CN110793954A - Portable Raman blood identification system based on echelle grating - Google Patents
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Abstract
The invention discloses a portable Raman blood identification system based on echelle grating, which comprises an excitation light path and a collection light path; the excitation light path comprises a laser, a collimating lens, a Rayleigh filter and a focusing objective lens which are sequentially arranged along the propagation direction of the excitation light; the collection light path comprises a focusing lens, a pinhole, a collimating mirror, a spectrum separation component, a focusing mirror and a CCD (charge coupled device), wherein the focusing lens, the pinhole, the collimating mirror, the spectrum separation component, the focusing mirror and the CCD are sequentially arranged along the propagation direction of the sample light, the spectrum separation component comprises an echelle grating and a prism, and the order of the echelle grating and the prism arranged along the sample light is interchangeable. The portable Raman blood identification system based on the echelle grating adopts a folding light path design scheme by introducing the echelle grating and the auxiliary dispersion element prism in the core dispersion element, realizes the coexistence of small volume and high resolution of a spectrometer, and has the characteristics of compact structure, high resolution, easy implementation and the like.
Description
Technical Field
The invention relates to the technical field of spectral analysis instruments, in particular to a portable Raman blood identification system based on echelle gratings.
Background
The blood is the most convenient and rapid gene carrier for human and animals, and carries a complete set of genetic information. In order to prevent the loss of national species resource information and the damage of intellectual property rights of related species and avoid the damage of ecological environment caused by the invasion of unknown foreign species, the customs import and export departments need to test and identify the species of blood products.
At present, the main methods for identifying human blood and animal blood at home and abroad comprise: microscopic observation, HPLC, NIR, GC-MS, LC-MS, immunological methods, DNA gene detection methods and the like. However, these methods all belong to contact detection, and all have the following disadvantages: on one hand, the sample basically needs to be pretreated, so that the sample is damaged; on the other hand, the detection time is long. Therefore, the conventional method is difficult to meet the actual demands of the import and export inspection and quarantine departments on non-damage and rapidity. Compared with the above technologies, the raman spectroscopy technology is more mature, has been widely applied to the fields of chemistry, biomedicine, food safety, aerospace, environmental protection and the like, and can realize non-contact detection, so that the raman spectroscopy technology is an ideal technical means for blood identification. The subject group of the applicant has carried out a number of years of research related to blood identification using raman techniques:
1、HaiYi BIAN,Peng WANG,NingWANG,et al.Dual-model analysis forimproving the discrimination performance of human and nonhuman blood based onRaman spectroscopy[J].Biomedical Optics Express,2018,9(8):3512.
2、Haiyi B,Jing G.Error analysis of the spectral shift for partialleast squares models in Raman spectroscopy[J].Optics Express,2018,26(7):8016)。
however, conventional raman spectrometers, such as the renishawa inVia and HORIBA, use conventional blazed gratings, and only low diffraction orders (e.g., -1 order or-2 order) can be used to avoid overlapping orders. If the high dispersion rate and spectral resolution of the spectrometer are to be realized, a blazed grating with high groove density is needed, so that the focal length of an imaging objective lens is increased, the size of the instrument is overlarge, and meanwhile, the high groove density grating has requirements on cleanliness, so that the traditional spectrometer can only be installed in an ultra-clean laboratory with constant temperature and humidity for use and is not suitable for field detection application.
Compared with the common blazed grating, the echelle grating can solve the problem that the echelle grating cannot simultaneously meet the actual requirements of small volume, wide spectrum, high resolution, transient measurement and the like. The echelle grating has the characteristics of low groove density, large blaze angle and high diffraction order, so the echelle grating has high dispersion rate and resolution, does not need to rotate the grating to search a spectrum surface for splicing, and can meet the requirement of measuring the wavelength of a full spectrum section by one-time exposure. Compared with the common blazed grating spectrometer, the echelle grating spectrometer has smaller volume and simpler structure and is beneficial to realizing high intellectualization and automation. The overlapping phenomenon existing in the spectrum order of the echelle grating can be solved only by using a transverse dispersion element to carry out order separation, the formed two-dimensional spectrum is reflected to a CCD to form a required spectrogram, then, the spectrogram is analyzed by using a chemometrics method, and a final identification result is given through a human-computer interaction interface.
The traditional Raman spectrometer has the defect that the small volume and high resolution of the spectrometer are difficult to coexist, and the echelle grating is expected to be applied to a Raman spectrometer to solve the problem and is used for realizing blood identification, but the prior art lacks a reliable scheme, so that a blood identification system utilizing the echelle grating is provided.
Disclosure of Invention
The technical problem to be solved by the present invention is to provide a portable raman blood identification system based on echelle grating, aiming at the above-mentioned deficiencies in the prior art.
In order to solve the technical problems, the invention adopts the technical scheme that: a portable Raman blood identification system based on echelle grating comprises an excitation light path and a collection light path;
the excitation light path comprises a laser, a collimating lens, a Rayleigh filter and a focusing objective lens which are sequentially arranged along the propagation direction of the excitation light;
the collection light path comprises a focusing lens, a pinhole, a collimating lens, a spectrum separation component, a focusing lens and a CCD (charge coupled device) which are sequentially arranged along the propagation direction of the sample light, the spectrum separation component comprises an echelle grating and a prism, and the order of the echelle grating and the prism arranged in sequence along the sample light can be interchanged;
exciting light emitted by the laser passes through the collimating lens, is reflected by the Rayleigh filter, passes through the focusing objective and then irradiates a sample; the Raman light generated by exciting the sample returns along the original light path, passes through the focusing objective lens, the Rayleigh filter, the focusing lens and the pinhole in sequence, is collimated into parallel light by the collimating lens, enters the spectrum separation component, and is reflected to the CCD through the focusing lens;
the light emitted by the collimating mirror firstly enters the echelle grating and then is reflected to the focusing mirror by the prism; or the light emitted by the collimating mirror firstly enters the prism, is transmitted by the prism and then reaches the echelle grating, and then enters the focusing mirror.
Preferably, the laser emits a laser wavelength of 532 nm.
Preferably, the rayleigh filter, the focusing objective lens and the focusing lens are plated with 537.7 nm-595.3 nm high-transmittance films on both sides.
Preferably, the collimating lens is a spherical collimating lens or an aspheric collimating lens with a concave surface, and a high reflective film of 537.7 nm-595.3 nm is plated on the optical surface of the collimating lens.
Preferably, the focusing lens is a spherical focusing lens or an aspheric focusing lens with a concave surface, and the optical surface of the focusing lens is plated with a high reflection film of 537.7 nm-595.3 nm.
Preferably, the prism is a reflective prism having an entrance face and a reflective face; parallel light emitted by the collimating mirror firstly enters the echelle grating, light with different wavelengths is diffracted and then is separated according to angles, and then the light firstly transmits the incident surface of the reflecting prism and then is reflected to the focusing mirror by the reflecting surface of the reflecting prism.
Preferably, the incidence surface of the reflecting prism is plated with a high-transmittance film of 537.7 nm-595.3 nm, and the reflection surface is plated with a high-reflection film of 537.7 nm-595.3 nm.
Preferably, the prism is a transmissive prism having two light-transmitting faces; parallel light emitted by the collimating lens sequentially transmits the two light transmitting surfaces of the transmission prism, then enters the echelle grating, light with different wavelengths is diffracted and then is separated according to an angle, and then the light transmits the two light transmitting surfaces of the transmission prism and then reaches the focusing lens.
Preferably, both light transmission surfaces of the transmission prism are plated with high-transmittance films of 537.7 nm-595.3 nm.
The invention has the beneficial effects that: the portable Raman blood identification system based on the echelle grating adopts a folding light path design scheme by introducing the echelle grating and the auxiliary dispersion element prism in the core dispersion element, realizes the coexistence of small volume and high resolution of a spectrometer, and has the characteristics of compact structure, high resolution, easy implementation and the like.
Drawings
Fig. 1 is a schematic structural diagram of a portable raman blood identification system based on echelle grating according to embodiment 1 of the present invention;
fig. 2 is a schematic structural diagram of a portable raman blood identification system based on echelle grating according to embodiment 2 of the present invention;
FIG. 3 shows Raman spectra of human blood and dog blood excited by 532nm laser according to the present invention.
Description of reference numerals:
1-laser, 2-collimating lens, 3-Rayleigh filter, 4-focusing objective lens, 5-blood sample, 6-focusing lens, 7-pinhole, 8-collimating lens, 9-echelle grating, 10-reflection/transmission prism, 11-focusing lens and 12-CCD; 20-excitation light path; and 30, collecting the light path.
Detailed Description
The present invention is further described in detail below with reference to examples so that those skilled in the art can practice the invention with reference to the description.
It will be understood that terms such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.
The portable raman blood identification system based on echelle grating 9 of the present embodiment includes an excitation light path 20 and a collection light path 30;
the excitation light path 20 comprises a laser 1, a collimating lens 2, a Rayleigh filter 3 and a focusing objective 4 which are sequentially arranged along the propagation direction of the excitation light;
the collecting light path 30 comprises a focusing lens 6, a pinhole 7, a collimating mirror 8, a spectrum separation component, a focusing mirror 11 and a CCD12 which are sequentially arranged along the propagation direction of the sample light, the spectrum separation component comprises an echelle grating 9 and a reflecting prism 10, and the order of the echelle grating 9 and the prism 10 arranged in sequence along the sample light can be interchanged;
exciting light emitted by the laser 1 passes through the collimating lens 2, is reflected by the Rayleigh filter 3, passes through the focusing objective 4 and then irradiates a sample (blood); the Raman light generated by the sample is returned along the original light path, passes through the focusing objective 4, the Rayleigh filter 3, the focusing lens 6 and the pinhole 7 in sequence, is collimated into parallel light by the collimating lens 8, enters the spectrum separation component, and is finally reflected to the CCD12 through the focusing lens 11 to form the blood Raman spectrum. Finally, by using these spectra, the blood species can be analyzed and identified by conventional techniques, such as chemometrics, and the final identification result can be obtained by a computer interface. The scheme adopted for performing the subsequent processing on the obtained spectrum to form the final identification result can be selected by those skilled in the art in a conventional way, and the invention is not limited in particular.
Wherein, the light emitted from the collimating mirror 8 firstly enters the echelle grating 9 and then is reflected to the focusing mirror 11 by the prism 10; or the light emitted by the collimating mirror 8 firstly enters the prism 10, is transmitted by the prism 10 and then reaches the echelle grating 9, and then enters the focusing mirror 11.
In a preferred embodiment, the laser 1 emits a laser wavelength of 532 nm. In the application, Raman experiments of 532nm, 633nm and 785nm laser excited blood are carried out at the earlier stage, and compared with that the 532nm laser excited blood sample 5 has the best effect, the experimental result shows that the spectral range for effectively distinguishing human blood from animal blood is 200-2000 cm-doped blood-1(ii) a According to the formula of Raman frequency shift
(in the formula:. lambda.0Is the wavelength of excitation light source, and is 532nm and lambdaiFor the output wavelength after exciting the sample) the wavelength range of the corresponding output raman optical signal can be calculated to be 537.7 nm-595.3 nm, the interval is in the visible light wave band, and the absorption by the prism 10 is avoided, so the design is simple (the conventional echelle grating 9 spectrometer generally requires the test range to be 200 nm-900 nm, and the absorption of the prism 10 to ultraviolet and infrared needs to be considered during the design).
In a preferred embodiment, in order to improve the utilization efficiency of the Raman signal, 537.7 nm-595.3 nm high-transmittance films are plated on two sides of the Rayleigh filter 3, the focusing objective 4 and the focusing lens 6.
In a preferred embodiment, in order to reduce the loss of the raman signal, the collimating mirror 8 is a spherical collimating mirror 8 or an aspheric collimating mirror 8 with a concave surface, and a high reflection film of 537.7 nm-595.3 nm is plated on the optical surface of the collimating mirror 8.
In a preferred embodiment, in order to reduce the loss of the raman signal, the focusing mirror 11 is a spherical focusing mirror 11 or an aspheric focusing mirror 11 with a concave surface, and a high reflective film of 537.7 nm-595.3 nm is plated on the optical surface of the focusing mirror 11.
The invention selects the echelle grating 9 as a core dispersion element, the prism 10 as an auxiliary dispersion element, selects two spherical reflectors or two aspheric reflectors as a collimating mirror 8 and a focusing mirror 11 respectively, and adopts a folding light path design scheme to realize the coexistence of small volume and high resolution of the spectrometer.
The collection light path 30 may adopt at least two forms as above, one of which is: the prism 10 selects a reflecting prism 10, light emitted by the collimating mirror 8 firstly enters the echelle grating 9 and then is reflected to the focusing mirror 11 by the prism 10; the other is as follows: the prism 10 is a transmission prism 10, light emitted by the collimating mirror 8 firstly enters the prism 10, is transmitted by the prism 10 and then reaches the echelle grating 9, and then enters the focusing mirror 11. On the basis of the above, the scheme of the collecting optical path 30 in fig. 2 is further described with reference to the specific embodiment.
Example 1
Referring to fig. 1, in the above embodiment, the prism 10 is a reflection prism 10 having an incident surface and a reflection surface; the parallel light emitted from the collimating mirror 8 firstly enters the echelle grating 9, the light with different wavelengths is diffracted and then is separated according to the angle, and then the light firstly transmits the incident surface of the reflecting prism 10 and then is reflected to the focusing mirror 11 by the reflecting surface of the reflecting prism 10. In order to reduce the loss of Raman signals, the incidence surface of the reflecting prism 10 is plated with a high-transmittance film of 537.7 nm-595.3 nm, and the reflection surface is plated with a high-reflection film of 537.7 nm-595.3 nm.
In this embodiment, the specific optical path process is as follows:
exciting light emitted by the laser 1 is collimated into parallel light through the collimating lens 2, enters the Rayleigh filter 3 at an angle of 45 degrees, is reflected by the Rayleigh filter 3, and is converged on a blood sample 5 through the focusing objective 4; in order to detect the blood in the blood sampling tube, the focusing objective 4 adopts a long-focus objective with long working distance;
blood is excited by laser to generate a blood Raman optical signal with frequency offset, the blood Raman optical signal returns according to an original optical path, Rayleigh scattered light with unchanged frequency is filtered by a Rayleigh filter 3, the blood is focused on a pinhole 7 by a focusing lens 6, the blood passes through the pinhole 7 and is collimated into parallel light by a collimating mirror 8 to be incident on a echelle grating 9, light with different wavelengths is diffracted by the echelle grating 9 and then is split according to angles and incident on a reflecting prism 10 for secondary separation (the incident surface of the reflecting prism 10 is firstly transmitted and then reflected by the reflecting surface of the reflecting prism 10), so that the overlapped-level spectra are completely separated, and then the separated Raman light is reflected to a CCD12 by a focusing mirror 11 to form the blood Raman spectrum. Finally, by using these spectra, the final result can be obtained by analyzing and identifying the blood species by conventional chemometric methods.
Wherein, the focusing lens 6 with reasonable curvature radius and aperture angle is selected, thereby ensuring the energy concentration of the incident light beam to the pinhole 7 and ensuring the function of the echelle grating 9 as the aperture diaphragm of the system.
In a further preferred embodiment, the system also comprises a conventional laser 1 driving and refrigerating circuit, a CCD12 driving and refrigerating circuit and a PC (personal computer) human-computer interaction interface, and the system controls and outputs the identification result through the PC human-computer interaction interface. The above elements are selected by the user according to the above scheme of the invention.
Example 2
Referring to fig. 2, a further preferred embodiment of the present invention is the following embodiment 1: the prism 10 is a transmission prism 10 having two light transmission surfaces; the parallel light emitted from the collimating lens 8 sequentially transmits through the two light-transmitting surfaces of the transmission prism 10, then enters the echelle grating 9, is diffracted and then separated according to the angle, and then returns to the two light-transmitting surfaces of the transmission prism 10 to reach the focusing lens 11. In order to reduce the loss of Raman signals, the two light transmission surfaces of the transmission prism 10 are respectively plated with 537.7 nm-595.3 nm high-transmission films.
In this embodiment, the specific optical path process is as follows:
exciting light emitted by the laser 1 is collimated into parallel light through the collimating lens 2, enters the Rayleigh filter 3 at an angle of 45 degrees, is reflected by the Rayleigh filter 3, and is converged on a blood sample 5 through the focusing objective 4; in order to detect the blood in the blood sampling tube, the focusing objective 4 adopts a long-focus objective with long working distance;
blood Raman optical signals with frequency shifted generated by laser excitation are returned according to an original optical path, Rayleigh scattering light with unchanged frequency is filtered by a Rayleigh filter 3, then is focused on a pinhole 7 by a focusing lens 6, passes through the pinhole 7 and is collimated into parallel light by a collimating mirror 8 to enter a transmission prism 10, then sequentially transmits two light transmission surfaces of the transmission prism 10, after primary dispersion is completed, enters an intermediate step grating 9 for primary dispersion, then returns to two light transmission surfaces of the transmission prism 10 again (the transmission sequence is opposite to that of the previous time), secondary dispersion is completed, complete separation of overlapped-level secondary spectra is realized, and then separated Raman light is reflected to a CCD12 through a focusing mirror 11 to form the blood Raman spectrum. Finally, by using these spectra, the final result can be obtained by analyzing and identifying the blood species by conventional chemometric methods.
Compared with embodiment 1, this embodiment uses a transmissive structure instead of the reflective structure of embodiment 1, and can further compress the volume without affecting the lateral dispersion resolution.
While embodiments of the invention have been disclosed above, it is not limited to the applications listed in the description and the embodiments, which are fully applicable in all kinds of fields of application of the invention, and further modifications may readily be effected by those skilled in the art, so that the invention is not limited to the specific details without departing from the general concept defined by the claims and the scope of equivalents.
Claims (9)
1. A portable Raman blood identification system based on echelle grating is characterized by comprising an excitation light path and a collection light path;
the excitation light path comprises a laser, a collimating lens, a Rayleigh filter and a focusing objective lens which are sequentially arranged along the propagation direction of the excitation light;
the collection light path comprises a focusing lens, a pinhole, a collimating lens, a spectrum separation component, a focusing lens and a CCD (charge coupled device) which are sequentially arranged along the propagation direction of the sample light, the spectrum separation component comprises an echelle grating and a prism, and the order of the echelle grating and the prism arranged in sequence along the sample light can be interchanged;
exciting light emitted by the laser passes through the collimating lens, is reflected by the Rayleigh filter, passes through the focusing objective and then irradiates a sample; the Raman light generated by exciting the sample returns along the original light path, passes through the focusing objective lens, the Rayleigh filter, the focusing lens and the pinhole in sequence, is collimated into parallel light by the collimating lens, enters the spectrum separation component, and is reflected to the CCD through the focusing lens;
the light emitted by the collimating mirror firstly enters the echelle grating and then is reflected to the focusing mirror by the prism; or the light emitted by the collimating mirror firstly enters the prism, is transmitted by the prism and then reaches the echelle grating, and then enters the focusing mirror.
2. The echelle grating-based portable raman blood discrimination system of claim 1, wherein the laser emits a laser wavelength of 532 nm.
3. The portable raman blood identification system according to claim 2 wherein the rayleigh filter, the focusing objective and the focusing lens are coated with 537.7 nm-595.3 nm high-transmittance films on both sides.
4. The portable raman blood identification system according to claim 3, wherein the collimating mirror is a spherical collimating mirror or an aspheric collimating mirror with a concave surface, and the optical surface of the collimating mirror is coated with a high reflective film of 537.7 nm-595.3 nm.
5. The portable Raman blood identification system based on echelle grating of claim 4, wherein the focusing mirror is a spherical focusing mirror or an aspheric focusing mirror with a concave surface, and the optical surface of the focusing mirror is plated with a high reflective film of 537.7 nm-595.3 nm.
6. The echelle grating-based portable raman blood discrimination system according to any one of claims 1 to 5 wherein the prism is a reflective prism having an entrance face and a reflective face; parallel light emitted by the collimating mirror firstly enters the echelle grating, light with different wavelengths is diffracted and then is separated according to angles, and then the light firstly transmits the incident surface of the reflecting prism and then is reflected to the focusing mirror by the reflecting surface of the reflecting prism.
7. The portable Raman blood identification system based on echelle grating of claim 6, wherein the incidence surface of the reflection prism is coated with a high transmittance film of 537.7 nm-595.3 nm, and the reflection surface is coated with a high reflectance film of 537.7 nm-595.3 nm.
8. The echelle grating-based portable raman blood discrimination system according to any one of claims 1 to 5 wherein the prism is a transmissive prism having two light transmissive faces; parallel light emitted by the collimating lens sequentially transmits the two light transmitting surfaces of the transmission prism, then enters the echelle grating, light with different wavelengths is diffracted and then is separated according to an angle, and then returns to the two light transmitting surfaces of the transmission prism and then reaches the focusing lens.
9. The portable raman blood discrimination system according to claim 8, wherein both light transmission surfaces of the transmission prism are coated with a high transmission film of 537.7 nm-595.3 nm.
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