CN106248643B - A kind of system of the scanning laser optical tomography method of high spectral resolution - Google Patents
A kind of system of the scanning laser optical tomography method of high spectral resolution Download PDFInfo
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- CN106248643B CN106248643B CN201610561213.0A CN201610561213A CN106248643B CN 106248643 B CN106248643 B CN 106248643B CN 201610561213 A CN201610561213 A CN 201610561213A CN 106248643 B CN106248643 B CN 106248643B
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/27—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using photo-electric detection ; circuits for computing concentration
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N2021/1738—Optionally different kinds of measurements; Method being valid for different kinds of measurement
- G01N2021/174—Optionally different kinds of measurements; Method being valid for different kinds of measurement either absorption-reflection or emission-fluorescence
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N2021/6417—Spectrofluorimetric devices
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Abstract
The present invention provides the system of a kind of scanning laser optical tomography method of high spectral resolution, including laser, the first reflecting mirror (M1), the second reflecting mirror (M2), third reflecting mirror (M3), scanning galvanometer, turntable, the first lens (L1), the second lens (L2), the third lens (L3), the 4th lens (L4), fluorescence filter plate, dispersion element, the first detector (1), the second detector (2), third detector (3), computer.The present invention organically combines high spectral resolution imaging and scanning laser optical tomography, photon utilization efficiency is high, has the advantages that carry out sample the rapidly and efficiently imaging of various dimensions fluorescence imaging, the imaging precision isotropism of 3 D stereo, the advantages that imaging system is compact, easy to use.
Description
Technical field
The present invention relates to photonics and biomedical skewing mechanism design fields more particularly to a kind of photonics and biology to cure
Learn the field for the scanning laser optical tomography method intersected.
Background technique
Life science and medicinal pharmacology research are one of the maximum hot spots of development in science and technology of world and mankind's concern now.Biology
The new branch of subject that medical photonics intersect as photonics with biomedicine, is related to the energy that biosystem is discharged with form of photons
It measures, the structure of related biosystem entrained by photon detection process and photon from biosystem and functional information etc.
Various aspects.Since the technology has the advantages that many protrusions, such as specific good, high sensitivity, resolution ratio height, therefore become
The important research tool that modern life science and biomedical research are increasingly relied on for counsel..
Biomedical research increasingly increases to better understand vital movement, disease development even pharmaceutically-active mechanism
It is added in the demand for studying biological functional information in complete biosystem in high precision.And traditional nuclear magnetic resonance (MRI), X-ray
Computed tomography (X-ray CT) and Positron emission computed tomography (PET) although etc. technologies can obtain three
Dimension imaging, but imaging object primary limitation is in the biomedical sample of macroscopic view.They are not optical image technology, Wu Fali simultaneously
Marker distribution is imaged with technologies such as dyeing/fluorescent markers extremely common on biologic medical.In biomedical light
Imaging field is learned, traditional copolymerization coke/multi-photon optical image technology is limited by light field and sweep speed limits, and is typically only capable to three
Tie up the sample of scanning imagery tens to several hundred μm, it is difficult to meet biomedical research to better understand vital movement, disease
Development even pharmaceutically-active mechanism increasingly increases the need for studying biological functional information in high precision in complete biosystem
It asks.Therefore, imaging depth in mm-cm range and there is novel Jie of high spatial resolution (μm magnitude) to see three-dimensional optical imaging
Technology is studied once proposition, and just the extensive concern by world biologic medical optical imagery researcher, is advanced by leaps and bounds
Development, is one of the forward position focus of current biologic medical optical imagery research field.
Currently, the approach for realizing the sight three-dimensional optical imaging that is situated between mainly includes optical projection tomoscan (Optical
Projection Tomography, OPT), scanning laser optical tomography (Scanning Laser Optical
Tomography, SLOT), mating plate frequency microscope (Light Sheet Microscopy).SLOT technology is optical computer tomography
A kind of form under the scope of (Optical Computed Tomography) is scanned, is excited and is detected using single-point, utilize laser
Scanning obtains a series of relevant intensity projection images of angles to form projection imaging, is then based on Computerized three-dimensional recombination and obtains
To the 3 D stereo strength information of sample, can collecting sample absorbing/fluorescent.But it is currently limited to intensity imaging, and intensity is surveyed
The influence of many factors such as the distributed density for being easy stimulated luminescence intensity, sample quenching and fluorescent dye or protein is measured, usually
It is only used as observational measurement, is difficult to carry out effectively quantitative measurment to sample, it is even more impossible to carry out functional imaging.
Fluorescence itself has many reference amounts characteristic in fact, other than intensity described above, the ginseng such as fluorescence spectrum, service life and polarization
Number all contains biological function information abundant.Since fluorescence emission wavelengths are related to the level structure of fluorogen, fluorescence
Spectral measurement can distinguish different fluorogens in conjunction with optical imagery, and to the biochemical function information imaging of spatial discrimination.
Current fluorescence spectrum imaging technique both domestic and external combines microscopic three-dimensional imaging technique more, and (such as confocal microscopic image, multi-photon are aobvious
Micro- imaging even super-resolution fluorescence micro-imaging), and it is mainly used in microcosmic sample (such as cell, unimolecule), what is utilized is
Be scanned imaging in transverse direction (x, y) and axial (z), imaging region is less than even much smaller than several hundred μm, can not suitable for pair
Biologic medical, which is situated between, sees the research of (mm-cm) sample (integrated biological systems: organ, tissue, embryo and model animal etc.).In addition,
In conjunction with these three-dimensional optical imaging techniques all by the anisotropic limitation of spatial accuracy, easily lead to laterally and axially resolution ratio
The fuzzy puzzlement of information extraction caused by inconsistent.These various dimensions Imaging-PAMs are not easy simply to be transplanted to biologic medical Jie
Three-dimensional optical imaging is seen, wherein a serious technical bottleneck is exactly that two kinds of technologies combine the effective rate of utilization of rear excitation photon not
It is high.
According to problem above, for the present invention compared to previous traditional approach, efficiently quick, system compact, behaviour is imaged in the present invention
Facilitate, imaging precision isotropism, the spectrally resolved of 3 D stereo information can be carried out to sample, suitable for tying to sample
Structure and functional analysis.The spectral information of 3 D stereo can be especially provided completely biological tissue's systematic sample, and photon utilizes
Rate is high, and therefore, the present invention has the advantages that more in terms of spectral matching factor.
Summary of the invention
The technical problem to be solved by the present invention is how to choose the fluorescence spectrum resolution techniques combination scanning laser light of high speed
Tomography is learned to form height original technical principle and scheme, realizes efficiently the quickly needle of spatial accuracy isotropic
High spectral resolution is carried out to the 3 D stereo information of sample, and realizes this fluorescence various dimensions quantitative imaging technique and system.This
Technology and systems device can be widely applied to fundamental biological knowledge, immuno-biology, anatomical structure biology, brain science, medicine and grind
Study carefully and the fields such as drug development, is a kind of novel various dimensions measuring technique for complete bio medical system sample, can send out
Exhibition is the important research tool that modern life science and medicinal pharmacology are studied.
To solve the above problems, the present invention is the following technical schemes are provided: a kind of scanning laser optical layer of high spectral resolution
The system for analysing imaging method, including laser, reflecting mirror M1, reflecting mirror M2, reflecting mirror M3, scanning galvanometer, turntable, lens
L1, lens L2, lens L3, lens L4, fluorescence filter plate, dispersion element, detector 1, detector 2, detector 3, computer；
Detector 1, detector 2, detector 3 are connect by electric circuit with computer；
Computer is connect with scanning galvanometer and turntable；
Lens L3, lens L4, fluorescence filter plate form the imaging system that can choose a specific wavelength, fluorescence filtering
Piece is for choosing specific wavelength；
Laser issues laser, and by reflecting mirror M1, laser is squeezed into scanning galvanometer by reflecting mirror M2, passes through scanning galvanometer
Reflection is focused on the sample under turntable by lens L1 again；Laser passes through spy by laser remaining after sample absorbance
Device 3 is surveyed, computer is acquired according to instruction；Laser passes through the fluorescence generated after sample can be by semi-transparent semi-reflecting lens BS, partly
The half of fluorescence is passed through lens L3 by saturating semi-reflective mirror BS, and fluorescence filter plate, after lens L4 squeezes into detector 2, computer is according to finger
Order is acquired；The other half of fluorescence focuses on the resolution that dispersion element carries out spectral signal by lens L2, further according to calculating
The instruction of machine carries out the acquisition of spectral signal using detector 1；
Computer issue instruction triggers scanning galvanometer do fast two-dimensional transversal scanning, thus to sample under each angle into
Row scanning, and intensity projection images and the spectrally resolved information projected image of be absorbed light or fluorescence；
Computer issues instruction control triggering turntable step angle, repeats the above steps and finally obtains angle relevant one
The intensity projection images of range of absorbent light fluorescence and spectrally resolved information projected image；
After computer carries out data destructing to the information of acquisition, the projection signal of multi-angle is recycled to pass through linear projection's layer
Reestablishment imaging is analysed, the 3 D stereo strength information of sample and the EO-1 hyperion 3 D stereo information of sample finally can be obtained；
Wherein, computer issues instruction, realizes scan control, turntable control, detector triggering, spectrum assignment, signal
Acquisition and storage function；Computer is connect with detector 1, detector 2, detector 3, and by trigger circuit and scanning galvanometer and
Turntable connection.
Detailed description of the invention
Fig. 1 is system assumption diagram of the invention；
Fig. 2 is work flow diagram of the invention；
Fig. 3 is the system assumption diagram of another embodiment of the present invention；
Specific embodiment
In order to which technical problems, technical solutions and advantages to be solved are more clearly understood, tie below
Accompanying drawings and embodiments are closed, the present invention will be described in detail.It should be noted that specific embodiment described herein is only used
It to explain the present invention, is not intended to limit the present invention, the product for being able to achieve said function belongs to equivalent replacement and improvement, includes
Within protection scope of the present invention.The specific method is as follows:
Specific embodiment 1:
The invention discloses the systems of a kind of scanning laser optical tomography method of high spectral resolution.Utilize spot scan
Laser excitation sample obtain fluorescence, high light spectrum image-forming is carried out by the fluorescence that issues to single-point, is carried out spectrally resolved；Using sweeping
It retouches system and two-dimensional transversal scanning is carried out to sample, sample is rotated in conjunction with turntable, obtains the bloom under all angles
The two-dimensional projection image differentiated is composed, by carrying out recombination operation to the projected image under these all angles, is realized to sample three
Tie up the high spectral resolution of steric information.
The present invention relates to the scanning laser optical tomography method and system dresses that a kind of pair of sample carries out high spectral resolution
It sets, including laser, reflecting mirror M1, reflecting mirror M2, reflecting mirror M3, scanning galvanometer, turntable, lens L1, lens L2, lens
L3, lens L4, fluorescence filter plate, dispersion element, detector 1, detector 2, detector 3, computer；Detector 1, detector 2,
Detector 3 is connect by electric circuit with computer；Computer is connect with scanning galvanometer and turntable；It is lens L3, lens L4, glimmering
Light filter plate forms the imaging system that can choose a specific wavelength, and fluorescence filter plate is for choosing specific wavelength；
Imaging method includes: to obtain fluorescence signal on the laser light incident to sample of (1) spot scan；(2) pass through dispersion element
Receive the spectral information of single-point fluorescence with detector；(3) manipulation laser carries out two-dimensional transversal scanning；(4) manipulation turntable into
The rotation of row angle；(5) after computer carries out data destructing to spectral information, the projection signal of multi-angle is recycled to throw by straight line
Shadow tomography；Described device by laser, dispersion element, detector, turntable, laser scanning system, external trigger signal source,
Data collecting card, computer composition；Detector, data collecting card, computer are successively electrically connected；External trigger signal source and detection
Device, data collecting card connection, and connect by trigger circuit with laser scanning system and turntable；
Laser issues laser, and by reflecting mirror M1, laser is squeezed into scanning galvanometer by reflecting mirror M2, passes through scanning galvanometer
Reflection is focused on the sample under turntable by lens L1 again；Laser passes through spy by laser remaining after sample absorbance
Device 3 is surveyed, computer is acquired according to instruction；Laser, which passes through the absorption fluorescence generated after sample, can pass through semi-transparent semi-reflecting lens
The half for absorbing fluorescence is passed through lens L3 by BS, semi-transparent semi-reflecting lens BS, and fluorescence filter plate after lens L4 squeezes into detector 2, calculates
Machine is acquired according to instruction；The other half absorption fluorescence focuses on point that dispersion element carries out spectral signal by lens L2
It distinguishes, detector 1 is recycled to carry out the acquisition of spectral signal according to the instruction of computer；
Computer issue instruction triggers scanning galvanometer do fast two-dimensional transversal scanning, thus to sample under each angle into
Row scanning, and the intensity projection images for the fluorescence that is absorbed and spectrally resolved information projected image；
Computer issues instruction control triggering turntable step angle, repeats the above steps and finally obtains angle relevant one
The intensity projection images of range of absorbent fluorescence and spectrally resolved information projected image；
After computer carries out data destructing to the information of acquisition, the projection signal of multi-angle is recycled to pass through linear projection's layer
Reestablishment imaging is analysed, the 3 D stereo strength information of sample and the 3 D stereo information of EO-1 hyperion finally can be obtained；
Wherein, computer issues instruction, realizes scan control, turntable control, detector triggering, spectrum assignment, signal
Acquisition and storage function；Computer is connect with detector 1, detector 2, detector 3, and by trigger circuit and scanning galvanometer and
Turntable connection.
Efficiently quick, system compact is imaged in the present invention, and easy to operate, imaging precision isotropism can carry out three to sample
The spectrally resolved of steric information is tieed up, is suitable for carrying out structure and function analysis to sample.It especially can be to completely biological tissue
Systematic sample provides the spectral information of 3 D stereo, and photon utilization rate is high, almost without phototoxicity and photobleaching, while can be to structure
Information and functional information detect, and can be used as the important research tool of fundamental biological knowledge medicinal pharmacology research, such as research form
Development and embry ogenesis, allelotaxis, anatomical structure tissue morphology, the architectural characteristic and functional characteristic of plant and marine organisms
Deng.
Specific embodiment 2:
Such as the workflow of Fig. 2 and the structural schematic diagram of Fig. 3, this device combination SLOT projection imaging system and spectrally resolved
System controls trigger circuit, scanning galvanometer, turntable and each detector by computer, to realize dependency number
According to acquisition.The laser generates the laser of spot scan, by reflecting mirror M1, reflecting mirror M2, scanning galvanometer, is incident on turntable
On sample on, obtain fluorescence signal；Fluorescence signal carries out spectrally resolved, letter of the generation with spectrum analysis by dispersion element
Number, computer allows detector 1 to acquire the signal for receiving fluorescence signal generation and having spectrum analysis by instruction；
Computer operating scanning galvanometer carries out two-dimensional transversal scanning；
Computer operating turntable carries out angle rotation；
After computer carries out data destructing to spectral information, the projection signal of multi-angle is recycled to chromatograph by linear projection
Imaging；
Wherein detector 2 is fluorescence intensity detector, and detector 3 is absorption intensity detector, this two-way optics can basis
Demand is added or cancels.
Design advantage mainly has: the present invention organically combines high spectral resolution imaging and scanning laser optical tomography, light
Sub- utilization efficiency is high, has the advantages that carry out sample the rapidly and efficiently imaging of various dimensions fluorescence imaging, the imaging of 3 D stereo
The advantages that precision isotropism, imaging system is compact, easy to use.
Claims (1)
1. the system of a kind of scanning laser optical tomography method of high spectral resolution, it is characterised in that: including laser,
One reflecting mirror (M1), the second reflecting mirror (M2), third reflecting mirror (M3), scanning galvanometer, turntable, the first lens (L1), second
Lens (L2), the third lens (L3), the 4th lens (L4), fluorescence filter plate, dispersion element, the first detector (1), the second detection
Device (2), third detector (3), computer；
First detector (1), second detector (2), the third detector (3) pass through electric circuit and the meter
The connection of calculation machine；
The computer is connect with the scanning galvanometer and the turntable；
The third lens (L3), the 4th lens (L4), fluorescence filter plate composition one can choose a certain wave
Long imaging system, the fluorescence filter plate is for choosing specific wavelength；
System includes following procedure: the laser issues laser, passes through first reflecting mirror (M1), second reflecting mirror
(M2) laser is squeezed into the scanning galvanometer, first lens (L1) is passed through by the reflection of the scanning galvanometer again and are focused on
On the sample under the turntable；The laser passes through third detector by remaining laser after the sample absorbance
(3), the computer is acquired according to instruction；The laser passes through the fluorescence generated after sample and passes through semi-transparent semi-reflecting lens
(BS), the half of the fluorescence is passed through the third lens (L3), the fluorescence filter plate, institute by the semi-transparent semi-reflecting lens (BS)
It states after the 4th lens (L4) squeeze into second detector (2), the computer is acquired according to instruction；The fluorescence it is another
Half focuses on the resolution that the dispersion element carries out spectral signal by second lens (L2), further according to the computer
Instruction using first detector (1) carry out spectral signal acquisition；
The computer issues scanning galvanometer described in instruction triggers and does fast two-dimensional transversal scanning, thus to the sample each
It is scanned under angle, and intensity projection images and the spectrally resolved information projected image of be absorbed light or fluorescence；
The computer issues instruction control triggering turntable step angle, repeats above procedure and finally obtains angle relevant one
Series intensity projection images and the spectrally resolved information projected image for absorbing light or fluorescence；
After the computer carries out data destructing to the information of acquisition, the projection signal of multi-angle is recycled to pass through linear projection's layer
Reestablishment imaging is analysed, the 3 D stereo strength information of sample and the EO-1 hyperion 3 D stereo information of sample finally can be obtained；
Wherein, the computer issues instruction, realizes scan control, turntable control, detector triggering, spectrum assignment, signal
Acquisition and storage function；The computer and first detector (1), second detector (2), the third detector
(3) it connects, and is connect by trigger circuit with the scanning galvanometer and the turntable.
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| CN106846296A (en) * | 2016-12-19 | 2017-06-13 | 深圳大学 | A kind of cell image tracks intelligent algorithm |
| CN107272014B (en) * | 2017-08-05 | 2023-12-12 | 广州市杜格科技有限公司 | Solid two-dimensional scanning laser radar and scanning method thereof |
| CN108836275A (en) * | 2018-07-10 | 2018-11-20 | 天津工业大学 | A kind of fluorescent molecular tomography system under free space |
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| US20110261367A1 (en) * | 2009-04-27 | 2011-10-27 | Gmitro Arthur F | Integrated Confocal and Spectral-Domain Optical Coherence Tomography Microscope |
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| US4329026A (en) * | 1978-03-22 | 1982-05-11 | Autologic, S.A. | Photocomposing machine and method |
| CN1276525A (en) * | 2000-07-07 | 2000-12-13 | 清华大学 | Parallel confocal detector based on Darman raster |
| CN102621116A (en) * | 2012-03-08 | 2012-08-01 | 华南农业大学 | Detection device and method for quality evaluation of American ginseng |
| CN103163111A (en) * | 2013-02-25 | 2013-06-19 | 天津大学 | Early stage cervical carcinoma detection system integrating fluorescent mesoscope imaging and optical coherence tomography (OCT) |
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Effective date of registration: 20190829 Address after: 518000 Lantian Road 3002, Shijing Street, Pingshan District, Shenzhen City, Guangdong Province Patentee after: Shenzhen Technical University Address before: Room 611, Science and Technology Building, Shenzhen University, 3688 Nanhai Avenue, Nanshan District, Shenzhen City, Guangdong Province, 518000 Patentee before: Chen Lingling |