CN105894537B - A kind of decaying of emission-type optical projection tomographic imaging and scatter correction method - Google Patents
A kind of decaying of emission-type optical projection tomographic imaging and scatter correction method Download PDFInfo
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- CN105894537B CN105894537B CN201610203306.6A CN201610203306A CN105894537B CN 105894537 B CN105894537 B CN 105894537B CN 201610203306 A CN201610203306 A CN 201610203306A CN 105894537 B CN105894537 B CN 105894537B
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Abstract
The invention discloses a kind of decaying of emission-type optical projection tomographic imaging and scatter correction methods, OPT imaging model under weak scattering situation is combined with OPT fluorescence algorithm for reconstructing, the correction of OPT fluorescent signal decay is carried out using the method based on normalization Born ratio, using the scattering component in Monte Carlo technology simulated measurement data, and it is removed it from detection data, correction for attenuation and scattering component are estimated into the iteration frame effectively combined, realize the quantitative three-dimensional reconstruction of OPT fluorescent dye concentration.The present invention has comprehensively considered the influence for absorbing and scattering in realizing the reconstruction of fluorescent dye concentration quantitative, and has been estimated the scattering component in signal using Monte Carlo technology and removed, and then realized the Quantitative Reconstruction of fluorescent dye concentration.
Description
Technical field
The invention belongs to medical image processing technical fields more particularly to a kind of emission-type optical projection tomographic imaging to decay
With scatter correction method.
Background technique
Optical projection tomographic imaging OPT is a kind of novel high-resolution three-dimension molecular image imaging technique, and imaging is former
It manages similar with the principle of X ray computer tomographic imaging.OPT, as irradiation source, is highly suitable for toy embryo using visible light
The imaging of transparent or semitransparent sample in the 1-10mm range scale such as tire, organ, drosophila, nematode.OPT technology is not necessarily to people
The integrality of loss biological tissue's organ can be obtained the three-dimensional structure picture of high-resolution, and can pass through fluorescent labelling techniques reality
Existing molecular specificity imaging.OPT has the characteristics that bimodulus fusion, this is met field of biomedical research just and is melted using multimode
Close the demand for more fully understanding Biont information.Therefore, OPT has received the extensive concern of each research unit, by
In research applied to gene expression, protein interaction, Preclinical Drug research and development etc., imaging object covers petty action sundries
Official, embryo, drosophila, nematode, zebra fish, locust, arabidopsis isotype biology, are integrally imaged for small sample and provide strong grind
Study carefully means, a series of biological basis progress of research will be promoted.
Some new problems are introduced to emission-type OPT in body bio-imaging, be excited generation in emission-type OPT imaging
Fluorescent photon is understood some photon in biological tissue's communication process and is absorbed by biological tissue, and traditional emission-type OPT weight
Construction method does not account for influence of this sink effect to decaying caused by fluorescence signal, therefore its fluorescence signal Quantitative Reconstruction is
Inaccuracy.Related experiment proves, even if sample interior places the fluorescent dye of same concentrations, since fluorescence information is from sample
Transmission comes out that signal attenuation degree is different, and also resulting in the fluorescence signal intensity of reconstruction, there are notable differences；It is dissipated simultaneously for weak
Sample is penetrated, the scattering effect of sample also will affect the quantitative accuracy of fluorescence signal.
Summary of the invention
The emission-type optical projection tomographic imaging decaying that the purpose of the present invention is to provide a kind of for weak scattering sample with
Scatter correction method, it is intended to solve traditional emission-type OPT method for reconstructing and not account for sink effect to caused by fluorescence signal
The influence of decaying, and the problem because of fluorescence signal Quantitative Reconstruction inaccuracy caused by weak scattering.
The invention is realized in this way a kind of emission-type optical projection tomographic imaging for weak scattering sample is decayed and is dissipated
Bearing calibration is penetrated, OPT under weak scattering situation is imaged for the emission-type optical projection tomographic imaging decaying and scatter correction method
Model is combined with OPT fluorescence algorithm for reconstructing, and ignoring scattering first influences, and directly utilizes detection data based on normalization Born
The method of ratio carries out the preliminary correction for attenuation of OPT fluorescence signal, is then calculated using filter back-projection reconstruction algorithm glimmering in sample
The concentration of photoinitiator dye, then based on the concentration for rebuilding obtained fluorescent dye, generated using the emulation of Monte Carlo technology corresponding
Scattering component, and removing scattering component from detection data, so further to the detection data of removal scattering component into
Row constitutes correction for attenuation and dissipates based on the sequence of operations such as the correction for attenuation of Born ratio, the concentration reestablishing of fluorescent dye are normalized
It penetrates component and estimates the iteration frame effectively combined, until the fluorescent dye concentration that front and back is rebuild twice is less than the threshold value of setting, most
The quantitative three-dimensional reconstruction of OPT fluorescent dye concentration is realized eventually.
Further, emission-type optical projection tomographic imaging decaying with scatter correction method the following steps are included:
Step 1 carries out correction for attenuation to measurement data using normalized Born ratio method；
Step 2 calculates the concentration of fluorescent dye in sample using filtered back projection's (FBP) algorithm for reconstructing；
Step 3, the scattering component in fluorescence data measured by Monte Carlo simulation estimate；
Step 4 removes scattering component, carries out the calculating of correction for attenuation and fluorescent dye concentration again；
Step 5, the difference and threshold comparison of reconstructed results, iteration carry out simulation calculation twice.
Further, described that measurement data progress correction for attenuation is specifically included using normalized Born ratio method:
According to the imaging model of emission-type OPT fluorescence imaging, exciting light fromPoint is incident on sample match liquid pool,Point
Sample is excited, and fromPlace projects sample cell, and the received fluorescence signal of CCD indicates are as follows:
WhereinIndicate CCD collected signal when excitation wavelength and filter plate mismatch,Indicate excitation point,
Indicate transmitted wave and filter plate contact point,Indicate the fluorescence signal intensity that excitation generates, ε and η respectively indicate extinction coefficient
And conversion quantum efficiency,The concentration for indicating fluorescent dye, to know:
The parameter meaning repeated in several formula herein is just the same, the visible context-sensitive part of concrete meaning.
When signal acquisition is using signal acquisition is carried out with the matched filter plate of excitation wavelength, signal that CCD is detected are as follows:
HaveWherein GfIt is exciting light along the direction of propagationRadon transformation,The collected signal of CCD camera under respectively two kinds excitation situations, other parameters have been introduced above.
Further, the concentration for calculating fluorescent dye in sample using FBP algorithm for reconstructing specifically includes:
Radon by obtaining fluorescent dye concentration is converted, i.e.,GfIt is exciting light along biography
Broadcast directionRadon transformation,The collected signal of CCD camera under respectively two kinds excitation situations, ε and η difference
Indicate extinction coefficient and conversion quantum efficiency,For the concentration of fluorescent dye.The Radon transformation of multiple directions can be rebuild
Fluorescent dye concentration outCollect 360 degree of measurement data GfAfterwards, fluorescent dye is calculated by inverse Radon transform
Concentration, i.e., calculating σ=FBP (Gf), obtain fluorescent dye concentration σ.
Further, the scattering component in the fluorescence data measured by Monte Carlo simulation estimate specifically wraps
It includes:
Using the fluorescent dye concentration σ reconstructed, while using information such as the absorption coefficient of sample and scattering coefficients, lead to
It crosses Monte Carlo emulation and establishes model, by adjusting simulation parameter to simulate actual experiment situation, can therefrom estimate glimmering
Scattering component in light data.
Further, the removal scattering component, the calculating for carrying out correction for attenuation and fluorescent dye concentration again specifically include:
Measurement data is further corrected by obtained scattering component, i.e., rejects scattering component from measurement data,
Then correction for attenuation and fluorescent dye concentration calculation again is carried out, that is, repeats following steps:
The concentration that fluorescent dye in sample is calculated using FBP algorithm for reconstructing, is specifically included:
Radon by obtaining fluorescent dye concentration is converted, i.e.,GfIt is exciting light along biography
Broadcast directionRadon transformation, multiple directions Radon transformation can reconstruct fluorescent dye concentrationCollect 360
The measurement data G of degreefIt afterwards, is the concentration for calculating fluorescent dye by inverse Radon transform, i.e. calculating σ=FBP (Gf), it obtains glimmering
Photoinitiator dye concentration σ；
Further, the difference and threshold comparison of reconstructed results, iteration carry out simulation calculation twice, and the fluorescent dye reconstructed is dense
Degree is compared with last reconstructed results, if the two difference is less than given threshold value, terminator, output is as a result, complete to decline
Subtract and scatter correction continues MonteCarlo emulation, remove the detection number of scattering component if being unsatisfactory for termination condition
The sequence of operations such as the concentration reestablishing according to the correction for attenuation, fluorescent dye that carry out based on normalization Born ratio terminate until meeting
Condition.
Emission-type optical projection tomographic imaging decaying provided by the invention and scatter correction method, are imaged with weak scattering OPT
Models coupling is realized and is rebuild in body OPT fluorescent dye concentration quantitative；By OPT imaging model under weak scattering situation and OPT fluorescence weight
Algorithm is built to combine, and using transmission-type OPT imaging in reconstruct come absorption coefficient and scattering coefficient, using based on normalize
The method of Born ratio carries out the correction of OPT fluorescent signal decay, using the scattering in Monte Carlo technology simulated measurement data point
Amount, and removed it from detection data, to eliminate the influence that fluorescent quantitation is rebuild in scattering, by correction for attenuation and scattering component
Estimate the iteration frame effectively combined, the removal of fluorescent signal decay correction and scattering component is completed by successive ignition, thus
Realize the quantitative three-dimensional reconstruction of OPT fluorescent dye concentration.
The present invention has comprehensively considered the influence for absorbing and scattering in realizing the reconstruction of fluorescent dye concentration quantitative, and uses
Monte Carlo technology has been estimated the scattering component in signal and has been removed, and then realizes the quantitative weight of fluorescent dye concentration
It builds.
Detailed description of the invention
Fig. 1 is emission-type optical projection tomographic imaging decaying provided in an embodiment of the present invention and scatter correction method process
Figure.
Fig. 2 is the flow chart of embodiment provided in an embodiment of the present invention.
Fig. 3 is emission-type OPT image acquisitions schematic diagram provided in an embodiment of the present invention.
Fig. 4 is fluorescence excitation provided in an embodiment of the present invention and signal decaying schematic diagram.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
The present invention combines OPT imaging model under weak scattering situation with OPT fluorescence algorithm for reconstructing, using based on normalization
The method of Born ratio carries out the correction of OPT fluorescent signal decay, using the scattering in Monte Carlo technology simulated measurement data point
Amount, and removed it from detection data, correction for attenuation and scattering component are estimated the iteration frame effectively combined by research, thus
Realize the quantitative three-dimensional reconstruction of OPT fluorescent dye concentration.
Application principle of the invention is explained in detail with reference to the accompanying drawing.
As shown in Figure 1, the emission-type optical projection tomographic imaging decaying of the embodiment of the present invention includes with scatter correction method
Following steps:
S101: correction for attenuation is carried out to measurement data using normalized Born ratio method；
S102: the concentration of fluorescent dye in sample is calculated using FBP algorithm for reconstructing；
S103: the scattering component in fluorescence data measured by Monte Carlo simulation estimate；
S104: removal scattering component carries out the calculating of correction for attenuation and fluorescent dye concentration again；
S105: the difference and threshold comparison of reconstructed results twice, iteration carry out simulation calculation.
Application principle of the invention is further described combined with specific embodiments below.
As shown in Fig. 2, the emission-type optical projection tomographic imaging decaying of the embodiment of the present invention includes with scatter correction method
Following steps:
Step 1, correction for attenuation is carried out to measurement data using normalized Born ratio method:
Firstly, the fluorescence signal portion that sample is generated through excitation is through bullet according to the imaging model of emission-type OPT fluorescence imaging
Road propagation is directly received by CCD, is received after being partially scattered by CCD, remaining photon direct projection or scatter to other directions can not be by
CCD is detected.Assuming that exciting light fromPoint is incident on sample match liquid pool,Point excites sample, and fromPlace is penetrated
Sample cell out.Relatively in view of the wavelength of exciting light and transmitting light, ignore difference of the two between optical coefficient, simultaneously
In view of fluorescent dye is likely distributed in any position that light passes through, then the received fluorescence signal of CCD can indicate are as follows:
WhereinIndicate that the fluorescence signal intensity that excitation generates, ε and η respectively indicate extinction coefficient and quantum conversion effect
Rate,The concentration for indicating fluorescent dye, to know:
It notices when using signal acquisition is carried out with the matched filter plate of excitation wavelength, the signal that CCD is detected are as follows:
To have
The above method for carrying out fluorescent signal decay correction is known as normalized Born and compares method.
Step 2, the concentration of fluorescent dye in sample is calculated using FBP algorithm for reconstructing:
The Radon transformation of fluorescent dye concentration is obtained by step 1, i.e.,From the above equation, we can see that
GfIt is exciting light along the direction of propagationRadon transformation, by Radon transformation and its inverse transformation theory it is found that measuring multiple sides
To Radon transformation can reconstruct fluorescent dye concentrationRealization of multidirectional DATA REASONING in OPT imaging can
Both may be used with driving sample rotates to carry out data acquisition using light source irradiation, CCD simultaneously using automatically controlled rotating device.When collecting
360 degree of measurement data GfAfterwards, the concentration of fluorescent dye can be calculated by inverse Radon transform.
Calculate σ=FBP (Gf), to obtain fluorescent dye concentration σ.
Step 3, the scattering component in fluorescence data measured by Monte Carlo simulation estimate:
Concrete methods of realizing is to utilize the fluorescent dye concentration σ reconstructed in step 2, while the absorption coefficient of utilization sample
With the information such as scattering coefficient, model is established by Monte Carlo emulation, by adjusting simulation parameter to simulate actual experiment feelings
Shape can therefrom estimate the scattering component in fluorescence data.
Calculation amount based on Monte Carlo simulation calculating is very big, while also having the characteristics that highly-parallel, also very
It is suitable for GPU acceleration, therefore is passed in biological tissues using the photon based on Monte Carlo method developed before this laboratory
Emulation platform, Molecular Optical Simulation Environment (MOSE) are broadcast, research is carried out using GPU technology
Algorithm accelerates, and realizes the acceleration of scattering component Monte Carlo simulation calculation.
Step 4, scattering component is removed, carries out the calculating of correction for attenuation and fluorescent dye concentration again:
The scattering component as obtained in step 3 further corrects measurement data, i.e., rejects and dissipate from measurement data
Component is penetrated, correction for attenuation and fluorescent dye concentration calculation again are then carried out, is i.e. repeatedly step 2 and 3.
Step 5, the difference and threshold comparison of reconstructed results, iteration carry out simulation calculation twice.
Reconstructed results and last reconstructed results compare, if the two difference is less than given threshold value, terminator,
Output is as a result, completion decaying and scatter correction continue Monte Carlo emulation etc. one if being unsatisfactory for termination condition
Sequence of maneuvers, until meeting termination condition.
With reference to the accompanying drawing 3,4 pairs of reconstructed results of the invention of attached drawing are described in detail.
Attached drawing 3 is emission-type OPT image acquisitions schematic diagram of the invention.The fluorescence signal portion that sample is generated through excitation
Lease making ballistic propagation is directly received by CCD, is received after being partially scattered by CCD, remaining photon direct projection or scatters to other directions
It can not be detected by CCD.Exciting light and transmitting light propagated in biological tissue will receive biological tissue decaying influence, this
It is irrespective in traditional OPT imaging, is also just the core content that the present invention is considered.
Attached drawing 4 is the decaying schematic diagram of fluorescence excitation and signal.Wherein figure (a) is to use and the matched filter of wavelength of transmitted light
Schematic diagram when wave plate progress signal acquisition, filter plate and excitation wavelength at this time mismatch, and figure (b) is use and exciting light
Wavelength matched filter plate carries out schematic diagram when signal acquisition.Assume exciting light from point in two width schematic diagramesIt is incident on sample
Product match liquid pool,Point excites sample, and fromProject sample cell in place.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (5)
1. a kind of emission-type optical projection tomographic imaging decaying and scatter correction method, which is characterized in that the emission-type optics
Projection tomographic imaging decaying mutually ties OPT imaging model under weak scattering situation and OPT fluorescence algorithm for reconstructing with scatter correction method
It closes, carries out the preliminary correction for attenuation of OPT fluorescence signal using the method based on normalization Born ratio to measurement data；Then it utilizes
Filter back-projection reconstruction algorithm calculates the concentration of fluorescent dye in sample, then based on the concentration for rebuilding obtained fluorescent dye, adopts
Corresponding scattering component is generated with the emulation of Monte Carlo technology, and removes scattering component from measurement data, removal is dissipated
The measurement data for penetrating component carries out the concentration reestablishing operation of correction for attenuation, fluorescent dye based on normalization Born ratio, and composition declines
Correction down and scattering component estimate the iteration frame effectively combined, until the fluorescent dye concentration that front and back is rebuild twice is less than setting
Threshold value；The final quantitative three-dimensional reconstruction for realizing OPT fluorescent dye concentration；
OPT imaging model under weak scattering situation is combined with OPT fluorescence algorithm for reconstructing, obtains the Radon of fluorescent dye concentration
Transformation, i.e., Indicate transmitted wave and filter plate contact point；Indicate laser light
Incidence point when irradiating sample；GfIt is exciting light along excitation pointThe direction of propagation Radon transformation,Respectively
It is mismatched for excitation wavelength with filter plate and excitation wavelength matches CCD camera under two kinds of excitation situations with filter plate and collects
Signal；Indicate CCD collected signal when excitation wavelength and filter plate mismatch；Indicate excitation wavelength with
CCD collected signal when filter plate matches, ε and η respectively indicate extinction coefficient and conversion quantum efficiency,For fluorescence
The concentration of dyestuff；The Radon transformation of multiple directions can reconstruct fluorescent dye concentrationCollect 360 degree of measurement
Data GfIt afterwards, is the concentration for calculating fluorescent dye by inverse Radon transform, i.e. calculating σ=FBP (Gf), it is dense to obtain fluorescent dye
Spend σ.
2. emission-type optical projection tomographic imaging decaying as described in claim 1 and scatter correction method, which is characterized in that adopt
Correction for attenuation is carried out to measurement data with normalized Born ratio method to specifically include:
According to the imaging model of emission-type OPT fluorescence imaging, exciting light fromPoint is incident on sample match liquid pool,Point pair
Sample is excited, and fromPlace projects sample cell, and the received fluorescence signal of CCD indicates are as follows:
WhereinIndicate CCD collected signal when excitation wavelength and filter plate mismatch,Indicate excitation point,Table
Show transmitted wave and filter plate contact point,Indicate the fluorescence signal intensity that excitation generates, ε and η respectively indicate extinction coefficient
And conversion quantum efficiency,The concentration for indicating fluorescent dye, to know:
When signal acquisition is using signal acquisition is carried out with the matched filter plate of excitation wavelength, signal that CCD is detected are as follows:
HaveWherein GfIt is exciting light along excitation pointRadon transformation,Respectively excitation wavelength is mismatched with filter plate and excitation wavelength matches under two kinds of excitation situations with filter plate
The collected signal of CCD camera；Indicate CCD collected signal when excitation wavelength and filter plate mismatch；It indicates
CCD collected signal when excitation wavelength is matched with filter plate.
3. emission-type optical projection tomographic imaging decaying as described in claim 1 and scatter correction method, which is characterized in that logical
The scattering component crossed in the fluorescence data that Monte Carlo simulation estimate measures specifically includes:
Using the fluorescent dye concentration σ reconstructed, while using the absorption coefficient and scattering coefficient information of sample, pass through Monte
Model is established in Carlo emulation, by adjusting simulation parameter to simulate actual experiment situation, can therefrom be estimated in fluorescence data
Scattering component.
4. emission-type optical projection tomographic imaging decaying as described in claim 1 and scatter correction method, which is characterized in that go
Except scattering component, the calculating for carrying out correction for attenuation and fluorescent dye concentration again is specifically included:
Measurement data is further corrected by obtained scattering component, i.e., rejects scattering component from measurement data, then
Correction for attenuation and fluorescent dye concentration calculation again is carried out, that is, repeats following steps:
The concentration that fluorescent dye in sample is calculated using FBP algorithm for reconstructing, is specifically included:
The Radon transformation of fluorescent dye concentration is obtained, i.e.,GfIt is exciting light along sharp
Hair pointRadon transformation,The collected signal of CCD camera under respectively two kinds excitation situations；It indicates to swash
Hair point,Indicate transmitted wave and filter plate contact point,Indicate incidence point when laser light irradiating sample；Multiple directions
Radon transformation can reconstruct fluorescent dye concentrationCollect 360 degree of measurement data GfAfterwards, pass through inverse Radon
Transformation calculates the concentration of fluorescent dye, i.e. calculating σ=FBP (Gf), obtain fluorescent dye concentration σ；Indicate excitation light wave
CCD collected signal when the long mismatch with filter plate；Indicate that CCD is collected when excitation wavelength is matched with filter plate
Signal；
The scattering component in fluorescence data measured by Monte Carlo simulation estimate, specifically includes:
Using the fluorescent dye concentration σ reconstructed, while using the absorption coefficient and scattering coefficient information of sample, pass through Monte
Model is established in Carlo emulation, by adjusting simulation parameter to simulate actual experiment situation, is therefrom estimated in fluorescence data
Scattering component.
5. emission-type optical projection tomographic imaging decaying as described in claim 1 and scatter correction method, which is characterized in that two
The difference and threshold comparison of secondary reconstructed results, iteration carry out simulation calculation, and reconstructed results and last reconstructed results compare, if two
Person's difference is less than given threshold value, then terminator, and output is as a result, complete decaying and scatter correction, if being unsatisfactory for terminating
Condition then continues Monte Carlo emulation, until meeting termination condition.
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| CN104586366A (en) * | 2015-01-27 | 2015-05-06 | 西安电子科技大学 | Fluorescence molecular tomography reconstruction method |
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| CN104586366A (en) * | 2015-01-27 | 2015-05-06 | 西安电子科技大学 | Fluorescence molecular tomography reconstruction method |
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