the premise of, control incident optical 4 and the output optical fiber 8 are moved respectively to the
Two position b, b ' places, spectrum acquisition device 7 gather incident optical 4 and the output optical fiber 8 relative second position b, b ' under transmitted light
Spectrum and fluorescence spectrum.
Wherein, the step of content of follow-up normalization, founding mathematical models and calculating free hemoglobin is with implementing
Example 1 is identical, and the embodiment of the present invention is not repeated this.
To the model of each device in addition to specified otherwise is done, the model of other devices is not limited the embodiment of the present invention,
As long as the device of above-mentioned function can be completed.
In summary, the embodiments of the invention provide a kind of double light path transmissions and fluorescence spectral measuring free hemoglobin to contain
The method of amount, transmitted light source change and the influence of blood bag are not only eliminated, and measurement is with strong points, greatly inhibits the non-of spectrum
Linear effect, improve the precision of free hemoglobin content analysis in blood bag.Realize in quick, free of contamination blood bag and dissociate
The measurement of hemoglobin, it is workable.
Embodiment 7
As different from Example 4, the embodiment only includes incident optical 4 to the embodiment of the present invention, described below：
Referring to Fig. 8, light source 3 is transmitted and excited to the blood sample in blood bag 5 by incident optical 4, is connect by spectrum
The acquisition of transmission spectrum of receiving apparatus 7 and fluorescence spectrum, incident optical 4 and the entrance slit of spectrum acquisition device 7 be close to respectively blood bag 5 and
Ensure coaxially, a positions are the first position of incident optical 4, and spectrum acquisition device 7 gathers incident optical 4 with respect to first position a
Under transmitted spectrum and fluorescence spectrum；Incident optical 4 and the slit position of spectrum acquisition device 7 are then being ensured by displacement platform 6
Put it is still coaxial on the premise of, control incident optical 4 is moved to the b of the second place at, the collection incident optical 4 of spectrum acquisition device 7
Transmitted spectrum and fluorescence spectrum under relative second position b.
Wherein, the step of content of follow-up normalization, founding mathematical models and calculating free hemoglobin is with implementing
Example 1 is identical, and the embodiment of the present invention is not repeated this.
To the model of each device in addition to specified otherwise is done, the model of other devices is not limited the embodiment of the present invention,
As long as the device of above-mentioned function can be completed.
In summary, the embodiments of the invention provide a kind of double light path transmissions and fluorescence spectral measuring free hemoglobin to contain
The method of amount, transmitted light source change and the influence of blood bag are not only eliminated, and measurement is with strong points, greatly inhibits the non-of spectrum
Linear effect, improve the precision of free hemoglobin content analysis in blood bag.Realize in quick, free of contamination blood bag and dissociate
The measurement of hemoglobin, it is workable.
Embodiment 8
As different from Example 4, the embodiment only includes the output optical fiber 8 to the embodiment of the present invention, described below：
Referring to Fig. 9, light source 3 is transmitted and excited to the blood sample in blood bag 5, by spectrum acquisition device 7 by going out
The acquisition of transmission spectrum of optical fiber 8 and fluorescence spectrum are penetrated, the light extraction optical port of light source 3 is close to blood bag 5 with the output optical fiber 8 and ensured same respectively
Axle, spectrum acquisition device 7 gather the transmitted spectrum and fluorescence spectrum under the light source 3 and relative first position a of the output optical fiber 8；Then
By displacement platform 6 ensure the light extraction optical port of light source 3 and the position of the output optical fiber 8 remain unchanged it is coaxial on the premise of, control light source 3 to move
To at the b of the second place, spectrum acquisition device 5 gathers light source 3 and the transmitted spectrum under the relative second position b of the output optical fiber 8 and glimmering
Light spectrum.
Wherein, the step of content of follow-up normalization, founding mathematical models and calculating free hemoglobin is with implementing
Example 1 is identical, and the embodiment of the present invention is not repeated this.
To the model of each device in addition to specified otherwise is done, the model of other devices is not limited the embodiment of the present invention,
As long as the device of above-mentioned function can be completed.
In summary, the embodiments of the invention provide a kind of double light path transmissions and fluorescence spectral measuring free hemoglobin to contain
The method of amount, transmitted light source change and the influence of blood bag are not only eliminated, and measurement is with strong points, greatly inhibits the non-of spectrum
Linear effect, improve the precision of free hemoglobin content analysis in blood bag.Realize in quick, free of contamination blood bag and dissociate
The measurement of hemoglobin, it is workable.
Embodiment 9
Select, the scheme in above-described embodiment 1-6 is further introduced, light source 3 with reference to specific device
Including：Transmitted light source and fluorescence excitation light source, transmitted light source can be super continuous wide range laser, the super continuous wide range laser covering
The combination of visible light wave range or near infrared light wave band or both；Excitation source is ultraviolet lamp, above-mentioned light source can directly light or
Conducted through incident optical 4.Displacement platform 6 is stepper motor, and spectrum acquisition device 7 is spectrometer, described below：
As shown in figure 5, above-mentioned light source 3 is transmitted and excited to the blood sample in blood bag 5 by incident optical 4, light
Spectrometer 7 passes through the acquisition of transmission spectrum of the output optical fiber 8 and fluorescence spectrum, incident optical 4 and the output optical fiber 8 be close to respectively blood bag 5 and
Coaxially, a positions are the first position of incident optical 4, and spectrometer 7 gathers incident optical 4 with the output optical fiber 8 with respect to the first position
Transmitted spectrum and fluorescence spectrum under a；Then ensureing that incident optical 4 and the position of the output optical fiber 8 are still same by stepper motor 6
On the premise of axle, control incident optical 4 is moved at the b of the second place, and it is relative with the output optical fiber 8 that spectrometer 7 gathers incident optical 4
Transmitted spectrum and fluorescence spectrum under the b of the second place.
Wherein, the step of content of follow-up normalization, founding mathematical models and calculating free hemoglobin is with implementing
Example 1 is identical, and the embodiment of the present invention is not repeated this.
To the model of each device in addition to specified otherwise is done, the model of other devices is not limited the embodiment of the present invention,
As long as the device of above-mentioned function can be completed.
In summary, the embodiments of the invention provide a kind of double light path transmissions and fluorescence spectral measuring free hemoglobin to contain
The method of amount, transmitted light source change and the influence of blood bag are not only eliminated, and measurement is with strong points, greatly inhibits the non-of spectrum
Linear effect, improve the precision of free hemoglobin content analysis in blood bag.Realize in quick, free of contamination blood bag and dissociate
The measurement of hemoglobin, it is workable.
Embodiment 10
As different from Example 9, transmitted light source is bromine tungsten filament lamp wide spectrum light source to the embodiment of the present invention, the bromine tungsten filament lamp wide spectrum optical
The combination of source covering visible light wave band or near infrared light wave band or both；Fluorescence excitation light source is ultraviolet laser tube, above-mentioned light source
It can directly light or be conducted through incident optical 4.
As shown in figure 5, above-mentioned light source 3 is transmitted and excited to the blood sample in blood bag 5 by incident optical 4, by
Spectrometer 7 passes through the acquisition of transmission spectrum of the output optical fiber 8 and fluorescence spectrum, and incident optical 4 is close to blood bag 5 respectively with the output optical fiber 8
And it is coaxial, spectrometer 7 gathers incident optical 4 with the output optical fiber 8 with respect to the transmitted spectrum and fluorescence spectrum under a of the first position；
Then by stepper motor 6 ensure incident optical 4 and the position of the output optical fiber 8 remain unchanged it is coaxial on the premise of, control incident optical 4
Be moved at the b of the second place, spectrometer 7 gather transmitted spectrum under incident optical 4 and the relative second position b of the output optical fiber 8 and
Fluorescence spectrum.
Wherein, the step of content of follow-up normalization, founding mathematical models and calculating free hemoglobin is with implementing
Example 1 is identical, and the embodiment of the present invention is not repeated this.
To the model of each device in addition to specified otherwise is done, the model of other devices is not limited the embodiment of the present invention,
As long as the device of above-mentioned function can be completed.
In summary, the embodiments of the invention provide a kind of double light path transmissions and fluorescence spectral measuring free hemoglobin to contain
The method of amount, transmitted light source change and the influence of blood bag are not only eliminated, and measurement is with strong points, greatly inhibits the non-of spectrum
Linear effect, improve the precision of free hemoglobin content analysis in blood bag.Realize in quick, free of contamination blood bag and dissociate
The measurement of hemoglobin, it is workable.
Embodiment 11
The embodiment of the present invention is unlike embodiment 9,10, and transmitted light source is xenon lamp wide spectrum light source, the xenon lamp wide spectrum optical
The combination of source covering visible light wave band or near infrared light wave band or both；Fluorescence excitation light source is ultra-violet light-emitting pipe, above-mentioned light source 3
It can directly light or be conducted through incident optical 4.Displacement platform 6 is magnet attraction device, described below：
As shown in figure 5, above-mentioned light source 3 is transmitted and excited to blood sample in blood bag 5 by incident optical 4, spectrum
Instrument 7 passes through the acquisition of transmission spectrum of the output optical fiber 8 and fluorescence spectrum, and incident optical 4 and the output optical fiber 8 are close to blood bag 5 and together respectively
Axle, spectrometer 7 gather incident optical 4 with the output optical fiber 8 with respect to the transmitted spectrum and fluorescence spectrum under a of the first position；Then
By magnet attraction device 6 ensure incident optical 4 and the position of the output optical fiber 8 remain unchanged it is coaxial on the premise of control incident optical 4
Be moved at the b of the second place, spectrometer 7 gather transmitted spectrum under incident optical 4 and the relative second position b of the output optical fiber 8 and
Fluorescence spectrum.
Wherein, the step of content of follow-up normalization, founding mathematical models and calculating free hemoglobin is with implementing
Example 1 is identical, and the embodiment of the present invention is not repeated this.
To the model of each device in addition to specified otherwise is done, the model of other devices is not limited the embodiment of the present invention,
As long as the device of above-mentioned function can be completed.
In summary, the embodiments of the invention provide a kind of double light path transmissions and fluorescence spectral measuring free hemoglobin to contain
The method of amount, transmitted light source change and the influence of blood bag are not only eliminated, and measurement is with strong points, greatly inhibits the non-of spectrum
Linear effect, improve the precision of free hemoglobin content analysis in blood bag.Realize in quick, free of contamination blood bag and dissociate
The measurement of hemoglobin, it is workable.
Embodiment 12
The embodiment of the present invention unlike above-described embodiment 9,10,11, may be used also by needs of the light source 3 in practical application
Can also use other mobile devices using light source, the displacement platform 6 of other models, spectrum acquisition device 7 can also be adopted
With other reception devices.During specific implementation, the embodiment of the present invention is not limited to the model of above-mentioned device.
The embodiment of the present invention is to position a, a '；Position b, b ' and move mode etc. be not restricted, as long as this hair can be realized
The function of bright embodiment, within the protection domain of the application.
In summary, the embodiments of the invention provide a kind of double light path transmissions and fluorescence spectral measuring free hemoglobin to contain
The method of amount, transmitted light source change and the influence of blood bag are not only eliminated, and measurement is with strong points, greatly inhibits the non-of spectrum
Linear effect, improve the precision of free hemoglobin content analysis in blood bag.Realize in quick, free of contamination blood bag and dissociate
The measurement of hemoglobin, it is workable.
Claims (10)
1. a kind of double light path transmissions and the method for fluorescence spectral measuring free hemoglobin content, it is characterised in that methods described
Comprise the following steps：
The light extraction optical port of light source is close to blood bag and coaxial with the entrance slit of spectrum acquisition device, and light source includes transmitted light source and glimmering
Phot-luminescence source, transmitted light source transmit to blood sample, and fluorescent exciting source excitation free hemoglobin produces fluorescence, light
Compose reception device acquisition of transmission spectrum and fluorescence spectrum；
Displacement platform is before ensureing that transmitted light source and fluorescence excitation light source light extraction optical port and spectrum acquisition device entrance slit are coaxial
Put, transmitted light source and fluorescence excitation light source movement are controlled, by spectrum acquisition device acquisition of transmission spectrum and fluorescence spectrum；
Ask logarithm to be absorbed spectrum light intensity ratio under each wavelength of two opening position transmitted spectrums, and combine two positions
Under fluorescence spectrum normalized, with reference to the data of chemical analysis, founding mathematical models；
The transmitted spectrum and fluorescence spectrum at unknown blood sample two positions are gathered, asks logarithm to obtain the ratio of two transmitted spectrums
Absorption spectrum and the normalization of two fluorescence spectrums after bring mathematical modeling into and calculated, obtain containing for free hemoglobin
Amount；
The transmitted spectrum and fluorescence spectrum are to cause the spectrum under the nonlinear factor collective effect of spectrum, and increase is free blood red
Protein Information amount；Eliminate transmitted light source change and the influence of blood bag；Suppress the non-linear effects of spectrum；Improve the blood that dissociates in blood bag
The precision of Lactoferrin content analysis.
2. a kind of double light path transmissions according to claim 1 and the method for fluorescence spectral measuring free hemoglobin content,
Characterized in that, displacement platform control transmitted light source and fluorescence excitation light source movement, by spectrum acquisition device acquisition of transmission spectrum
And the step of fluorescence spectrum, is specially：
Transmitted light source and fluorescence excitation light source are transmitted and excited respectively to blood sample at a of position, are filled by spectrum acquisition
Put acquisition of transmission spectrum and fluorescence spectrum；
Displacement platform controls transmitted light source and fluorescence excitation light source to be moved to position b, by spectrum acquisition device acquisition of transmission spectrum
And fluorescence spectrum；
Or,
Transmitted light source and fluorescence excitation light source are transmitted and excited respectively to the blood sample in blood bag, by spectrum acquisition device
Acquisition of transmission spectrum and fluorescence spectrum at a of position；
Displacement platform control spectrum acquisition device is moved to position b, gathers transmitted spectrum and fluorescence spectrum at the b of position；
Or,
The blood sample in blood bag is transmitted and excited respectively by transmitted light source and fluorescence excitation light source at a of position,
Position a ' places are by spectrum acquisition device acquisition of transmission spectrum and fluorescence spectrum；
Displacement platform controls transmitted light source and fluorescence excitation light source to be moved to position b, and spectrum acquisition device is moved to position b ' places,
By spectrum acquisition device acquisition of transmission spectrum and fluorescence spectrum.
3. a kind of double light path transmissions according to claim 1 or 2 and the side of fluorescence spectral measuring free hemoglobin content
Method, it is characterised in that methods described also includes：
One optical fiber is set at light source, as incident optical, and ensures that incident optical is close to spectrum acquisition device entrance slit
Blood bag and coaxial；
Or,
One optical fiber is set at spectrum acquisition device, as the output optical fiber, and ensures that the output optical fiber is close to light source light extraction optical port
Blood bag and coaxial；
Or,
Incident optical and the output optical fiber are set respectively at light source and spectrum acquisition device, and ensure incident optical and the output optical fiber
It is close to blood bag and coaxial.
4. a kind of double light path transmissions according to claim 3 and the method for fluorescence spectral measuring free hemoglobin content,
Characterized in that,
A positions are the first position of incident optical, are gathered by spectrum acquisition device under incident optical and the output optical fiber relative position a
Transmitted spectrum and fluorescence spectrum；Displacement platform is on the premise of ensureing that incident optical and the output optical fiber are coaxial, control incident light
Fibre is moved at the b of position, and incident optical and the transmitted spectrum under the output optical fiber relative position b and glimmering are gathered by spectrum acquisition device
Light spectrum.
5. a kind of double light path transmissions according to claim 3 and the method for fluorescence spectral measuring free hemoglobin content,
Characterized in that,
A positions are the first position of the output optical fiber, are gathered by spectrum acquisition device under incident optical and the output optical fiber relative position a
Transmitted spectrum and fluorescence spectrum；Displacement platform is on the premise of ensureing that incident optical and the output optical fiber are coaxial, control emergent light
Fibre is moved at the b of position, and incident optical and the transmitted spectrum under the output optical fiber relative position b and glimmering are gathered by spectrum acquisition device
Light spectrum.
6. a kind of double light path transmissions according to claim 3 and the method for fluorescence spectral measuring free hemoglobin content,
Characterized in that,
A, a ' is respectively the first position of incident optical and the output optical fiber, and incident optical and emergent light are gathered by spectrum acquisition device
Transmitted spectrum and fluorescence spectrum under relative position a, a ' of fibre；Displacement platform is ensureing that incident optical and the output optical fiber are coaxial
Under the premise of, control incident optical and the output optical fiber to be moved respectively to position b, b ' place, incident optical is gathered by spectrum acquisition device
The position b, b relative with the output optical fiber ' under transmitted spectrum and fluorescence spectrum.
7. a kind of double light path transmissions according to claim 3 and the method for fluorescence spectral measuring free hemoglobin content,
Characterized in that, the lit transmissive light source is super continuous wide range laser, the super continuous wide range laser covering visible light wave band or
Near infrared light wave band, or both combination；Fluorescence excitation light source is ultraviolet lamp, and above-mentioned light source can directly light or through incident light
Fibre conduction.
8. a kind of double light path transmissions according to claim 3 and the method for fluorescence spectral measuring free hemoglobin content,
Characterized in that, institute's translation stage is stepper motor；The spectrum acquisition device is spectrometer.
9. a kind of double light path transmissions according to claim 3 and the method for fluorescence spectral measuring free hemoglobin content,
Characterized in that, the transmitted light source is xenon lamp wide spectrum light source or bromine tungsten filament lamp wide spectrum light source, covering visible light wave band or near-infrared
Optical band, or both combination；The fluorescence excitation light source is ultraviolet laser tube or ultra-violet light-emitting pipe, and above-mentioned light source can directly be sent out
Light conducts through incident optical；Institute's translation stage is magnet attraction device.
The blood 10. a kind of double light path transmissions and fluorescence spectral measuring according to any claim in claim 1-3 dissociate
The method of Lactoferrin content, it is characterised in that
The mathematical modeling utilizes principal component analysis, artificial neural network, PLS, SVMs, signal point
Analysis or statistical method are established.
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