CN101785662A - Bimodal system and method integrating photoacoustic imaging and fluorescence imaging - Google Patents
Bimodal system and method integrating photoacoustic imaging and fluorescence imaging Download PDFInfo
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Abstract
The invention discloses a bimodal system and method integrating photoacoustic imaging and fluorescence imaging. In the invention, the complementarity between photoacoustic imaging and fluorescence imaging on the imaging theory is used, and through a common scanning light path system, photoacoustic imaging and fluorescence imaging are effectively integrated into a whole. The device for realizing the method of the invention comprises an photoacoustic imaging subsystem, a fluorescence imaging subsystem and a computer, wherein the photoacoustic imaging subsystem and the fluorescence imaging subsystem are integrated through the scanning light path system; the computer is respectively connected with the photoacoustic imaging subsystem, the fluorescence imaging subsystem and the scanning light path system; and the computer not only controls the scanning mode of the scanning light path system, but also processes the data of the photoacoustic imaging subsystem and the fluorescence imaging subsystem, combines the photoacoustic information and fluorescence information, and reconstructs photoacoustic and fluorescence bimodal images. The method of the invention has accurate positioning and high resolution, and the imaging system has lower cost and easy popularization.
Description
Technical field
The present invention relates to a kind of multi-modality imaging method, the imaging system and the formation method of particularly a kind of integrating photoacoustic and fluorescent dual module attitude.
Background technology
The invention of medical image physics and development of technology and medical imaging device and development promoted widely noinvasive or less under the wound condition to the diagnosis and the treatment of human diseases, thereby prolonged people's life-span greatly and created condition for the research of carrying out the bodily fuctions in the noinvasive condition.Though traditional medical image technology respectively has characteristics on the diagnosis of disease and auxiliary treatment, still there are many deficiencies in single image means.The resolution of nuclear magnetic resonance, NMR (MRI) is higher, can obtain simultaneously to dissect and physiologic information.But the weakness of MRI is, sensitivity lower (micromolar level) not only, and costing an arm and a leg is added the anemia of pregnant woman, patient's forbidding of cardiac pacemaker is housed, and makes the application of MRI be restricted.Nucleus medical image learn (PET) in iconography research at present in occupation of extremely important status, but because existing tracer kind development is limited, PET equipment complexity, and image taking speed is slower, its equipment and operation need a large amount of expense and personnel, cause the popularization of PET to be restricted.Ultra sonic imaging depends on the acoustic impedance of biological tissue, and poor contrast is difficult to find early stage canceration.The X-ray imaging depends on the density of biological tissue, and when density variation was little, the X-ray imaging then can't be judged, and induce tissue produces canceration easily.Pure optical image technology not only has higher spatial and temporal resolution and cost than MRI and the cheap advantage of PET, also have characteristics such as volume is little, in light weight and easy to use, but because biological tissue is to the strong scattering and the absorption of light, the pure optical image technology penetrate tissue degree of depth is limited, so can not carry out accurate imaging to deep tissues with it, along with the increase of imaging depth, the spatial resolution of pure optical image technology significantly reduces in addition.Therefore, want to need development multi-modality imaging method and technology observing intravital 26S Proteasome Structure and Function change simultaneously than no wound ground under the high-spatial and temporal resolution.
The multi-modality imaging technology is integrated into same imaging system with different image modes, because each image mode all has based on predetermined substance ripple and the interactional rule of human body, the information of the human body that obtains during based on the imaging technique of these different rules developments and device observes human body is also just incomplete same.Therefore, multi-modality imaging can provide the 26S Proteasome Structure and Function information of tissue simultaneously, is the development trend of present biomedical imaging.
Photoacoustic imaging (Photoacoustic imaging) is based on the novel harmless medical imaging method of optoacoustic effect.When pulsed laser irradiation is in biological tissue, organize the absorbing light energy to cause temperature rise, temperature rise causes thermal expansion and produces photoacoustic signal.The photoacoustic signal of biological tissue has carried the light absorption characteristics information of tissue, can reconstruct light absorption distributed image in the tissue by the measuring light acoustical signal.Because the malignant tumor of growth needs more blood supply fast, malignant tumor tissue is attended by more blood capillary hypertrophy, so the difference of cancerous issue and normal structure light absorption on every side has at least more than 5 times, thereby photoacoustic imaging can provide the tumor blood capillary image of high-resolution and high-contrast.
Fluorescence molecule imaging (Fluorescent Molecular imaging) adopts fluorescence report group (GFP, YFP, Cyt and Dyes etc.) to carry out labelling, utilize the very sensitive optical detecting instrument of a cover, can directly monitor the movable and gene behavior of active somatic cell in the organism.Can observe the growth and the biological processes such as transfer, infectious disease evolution and specific gene expression of in-vivo tumour by the fluorescence molecule imaging.
Therefore, start optoacoustic and fluorescent dual module attitude formation method, have the novelty on the principle.On physical principle, what optoacoustic reflected is radiationless transition process after the biomolecule light absorption; What fluorescence reflected is the radiation transistion process just, and both are complementary on principle.From imaging means, photoacoustic imaging can provide high-resolution tumor blood capillary image; Fluorescence imaging can provide highly sensitive tumor molecular information.Therefore, how photoacoustic imaging and two kinds of image modes of fluorescence imaging are integrated in same system, the characteristic of effectively utilizing two kinds of image modes to have complementary advantages is in the research tumor growth process tumor development and neovascularization thereof being had very important realistic meaning.
Summary of the invention
Primary and foremost purpose of the present invention is to overcome the shortcoming of single image mode of prior art with not enough, and the formation method of a kind of integrating photoacoustic and fluorescent dual module attitude is provided.
Another object of the present invention is to provide the imaging system that realizes described formation method.
Purpose of the present invention is achieved through the following technical solutions: the formation method of a kind of integrating photoacoustic and fluorescent dual module attitude comprises following steps:
(1) by the scanning light path system pulse laser focusing is arrived object under test, obtain exciting the photoacoustic signal of generation, receive by detector of sound again, send into signal amplifier, gather the photoacoustic signal data after amplifying;
(2) by the scanning light path system continuous laser is focused on object under test, obtains exciting the fluorescence signal of generation, then fluorescence signal after filtration mating plate filter and received by photomultiplier tube, gather the fluorescence signal after receiving;
(3) fluorescence signal after the photoacoustic signal data of the amplification that step (1) is obtained by data processing software and the reception that step (2) obtains merges and rebuilds, and obtains the image of optoacoustic and fluorescent dual module attitude.
The wavelength of described pulse laser is 400～2500nm;
Described detector of sound is a unit non-focusing detector, dominant frequency 20KHz～100MHz;
The wavelength of described continuous laser is 200～900nm;
Realize the imaging system of the formation method of integrating photoacoustic and fluorescent dual module attitude, form by photoacoustic imaging subsystem, fluorescence imaging subsystem and computer; Photoacoustic imaging subsystem and fluorescence imaging subsystem are by scanning the light path system integration in one; Computer is connected with the fluorescence imaging subsystem with the photoacoustic imaging subsystem respectively; Computer is the scan mode of gated sweep light path system not only, also handles the data of photoacoustic imaging subsystem and fluorescence imaging subsystem, merges optoacoustic and fluorescence information, reconstructs optoacoustic and fluorescent dual module attitude image simultaneously.
Described photoacoustic imaging subsystem comprises laser instrument, scanning light path system, detector of sound and signal amplifier; Laser instrument, scanning light path system, detector of sound are connected successively with signal amplifier; Laser instrument produces pulse laser as the photo-acoustic excitation light source, enter the scanning light path system, the scanning area that focuses on testing sample excites the photoacoustic signal of generation, photoacoustic signal is received by detector of sound, sending into signal amplifier amplifies, gather by computer-internal high-speed signal acquisition card, the signal of collection is stored into the medium pending data process software of Computer Cache and carries out image reconstruction again, wherein:
Laser instrument, the output pulse laser wavelength is 400～2500nm;
Described output laser preferably carries out the light path adjustment by reflecting mirror, thereby enters the scanning light path system;
Detector of sound is unit non-focusing detector, dominant frequency 20KHz～100MHz;
The self-control multi-channel parallel real-time acquisition system of described collection by the LABVIEW controlling platform realize, the effect of the self-control multi-channel parallel real-time acquisition system of LABVIEW controlling platform is to be digital signal with analog signal conversion;
Described image reconstruction is for to utilize filter back-projection algorithm to carry out image reconstruction by the MATLAB program, thereby obtains the photoacoustic image of detected part;
Described fluorescence imaging subsystem comprises laser instrument, scanning light path system, optical filter and photomultiplier tube (PMT); Laser instrument, scanning light path system, optical filter, photomultiplier tube (PMT) are connected successively with photomultiplier tube; Laser instrument is connected by excitation fiber with the scanning light path system, and the scanning light path system is connected by receiving optical fiber with optical filter; Laser instrument, light source as fluorescence excitation, exciting light enters the scanning light path system by excitation fiber, focus on scanning area, excite the fluorescence signal of generation to receive by receiving optical fiber, through the optical filter laggard photoelectricity multiplier tube (PMT) of going into that filters, carry out image reconstruction by being stored into the medium pending data process software of Computer Cache after the data collecting card collection, wherein:
Laser instrument, output continuous laser wavelength is 200～900nm;
Described excitation fiber preferably is made of the multichannel multimode fibre;
Described reception optical fiber preferably is made of the multichannel multimode fibre;
Described fluorescence signal is from vivo the molecule of labelling or the fluorescent probe stimulated luminescence of cell excite the back fluorescence that is sent;
Described fluorescence imaging subsystem preferably also contains avalanche photo diode (APD), and avalanche photodide is connected by receiving optical fiber with the scanning light path system, can be used for the fluorescence signal that receives is proofreaied and correct and normalization; The ratio of the fluorescence signal intensity that fluorescence signal intensity that APD accepts and PMT receive is preferably 1: 99;
Described scanning light path system contains control and scanning light path, wherein:
Control is connected with computer, thereby computer can be adjusted the scan mode of scanning light path;
Scan light route such as lower member are formed: reflecting mirror, filter lens, beam expanding lens, two dimension shake mirror scanning device and achromat; Each parts is separate; The parts that relate to photoacoustic imaging have reflecting mirror, two dimension shake mirror scanning device and achromat, and the parts that relate to fluorescence imaging have filter lens, beam expanding lens, two dimension shake mirror scanning device and achromat; Reflecting mirror, two dimension shake mirror scanning device and achromat are arranged from top to bottom; With the horizontal line is benchmark, and reflecting mirror is 45 ° of settings, and shake mirror and reflecting mirror in the two dimension shake mirror scanning device are parallel relation, the achromat horizontal positioned, beam expanding lens vertically is positioned at two dimension shake mirror scanning device top, and filter lens and beam expanding lens are positioned at same horizontal level, are 45 ° of settings; Pulse laser vertically enters light path by 45 ° of mirror reflects, incides two dimension shake mirror scanning device, utilizes achromat to focus on scanning area; Continuous laser incides filter lens by beam expanding lens after the horizontal outgoing of optical fiber, and vertically enters into two dimension shake mirror scanning device after 90 ° of reflections, utilizes achromat to focus on scanning area;
Described computer contains high-speed data acquisition card, scanning light path control software and image reconstruction software;
The self-control multi-channel parallel real-time acquisition system of described collection by the LABVIEW controlling platform realize, the effect of the self-control multi-channel parallel real-time acquisition system of LABVIEW controlling platform is to be digital signal with analog signal conversion;
Described image reconstruction is for to utilize filter back-projection algorithm to carry out image reconstruction by the MATLAB program, thereby obtains the fluoroscopic image of detected part;
The generation development that described imaging system not only can be tumor provides the tracking and monitoring data, but also can be used for observing other relevant issues researchs such as protein fluorescence quantum yield of relating to optoacoustic and fluorescent effect etc.
Principle of the present invention is: what optoacoustic reflected is radiationless transition process after the biomolecule light absorption, and what fluorescence reflected is the radiation transistion process just, realizes highly sensitive multi-modal molecular imaging.Therefore, utilize photoacoustic imaging and the fluorescence imaging complementarity on image-forming principle, can obtain the imaging of sufficient optoacoustic of information and fluorescent dual module attitude.The present invention is integrated in one by common light path scanning system and distinctive frame for movement layout, can realize bimodal imaging simultaneously.
The present invention has following advantage and effect with respect to prior art:
(1) formation method of the present invention can obtain complementary high-contrast optoacoustic and fluorescent dual module attitude image on the principle, has effectively overcome the insufficient shortcoming of single image mode information, can be used for studying the tumor development mechanism research in the tumor growth process.
(2) imaging system of the present invention structurally organically is integrated into a system with photoacoustic imaging and fluorescence imaging, can carry out optoacoustic and fluorescence imaging synchronously, and is easy to use.
(3) light harvesting acoustic imaging of the present invention and fluorescence imaging are the common scanning light paths in the critical component of one, help the information fusion and the image reconstruction location of bimodal imaging.
(4) cost of each assembly of imaging system of the present invention is lower, so the cost of single unit system is also relatively low, does not have particular restriction, is widely used.
Description of drawings
Fig. 1 is the frame diagram of the formation method of integrating photoacoustic of the present invention and fluorescent dual module attitude.
Fig. 2 is the structure chart of imaging system of the formation method of realization integrating photoacoustic of the present invention and fluorescent dual module attitude.
Fig. 3 is the structure chart of scanning light path of the present invention.
Fig. 4 is optoacoustic and the fluoroscopic image that embodiment 2 utilizes imaging system shown in Figure 2 to obtain:
(a) be single fluoroscopic image;
(b) be single photoacoustic image;
(c) be optoacoustic and fluorescence fusion image;
(d) be the laboratory sample photo.
The specific embodiment
The present invention is described in further detail below in conjunction with embodiment and accompanying drawing, but embodiments of the present invention are not limited thereto.
Embodiment 1
Fig. 1 is the frame diagram of the formation method of integrating photoacoustic of the present invention and fluorescent dual module attitude.Fig. 2 is a structure chart of realizing the imaging system of the described formation method of Fig. 1, is made up of photoacoustic imaging subsystem, fluorescence imaging subsystem and computer; In one, computer is connected with photoacoustic imaging subsystem, fluorescence imaging subsystem and scanning light path system respectively by the system integration of scanning light path for photoacoustic imaging subsystem and fluorescence imaging subsystem; Computer is the scan mode of gated sweep light path system not only, also handles the data of photoacoustic imaging subsystem and fluorescence imaging subsystem, merges optoacoustic and fluorescence information, reconstructs the image that has optoacoustic and fluorescent dual module attitude.
The process of photoacoustic imaging is (as shown in Figures 2 and 3): laser instrument 1-1 produces pulse laser as the photo-acoustic excitation light source, after 45 ° of mirror reflects, vertically enters scanning light path 3-2, focuses on scanning area 4-3; Pulse laser excites the generation photoacoustic signal, through acoustical coupling liquid 4-2, receive by detector of sound 1-2, send into signal amplifier 1-3, amplifying signal is realized A/D conversion and data acquisition by the high-speed signal acquisition card of computer 5 inside, and the signal of collection is stored into waits in the Computer Cache that image reconstruction software carries out image reconstruction.Wherein, laser instrument 1-1 select for use the Nd:YAG pumping the OPO laser instrument (Vibrant 532 I, Opotek, Carlsbad, Calif.), output optical maser wavelength is 532nm, 690～960nm, pulsewidth is 10ns, repetition rate is 10HZ.Detector of sound 1-2 is that (UK), the array element diameter is 1mm to unit non-focusing ultrasonic detector for Precision Acoustics Ltd, Dorchester, and sensitivity is 850nv/Pa, and bandwidth is 200～15MHz.The acquisition controlling program realizes that with LABVIEW software digital graphics signal is handled with MATLAB software and realized; Utilize low-angle filtered back projection to obtain the optoacoustic two dimensional image in the imaging.
The process of fluorescence imaging is (as shown in Figures 2 and 3): laser instrument 2-1 produces continuous laser as the light source of fluorescence excitation, enter scanning light path system 3-2 by excitation fiber 2-2, focus on scanning area 4-3, the fluorescence signal that excites generation is by receiving the fluorescence signal that optical fiber 2-3 receives the different scanning position, through beam split, a part enters avalanche photo diode (APD) 2-6, part fluorescence mating plate 2-4 optical filtering is after filtration received by photomultiplier tube (PMT) 2-5, after the data collecting card 2-7 image data signal sent into computer 5 buffer memorys, data collecting card can be positioned at the computer outside, also can be positioned at the inside of computer.Because the fluorescence signal that APD and PMT receive from same excitaton source, utilizes both proportionate relationships to proofread and correct and normalization the fluorescence signal that receives, to eliminate the error of being brought by excitation source shake, explorer response etc.
The integrated critical component of optoacoustic and the imaging of fluorescent dual module attitude is the common scanning light path.Scanning light path sketch map is seen Fig. 3.Wherein, control 3-1 is by the scan mode of computer 5 by LABVIEW control software control scanning light path 3-2, and optoacoustic is identical with the fluorescent scanning mode.Pulse laser is reflected into into light path by reflecting mirror 3-2-1 in the photoacoustic imaging system, incides two dimension shake mirror 3-2-3, utilizes achromat 3-2-4 to focus on scanning area 4-3 at last.Continuous laser by beam expanding lens 3-2-6 incident filter lens 3-2-2, utilizes achromat 3-2-4 to focus on scanning area 4-3 through launching fiber 2-2 at last in the fluorescence imaging system.
Application Example 1 described imaging system is carried out the optoacoustic and the two imagings of fluorescence of analog sample.
The lung adenocarcinoma cell of in advance 1 simulated blood vessel and YFP being stablized high expressed is embedded in organize (shown in Fig. 4 (d)) in the analog sample that an agar makes.Wherein simulated blood vessel is the venous blood that mouse is housed in the glass tubing.Laser instrument 1-1 in the experiment, the output pulse laser wavelength is 532nm, and pulsewidth is 10ns, and repetition rate is 10Hz, enters scanning light path system 3-2 by the light path adjustment, focuses on the scanning area 4-3 of testing sample; Pulse laser excites the photoacoustic signal of generation to pass through through acoustical coupling liquid (for water) 4-2, receive by detector of sound 1-2, send into signal amplifier 1-3, high-speed signal acquisition card by computer 5 inside is gathered, the gated sweep light path scans scanning area, utilize MATLAB software to realize image reconstruction to the signal of gathering, can obtain the photoacoustic image shown in Fig. 4 (b).Wherein be contained among the tank 4-1 through acoustical coupling liquid (for water) 4-2, scanning area 4-3 is on testing sample.Fig. 4 (b) has provided the horizontal tomographic map of sample drawing 4 (d), and can finely coincide with the sample photo.Method of the present invention as can be seen and imaging system can access the horizontal tomographic map of optoacoustic of high-resolution and high-contrast.During fluorescence imaging, laser instrument 2-1, the output excitation wavelength is 488nm, exciting light enters scanning light path system 3-2 by excitation fiber 2-2, focus on scanning area 4-3, the fluorescence signal that excites generation is by receiving the fluorescence signal that optical fiber 2-3 receives the different scanning position.The fluorescence mating plate 2-4 fluorescence obtain about 530nm that filters is after filtration received by photomultiplier tube (PMT) 2-5, after the data collecting card 2-7 image data signal is sent into computer 5 buffer memorys.Simultaneously, reception optical fiber 2-3 enters avalanche photo diode (APD) 2-6 with the fluorescence signal of gathering through 1% beam split and is used for the fluorescence signal that receives is proofreaied and correct and normalization, can obtain fluoroscopic image shown in Fig. 4 (a).Utilize computer 5 by image co-registration with rebuild software can be at the image that obtains optoacoustic and fluorescent dual module attitude in the same image simultaneously (shown in Fig. 4 (c)), thereby it is abundanter to obtain organizational information, more comprehensively.
The foregoing description is a preferred implementation of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (10)
1. the formation method of integrating photoacoustic and fluorescent dual module attitude is characterized in that comprising following steps:
(1) by the scanning light path system pulse laser focusing is arrived object under test, obtain exciting the photoacoustic signal of generation, receive by detector of sound again, send into signal amplifier, gather the photoacoustic signal data of amplifying;
(2) by the scanning light path system continuous laser is focused on object under test, obtain exciting the fluorescence signal of generation, receive optical fiber and receive fluorescence signal, filtering through optical filter is received by photomultiplier tube, gathers the fluorescence signal after receiving;
(3) fluorescence signal after the photoacoustic signal data of the amplification that step (1) is obtained by data processing software and the reception that step (2) obtains merges and rebuilds, and obtains the image of optoacoustic and fluorescent dual module attitude.
2. formation method according to claim 1 is characterized in that: the wavelength of described pulse laser is 400～2500nm.
3. formation method according to claim 1 is characterized in that: described detector of sound is a unit non-focusing detector, dominant frequency 20KHz～100MHz.
4. formation method according to claim 1 is characterized in that: the wavelength of described continuous laser is 200～900nm.
5. realize the imaging system of each described formation method of claim 1～4, it is characterized in that: described imaging system is made up of photoacoustic imaging subsystem, fluorescence imaging subsystem and computer; In one, computer is connected with photoacoustic imaging subsystem, fluorescence imaging subsystem and scanning light path system respectively by the system integration of scanning light path for photoacoustic imaging subsystem and fluorescence imaging subsystem.
6. imaging system according to claim 5 is characterized in that: described photoacoustic imaging subsystem comprises laser instrument, scanning light path system, detector of sound and signal amplifier; Laser instrument, scanning light path system, detector of sound are connected successively with signal amplifier.
7. imaging system according to claim 5 is characterized in that: described fluorescence imaging subsystem comprises laser instrument, scanning light path system, optical filter and photomultiplier tube; Laser instrument is connected by excitation fiber with the scanning light path system, and the scanning light path system is connected by receiving optical fiber with optical filter, and optical filter is connected with photomultiplier tube.
8. imaging system according to claim 6 is characterized in that:
Described excitation fiber is made of the multichannel multimode fibre;
Described reception optical fiber is made of the multichannel multimode fibre.
9. imaging system according to claim 6 is characterized in that: described fluorescence imaging subsystem also contains avalanche photodide, and avalanche photodide is connected by receiving optical fiber with the scanning light path system.
10. imaging system according to claim 3 is characterized in that: described scanning light path system contains control and scanning light path, and wherein: control is connected with computer; Scan light route such as lower member are formed: reflecting mirror, filter lens, beam expanding lens, two dimension shake mirror scanning device and achromat; Each parts is separate; The parts that relate to photoacoustic imaging have reflecting mirror, two dimension shake mirror scanning device and achromat, and the parts that relate to fluorescence imaging have filter lens, beam expanding lens, two dimension shake mirror scanning device and achromat; Reflecting mirror, two dimension shake mirror scanning device and achromat are arranged from top to bottom; With the horizontal line is benchmark, reflecting mirror is 45 ° of settings, shake mirror and reflecting mirror in the two dimension shake mirror scanning device are parallel relation, the achromat horizontal positioned, beam expanding lens vertically is positioned at two dimension shake mirror scanning device top, filter lens and beam expanding lens are positioned at same horizontal level, are 45 ° of settings, and wherein reflecting mirror and filter lens are dislocatedly distributed;
Described computer contains high-speed data acquisition card, scanning light path control software and image reconstruction software.
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