CN103389287B - A kind of high-resolution optics system being applicable to the surface imaging of live body liver - Google Patents
A kind of high-resolution optics system being applicable to the surface imaging of live body liver Download PDFInfo
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- CN103389287B CN103389287B CN201310298452.8A CN201310298452A CN103389287B CN 103389287 B CN103389287 B CN 103389287B CN 201310298452 A CN201310298452 A CN 201310298452A CN 103389287 B CN103389287 B CN 103389287B
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Abstract
The present invention relates to a kind of high-resolution optics system being applicable to the surface imaging of live body liver, comprise a Ti∶Sapphire laser locked mode femto-second laser, an acousto-optic modulator, an optical scanning device, a variable mirror, a microcobjective, an objective table, a dichroic beam splitter, a near infrared filter plate, a reflection grating, a photomultiplier tube array, a computing machine controlled processing unit, semiconductor laser instrument and an imageing sensor.The present invention is reasonable in design, is skillfully constructed, and has vast potential for future development and larger dissemination.
Description
Technical field
The present invention relates to a kind of high-resolution optics system being applicable to the surface imaging of live body liver.
Background technology
As everyone knows, liver surface is mainly made up of extracellular matrix (collagenous fibres and snapback fibre), liver cell and microcirculqtory system.These compositions have great importance to the support structure of liver and function.When liver is impaired or pathology time, also can there is corresponding change in these compositions, as the hyperplasia of extracellular matrix, liver cell form and metabolic change and Microvascular architecture and micro-blood flow change dynamically.In addition, because these changes all occur in micro-meter scale.Therefore can realize to these information such as composition and structure thereof micro-meter scale extraction and visual to liver physiological Study and liver disease diagnosis there is great value.
But, current image technology, such as traditional endoscope, x-ray imaging, magnetic resonance imaging, CT scan, ultrasonic imaging, positron emission tomography and optical coherent chromatographic imaging etc., because its precision and resolution can't reach micro-meter scale, thus the detection to these changes cannot be realized.
In recent years, multi-photon imaging technique, owing to can realize biological tissue at the information integration of micro-meter scale and visual, shows huge potentiality in the application of live body high-resolution imaging.This technology utilizes laser and second_harmonic generation (the Second Harmonic Generation organizing inherent interaction between component to occur, and two-photon fluorescence excitation (Two-Photon Excited Fluorescence SHG), the nonlinear optical effect such as TPEF), reduction pyridine nucleotide (the nicotinamide adenine dinucleotide to cell source can be realized, NADH) and oxidation flavoprotein (falvin adenine dinucleotide, FAD), the collagenous fibres in extracellular matrix source and the high-resolution imaging of snapback fibre.Meanwhile, the fluorescence intensity ratio of two of cell source different TPEF signal in band (430-490 nm and 500-560 nm) can be used as the Endogenous index evaluating cell metabolism.In addition, laser speckle imaging technology can carry out the real-time whole audience imaging of high-spatial and temporal resolution to living body biological microcirculation blood flow.Owing to having noncontact, hurtless measure, the advantages such as fast imaging, laser speckle imaging technology is highly suitable for the measurement of blood microcirculation.Use laser speckle technique can measure blood vessels caliber, vessel density, the microcirculation parameter such as velocity of blood flow and blood perfusion.Can infer, laser speckle imaging technology also has very large application potential in the monitoring of the microcirculqtory system on live body liver surface.
Therefore, develop a kind of high-resolution optics system combined with laser speckle imaging technology based on multi-photon imaging technique, for realizing the live body liver surface information such as principal ingredient and structure thereof in the extraction of micro-meter scale with visually provide new technology, liver physiological Study and liver disease diagnosis can be had great importance.
Summary of the invention
In view of this, the object of this invention is to provide a kind of high-resolution optics system being applicable to the surface imaging of live body liver.
The present invention adopts following scheme to realize: a kind of high-resolution optics system being applicable to the surface imaging of live body liver, comprise a Ti∶Sapphire laser locked mode femto-second laser, it is characterized in that: the near infrared ultrashort pulse light that described Ti∶Sapphire laser locked mode femto-second laser inspires carries out power attenuation through an acousto-optic modulator, again through a dichroic beam splitter, arrived the live body liver surface on objective table by a microcobjective after one optical scanning device and a variable mirror, excite and produce endogenous non-linearity luminous signs, described light signal is through described microcobjective, variable mirror and optical scanning device arrive a dichroic beam splitter, a reflection grating is incided again through the isolated light signal of a near infrared filter plate, light signal is separated by different-waveband described reflection grating and the photomultiplier tube array that leads detects, and the signal detected is inputted a computing machine controlled processing unit, realize the imaging of different-waveband endogenous non-linearity luminous signs, then, the live body liver providing semiconductor laser instrument to incide on described objective table is surperficial, produce a scattered light signal, described scattered light signal detects through described microcobjective and the variable mirror imageing sensor that leads, and the signal detected is inputted described computing machine controlled processing unit, realize laser speckle blood current imaging.
In an embodiment of the present invention, described semiconductor laser incides on described objective table with level 30 °.
In an embodiment of the present invention, described variable mirror can realize the switching of catoptron and empty mirror.
In an embodiment of the present invention, described Ti∶Sapphire laser locked mode femto-second laser is the ultrashort pulse laser of high repetition frequency, and frequency reaches 84 MHz, and ultrashort pulse is 10 fs, and wavelength coverage is 730-980 nm, and output power is 1.8 W.
In an embodiment of the present invention, described reflection grating is a high-quality reflection grating, plays a point light action, light signal can be separated by different-waveband, interval 6 nm.
In an embodiment of the present invention, described photomultiplier tube array is made up of 30 photomultipliers, the wavelength coverage 380-560 nm of detection.
In an embodiment of the present invention, described semiconductor laser is mini semiconductor laser, wavelength 650nm, and output power is 30 mW.
Remarkable advantage of the present invention is: utilize the inherent interaction between component of femtosecond laser and liver surface to produce endogenic non linear optical signal to NADH and FAD of cell source, the collagenous fibres in extracellular matrix source and the high-resolution imaging of snapback fibre, realizes hepatocellular form and metabolism, the extraction of the change information such as 26S Proteasome Structure and Function of extracellular matrix and visual; Utilize semiconductor laser to incide liver surface and the scattered light signal that produces to the imaging of liver microcirculqtory system, thus realize the Microvascular architecture of liver surface and the monitoring of micro-blood flow multidate information and visual; The optical system combined with laser speckle imaging technology based on multi-photon imaging technique is that the high-resolution imaging realizing live body liver surface provides new method and new technology, has great value to liver physiological Study and liver disease diagnosis.The present invention is reasonable in design, is skillfully constructed, and has vast potential for future development and larger dissemination.
For making object of the present invention, technical scheme and advantage clearly understand, below by specific embodiment and relevant drawings, the present invention will be described in further detail.
Accompanying drawing explanation
Fig. 1 is system architecture schematic diagram of the present invention.
Embodiment
The present invention utilizes the inherent interaction between component of femtosecond laser and liver surface to produce endogenic non linear optical signal to NADH and FAD of cell source, the collagenous fibres in extracellular matrix source and the high-resolution imaging of snapback fibre, realize hepatocellular form and metabolism, the extraction of the change information such as 26S Proteasome Structure and Function of extracellular matrix and visual, then utilize semiconductor laser to incide liver surface and the scattered light signal that produces to the imaging of liver microcirculqtory system, thus to realize the Microvascular architecture of liver surface and the monitoring of micro-blood flow multidate information and visual.
As shown in Figure 1, the invention provides a kind of high-resolution optics system being applicable to the surface imaging of live body liver, comprise a Ti∶Sapphire laser locked mode femto-second laser 1, the near infrared ultrashort pulse light that described Ti∶Sapphire laser locked mode femto-second laser 1 inspires carries out power attenuation through an acousto-optic modulator 2, again through a dichroic beam splitter 7, arrived the live body liver surface on objective table 6 by a microcobjective 5 after one optical scanning device 3 and a variable mirror 4, excite and produce endogenous non-linearity luminous signs, described light signal is through described microcobjective 5, variable mirror 4 and optical scanning device 3 arrive a dichroic beam splitter 7, a reflection grating 9 is incided again through the isolated light signal of a near infrared filter plate 8, light signal is separated by different-waveband described reflection grating 9 and the photomultiplier tube array 10 that leads detects, and the signal detected is inputted a computing machine controlled processing unit 15, realize different-waveband (SHG:390-410 nm, TPEF:430-490 nm and TPEF:500-560 nm) imaging 11 of endogenous non-linearity luminous signs, thus NADH and FAD realized cell source, the collagenous fibres in extracellular matrix source and the high-resolution imaging of snapback fibre, liver cell form and metabolism can be extracted by graphical analysis, the information such as collagenous fibres and snapback fibre fine structure, then, the live body liver providing semiconductor laser instrument 12 to incide on described objective table 6 is surperficial, produce a scattered light signal, described scattered light signal to lead an imageing sensor 13(CCD through described microcobjective 5 and variable mirror 4) detect, and the signal detected is inputted described computing machine controlled processing unit, realize laser speckle blood current imaging 14, the microcirculation information such as blood vessels caliber, vessel density, velocity of blood flow and blood perfusion can be extracted by graphical analysis.
Preferably, described semiconductor laser incides on described objective table with level 30 °; Described Ti∶Sapphire laser locked mode femto-second laser is the ultrashort pulse laser of high repetition frequency, and frequency reaches 84 MHz, and ultrashort pulse is 10 fs, and wavelength coverage is 730-980 nm, and output power is 1.8 W; Described reflection grating is a high-quality reflection grating, plays a point light action, light signal can be separated by different-waveband, interval 6 nm; Described photomultiplier tube array is made up of 30 photomultipliers, the wavelength coverage 380-560 nm of detection; Described semiconductor laser is mini semiconductor laser, wavelength 650nm, and output power is 30 mW.
It is worth mentioning that, described variable mirror is a mirror with switching effect, it can make light produce reflection or transmission, catoptron and empty mirror (i.e. diaphotoscope can be realized, light can directly be propagated through empty mirror) switching, the near infrared ultrashort pulse light sent when Ti∶Sapphire laser locked mode femto-second laser by optical scanning device after, variable mirror switches to catoptron to be beneficial near infrared ultrashort pulse light to incide in microcobjective, when scattered light signal is by after microcobjective, variable mirror switches to sky mirror to be beneficial to scattered light signal and incides in imageing sensor.
Above-listed preferred embodiment; the object, technical solutions and advantages of the present invention are further described; be understood that; the foregoing is only preferred embodiment of the present invention; not in order to limit the present invention; within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (7)
1. one kind is applicable to the high-resolution optics system of live body liver surface imaging, comprise a Ti∶Sapphire laser locked mode femto-second laser, it is characterized in that: the near infrared ultrashort pulse light that described Ti∶Sapphire laser locked mode femto-second laser inspires carries out power attenuation through an acousto-optic modulator, again through a dichroic beam splitter, arrived the live body liver surface on objective table by a microcobjective after one optical scanning device and a variable mirror, excite and produce endogenous non-linearity luminous signs, described light signal is through described microcobjective, variable mirror and optical scanning device arrive a dichroic beam splitter, a reflection grating is incided again through the isolated light signal of a near infrared filter plate, light signal is separated by different-waveband described reflection grating and the photomultiplier tube array that leads detects, and the signal detected is inputted a computing machine controlled processing unit, realize the imaging of different-waveband endogenous non-linearity luminous signs, then, the live body liver providing semiconductor laser instrument to incide on described objective table is surperficial, produce a scattered light signal, described scattered light signal detects through described microcobjective and the variable mirror imageing sensor that leads, and the signal detected is inputted described computing machine controlled processing unit, realize laser speckle blood current imaging.
2. a kind of high-resolution optics system being applicable to the surface imaging of live body liver according to claim 1, is characterized in that: described semiconductor laser incides on described objective table with level 30 °.
3. a kind of high-resolution optics system being applicable to the surface imaging of live body liver according to claim 1, is characterized in that: described variable mirror can realize the switching of catoptron and empty mirror.
4. a kind of high-resolution optics system being applicable to the surface imaging of live body liver according to claim 1, it is characterized in that: described Ti∶Sapphire laser locked mode femto-second laser is the ultrashort pulse laser of high repetition frequency, frequency reaches 84 MHz, ultrashort pulse is 10 fs, wavelength coverage is 730-980 nm, and output power is 1.8 W.
5. a kind of high-resolution optics system being applicable to the surface imaging of live body liver according to claim 1, it is characterized in that: described reflection grating is a high-quality reflection grating, play a point light action, light signal can be separated by different-waveband, interval 6 nm.
6. a kind of high-resolution optics system being applicable to the surface imaging of live body liver according to claim 1, is characterized in that: described photomultiplier tube array is made up of 30 photomultipliers, the wavelength coverage 380-560 nm of detection.
7. a kind of high-resolution optics system being applicable to the surface imaging of live body liver according to claim 1, it is characterized in that: described semiconductor laser is mini semiconductor laser, wavelength 650nm, output power is 30 mW.
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| US9657290B2 (en) | 2012-07-03 | 2017-05-23 | The Board Of Trustees Of The Leland Stanford Junior University | Scalable bio-element analysis |
| ES2712200T3 (en) * | 2014-12-23 | 2019-05-09 | Max Planck Gesellschaft | Method for measuring a spectral sample response |
| CN107688014B (en) * | 2016-09-21 | 2020-06-30 | 北京大学 | Cell imaging method |
| WO2018089953A1 (en) * | 2016-11-14 | 2018-05-17 | Orca Biosystems, Inc. | Methods and apparatuses for sorting target particles |
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| CN101548153A (en) * | 2006-05-12 | 2009-09-30 | 西北大学 | Systems, methods, and apparatuses of low-coherence enhanced backscattering spectroscopy |
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