CN109497959A - A kind of fluoroscopic imaging systems and method of quantitative detection photosensitizer spatial distribution - Google Patents
A kind of fluoroscopic imaging systems and method of quantitative detection photosensitizer spatial distribution Download PDFInfo
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- 230000003287 optical effect Effects 0.000 claims abstract description 15
- 238000000799 fluorescence microscopy Methods 0.000 claims abstract description 6
- 230000001575 pathological effect Effects 0.000 claims description 33
- 230000005284 excitation Effects 0.000 claims description 16
- 238000007493 shaping process Methods 0.000 claims description 13
- 238000010521 absorption reaction Methods 0.000 claims description 3
- 230000008033 biological extinction Effects 0.000 claims description 3
- 238000006862 quantum yield reaction Methods 0.000 claims description 3
- 230000015572 biosynthetic process Effects 0.000 claims description 2
- 230000000694 effects Effects 0.000 claims description 2
- 230000003902 lesion Effects 0.000 claims 1
- 238000012632 fluorescent imaging Methods 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 8
- 201000010099 disease Diseases 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 238000002560 therapeutic procedure Methods 0.000 description 4
- 206010028980 Neoplasm Diseases 0.000 description 3
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- 238000004980 dosimetry Methods 0.000 description 1
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- 238000012545 processing Methods 0.000 description 1
- 238000011155 quantitative monitoring Methods 0.000 description 1
- 238000009738 saturating Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
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- A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
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Abstract
本发明涉及一种定量检测光敏剂空间分布的荧光成像系统，其特征在于：包括第一LED光源、第一自由曲面全反射透镜、第一复眼透镜、第一积分透镜、第一反射镜、第二LED光源、第二自由曲面全反射透镜、第二复眼透镜、第二积分透镜、第二反射镜、第一半透半反镜、第二半透半反镜、激光光源、扩束镜、第三复眼透镜、数字微镜、投影镜头、聚光透镜、CMOS相机和计算机；本发明采用漫反射图像定量校正组织光学性质对荧光图像的影响，实现对光敏剂空间分布进行定量荧光成像，获得光敏剂在病变组织中的浓度空间分布及变化情况。
The invention relates to a fluorescence imaging system for quantitatively detecting the spatial distribution of photosensitizers, which is characterized by comprising a first LED light source, a first free-form surface total reflection lens, a first fly-eye lens, a first integrating lens, a first reflecting mirror, a first Two LED light sources, the second free-form surface total reflection lens, the second fly-eye lens, the second integrator lens, the second mirror, the first half mirror, the second half mirror, the laser light source, the beam expander, A third fly-eye lens, a digital micromirror, a projection lens, a condenser lens, a CMOS camera and a computer; the invention adopts the diffuse reflection image to quantitatively correct the influence of the optical properties of the tissue on the fluorescent image, and realizes quantitative fluorescent imaging of the spatial distribution of the photosensitizer, and obtains Concentration spatial distribution and changes of photosensitizers in diseased tissues.
Description
Technical field
The present invention relates to biomedical imaging fields, are related to a kind of fluorescence imaging system of quantitative detection photosensitizer spatial distribution
System and method.
Background technique
Using photosensitizer can be formed in tumor tissues rather high concentration accumulation and the light of appropriate wavelength irradiation after can
The characteristic of fluorescence is generated, detection technique of fluorescence can be clinically applied, pathological tissues can not only be determined by analysis of fluorescence intensity
Growth position, pathological tissues and normal tissue boundary, and the region for the treatment of can also be real during optical dynamic therapy
When monitor the variation of photosensitizer concentration, instruct real-time dosage to adjust, fluorescence diagnosis, the operation of the diseases such as tumour referred to realize
It leads and Treatment monitoring, there is extensive clinical application range.
Detection technique of fluorescence includes fluorescence spectroscopy technique and Imaging-PAM.Currently, generalling use fluorescence in clinic
Spectral technique carries out fluorescence detection and combines the Quantitative Monitoring of correcting algorithm realization photosensitizer concentration, although this method sensitivity
It is higher, it is easy to implement stable state and Transient detection, but the measurement method that contacts with tissue surface of fibre-optical probe is often generated and contacted
Pressure changes the optical property parameter of tissue local, influences the accuracy of measurement；In addition, spectroscopic way can only obtain list
The information of point, can not reflect photosensitizer concentration distribution situation in the biggish target tissue of area.Imaging-PAM can then be imaged
Mode realize in the biggish target tissue of area photosensitizer concentration distribution detect, however, in imaging mode on imaging surface
The average value of sampled signal of each pixel light detected in certain volume in tissue；Imaging surface is also possible to receive
To the photon outside target tissue, so that the fluorescence signal correction based on imaging technique is more more than the correction based on spectral technique
It is intractable.Existing fluoroscopic imaging systems often use semiempirical ratio algorithm, i.e., compare fluorescent image and reflected light data
Be worth operation, not only structure is complicated for such fluoroscopic imaging systems, expensive, and cannot parse respectively tissue absorption and
Scattering properties parameter may be only available for specific tissue sample.At the same time, the equally distributed exciting light of intensity of illumination reaches special
Fixed tissue depth is only capable of exciting the photosensitizer of the depth, is unable to the photosensitizer concentration of correct response different depth around it
Distributed intelligence.
Summary of the invention
In view of this, the purpose of the present invention is to provide a kind of fluoroscopic imaging systems of quantitative detection photosensitizer spatial distribution
And method, it realizes to the quantitative detection of photosensitizer concentration spatial distribution in different depth pathological tissues, is conducive to tumour etc.
Disease carries out the foundation that dosimetry parameter adjustment is provided when fluorescence diagnosis, surgical guidance and optical dynamic therapy monitor, and effectively improves
The accuracy rate and optical dynamic therapy curative effect of diagnosis.
To achieve the above object, the present invention adopts the following technical scheme:
A kind of fluoroscopic imaging systems of quantitative detection photosensitizer spatial distribution, including the first LED light source, the first free form surface
Total reflection lens, first fly's-eye lens, first integral lens, the first reflecting mirror, the second LED light source, the second free form surface are all-trans
It penetrates lens, the second fly's-eye lens, second integral lens, the second reflecting mirror, the first semi-transparent semi-reflecting lens, the second semi-transparent semi-reflecting lens, swash
Radiant, beam expanding lens, third fly's-eye lens, digital micro-mirror, projection lens, collector lens, CMOS camera and computer；Described
One LED light source issues the light beam that light forms collimation by the total reflection of the first freely curved face total reflection lens, the light beam warp of collimation
It crosses first fly's-eye lens shaping and homogenizes to form the first uniform rectangular hot spot and be emitted to the first reflecting mirror, be emitted through the first reflecting mirror
To the first semi-transparent semi-reflecting lens；Second LED light source issues light and is formed by the total reflection of the second freely curved face total reflection lens
The light beam of collimation, the light beam of collimation is by the second fly's-eye lens shaping and homogenizes and to form the second uniform rectangular hot spot and be emitted to second
Semi-transparent semi-reflecting lens；The Gaussian beam of the laser light source transmitting is expanded by beam expanding lens, and the laser after expanding passes through third
It the shaping of fly's-eye lens and homogenizes, forms third uniform rectangular hot spot and be emitted to the first semi-transparent semi-reflecting lens, through second semi-transparent half
Anti- mirror outgoing；It is incident upon digital micro-mirror by the light beam of the second semi-transparent semi-reflecting lens, and successively saturating by pathological tissues to be measured and optically focused
Mirror enters CMOS camera；Acquired image information is transferred to computer by the CMOS camera.
Further, the LED light source is made of the light source of two kinds of different wave lengths, and wavelength corresponds respectively to photosensitizer
Maximum excitation wavelength and launch wavelength.
Further, the beam expanding lens is 5 power beam expansion lens.
Further, the digital micro-mirror is made of 1024 × 768 micro mirrors, and the surface area of each micro mirror is 7.56 μ ms
7.56μm.Under electronic switching, micro mirror is overturn within the scope of ± 12 °, not by load computer programming Software Create
The structure light that the rectangular light spot of irradiation on it forms different space frequency is reflexed to projection object by isospace frequency grayscale image
Mirror；Projection objective, which shines structure light, to be projected on pathological tissues surface to excite the photosensitizer in different depth pathological tissues dense
Degree.
Further, the CMOS camera uses high pixel high-resolution camera.
Further, a kind of fluorescent quantitation method of the fluoroscopic imaging systems of quantitative detection photosensitizer spatial distribution, it is special
Sign is, comprising the following steps:
Step S1: according to photosensitizer, being arranged the first LED light source and the second LED light source, respectively as corresponding to photosensitizer most
The LED light source of good excitation wavelength and LED light source corresponding to the best launch wavelength of photosensitizer；
Step S2: the LED light source for corresponding to photosensitizer maximum excitation wavelength is opened, using particular space frequency (fx, fy) no
Same-phase θ=[0,2 π/3,4 π/3] structure light irradiates simulated solution known to optical parameter, obtains the three of simulated solution by camera
Width diffuses image I1, I2, I3；
Step S2: using the identical method of step S1, obtains M to pathological tissues to be measuredAC, sample(x, y, fx, fy), by
The modulation function M of pathological tissuesAC, sample(x, y, fx, fy) modulation function M with the simulated solution of known optical characteristicAC, ref(x, y,
fx, fy) the ratio between multiplied by known diffusing reflection rate RD, ref, tissue real reflectance R after being calibratedD, ex:
Step S3: closing the LED light source for corresponding to photosensitizer maximum excitation wavelength, opens and most preferably emits corresponding to photosensitizer
The LED light source of wavelength obtains the diffusing reflection rate R of pathological tissues using identical stepD, emFor
Step S4: closing the LED light source for corresponding to the best launch wavelength of photosensitizer, and photosensitizer is gathered in pathological tissues, opened
LD light source is opened, fluorescence imaging is carried out to pathological tissues, obtains fluorescent image Fmeasured, the intrinsic fluorescence information of pathological tissues
FintrinsicFor
Wherein, μA, exTo organize the absorption coefficient to excitation wavelength.
The photosensitizer concentration [PS] and intrinsic fluorescence intensity of pathological tissues are linear relationship, are indicated are as follows:
Wherein QEx, emIt is the fluorescence quantum yield that respective excitation wavelength excites lower photosensitizer, εEx, emIt is respective excitation wavelength
Excite the molar extinction coefficient of lower photosensitizer.
Compared with the prior art, the invention has the following beneficial effects:
Present invention sequence capture spatial frequency domain diffusing reflection image and fluorescent image, using diffusing reflection image quantitative correction tissue
Influence of the optical property to fluorescent image is realized and carries out quantitative fluorescence imaging to photosensitizer spatial distribution, obtains photosensitizer in disease
Become concentration space distribution and the situation of change in tissue, can be not only used for the boundary and the disease that determine pathological tissues and normal tissue
Become the size in region, growth position and treatment region to pathological tissues are accurately positioned, can also be to optical dynamic therapy
Photosensitizer doses detection is carried out in the process.
Detailed description of the invention
Fig. 1 is fluoroscopic imaging systems structural schematic diagram of the present invention；
Fig. 2 is the structural schematic diagram that free lens are totally reflected in one embodiment of the invention；
Fig. 3 is LED light source structure figure in one embodiment of the invention；
Fig. 4 is LED light source collimation, shaping and homogenized optical path simulation drawing in one embodiment of the invention；
Fig. 5 be in one embodiment of the invention LD light source expand, shaping and the optical path simulation drawing after homogenizing；
Fig. 6 is that 620nm LED light source is radiated at the surface of intensity distribution on DMD in one embodiment of the invention；
Fig. 7 is the surface of intensity distribution of 405nm LD light source analogy in one embodiment of the invention；
In figure: the first LED light source of 1-, the first freely curved face total reflection of 2- lens, 3- first fly's-eye lens, 4- first integral
Lens, the first reflecting mirror of 5-, 6- laser light source, 7- beam expanding lens, 8- third fly's-eye lens, the first semi-transparent semi-reflecting lens of 9-, 10- second
Semi-transparent semi-reflecting lens, the second LED light source of 11-, the second freely curved face total reflection of 12- lens, the second fly's-eye lens of 13-, the product of 14- second
Point lens, the second reflecting mirror of 15-, 16- digital micro-mirror, 17- projection lens, 18- tissue, 19- collector lens, 20-CMOS camera,
21- computer.
Specific embodiment
The present invention will be further described with reference to the accompanying drawings and embodiments.
Fig. 1 is please referred to, the present invention provides a kind of fluoroscopic imaging systems of quantitative detection photosensitizer spatial distribution, including first
LED light source 1, the first freely curved face total reflection lens 2, first fly's-eye lens 3, first integral lens 4, the first reflecting mirror 5, second
LED light source 11, the second freely curved face total reflection lens 12, the second fly's-eye lens 13, second integral lens 14, the second reflecting mirror
15, the first semi-transparent semi-reflecting lens 9, the second semi-transparent semi-reflecting lens 10, laser light source 6, beam expanding lens 7, third fly's-eye lens 8, digital micro-mirror
16, projection lens 17, collector lens 19, CMOS camera 20 and computer 21.First LED light source 1 issues light and passes through first
The total reflection of freely curved face total reflection lens 2 forms the light beam of collimation, and the light beam of collimation is by 3 shaping of first fly's-eye lens and even
Change the first uniform rectangular hot spot of formation and be emitted to the first reflecting mirror 5, is emitted to the first semi-transparent semi-reflecting lens 9 through the first reflecting mirror 5, passes through
The outgoing of second semi-transparent semi-reflecting lens 10；；Second LED light source 11 issues light by the complete of the second freely curved face total reflection lens 12
The light beam of collimation is reflected to form, the light beam of collimation passes through 13 shaping of the second fly's-eye lens and homogenize to form the second uniform rectangular hot spot
It is emitted to the second reflecting mirror 15, is emitted through the second semi-transparent semi-reflecting lens 10；The Gaussian beam that the laser light source 6 emits is by expanding
Mirror 7 is expanded, and the laser after expanding passes through the shaping of third fly's-eye lens 8 and homogenizes, and is formed third uniform rectangular hot spot and is gone out
The first semi-transparent semi-reflecting lens 9 are incident upon, are emitted through the second semi-transparent semi-reflecting lens 10；Number is incident upon by the light beam of the second semi-transparent semi-reflecting lens 10
Micro mirror 16, and successively CMOS camera 20 is injected by pathological tissues 18 to be measured and collector lens 19；The CMOS camera 20 will be adopted
The image information collected is transferred to computer 21.
The specific implementation of the present embodiment progress quantitative fluorescence imaging are as follows:
The LED light source (LB-H9GP, OSRAM, German) of 405nm is opened, LED light source forms standard by the total reflection of TIR
Straight light beam, the light beam of collimation by fly's-eye lens shaping and homogenizing formed on DMD (0.45WXGA, TI, USA) 12mm ×
The rectangular light spot of 9mm；There is the structure light image of the sinusoidal model as shown in (1) formula to load computer programming Software Create simultaneously
Onto DMD.
Wherein, wherein S0It is illumination source intensities, fxAnd fyIt is the spatial frequency on the direction x and y, M respectively0It is modulation depth
It is space phase with φ, φ=0 when being imaged for the first time.DMD surface reflection provides the structure light of certain frequency via projecting subassembly
Be radiated on the simulated solution of known optical characterisitic parameter, projection lens range simulation liquid sample 150mm, projected area be 40mm ×
The image I that diffuses of 30mm, CMOS camera (PC0.EDGE5.5, PCO, German) acquisition sample1, send to computer and carry out
Processing.Changing phase respectively is 2 π/3 and 4 π/3, and the acquisition that repeats the above steps diffuses image I2And I3.It is collected unrestrained anti-
Penetrating image includes direct current and AC portion, and AC portion can indicate are as follows:
Wherein, MACIt is modulation amplitude, can be calculated and be obtained according to formula (3).
Using identical step, M is obtained to pathological tissues to be measuredAC, sample(x, y, fx, fy), by the modulation of pathological tissues
Function MAC, sample(x, y, fx, fy) modulation function M with the simulated solution of known optical characteristicAC, ref(x, y, fx, fy) the ratio between multiplied by
With known diffusing reflection rate RD, ref, tissue real reflectance R after being calibratedD, 405。
The LED light source for closing 405nm, opens the LED light source (HPWS-TH77, OSRAM, German) of 620nm, using phase
Same step obtains the diffusing reflection rate R of pathological tissuesD, 620
LED light source is closed, injects photosensitizer HMME in the tissue, after waiting HMME to gather in pathological tissues, opens LD
The Gaussian beam of light source (LaserBoxx405, Oxxius, France), laser light source transmitting is expanded with 5 power beam expansion lens, is expanded
The laser of beam passes through the shaping of fly's-eye lens and homogenizes, and forming size is that 12mm × 9mm uniform rectangular hot spot is emitted in DMD
On.The structure light that DMD surface reflection provides certain frequency is radiated on pathological tissues via projecting subassembly, the acquisition of CMOS camera
Fluorescent image Fmeasured, send to computer, the intrinsic fluorescence information F in pathological tissues calculated according to formula (6)intrinsicFor
Photosensitizer concentration [HMME] and intrinsic fluorescence intensity in pathological tissues are linear relationship, are indicated are as follows:
Wherein Q405,620It is the fluorescence quantum yield of the HMME under the excitation of 405nm excitation wavelength, ε405,620It is to swash in 405nm
Send out the molar extinction coefficient that wavelength excites lower HMME.
Change spatial frequency (fx, fy), it repeats the above steps, successively acquisition diffuses image and fluorescent image, and according to
The above method carries out calculating the fluorescence information obtained apart from surface different depth pathological tissues, realizes photosensitizer HMME spatial distribution
Quantitative detection.
The foregoing is merely presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, is all covered by the present invention.
Claims (6)
1. a kind of fluoroscopic imaging systems of quantitative detection photosensitizer spatial distribution, it is characterised in that: including the first LED light source,
One freely curved face total reflection lens, first fly's-eye lens, first integral lens, the first reflecting mirror, the second LED light source, second are certainly
By curved face total reflection lens, the second fly's-eye lens, second integral lens, the second reflecting mirror, the first semi-transparent semi-reflecting lens, second semi-transparent
Semi-reflective mirror, laser light source, beam expanding lens, third fly's-eye lens, digital micro-mirror, projection lens, collector lens, CMOS camera and calculating
Machine；First LED light source issues the light beam that light forms collimation by the total reflection of the first freely curved face total reflection lens, collimation
Light beam by first fly's-eye lens shaping and homogenize and to form the first uniform rectangular hot spot and be emitted to the first reflecting mirror, instead through first
It penetrates mirror and is emitted to the first semi-transparent semi-reflecting lens, emergent light is incident upon the second semi-transparent semi-reflecting lens；Second LED light source issues light by the
The total reflection of two freely curved face total reflection lens forms the light beam of collimation, and the light beam of collimation is by the second fly's-eye lens shaping and even
Change the second uniform rectangular hot spot of formation and is emitted to the second semi-transparent semi-reflecting lens；The Gaussian beam of the laser light source transmitting is by expanding
Mirror is expanded, and the laser after expanding passes through the shaping of third fly's-eye lens and homogenizes, and forms the outgoing of third uniform rectangular hot spot
To the first semi-transparent semi-reflecting lens, it is emitted through the second semi-transparent semi-reflecting lens；It is incident upon digital micro-mirror by the light beam of the second semi-transparent semi-reflecting lens, and
Successively CMOS camera is injected by pathological tissues to be measured and convergent lens；The CMOS camera transmits acquired image information
To computer.
2. a kind of fluoroscopic imaging systems of quantitative detection photosensitizer spatial distribution according to claim 1, it is characterised in that:
The LED light source is made of the light source of two kinds of different wave lengths, and wavelength corresponds respectively to the maximum excitation wavelength and hair of photosensitizer
Ejected wave is long.
3. a kind of fluoroscopic imaging systems of quantitative detection photosensitizer spatial distribution according to claim 1, it is characterised in that:
The beam expanding lens is 5 power beam expansion lens.
4. a kind of fluoroscopic imaging systems of quantitative detection photosensitizer spatial distribution according to claim 1, it is characterised in that:
The digital micro-mirror is made of 1024 × 768 micro mirrors, and the surface area of each micro mirror is 7.56 μm of 7.56 μ m；In electronic switch
Under effect, micro mirror is overturn within the scope of ± 12 °, by loading the different space frequency grayscale image of computer programming Software Create,
The structure light that the rectangular light spot of irradiation on it forms different space frequency is reflexed into projection objective；Projection objective is by structure light
According to being projected in the photosensitizer excited in different depth pathological tissues on pathological tissues surface.
5. a kind of fluoroscopic imaging systems of quantitative detection photosensitizer spatial distribution according to claim 1, it is characterised in that:
The CMOS camera uses high pixel high-resolution camera.
6. a kind of fluorescent quantitation of the fluoroscopic imaging systems of quantitative detection photosensitizer spatial distribution described in -5 according to claim 1
Method, which comprises the following steps:
Step S1: according to photosensitizer, being arranged the first LED light source and the second LED light source, most preferably swashs respectively as photosensitizer is corresponded to
Send out the LED light source of wavelength and the LED light source corresponding to the best launch wavelength of photosensitizer；
Step S2: the LED light source for corresponding to photosensitizer maximum excitation wavelength is opened, using particular space frequency (fx, fy) difference phase
Simulated solution known to θ=[0,2 π/3,4 π/3] the structure light irradiation optical parameter of position, three width for obtaining simulated solution by camera are unrestrained
Reflected light image I1, I2, I3；
Step S2: using the identical method of step S1, obtains M to pathological tissues to be measuredAC, sample(x, y, fx, fy), by lesion
The modulation function M of tissueAC, sample(x, y, fx, fy) modulation function M with the simulated solution of known optical characteristicAC, ref(x, y, fx,
fy) the ratio between multiplied by known diffusing reflection rate RD, ref, tissue real reflectance R after being calibratedD, ex:
Step S3: closing the LED light source for corresponding to photosensitizer maximum excitation wavelength, opens and corresponds to the best launch wavelength of photosensitizer
LED light source, the diffusing reflection rate R of pathological tissues is obtained using identical stepD, emFor
Step S4: closing the LED light source for corresponding to the best launch wavelength of photosensitizer, and photosensitizer is gathered in pathological tissues, opens LD
Light source carries out fluorescence imaging to pathological tissues, obtains fluorescent image Fmeasured, the intrinsic fluorescence information F of pathological tissuesintrinsic
For
Wherein, μA, exTo organize the absorption coefficient to excitation wavelength.
The photosensitizer concentration [PS] and intrinsic fluorescence intensity of pathological tissues are linear relationship, are indicated are as follows:
Wherein QEx, emIt is the fluorescence quantum yield that respective excitation wavelength excites lower photosensitizer, εEx, emIt is under respective excitation wavelength excitation
The molar extinction coefficient of photosensitizer.
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| CN115519138A (en) * | 2022-09-23 | 2022-12-27 | 华南理工大学 | A low melting point metal printing device and method based on DMD micromirror group |
| CN115868927A (en) * | 2022-11-15 | 2023-03-31 | 哈尔滨工业大学 | A device for measuring the concentration of photosensitizer in tissue with high precision and its application method |
| CN119043213A (en) * | 2024-10-30 | 2024-11-29 | 四川川大智胜软件股份有限公司 | Three-dimensional measuring instrument based on optical fiber coding structured light projection |
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| CN115868927A (en) * | 2022-11-15 | 2023-03-31 | 哈尔滨工业大学 | A device for measuring the concentration of photosensitizer in tissue with high precision and its application method |
| CN115868927B (en) * | 2022-11-15 | 2024-07-23 | 哈尔滨工业大学 | High-precision device for measuring concentration of photosensitizer in tissue and application method thereof |
| CN119043213A (en) * | 2024-10-30 | 2024-11-29 | 四川川大智胜软件股份有限公司 | Three-dimensional measuring instrument based on optical fiber coding structured light projection |
| CN119043213B (en) * | 2024-10-30 | 2025-02-07 | 四川川大智胜软件股份有限公司 | Three-dimensional measuring instrument based on optical fiber coding structured light projection |
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