CN116509339A - Low-power Raman spectrum assisted unmarked diffuse reflection hyperspectral imaging system - Google Patents
Low-power Raman spectrum assisted unmarked diffuse reflection hyperspectral imaging system Download PDFInfo
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Abstract
The invention discloses a low-power Raman spectrum assisted unmarked diffuse reflection hyperspectral imaging system. The device consists of an objective lens, a beam splitter, an adapter with a collimating lens, a sleeve, a laser, an LED, a long-wave-pass dichroic mirror, an adapter with a long-wave-pass filter and a focusing lens, an aberration-eliminating Raman spectrometer, a high-sensitivity camera, an adapter with a focusing lens, a diffuse reflection hyperspectral spectrometer and a CMOS camera. The diagnosis and identification are carried out by using the low-power (lower than the human body safety threshold) laser to excite the Raman spectrum to assist the unmarked diffuse reflection hyperspectral, so that the information acquisition amount can be greatly increased, and the diagnosis precision is improved. The system can also be used in combination with an endoscope, and simultaneously obtains the space information in the patient and the corresponding hyperspectral information, thereby having important significance for rapidly and accurately judging the tumor position and guiding the incisional margin in the operation.
Description
Technical Field
The invention belongs to the technical field of hyperspectrum, and relates to a low-power Raman spectrum assisted unmarked diffuse reflection hyperspectral imaging system.
Background
Tumor diagnosis is a detection method which needs to be rapid, accurate and noninvasive so as to discover and treat cancers in time. Traditional tumor diagnosis methods, such as biopsy, pathological section and the like, require steps of sampling, preparing, staining and the like on a sample, and consume long time. The hyperspectral imaging technology is a label-free in-situ imaging technology capable of simultaneously acquiring 2-dimensional spatial information and 1-dimensional spectral information of biological tissues, covers the spectral ranges of visible light, infrared, ultraviolet and the like, has higher spectral resolution and spatial resolution, can provide diagnostic information about physiological, morphological and biochemical components of the tissues, realizes rapid identification of tumors, and improves the accuracy and sensitivity of diagnosis.
The hyperspectral characteristics of normal tissues and cancerous tissues are different due to a certain difference in molecular structure. The diffuse reflection hyperspectral has good morphological information and is not influenced by fluorescence, but the spectrum specificity is inferior to that of a Raman spectrum, a low-power Raman spectrum assisted unmarked diffuse reflection hyperspectral imaging system is built, a Raman spectrum image and an unmarked diffuse reflection hyperspectral image can be measured simultaneously, the damage of high-power laser to a living body is avoided while the Raman fingerprint information is maintained, artificial intelligence is combined, the in-vitro sample is subjected to benign and malignant properties and differentiation degree rapid identification, and the technology can realize the following two functions simultaneously: (1) The method is applied to rapid detection and intraoperative guidance of incisional edge and infiltration depth, and ensures that cancer tissues are not missed; (2) For the ultra-low rectal resection operation, unnecessary tissue resection is reduced, and the anus protection rate is improved.
The combination of hyperspectral and laparoscope can improve the accuracy and reliability of disease diagnosis, provide spectral information with more wavelengths, and present micro focus or lymph node metastasis which cannot be seen under common light in the visual field of doctors; tumor boundaries, blood perfusion, lymph and the like can be observed in real time, the boundaries and the cutting edges of the tumor can be more accurately determined, and residual tumor tissues or excessive normal tissues are avoided; can improve the stage accuracy of colorectal cancer, more clearly judge the infiltration depth of tumor, lymph node metastasis and distant metastasis, and provide basis for surgical scheme selection and prognosis evaluation. For in vivo label-free detection, the security of the excitation light source, the imaging signal-to-noise ratio, and the rapidity of scanning imaging are of most concern. The low-power Raman spectrum assisted label-free diffuse reflection hyperspectral imaging can provide spatial information and spectral information in a patient, and can avoid the trauma of high-power laser to a living body while keeping Raman fingerprint information, so that the method has important significance for clinical applications such as tumor diagnosis, intraoperative cutting edge guidance and the like.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention provides a low-power Raman spectrum assisted unmarked diffuse reflection hyperspectral imaging system, which is characterized by comprising a front-end module, a light source module, a Raman imaging module and a diffuse reflection hyperspectral imaging module; the front module comprises an objective lens 1, a beam splitter 2, an adapter 3 with a collimating lens, a sleeve 4 and a long-wave-pass dichroic mirror 7; the light source module comprises a laser 5 and an LED 6; the Raman imaging module comprises an adapter 8 with a long-pass filter and a focusing lens, an aberration-eliminating Raman spectrometer 9 and a high-sensitivity camera 10; the diffuse reflection hyperspectral imaging module comprises an adapter 11 with a focusing lens, a diffuse reflection hyperspectral spectrometer 12 and a CMOS camera 13;
the detection light of the light source module is focused on a sample to be detected, the long-wave-pass dichroic mirror 7 splits scattered/reflected signals, and the Raman imaging module and the diffuse reflection hyperspectral imaging module perform conjugate imaging, so that simultaneous measurement of a Raman spectrum and a diffuse reflection spectrum is realized.
The low-power Raman spectrum assisted label-free diffuse reflection hyperspectral imaging system selects the wavelength of the LED 6 to be 400-700nm, and selects the wavelength of the laser 5 to be 785nm, so that the diffuse reflection signal wave band is 400-700nm, the Raman signal wavelength is greater than 785nm, the two paths of signals have no wave band overlapping, and the simultaneous measurement of a diffuse reflection spectrum and a Raman spectrum can be realized.
The low-power Raman spectrum assisted unmarked diffuse reflection hyperspectral imaging system can be provided with a microprobe module in front of a front-end module, wherein the microprobe module consists of a front-end probe, an image transmission optical fiber and a signal output end, the microprobe module enters a patient body through a biopsy pore canal of a standard endoscope, the front-end probe couples signal light into the image transmission optical fiber, the signal light is transmitted to the signal output end through the image transmission optical fiber, and the signal output end focuses signals to an object plane of an objective lens; the low-power laser excitation Raman spectrum is utilized to assist diffuse reflection hyperspectral to carry out endoscopic diagnosis and identification, and the method can also be used for rapidly and accurately judging the tumor position and guiding the incisal margin in operation, and improves the diagnosis precision.
The low-power Raman spectrum assisted label-free diffuse reflection hyperspectral imaging system adopts a low-power laser 5 and a high-sensitivity camera 10, so that the thermal damage and the illumination damage to a sample can be reduced while the signal-to-noise ratio of a Raman signal is ensured, and the system can be applied to an endoscopic system.
The diffuse reflection hyperspectral spectrometer 12 can adopt a wedge prism-grating-wedge prism (PGP) structure or a Liquid Crystal Tunable Filter (LCTF) to realize light splitting.
The Raman and diffuse reflection hyperspectral spectrum data acquired by the low-power Raman spectrum auxiliary unmarked diffuse reflection hyperspectral imaging system are sent into a computer in a digital mode, space features are extracted by using a deep Lab v3+ neural network segmentation frame and are fused with the spectrum features, and a Support Vector Machine (SVM) classifier is used for classification.
The low-power Raman spectrum assisted unmarked diffuse reflection hyperspectral imaging system can change a laser 5 into linear laser, and realize hyperspectral imaging under the depth of a plurality of millimeters by using a space shift Raman technology.
The front end probe of the micro probe module in the low-power Raman spectrum assisted non-marking diffuse reflection hyperspectral endoscopic imaging system adopts a fiber probe which is offset in space, namely, a collecting fiber is arranged in an inner ring, and an excitation fiber is arranged in an outer ring, so that detection of deeper tissues can be realized, and the tumor infiltration depth can be diagnosed.
The beneficial effects of the invention are that
The low-power Raman spectrum assisted non-marking diffuse reflection hyperspectral imaging system provided by the invention can be used for simultaneously measuring Raman spectrum and diffuse reflection hyperspectral, performing non-marking hyperspectral detection on a laparoscopic excised sample, combining with artificial intelligence, realizing rapid identification of benign and malignant and differentiation degrees of the excised sample, and also can be integrated into a laparoscope, simultaneously obtaining spatial information and corresponding hyperspectral information in a patient, and rapidly and accurately judging the position of a tumor in an operation, predicting the positive condition of metastasis of a lateral lymph node and the outside of a cleaning range, thereby having great significance on medical diagnosis.
Drawings
Fig. 1 is a schematic diagram of a low-power raman spectroscopy-assisted label-free diffuse reflectance hyperspectral imaging system.
Fig. 2 is a low power raman spectroscopy assisted label free diffuse reflectance hyperspectral imaging system.
Fig. 3 is a low power raman spectroscopy assisted label-free diffuse reflectance hyperspectral endoscopic imaging system.
Meaning of each number in the figure: 1. an objective lens; 2. a beam splitter; 3. an adapter with a collimating lens; 4. a sleeve; 5. a laser; 6. an LED; 7. a long-wave pass dichroic mirror; 8. an adapter with a long pass filter and a focusing lens; 9. aberration-eliminating raman spectrometer; 10. a high sensitivity camera; 11. an adapter with a focusing lens; 12. diffuse reflection hyperspectral spectrometer; 13. a CMOS camera; 14. an adjustable lens; 15. a liquid crystal tunable filter; 16. an adapter with an objective lens; 17. a slit; 18. an achromatic lens; 19. wedge prism-grating-wedge prism (PGP) structures; 20. an achromatic imaging lens; 21. an imaging optical fiber; 22. a front end probe; 23. biopsy channel of standard endoscope.
Detailed Description
The present invention will be described in further detail with reference to the drawings and examples, in order to make the objects, technical solutions and advantages of the present invention more apparent.
Example 1:
as shown in fig. 1, the low-power raman spectrum assisted unmarked diffuse reflection hyperspectral imaging system comprises a front-end module, a light source module, a raman imaging module and a diffuse reflection hyperspectral imaging module; the front module comprises an objective lens 1, a beam splitter 2, an adapter 3 with a collimating lens, a sleeve 4 and a long-wave-pass dichroic mirror 7; the light source module comprises a laser 5 and an LED 6; the Raman imaging module comprises an adapter 8 with a long-pass filter and a focusing lens, an aberration-eliminating Raman spectrometer 9 and a high-sensitivity camera 10; the diffuse reflection hyperspectral imaging module comprises an adapter 11 with a focusing lens, a diffuse reflection hyperspectral spectrometer 12 and a CMOS camera 13.
In this embodiment, the wavelength of the LED 6 is 400-700nm, the wavelength of the laser 5 is 785nm, the laser power is 0.3mw, and the unmarked diffuse reflection hyperspectral imaging module uses a tunable liquid crystal filter 15 structure to realize light splitting, as shown in fig. 2. The detection light of the light source module is focused on a sample to be detected, the long-wave-pass dichroic mirror 7 splits scattered/reflected signals, and the Raman imaging module and the diffuse reflection hyperspectral imaging module perform conjugate imaging, so that simultaneous measurement of a Raman spectrum and a diffuse reflection spectrum is realized.
In this embodiment, the deep neural network is trained by collecting and labeling a large number of hyperspectral normal and tumor tissue images, and the design of the network algorithm and structure is extracted by iterative experiment features, so as to effectively utilize the rich information contained in the hyperspectral images. Finally, the in-vitro sample can be rapidly identified in benign and malignant and differentiation degree, and a foundation is laid for rapid detection and intraoperative guidance of incisional edge and infiltration depth, and no missed incision of cancer tissues is ensured.
Example 2:
as shown in fig. 3, the low-power raman spectroscopy-assisted label-free diffuse reflection hyperspectral endoscopic imaging system consists of an imaging optical fiber 21, a front-end probe 22, a laser 5, an led 6, an adapter with an objective lens 16, a long-pass dichroic mirror 7, an adapter with a long-pass filter and a focusing lens 8, an aberration-eliminating raman spectrometer 9, a high-sensitivity camera 10, an adapter with a focusing lens 11, a slit 17, an achromatic lens 18, a wedge prism-grating-wedge prism (PGP) structure 19, an achromatic imaging lens 20, a cmos camera 13, and a biopsy tunnel 23 of a standard endoscope.
In this embodiment, the wavelength of the LED 6 is 400-700nm, the wavelength of the laser 3 is 785nm, the laser power is 0.3mw, and the front-end probe 22 adopts a spatially offset optical fiber probe, i.e. the collecting optical fiber is in the inner ring and the exciting optical fiber is in the outer ring, so that the detection of deeper tissues can be realized. The micro probe module formed by the imaging optical fiber 21 and the front end probe 22 enters the patient through a biopsy pore 23 of a standard endoscope of the alimentary canal or the respiratory tract, such as a enteroscope, a gastroscope, a bronchoscope or a respiratory tract endoscope, so as to realize the detection of hyperspectral spectrogram in the body; the back/scattered signal is coupled to the imaging optical fiber 21 by the probe 22, is conducted to the signal output end through the imaging optical fiber, and is focused to the object plane of the objective lens, and the long-wave-pass dichroic mirror 7 splits the scattered/reflected signal, so that the raman imaging module and the diffuse reflection hyperspectral imaging module perform conjugate imaging, and simultaneous measurement of the raman spectrum and the diffuse reflection spectrum is realized.
In the embodiment, the laser excitation Raman spectrum with low power (lower than the safe laser power of a human body) assists in marking-free diffuse reflection hyperspectral imaging, and meanwhile, spatial information in a patient and corresponding hyperspectral information are obtained, so that the accuracy and reliability of disease diagnosis can be improved, and tiny focus or lymph node metastasis which cannot be seen under common light is displayed in the visual field of doctors; tumor boundaries, blood perfusion, lymph and the like can be observed in real time, the boundaries and the cutting edges of the tumor can be more accurately determined, and residual tumor tissues or excessive normal tissues are avoided; can improve the stage accuracy of colorectal cancer, more clearly judge the infiltration depth of tumor, lymph node metastasis and distant metastasis, and provide basis for surgical scheme selection and prognosis evaluation.
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The above examples illustrate only a few embodiments of the invention, which are described in detail and are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.
Claims (8)
1. The low-power Raman spectrum assisted unmarked diffuse reflection hyperspectral imaging system is characterized by comprising a front-end module, a light source module, a Raman imaging module and a diffuse reflection hyperspectral imaging module; the front module comprises an objective lens (1), a beam splitter (2), an adapter (3) with a collimating lens, a sleeve (4) and a long-wave-pass dichroic mirror (7); the light source module comprises a laser (5) and an LED (6); the Raman imaging module comprises an adapter (8) with a long-pass filter and a focusing lens, an aberration-eliminating Raman spectrometer (9) and a high-sensitivity camera (10); the diffuse reflection hyperspectral imaging module comprises an adapter (11) with a focusing lens, a diffuse reflection hyperspectral spectrometer (12) and a CMOS camera (13);
the detection light of the light source module is focused on a sample to be detected, the long-wave-pass dichroic mirror (7) splits scattered/reflected signals, and the Raman imaging module and the diffuse reflection hyperspectral imaging module perform conjugate imaging, so that simultaneous measurement of a Raman spectrum and a diffuse reflection spectrum is realized.
2. The low-power Raman spectrum assisted label-free diffuse reflection hyperspectral imaging system is characterized in that the wavelength of an LED (6) is selected to be 400-700nm, the wavelength of a selected laser (5) is selected to be 785nm, the wavelength of a diffuse reflection signal wave band is 400-700nm, the wavelength of a Raman signal is larger than 785nm, the wave bands of the two signals are not overlapped, and simultaneous measurement of a diffuse reflection spectrum and a Raman spectrum can be realized.
3. The low-power Raman spectrum assisted label-free diffuse reflection hyperspectral imaging system of claim 1, wherein a microprobe module can be added in front of the front-end module, the microprobe module consists of a front-end probe, an image transmission optical fiber and a signal output end, the microprobe module enters a patient body through a biopsy duct of a standard endoscope, the front-end probe couples signal light into the image transmission optical fiber, the signal light is transmitted to the signal output end through the image transmission optical fiber, and the signal output end focuses the signal at an object plane of an objective lens; the low-power laser excitation Raman spectrum is utilized to assist diffuse reflection hyperspectral to carry out endoscopic diagnosis and identification, and the method can also be used for rapidly and accurately judging the tumor position and guiding the incisal margin in operation, and improves the diagnosis precision.
4. The low-power raman spectroscopy-assisted label-free diffuse reflectance hyperspectral imaging system of claim 1 wherein a low-power laser (5) and a high-sensitivity camera (10) are employed to reduce thermal and optical damage to the sample while guaranteeing raman signal to noise ratio for use in endoscopic systems.
5. The low-power raman spectroscopy assisted markerless diffuse reflectance hyperspectral imaging system according to claim 1 wherein the diffuse reflectance hyperspectral spectrometer (12) can be a wedge prism-grating-wedge prism (PGP) structure or a Liquid Crystal Tunable Filter (LCTF) for spectroscopic purposes.
6. The low-power raman spectroscopy assisted markerless diffuse reflectance hyperspectral imaging system according to claim 1 wherein the collected hyperspectral spectrum data is digitally fed into a computer, spatial features are extracted by a deep lab v3+ neural network segmentation framework and fused with spectral features, and a Support Vector Machine (SVM) classifier is used for classification.
7. The low-power raman spectroscopy assisted markerless diffuse reflection hyperspectral imaging system according to claim 1 wherein the laser (5) can be switched to a line laser and hyperspectral imaging at a depth of several millimeters is achieved using spatially offset raman techniques.
8. The low-power raman spectroscopy assisted label-free diffuse reflection hyperspectral imaging system of claim 1, wherein a microprobe module can be added in front of the front module, and a spatial offset optical fiber probe is adopted as a front end probe of the microprobe module, namely a collecting optical fiber is arranged in an inner ring, and an excitation optical fiber is arranged in an outer ring, so that the tumor infiltration depth can be diagnosed.
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