CN105873501A - A fluorescent scattering optical imaging system and method - Google Patents
A fluorescent scattering optical imaging system and method Download PDFInfo
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- CN105873501A CN105873501A CN201680000101.6A CN201680000101A CN105873501A CN 105873501 A CN105873501 A CN 105873501A CN 201680000101 A CN201680000101 A CN 201680000101A CN 105873501 A CN105873501 A CN 105873501A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
- A61B5/0071—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence by measuring fluorescence emission
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
- A61B5/0073—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence by tomography, i.e. reconstruction of 3D images from 2D projections
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/48—Other medical applications
- A61B5/4887—Locating particular structures in or on the body
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B2503/00—Evaluating a particular growth phase or type of persons or animals
- A61B2503/40—Animals
Abstract
The invention provides a fluorescent scattering optical imaging system and method. The system comprises a laser, a micro-displacement table, an object table, at least one planar reflective mirror, a filter, a CCD camera and a processor. An optical fiber head of the laser is placed on the micro-displacement table; the reflecting surface of the planar reflective mirror faces a to-be-tested object on the object table; the micro-displacement table and the CCD camera are electrically connected with the processor; the micro-displacement table moves in a two-dimensional plane according to control signals of the processor; the laser is used for scanning a to-be-tested area of the to-be-tested object containing fluorescent materials; the CCD camera obtains fluorescent images and laser images directly or based on the reflection of the planar reflective mirror; the processor acquires the position information of the CCD camera, the position information of the optical fiber head, the position information of the planar reflective mirror, the CT images or MRI images of the to-be-tested object, the fluorescent images and the laser images and generates a three-dimensional fluorescent image of the to-be-tested area. The imaging quality if improved and the systematic structure is simplified.
Description
Technical field
The present invention relates to medical imaging technology field, particularly relate to a kind of fluorescent scattering optical imaging system and method.
Background technology
Fluorescent scattering optical sectioning imaging technology (Fluorescence Diffuse Optical Tomography, FDOT)
Operation principle be, in advance at toy et al. Ke tumor and corresponding targeting fluorometric reagent, use laser at toy
The a certain plane interscan of region, fluorometric reagent Stimulated Light excites, and launches near infrared light, passes through detector afterwards
Obtain the picture of exciting light, determine tumor position in animal body and distribution situation finally by three-dimensional reconstruction.With core
Nuclear magnetic resonance (Magnetic Resonance Imaging, MRI), Computed tomography (Computed
Tomography, CT), positron emission tomography (PET) (Positron Emission Tomography, PET) compares, FDOT
Imaging has cost operation low, easy, the advantage such as radiationless, is usually used in small animal living body imaging.
Now there are some researches show that the FDOT/PET bimodal imaging that FDOT with PET combines can provide test articles not
Same physiological function process relevant information.This combination can help to improve puts, in preset time, the molecular level provided
Information.Relatively both modalities which imaging system characteristic, PET system stand can gather signal from each possible angle recordings.
But, this FDOT/PET bimodal imaging system structure is complicated, and cost is high.
In the FDOT system of existing single angle imaging, CCD (Charge-coupled Device, charge coupled cell)
Camera is integrally fixed at the top of determinand, and excitation source performs flat scanning at the right opposite of CCD camera, thus existing
There is common FDOT system in single angle imaging acquisition structure.Compared with PET, the FDOT system of single angle imaging
The collection mode of system is few, gathers quantity of information few, and collection period is long, and it is bigger to rebuild difficulty.It addition, single angle acquisition
Image makes FDOT imaging be longitudinal tomography imaging in poor quality along light source-CCD camera line direction.In normal conditions
Under, the geometry framework meeting of image capturing system and then the reconstruction quality of impact output image.
Summary of the invention
The present invention provides a kind of fluorescent scattering optical imaging system and method, to improve fluorescent scattering optical sectioning imaging matter
Amount, shortens imaging time.
The present invention provides a kind of fluorescent scattering optical imaging system, including: laser instrument, micro-displacement platform, object stage, extremely
A few plane mirror, optical filter, CCD camera and processor；The optical fiber head of described laser instrument is mounted in described
On micro-displacement platform；The reflecting surface of described plane mirror is towards the determinand on described object stage；Described micro-displacement platform and
Described CCD camera electrically connects with described processor respectively；Described micro-displacement platform is for the control according to described processor
Signal moves in the setting plane domain below described object stage；Described laser instrument is for scanning built-in fluorescent material
The district to be measured of described determinand is to inspire fluorescence；Described CCD camera is for obtaining fluorescence above described object stage
Image and laser image, acquisition mode includes: directly from determinand collection and reflection based on described plane mirror
Gather；Described processor is for obtaining the positional information of CCD camera, the positional information of optical fiber head, plane mirror
Positional information, the CT image of determinand or MRI image, described fluoroscopic image and laser image, and take this raw
Become the three-dimensional fluorescence image in described district to be measured.
In one embodiment, an edge of described plane mirror fits on described object stage.
In one embodiment, described system includes two described plane mirrors；Two described plane mirrors each with
The limit of described object stage laminating is parallel to each other, and the corner dimension phase of two described plane mirrors and described object stage
With.
In one embodiment, described optical filter includes the fluorescent optical filter for filtering fluorescence and for filtering swashing of laser
Light optical filter；Described fluorescent optical filter is 488nm narrow-band pass filter, and described Excitation Filter with High is more than 600nm
Long pass filter sheet.
The present invention also provides for a kind of fluorescent scattering optical imaging method, including: micro-displacement platform is believed according to the control of processor
Move in number driving the optical fiber head setting plane domain below object stage carrying laser instrument thereon；Described laser instrument
The district to be measured of determinand is carried out two dimensional laser scanning to induce the fluorescent material in described district to be measured to send fluorescence；CCD
Camera gathers composite fluorescence image and recombination laser image above described object stage, and acquisition mode includes: directly from treating
Survey thing is acquired and reflection based on described plane mirror is acquired；Processor obtains the position of CCD camera
Confidence breath, the CT image of the positional information of optical fiber head, the positional information of plane mirror, determinand or MRI figure
Picture, described composite fluorescence image and recombination laser image, and take this to generate the three-dimensional fluorescence image in described district to be measured.
In one embodiment, the processor acquisition positional information of CCD camera, the positional information of optical fiber head, plane are anti-
The positional information of light microscopic, the CT image of determinand or MRI image, described composite fluorescence image and recombination laser figure
Picture, and take this to generate the three-dimensional fluorescence image in described district to be measured, including: by described recombination laser image and composite fluorescence
Image is cut into multiple single width laser image and multiple single width fluoroscopic image respectively；Position letter according to described optical fiber head
Breath, the positional information of described CCD camera, the positional information of described plane mirror, the CT of described determinand or
MRI image, described single width laser image and described single width fluoroscopic image, treat described in three-dimensional reconstruction Software Create
Survey the three-dimensional fluorescence image in district.
In one embodiment, the method also includes: arranges fluorescent optical filter before described CCD camera and filters described glimmering
The fluorescence that stimulative substance sends；CCD camera gathers recombination laser image above described object stage, including: described CCD
Camera directly gathers by described laser fiber head outgoing and through the laser of described determinand, generates the first laser figure
Picture, gathers through described determinand the laser that reflects through described plane mirror simultaneously, generates the second laser image,
Described first laser image and the second laser image constitute described recombination laser image.
In one embodiment, the method also includes: arranges Excitation Filter with High before described CCD camera and filters described sharp
The laser of light device optical fiber head outgoing；CCD camera gathers composite fluorescence image above described object stage, including: institute
State CCD camera and directly gather the fluorescence sent by the fluorescent material in described district to be measured, generate the first fluoroscopic image,
Gather the fluorescence of that the fluorescent material in described district to be measured sends and through described plane mirror reflection simultaneously, generate the
Two fluoroscopic images, described first fluoroscopic image and the second fluoroscopic image constitute described composite fluorescence image.
In one embodiment, according to the positional information of described optical fiber head, the positional information of described CCD camera, described
The positional information of plane mirror, the CT of described determinand or MRI image, described single width laser image and described
Single width fluoroscopic image, by the three-dimensional fluorescence image in district to be measured described in three-dimensional reconstruction Software Create, including: by body net
Lattice generate software and described CT image or MRI image are carried out stress and strain model, generate the body surface veil in described district to be measured
Lattice data；Positional information according to described CCD camera and the positional information of described plane mirror, utilize minute surface anti-
Penetrate the positional information of the CCD camera picture that principle is calculated in described plane mirror；Position by described optical fiber head
Information, the positional information of described CCD camera, the positional information of described CCD camera picture, described single width laser image,
In described single width fluoroscopic image and described surface grid data input extremely described three-dimensional reconstruction software, it is calculated described
Three-dimensional fluorescence image.
In one embodiment, by the positional information of described optical fiber head, the positional information of described CCD camera, described CCD
The positional information of camera picture, described single width laser image, described single width fluoroscopic image and described surface grid data are defeated
Enter to described three-dimensional reconstruction software, be calculated described three-dimensional fluorescence image, including: by described laser image and institute
State fluoroscopic image scaling and be matched to the actual size in described district to be measured；By the described laser image after scaling coupling, scaling
Described fluoroscopic image after coupling, the positional information of described optical fiber head, the positional information of described CCD camera, described
In the positional information of CCD camera picture and described surface grid data input extremely described three-dimensional reconstruction software, calculate
To described three-dimensional fluorescence image.
In one embodiment, fitting on described object stage of described plane mirror；By described recombination laser figure
Picture and composite fluorescence image are cut into multiple single width laser image and multiple single width fluoroscopic image respectively, including: along described
Described recombination laser image cutting-out is become multiple with the intersection of described object stage place plane by plane mirror place plane
Described single width laser image；Along the intersection of described plane mirror place plane and described object stage place plane by described
Composite fluorescence image cutting-out becomes multiple described single width fluoroscopic image.
The fluorescent scattering optical imaging system of the embodiment of the present invention and method, by plane mirror reflection laser and glimmering
Light, from being different from the angle acquisition of real CCD camera from determinand laser out and fluorescence, thus can obtain
The two-dimensional fluoroscopic image of more rich determinand and two-dimensional laser image information, can improve image reconstruction accuracy accordingly,
Improve the intensity of reconstruction signal, image quality can be obtained than existing single angle FDOT system imaging quality higher three
Dimension fluoroscopic image.The imaging system of the embodiment of the present invention only needs a real CCD camera just can realize multi-angle bat
Take the photograph, compared with the FDOT system of multi-angle imaging, there is the advantage that equipment manufacturing cost is low, and real CCD camera
With laser image and the fluoroscopic image that at least one CCD camera picture shoots determinand simultaneously, the one-tenth system of the present invention
Image taking speed is faster.
Accompanying drawing explanation
In order to be illustrated more clearly that the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
In having technology to describe, the required accompanying drawing used is briefly described, it should be apparent that, the accompanying drawing in describing below is only
It is some embodiments of the present invention, for those of ordinary skill in the art, in the premise not paying creative work
Under, it is also possible to other accompanying drawing is obtained according to these accompanying drawings.In the accompanying drawings:
Fig. 1 is the structural representation of the fluorescent scattering optical imaging system of one embodiment of the invention；
Fig. 2 is CCD camera schematic diagram of imaging in plane mirror in one embodiment of the invention；
Fig. 3 is the composite fluorescence image that generated of fluorescent scattering optical imaging system shown in Fig. 1 and cuts out schematic diagram；
Fig. 4 is that the position of one embodiment of the invention midplane illuminator arranges schematic diagram；
Fig. 5 is the schematic flow sheet of the fluorescent scattering optical imaging method of one embodiment of the invention；
Fig. 6 is the method flow schematic diagram of three-dimensional fluorescence image imaging in one embodiment of the invention；
Fig. 7 is the schematic flow sheet of the method that combination picture is cut in one embodiment of the invention single image；
Fig. 8 is the method flow schematic diagram of three-dimensional fluorescence image imaging in one embodiment of the invention；
Fig. 9 is the method flow schematic diagram carrying out three-dimensional fluorescence image imaging in one embodiment of the invention.
Detailed description of the invention
For making the purpose of the embodiment of the present invention, technical scheme and advantage clearer, below in conjunction with the accompanying drawings to this
Bright embodiment is described in further details.Here, the schematic description and description of the present invention is used for explaining the present invention,
But it is not as a limitation of the invention.
The FDOT system of existing single angle imaging has the advantages such as cost is low, radiationless, but exists and gather mode
Less, collection quantity of information is few, it is big to rebuild difficulty and becomes the shortcomings such as quality longitudinal direction deterioration.In order to utilize current FDOT system
Advantage, inventor is in view of the impact on its output image quality of the geometry framework of existing FDOT system, Jing Guochuan
The geometry framework of the FDOT system of existing single angle imaging is improved by the work of the property made, to ensure image taking speed
While improve image quality.
Fig. 1 is the structural representation of the fluorescent scattering optical imaging system of one embodiment of the invention.As it is shown in figure 1, this
The fluorescent scattering optical imaging system of inventive embodiments comprise the steps that laser instrument 101, micro-displacement platform 102, object stage 103,
At least one plane mirror 104, optical filter 105, CCD camera 106 and processor 107.Laser instrument 101
Optical fiber head is mounted on micro-displacement platform 102.The reflecting surface of plane mirror 104 is towards the determinand on object stage 103.
Micro-displacement platform 102 and CCD camera 103 electrically connect with processor 107 respectively.
Laser instrument 101 is for scanning the district to be measured of the determinand of built-in fluorescent material to inspire fluorescence.This determinand can
To be living small animal, this district to be measured can be tissue or the organ of toy, such as tumor region.This laser instrument
101 laser sent can be near-infrared laser.As it is shown in figure 1, laser launched from the bottom to top by laser instrument 101, should
The fluorescent material in determinand on induced with laser object stage 103 sends fluorescence, and this laser and this fluorescence all can be by CCD
Camera 106 receives.In other embodiments, laser instrument 101 can irradiate determinand from from top to bottom, correspondingly,
CCD camera 106 can be at the received down laser of determinand and fluorescence.
Micro-displacement platform 102 is used for the setting plane domain below object stage 103 of the control signal according to processor 107
Interior movement.Under the drive of micro-displacement platform 102, such as, optical fiber head is clamped on micro-displacement platform, laser instrument 101
Optical fiber head can move along set path in x/y plane, to carry out two dimensional laser scanning.Such as, laser scanning
Position can be to move a position along x-axis every a setpoint distance, move n times altogether, set every one along y-axis
Set a distance moves a position, moves n times altogether, forms (N+1) * (N+1) array of laser.The most such as, laser is swept
Retouch can be with a certain set point as the center of circle, circumferentially moves a position every a set angle, mobile
M time, form laser array.
Owing to above-mentioned laser and two kinds of light of fluorescence exist the most simultaneously, so individually gathering laser image or individually adopting
During collection fluoroscopic image, need to utilize optical filter 105 to be acquired by CCD camera 106 again after filtering unwanted light.
Optical filter 105 can include the fluorescent optical filter for filtering fluorescence and for filtering the Excitation Filter with High of laser.This fluorescence
Optical filter can be 488nm narrow-band pass filter, can only allow CCD camera 106 collect the light of 488nm, is suitable for
In the laser gathering respective wavelength.This Excitation Filter with High can be more than 600nm long pass filter sheet, can only allow CCD phase
Machine 106 collects the light of more than 600nm, and owing to, under the exciting of 488nm laser, determinand generally can be sent out
Go out the fluorescence of 600～700nm, so this more than 600nm long pass filter sheet can preferably gather fluorescence.
CCD camera 106 is for obtaining fluoroscopic image and laser image above object stage 103, and acquisition mode includes:
Directly from determinand collection and reflection collection based on plane mirror 104.Wherein, utilization is directly adopted from determinand
When the mode of collection gathers fluoroscopic image and laser image, laser and fluorescence are without plane mirror 104, specifically,
Laser through determinand is directly entered CCD camera 106, fluorescent material in determinand the fluorescence sent directly enters
Enter CCD camera 106.The fundamental difference of two kinds of acquisition modes be directly when determinand gathers from determinand out
Laser and fluorescence subsequent propagation to the path of CCD camera 106 be not changed, so the present invention does not limit directly
When determinand gathers image, between determinand and CCD camera 106, there is other elements not changing light path.
In one embodiment, CCD camera 106 can be electron multiplication CCD (Electron-Multiplying CCD,
EMCCD) camera or liquid refrigerating CCD, can have more image acquisition effect.
Processor 107 is for obtaining the position letter of the optical fiber head of the positional information of CCD camera 106, laser instrument 101
The CT image of determinand or MRI image on breath, the positional information of plane mirror 104, object stage 103, glimmering
Light image and laser image, and take this to generate the three-dimensional fluorescence image in above-mentioned district to be measured.Processor 107 can be various
The equipment of above-mentioned three-dimensional fluorescence image, such as computer can be calculated according to the information of above-mentioned input.Plane is reflective
The positional information of mirror 104 can be by inputting to processor 107 manually.Utilize the position of plane mirror 104
The positional information of information and CCD camera 106 can be calculated CCD camera 106 in plane mirror 104
The positional information of CCD camera picture, utilizes the positional information of CCD camera picture to may be used for being calculated treating of determinand
Survey the three-dimensional fluorescence image in district.On object stage 103, CT image or the MRI image of determinand can be determinands
The 3-D view in district to be measured, can collect beforehand through corresponding equipment.
Fig. 2 is CCD camera schematic diagram of imaging in plane mirror in one embodiment of the invention.As in figure 2 it is shown,
Plane mirror 104 for by laser and fluorescent reflection to real CCD camera 106, to form CCD camera picture
106a, 106b, and from be different from this real CCD camera 106 shooting angle CCD camera as 106a,
The shooting angle of 106b shoots two-dimensional laser image and two kinds of images of two-dimensional fluoroscopic image in district to be measured.Can be clear by Fig. 2
Chu get Zhi, CCD camera 106 can become a CCD camera picture in each plane mirror 104.It is being provided with
In the fluorescent scattering optical imaging system of multiple plane mirrors 104, can correspondingly become multiple CCD camera picture,
The district to be measured of each CCD camera picture determinand from object stage 103 gathers the angle of fluorescence and laser can be different, from
And at least treating from two different angles shootings can be obtained by above-mentioned CCD camera 106 and each CCD camera picture
Survey two-dimensional fluoroscopic image and the two-dimensional laser image in district.
As depicted in figs. 1 and 2, being provided with two plane mirrors 104 in this system, such structure is equivalent to
The CCD camera of three same optical properties is (virtual on the left of the real CCD camera in object top 106 and object
Virtual CCD camera 106a of CCD camera 106b and right side), send in three positions record object simultaneously
Fluorescence signal.Compared with existing multi-angle FDOT system, the multi-angle imaging system formed by plane mirror
While significantly simplied system structure, owing to remaining former single angle system structure, often the most still go for
The FDOT image acquisition scheme of rule, applied widely.
In the fluorescent scattering optical imaging system of the embodiment of the present invention can the real CCD camera of only one of which 106, as
This one, each two-dimensional fluoroscopic image can be on a width composite fluorescence image, and each two-dimensional laser image can be multiple at a width
Close on laser image.
Fig. 3 is the composite fluorescence image that generated of fluorescent scattering optical imaging system shown in Fig. 1 and cuts out schematic diagram.As
Shown in Fig. 1 and Fig. 3, the fluorescent scattering optical imaging system of this embodiment is provided with two plane mirrors 104,
By being arranged on two plane mirrors 106 of mice both sides, a width can be obtained and comprise three width two-dimensional fluoroscopic image
Composite fluorescence image 200.This composite fluorescence image 200 comprises, the top of mice (determinand) on object stage 103
CCD camera 106 photograph the fluoroscopic image 201 at mice top, and CCD camera 106 is reflective on two planes
CCD camera picture in mirror 106 photographs the fluoroscopic image 202 and 203 of mice side respectively.
Fig. 4 is that the position of one embodiment of the invention midplane illuminator arranges schematic diagram.As shown in Figure 4, plane is reflective
Mirror 104 can be arranged in many ways, can be located at various position, itself and the angle (such as angle α, β) of horizontal plane
Can be various angle, distance L between plane mirror 104 and determinand can be various numerical value, if plane
Illuminator 104 can be by the laser in the district to be measured from determinand and fluorescent reflection to CCD camera 106.
In one embodiment, plane mirror 104 can be arranged on object stage 103, specifically, plane can be made anti-
One edge of light microscopic 104 fits on object stage 103, consequently, it is possible to plane mirror 104 is arranged conveniently, and can
To collect the image of determinand side.
In one embodiment, this fluorescent scattering optical imaging system can include two plane mirrors 104.The two is put down
The limit that face illuminator 104 is each fitted with object stage 103 can be parallel to each other, and i.e. two plane mirror 104 places are put down
Face each intersection with object stage 103 place plane can be parallel to each other.The two plane mirror 104 and object stage
The corner dimension of 103 can be identical, i.e. at plane mirror 104 towards in the case of above-mentioned determinand, and the two plane
The corner dimension of illuminator 104 place plane and object stage 103 place plane can be identical.As it is shown in figure 1, two are put down
Face illuminator 104 is identical with the big I of the angle α of object stage 103 and β, in other embodiments, angle α and
The big I of β is different.Angle α, the size of β can be multiple value, and inventor is by calculating CCD camera 106
Plane mirror 104 finds after the shooting angle of CCD camera picture, it is preferred that the size of angle α, β exists
30 °～40 ° of scopes, consequently, it is possible to the image information of more determinand can be obtained.
The fluorescent scattering optical imaging system of the embodiment of the present invention, by plane mirror reflection laser and fluorescence, permissible
From being different from the angle acquisition of real CCD camera from determinand laser out and fluorescence, thus obtain more rich
The two-dimensional fluoroscopic image of determinand and two-dimensional laser image information, can obtain image quality accordingly than existing single angle
FDOT system imaging quality higher three-dimensional fluorescence image.The imaging system of the embodiment of the present invention only needs one truly
CCD camera just can realize multi-angled shooting, compared with the FDOT system of multi-angle imaging, has equipment manufacturing cost
Low advantage, and real CCD camera and at least one CCD camera picture shoot simultaneously determinand laser image and
Fluoroscopic image, the image taking speed of the one-tenth system of the present invention is faster.
Based on the inventive concept identical with the fluorescent scattering optical imaging system shown in Fig. 1, the embodiment of the present application also provides for
A kind of fluorescent scattering optical imaging method, as described in example below.Due to this fluorescent scattering optical imaging method solution
Certainly the principle of problem is similar to fluorescent scattering optical imaging system, and therefore the enforcement of this fluorescent scattering optical imaging method can
To see the enforcement of fluorescent scattering optical imaging system, repeat no more in place of repetition.
Fig. 5 is the schematic flow sheet of the fluorescent scattering optical imaging method of one embodiment of the invention.As it is shown in figure 5, this
The fluorescent scattering optical imaging method of inventive embodiments, it may include step:
S310: micro-displacement platform drives the optical fiber head carrying laser instrument thereon in object stage according to the control signal of processor
Move in the setting plane domain of lower section；
It is glimmering with induce in above-mentioned district to be measured that S320: above-mentioned laser instrument carries out two dimensional laser scanning to the district to be measured of determinand
Stimulative substance sends fluorescence；
S330:CCD camera gathers composite fluorescence image and recombination laser image, collection side above above-mentioned object stage
Formula includes: be directly acquired from determinand and reflection based on above-mentioned plane mirror is acquired；
S340: processor obtains the positional information of CCD camera, the positional information of optical fiber head, the position of plane mirror
Confidence breath, the CT image of determinand or MRI image, above-mentioned composite fluorescence image and recombination laser image, and nationality
This generates the three-dimensional fluorescence image in above-mentioned district to be measured.
In above-mentioned steps S310, under the drive of micro-displacement platform, such as, optical fiber head is clamped on micro-displacement platform,
The optical fiber head of laser instrument can move along set path, to carry out two dimensional laser scanning in setting two dimensional surface.Such as,
As it has been described above, the position of laser scanning can be to move a position along x-axis every a setpoint distance, move N altogether
Secondary, move a position along y-axis every a setpoint distance, move n times altogether, form (N+1) * (N+1) battle array of laser
Row.The most such as, laser scanning can be with a certain set point as the center of circle, circumferentially every a set angle
A mobile position, mobile M time, forms laser array.The laser that this laser instrument sends can be near-infrared laser.
In above-mentioned steps S320, this determinand can be living small animal, and this district to be measured can be the group of toy
Knit or organ, such as tumor region.Determinand is irradiated, in determinand for example with the laser that wavelength is 488nm
Fluorescent material can send the fluorescence of 600～700nm.
In above-mentioned steps S330, utilize and directly gather composite fluorescence image from the mode of determinand collection and be combined sharp
During light image, laser and fluorescence reflect without plane mirror, and specifically, the laser through determinand is directly entered
CCD camera, fluorescent material in determinand the fluorescence sent is directly entered CCD camera.The root of two kinds of acquisition modes
This difference be directly when determinand gathers from determinand laser out and fluorescence subsequent propagation to CCD camera
Path is not changed.Wherein, recombination laser image can include several two-dimensional laser images, can in composite fluorescence image
Including several two-dimensional fluoroscopic image, as it is shown on figure 3, composite fluorescence image 200 include three width two-dimensional fluoroscopic image 201,
202 and 203.
In above-mentioned steps S340, the CT image of this determinand or MRI image can be the districts to be measured of determinand
3-D view, can collect beforehand through corresponding equipment.
In the embodiment of the present invention, FDOT imaging system acquired image can be with CT image co-registration, or and MRI
Image co-registration, in order to the function image making FDOT be provided can compare with CT image or MRI image
With process during, by the space structure provided by CT image or MRI image more intuitively and accurately by
Present.
The fluorescent scattering optical imaging system of the embodiment of the present invention is by plane mirror reflection laser and fluorescence, Ke Yicong
It is different from the angle acquisition of real CCD camera from determinand laser out and fluorescence, thus obtains more rich treating
Survey two-dimensional fluoroscopic image and the two-dimensional laser image information of thing, image quality can be obtained accordingly than existing single angle
FDOT system imaging quality higher three-dimensional fluorescence image.
Fig. 6 is the method flow schematic diagram of three-dimensional fluorescence image imaging in one embodiment of the invention.As shown in Figure 6, exist
In above-mentioned steps S340, the processor acquisition positional information of CCD camera, the positional information of optical fiber head, plane are reflective
The positional information of mirror, the CT image of determinand or MRI image, above-mentioned composite fluorescence image and recombination laser image,
And the method taking this to generate the three-dimensional fluorescence image in above-mentioned district to be measured, it may include step:
S341: above-mentioned recombination laser image and composite fluorescence image are cut into respectively multiple single width laser image and multiple
Single width fluoroscopic image；
S342: reflective according to the positional information of above-mentioned optical fiber head, the positional information of above-mentioned CCD camera, above-mentioned plane
The positional information of mirror, the CT of above-mentioned determinand or MRI image, above-mentioned single width laser image and above-mentioned single width fluorescence
Image, by the three-dimensional fluorescence image in the above-mentioned district to be measured of three-dimensional reconstruction Software Create.
In above-mentioned steps S341, under the drive of micro-displacement platform, laser is equal when diverse location spot scan determinand
Collection ccd image, and save as laser image sequence and the fluoroscopy image sequence of laser excitation, then by above-mentioned compound
When laser image and composite fluorescence image are cut into multiple single width laser image and multiple single width fluoroscopic image respectively, the most right
Above-mentioned laser image sequence and fluoroscopy image sequence are cut out processing.Wherein, above-mentioned single width laser image and list
Width fluoroscopic image may each be the image in above-mentioned district to be measured.
In the present embodiment, recombination laser image and composite fluorescence image are cut into respectively multiple single width laser image and many
Individual single width fluoroscopic image, can only retain the image in district to be measured, when generating three-dimensional fluorescence image, and can be only for district to be measured
Image is rebuild, it is not necessary to rebuild non-targeted imaging region, when contributing to the reconstruction saving three-dimensional fluorescence image
Between, and then improve image taking speed.
Fig. 7 is the schematic flow sheet of the method that combination picture is cut in one embodiment of the invention single image.Above-mentioned
Plane mirror while fitting on above-mentioned object stage, now, as it is shown in fig. 7, in above-mentioned steps S341,
Above-mentioned recombination laser image and composite fluorescence image are cut into respectively multiple single width laser image and multiple single width fluorescence
The method of image, it may include step:
S3411: along above-mentioned plane mirror place plane with the intersection of above-mentioned object stage place plane by above-mentioned compound sharp
Light image is cut into multiple above-mentioned single width laser image；
S3412: along above-mentioned plane mirror place plane with the intersection of above-mentioned object stage place plane by above-mentioned compound glimmering
Light image is cut into multiple above-mentioned single width fluoroscopic image.
In the present embodiment, with the intersection of object stage place plane, combination picture is (compound along plane mirror place plane
Laser image, composite fluorescence image) it is cut into multiple single image (single width laser image, single width fluoroscopic image).
In one embodiment, as it is shown on figure 3, composite fluorescence image 200 is cut into three width lists along intersection 2021,2031
Width fluoroscopic image 201,202 and 203.Wherein, the cut out areas 2022 of single width fluoroscopic image 202 and single width fluorescence
The cut out areas 2032 of image 203 can be selected as required, such as, can be cut out square region as shown in Figure 3
2022,2032, then cut out the image-region corresponding to district to be measured obtaining determinand, or, only pass through single treatment
Just it is cut out fluoroscopic image and the laser image of the two dimension in district to be measured, specifically can optionally select.
In the present embodiment, by combination picture being cut along the intersection of plane mirror place plane with object stage place plane
It is cut into single image, complete single image can be readily derived, be difficult to occur cutting out error.
In other embodiments, the method shown in Fig. 7 is utilized to cut out the recombination laser image and composite fluorescence figure obtained
Picture, before being used for generating three-dimensional fluorescence image, it is also possible to is cut into above-mentioned to be measured by this single width laser image further
The laser image in district, is cut into the fluoroscopic image in this district to be measured by this single width fluoroscopic image, to reduce the data of processor
Treating capacity, improves the imaging time of three-dimensional fluorescence image.
In one embodiment, arrange before above-mentioned CCD camera that fluorescent optical filter filters that above-mentioned fluorescent material sends is glimmering
Light, such as, determinand is excited under 488nm, sends the fluorescence of 600-700nm, the optical filtering first placed
Sheet is the optical filter of 488nm narrow bandpass (passband 10nm), only allows CCD collect the light of 488nm.Now,
In above-mentioned steps S330, CCD camera gathers the method for recombination laser image above above-mentioned object stage, it may include
Step:
S331: above-mentioned CCD camera directly gathers by above-mentioned laser fiber head outgoing swashing through above-mentioned determinand
Light, generates the first laser image, gathers through above-mentioned determinand the laser that reflects through above-mentioned plane mirror simultaneously,
Generate the second laser image, above-mentioned first laser image and the second laser image and constitute above-mentioned recombination laser image.
In one embodiment, Excitation Filter with High is set before above-mentioned CCD camera and filters above-mentioned laser fiber head outgoing
Laser, specifically, such as, determinand is excited under 488nm, sends the fluorescence of 600-700nm, more
Changing optical filter is the long logical optical filter of more than 600nm, makes CCD camera collect fluoroscopic image.Now, CCD
Camera gathers the method for composite fluorescence image above above-mentioned object stage, it may include step:
S332: above-mentioned CCD camera directly gathers the fluorescence sent by the fluorescent material in above-mentioned district to be measured, generates the
One fluoroscopic image, gathers that the fluorescent material in above-mentioned district to be measured sends and through the reflection of above-mentioned plane mirror simultaneously
Fluorescence, generates the second fluoroscopic image, above-mentioned first fluoroscopic image and the second fluoroscopic image and constitutes above-mentioned composite fluorescence image.
Fig. 8 is the method flow schematic diagram of three-dimensional fluorescence image imaging in one embodiment of the invention.As shown in Figure 8, exist
In above-mentioned steps S342, according to the positional information of above-mentioned optical fiber head, the positional information of above-mentioned CCD camera, above-mentioned flat
The positional information of face illuminator, the CT of above-mentioned determinand or MRI image, above-mentioned single width laser image and above-mentioned list
Width fluoroscopic image, by the method for the three-dimensional fluorescence image in the above-mentioned district to be measured of three-dimensional reconstruction Software Create, it may include step:
S3421: generate software by volume mesh and above-mentioned CT image or MRI image are carried out stress and strain model, in generation
State the surface grid data in district to be measured；
S3422: according to positional information and the positional information of above-mentioned plane mirror of above-mentioned CCD camera, utilize minute surface
The positional information of the CCD camera picture that principle of reflection is calculated in above-mentioned plane mirror；
S3423: by the positional information of above-mentioned optical fiber head, the positional information of above-mentioned CCD camera, above-mentioned CCD camera
The positional information of picture, above-mentioned single width laser image, above-mentioned single width fluoroscopic image and above-mentioned surface grid data input extremely
In above-mentioned three-dimensional reconstruction software, it is calculated above-mentioned three-dimensional fluorescence image.
In above-mentioned steps S3421, it can be multiple different stress and strain model software that volume mesh generates software, such as
Iso2mesh software.In above-mentioned steps S3421, three-dimensional reconstruction software can be multiple different reconstruction software, such as
Toast software.
When generating three-dimensional fluorescence image utilizing three-dimensional reconstruction software to rebuild, first, utilize finite element benchmark iterative algorithm
Generating fluoroscopic image, wherein, the exciting light in district to be measured that describes of this benchmark iterative algorithm institute foundation is propagated and scattered light
Coupling diffusion equation is:
Wherein, r represents location variable, φxIt is the photon density of exciting light x, φmIt is the photon density of scattered light m,It is the diffusion coefficient of exciting light x,It it is diffusion system
Number,It is the absorptance of exciting light x,It is the absorptance of scattered light m,It it is the decay of exciting light x
Scattering coefficient,Being the scattering coefficient of the decay of scattered light m, a is the border correlation coefficient of internal reflection,
Sx(r)=S0δ(r-r0) it is the excitation source item of exciting light x point source, S0Represent the intensity of point source, δ (r-r0) it is with position r0
Point source centered by Dirac-delta function, η is edge-diffusion coefficient,Absorptance for exciting light
Or the absorptance of scattered light
The equation matrix such as formula (1) and (2) utilizing finite element discretization relation to obtain is shown, and obtaining one further is
Row are for solving the equation of inverse problem:
[Ax,m]{φx,m}={ bx,m, (3)
Wherein, matrix [Ax,m] parameter and column vector { bx,mBy one group of spatial variations, item in } can represent that glug is bright
Day basic function；Jx,mIt is by φx,mDerivant observe node on each border for the Jacobian matrix of χ；Δ χ is
Optics and fluorescent characteristic distributed update vector；I is unit matrix；λ can be a yardstick or diagonal matrix；It is
Matrix Jx,mTransposed matrix；χ is fluorescent characteristic distributing vector, have expressed Dx、Orφx,mIt is sharp
Luminous x or the photon density of scattered light m；It is the exciting light x or the photon density of scattered light m being observed；It is calculated exciting light x or the photon density of scattered light m.Laser image and fluoroscopic image are by formula (3)
～(5) solve formation iteratively, and from the probably uniformly preresearch estimates renewal optical fluorescence characteristic distribution of these attributes.
Fig. 9 is the method flow schematic diagram carrying out three-dimensional fluorescence image imaging in one embodiment of the invention.As it is shown in figure 9,
In above-mentioned steps S3423, by the positional information of above-mentioned optical fiber head, the positional information of above-mentioned CCD camera, above-mentioned
The positional information of CCD camera picture, above-mentioned single width laser image, above-mentioned single width fluoroscopic image and above-mentioned surface grid
In data input extremely above-mentioned three-dimensional reconstruction software, the method being calculated above-mentioned three-dimensional fluorescence image, it may include step:
S34231: above-mentioned laser image and above-mentioned fluoroscopic image are scaled the actual size being matched to above-mentioned district to be measured；
S34232: by the above-mentioned laser image after scaling coupling, the above-mentioned fluoroscopic image after scaling coupling, above-mentioned optical fiber
Positional information, the positional information of above-mentioned CCD camera, the positional information of above-mentioned CCD camera picture and the above-mentioned body surface of head
In surface grids data input extremely above-mentioned three-dimensional reconstruction software, it is calculated above-mentioned three-dimensional fluorescence image.
In one embodiment, fluorescent scattering optical imaging method includes step:
1) placing on object stage and need rebuilt object, object is contained within can inspiring under corresponding excitation source glimmering
The fluorescent material of light, adjusts the visual field of CCD camera to covering whole object；
2) transfer upper filter in CCD camera, filter the fluorescence that object sends.Such as object is under 488nm laser
Excited, sent the fluorescence that wavelength is 600-700nm.First the optical filter placed is that 488nm narrow bandpass is (logical
Band 10nm) optical filter, only allow CCD collect the light of 488nm.
3) operation two-dimensional micro-displacement platform, the program by finishing in processor that is allowed to moves, and i.e. moves the position of laser fiber head
Put, be allowed to launch laser in different positions；
4) on laser different scanning position, gather ccd image simultaneously, save as the image sequence of laser excitation；
5) change optical filter, such as object is excited under 488nm laser, when sending the fluorescence of 600-700nm,
Can change optical filter is the long logical optical filter of more than 600nm, makes CCD collect fluoroscopic image；
6) the recombination laser image and the composite fluorescence image that photographed CCD camera are cut out, front and both sides
Subject image is cut out out, and can be cut out with the intersection of plane mirror with horizontal plane for cut-off rule.
7) scan the CT image of object, and generated object by the software (such as iso2mesh) generating volume mesh
Surface volume mesh data grids mesh；
8) laser image and fluoroscopic image, and CCD camera (real CCD camera and CCD camera exist
Picture in plane mirror) and the positional information of lasing light emitter, the mesh information of object is believed as the input of reconstruction software
Breath, by three-dimensional reconstruction software (such as toast), generates the three-dimensional data comprising fluorescence distribution.
In step 8) in, this fluoroscopic image includes the fluorescence of fluoroscopic image and the side splitting the front of the object obtained
Image.Specifically cut out the pulmonary that step comprises the steps that certain region of interest ROI region, such as mice of selected object
(the most a size of 1.2cm*1.2cm), by all equal to the laser image after selected ROI region, fluoroscopic image and object
Actual size match, be then cut out ROI region, by three-dimensional reconstruction Software Create Jacobian matrix.
Positional information includes: (such as CCD camera positional information is " 12 14 40 0 0-1 " to the positional information of CCD, depends on
Secondary expression x-axis, y-axis, the coordinate of z-axis, unit is mm, and 0 0-1 represents that CCD is to gather downwards), laser
The positional information in source (such as lasing light emitter positional information is such as " 12 14-5 001 ", represent successively x-axis, y-axis,
The coordinate of z-axis, unit is mm, 001 represent lasing light emitters upwards excite), mesh information (i.e. surface grid three
Dimension coordinate, for example, 25.595,60.6565,20.565, represent the coordinate of x-axis, y-axis, z-axis respectively), pass through
Three-dimensional reconstruction software (such as call toastMapSolToMesh, toastSolutionMask, IWT2_P0,
FDOTAdj0p or tostQvec etc.) rebuild the distributed intelligence obtaining fluorescent material in object, i.e. three-dimensional fluorescence
Image.
The fluorescent scattering optical imaging method of the embodiment of the present invention, by plane mirror reflection laser and fluorescence, permissible
From being different from the angle acquisition of real CCD camera from determinand laser out and fluorescence, thus obtain more rich
The two-dimensional fluoroscopic image of determinand and two-dimensional laser image information, can improve image reconstruction accuracy accordingly, improves and rebuilds
The intensity of signal, can obtain image quality than existing single angle FDOT system imaging quality higher three-dimensional fluorescence figure
Picture.The imaging system of the embodiment of the present invention only needs a real CCD camera just can realize multi-angled shooting, with many
The FDOT system of angle imaging is compared, and has an advantage that equipment manufacturing cost is low, and real CCD camera and at least
Individual CCD camera picture shoots laser image and the fluoroscopic image of determinand simultaneously, the image taking speed of the one-tenth system of the present invention
Faster.
In the description of this specification, reference term " embodiment ", " specific embodiment ", " some realities
Execute example ", " such as ", " example ", the description of " concrete example " or " some examples " etc. mean to combine this embodiment
Or specific features, structure, material or the feature of example description is contained at least one embodiment or the example of the present invention
In.In this manual, the schematic representation to above-mentioned term is not necessarily referring to identical embodiment or example.And
And, the specific features of description, structure, material or feature can be in any one or more embodiments or example
Combine in an appropriate manner.
Those skilled in the art are it should be appreciated that embodiments of the invention can be provided as method, system or computer journey
Sequence product.Therefore, the present invention can use complete hardware embodiment, complete software implementation or combine software and hardware
The form of the embodiment of aspect.And, the present invention can use and wherein include computer available programs one or more
The computer-usable storage medium (including but not limited to disk memory, CD-ROM, optical memory etc.) of code
The form of the computer program of upper enforcement.
The present invention is with reference to method, equipment (system) and the stream of computer program according to embodiments of the present invention
Journey figure and/or block diagram describe.It should be understood that can be by computer program instructions flowchart and/or block diagram
Flow process in each flow process and/or square frame and flow chart and/or block diagram and/or the combination of square frame.These can be provided
Computer program instructions processes to general purpose computer, special-purpose computer, Embedded Processor or other programmable datas and sets
Standby processor is to produce a machine so that held by the processor of computer or other programmable data processing device
The instruction of row produces for realizing one flow process of flow chart or multiple flow process and/or one square frame of block diagram or multiple side
The device of the function specified in frame.
These computer program instructions may be alternatively stored in and computer or other programmable data processing device can be guided with spy
Determine in the computer-readable memory that mode works so that the instruction being stored in this computer-readable memory produces bag
Including the manufacture of command device, this command device realizes at one flow process of flow chart or multiple flow process and/or block diagram one
The function specified in individual square frame or multiple square frame.
These computer program instructions also can be loaded in computer or other programmable data processing device so that at meter
On calculation machine or other programmable devices, execution sequence of operations step is to produce computer implemented process, thus is calculating
The instruction performed on machine or other programmable devices provide for realizing in one flow process of flow chart or multiple flow process and/or
The step of the function specified in one square frame of block diagram or multiple square frame.
Particular embodiments described above, has been carried out the purpose of the present invention, technical scheme and beneficial effect the most in detail
Describe in detail bright, be it should be understood that the specific embodiment that the foregoing is only the present invention, be not used to limit this
Bright protection domain, all within the spirit and principles in the present invention, any modification, equivalent substitution and improvement etc. done,
Should be included within the scope of the present invention.
Claims (11)
1. a fluorescent scattering optical imaging system, it is characterised in that including: laser instrument, micro-displacement platform, loading
Platform, at least one plane mirror, optical filter, CCD camera and processor；
The optical fiber head of described laser instrument is mounted on described micro-displacement platform；The reflecting surface of described plane mirror is towards described
Determinand on object stage；Described micro-displacement platform and described CCD camera electrically connect with described processor respectively；
Described micro-displacement platform is used for the setting plane area below described object stage of the control signal according to described processor
Move in territory；
Described laser instrument is for scanning the district to be measured of the described determinand of built-in fluorescent material to inspire fluorescence；
Described CCD camera is for obtaining fluoroscopic image and laser image above described object stage, and acquisition mode includes:
Directly from determinand collection and reflection collection based on described plane mirror；
Described processor is for obtaining the positional information of CCD camera, the positional information of optical fiber head, plane mirror
Positional information, the CT image of determinand or MRI image, described fluoroscopic image and laser image, and take this to generate
The three-dimensional fluorescence image in described district to be measured.
2. fluorescent scattering optical imaging system as claimed in claim 1, it is characterised in that described plane mirror
An edge fit on described object stage.
3. fluorescent scattering optical imaging system as claimed in claim 2, it is characterised in that described system includes two
Individual described plane mirror；Two described plane mirrors each limit with the laminating of described object stage is parallel to each other, and two
Individual described plane mirror is identical with the corner dimension of described object stage.
4. fluorescent scattering optical imaging system as claimed in claim 1, it is characterised in that described optical filter includes
For filtering the fluorescent optical filter of fluorescence and for filtering the Excitation Filter with High of laser；Described fluorescent optical filter is 488nm
Narrow-band pass filter, described Excitation Filter with High is more than 600nm long pass filter sheet.
5. a fluorescent scattering optical imaging method, it is characterised in that including:
Micro-displacement platform drives the optical fiber head carrying laser instrument thereon below object stage according to the control signal of processor
Setting plane domain in move；
Described laser instrument carries out two dimensional laser scanning with the fluorescence in the described district to be measured of induction to the district to be measured of determinand
Matter sends fluorescence；
CCD camera gathers composite fluorescence image and recombination laser image above described object stage, and acquisition mode includes:
Directly it is acquired from determinand and reflection based on described plane mirror is acquired；
Processor obtain the positional information of CCD camera, the positional information of optical fiber head, the positional information of plane mirror,
The CT image of determinand or MRI image, described composite fluorescence image and recombination laser image, and take this to generate institute
State the three-dimensional fluorescence image in district to be measured.
6. fluorescent scattering optical imaging method as claimed in claim 5, it is characterised in that processor obtains CCD
The positional information of camera, the positional information of optical fiber head, the positional information of plane mirror, the CT image of determinand or
MRI image, described composite fluorescence image and recombination laser image, and take this to generate the three-dimensional fluorescence in described district to be measured
Image, including:
Described recombination laser image and composite fluorescence image are cut into multiple single width laser image and multiple single width respectively
Fluoroscopic image；
Positional information according to described optical fiber head, the positional information of described CCD camera, the position of described plane mirror
Confidence breath, the CT of described determinand or MRI image, described single width laser image and described single width fluoroscopic image,
Three-dimensional fluorescence image by district to be measured described in three-dimensional reconstruction Software Create.
7. fluorescent scattering optical imaging method as claimed in claim 5, it is characterised in that also include: described
Fluorescent optical filter is set before CCD camera and filters the fluorescence that described fluorescent material sends；CCD camera is from described object stage
Top gathers recombination laser image, including:
Described CCD camera directly gathers by described laser fiber head outgoing and through the laser of described determinand, raw
Become the first laser image, gather through described determinand the laser that reflects through described plane mirror simultaneously, generate the
Dual-laser image, described first laser image and the second laser image constitute described recombination laser image.
8. fluorescent scattering optical imaging method as claimed in claim 5, it is characterised in that also include: described
Excitation Filter with High is set before CCD camera and filters the laser of described laser fiber head outgoing；CCD camera is from described load
Composite fluorescence image is gathered above thing platform, including:
Described CCD camera directly gathers the fluorescence sent by the fluorescent material in described district to be measured, generates the first fluorescence
Image, gathers the fluorescence of that the fluorescent material in described district to be measured sends and through described plane mirror reflection simultaneously,
Generate the second fluoroscopic image, described first fluoroscopic image and the second fluoroscopic image and constitute described composite fluorescence image.
9. fluorescent scattering optical imaging method as claimed in claim 6, it is characterised in that according to described optical fiber head
Positional information, the positional information of described CCD camera, the positional information of described plane mirror, described determinand
CT or MRI image, described single width laser image and described single width fluoroscopic image, raw by three-dimensional reconstruction software
Become the three-dimensional fluorescence image in described district to be measured, including:
Generate software by volume mesh and described CT image or MRI image are carried out stress and strain model, generate described to be measured
The surface grid data in district；
Positional information according to described CCD camera and the positional information of described plane mirror, utilize direct reflection former
The positional information of the CCD camera picture that reason is calculated in described plane mirror；
By the positional information of described optical fiber head, the positional information of described CCD camera, the position of described CCD camera picture
Information, described single width laser image, described single width fluoroscopic image and the input of described surface grid data are to described three-dimensional
In reconstruction software, it is calculated described three-dimensional fluorescence image.
10. fluorescent scattering optical imaging method as claimed in claim 9, it is characterised in that by described optical fiber head
Positional information, the positional information of described CCD camera, the positional information of described CCD camera picture, described single width laser
In image, described single width fluoroscopic image and described surface grid data input extremely described three-dimensional reconstruction software, calculate
To described three-dimensional fluorescence image, including:
Described laser image and described fluoroscopic image are scaled the actual size being matched to described district to be measured；
By the described laser image after scaling coupling, the described fluoroscopic image after scaling coupling, the position of described optical fiber head
Information, the positional information of described CCD camera, the positional information of described CCD camera picture and described surface grid number
According in input to described three-dimensional reconstruction software, it is calculated described three-dimensional fluorescence image.
11. fluorescent scattering optical imaging methods as claimed in claim 6, it is characterised in that described plane mirror
While fitting on described object stage；
Described recombination laser image and composite fluorescence image are cut into multiple single width laser image and multiple single width respectively
Fluoroscopic image, including:
Along the intersection of described plane mirror place plane and described object stage place plane by described recombination laser image
It is cut into multiple described single width laser image；
Along the intersection of described plane mirror place plane and described object stage place plane by described composite fluorescence image
It is cut into multiple described single width fluoroscopic image.
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