CN105675568B - A kind of multicomponent fluorescence lifetime and component ratio estimation method for Single Photon Counting - Google Patents
A kind of multicomponent fluorescence lifetime and component ratio estimation method for Single Photon Counting Download PDFInfo
- Publication number
- CN105675568B CN105675568B CN201610052282.9A CN201610052282A CN105675568B CN 105675568 B CN105675568 B CN 105675568B CN 201610052282 A CN201610052282 A CN 201610052282A CN 105675568 B CN105675568 B CN 105675568B
- Authority
- CN
- China
- Prior art keywords
- fluorescence lifetime
- data
- component
- histogram
- tcspc
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 238000000034 method Methods 0.000 title claims abstract description 22
- 239000011159 matrix material Substances 0.000 claims abstract description 25
- 238000001161 time-correlated single photon counting Methods 0.000 claims abstract description 19
- 238000000354 decomposition reaction Methods 0.000 claims abstract description 5
- 239000000470 constituent Substances 0.000 claims abstract description 4
- 238000001228 spectrum Methods 0.000 claims abstract description 4
- 238000012545 processing Methods 0.000 claims description 5
- 238000006116 polymerization reaction Methods 0.000 claims description 3
- 241001269238 Data Species 0.000 claims description 2
- 230000017105 transposition Effects 0.000 claims description 2
- 238000009499 grossing Methods 0.000 claims 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 201000009310 astigmatism Diseases 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000013178 mathematical model Methods 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 230000002123 temporal effect Effects 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
Abstract
The present invention relates to a kind of multicomponent fluorescence lifetimes and component ratio estimation method applied in Single Photon Counting TCSPC.First collected histogram data is combined to form the lesser new histogram in section, to eliminate the influence of partial noise and algorithm operation quantity can be greatly reduced.Auto-correlation computation is carried out to new histogram interval censored data, the characteristics of according to TCSPC, autocorrelation matrix is smoothed.Eigenvalues Decomposition is carried out to its autocorrelation matrix, obtains signal subspace.According to possible fluorescence lifetime range, locating vector is formed, the estimation of fluorescence lifetime is obtained according to the spectrum peak position of search, least square method is recycled, estimates each component content.This method requires no knowledge about multicomponent substantially fluorescence lifetime and substantially constituent content information, can once calculate the fluorescence lifetime information of all components.
Description
Technical field
The present invention is a kind of multicomponent fluorescence lifetime and component ratio estimation method for Single Photon Counting,
The major function of realization is that each component fluorescence lifetime and each component are estimated from the data of time correlation single photon technique system
Ratio.
Background technique
Single Photon Counting time-correlated single photon counting t (TCSPC) first by Bollinger,
Tri- people of Bennett, Koechlin the sixties be the scintillator that excite by ray of detection shine and foundation, gradually answered later
Use the measurement of fluorescence lifetime.The advantages of TCSPC technology, is as follows：Temporal resolution is high, high sensitivity, and measurement accuracy is high, dynamic
Range is big, output data digitlization, convenient for computer storage and processing etc..It is obtained in the fields such as Contemporary Physics, chemistry, biology
It is widely applied, especially expands in time resolution scan laser microphotograph, unimolecule spectrum, Photon Correlation experiment and biological tissue
More there is its specific use in astigmatism chromatography.
On fluorescence lifetime algorithm for estimating, data fitting method is usually used.Since approximating method operand is huge, and hold very much
Local extremum easily is converged to, causes estimated result confidence level poor.Prony method, IEM method, CMM are developed recent years
The operands such as method, Phasor method are compared with submethod, but these methods are suitable for greatly one pack system, for multicomponent, or need
Will using approximating method carry out technology, and some algorithm use approximate method, estimated result there are deviation, need to result into
Row correction.
Summary of the invention
The present invention efficiently uses data, requires no knowledge about multicomponent substantially glimmering on the basis of analyzing TCSPC mathematical model
Light service life and substantially constituent content information can once calculate the fluorescence lifetime information of all components, in fluorescence lifetime base
On plinth, each component content information can be once calculated.Collected histogram data is combined first to be formed section compared with
Small new histogram, to eliminate the influence of partial noise and algorithm operation quantity can be greatly reduced.To new histogram interval censored data into
Row auto-correlation computation the characteristics of according to TCSPC, is smoothed autocorrelation matrix.Feature is carried out to its autocorrelation matrix
Value is decomposed, and signal subspace is obtained.According to possible fluorescence lifetime range, locating vector is formed, according to the spectrum peak position of search
The estimation of fluorescence lifetime is obtained, least square method is recycled, estimates each component content.
Assuming that shared P kind component, the data of TCSPC at a time t output can model as follows
Wherein fDjFor the content of j-th of component, τjFor the fluorescence lifetime of j-th of component, n (t) is additivity shot noise.Then
The output data in the mh section is in the histogram of TCSPC output
Wherein h is the resolution ratio of TCSPC, and m is the serial number of discretization sequence；
The data in all sections of histogram can be listed in together
Wherein M is the section number of histogram；
Its autocorrelation matrix is
RI=E [YYH]=E [(AS+N) (AS+N)H]=ARSAH+σ2∑0,
Wherein ()HIndicate conjugate transposition, ∑0For the autocorrelation matrix of noise, RS=E [SSH]；
Theorem：Enable λiAnd ui(i=1,2 ..., M) is equation RxU=λ ∑0The solution of u, wherein λiIt is arranged according to descending, then λi
And uiFor pencil of matrix (Rx, ∑0) characteristic value and feature vector.If RSFor non-singular matrix, then each column of matrix A are all orthogonal
In matrix Un=[uP+1 uP+2 … uM]；
Therefore, matrix U is being obtainedn, enable
Wherein A (τ)=| 1 eh/τ … e(M-1)h/τ|.Then QsearchThe maximum P peak value corresponding time of (τ) is exactly glimmering
The estimation time in light service life.
In a model, since each component is constant, the R in multicomponentSBy scarce order, directly it is with above-mentioned formula
It cannot obtain P peak value.Meanwhile above-mentioned formula is directly used in one-component, due to the influence of noise, performance can also compare
Difference；
Firstly, we carry out data aggregate to the histogram data of TCSPC output, b adjacent data are added up, it is false
If after polymerization, sharing K section.Data are as follows
Wherein：
In order to eliminate RSThe case where by scarce order, we are smoothed autocorrelation matrix.It can be put down using following
Sliding processing：
Wherein, XiFor a series of continuous datas taken out from X, each XiIt is isometric；
Eigenvalues Decomposition is carried out to R, the estimation of each fluorescence lifetime can be solved according to the method described above；
After the estimated result for obtaining fluorescence lifetime, we can obtain the content information of each component with following formula：
S=(BHB)-1BHX。
This method advantage is：(1) blind analysis.The fluorescence lifetime and component of any ingredient are required no knowledge about in advance.(2) it fits
For multiple component analyses, and primary search can complete multiple component fluorescence lifetime estimations.(3) in each component fluorescence lifetime
On the basis of estimated result, using the calculation formula of the constituent content derived by strict mathematical, once-through operation is it can be concluded that institute
There is the content of component.(4) precision of estimation result is high, and operand is moderate, is convenient for embedded system real-time implementation.(5) defeated to TCSPC
The group number of histogram requires smaller out, greatly reduces the data volume that acquisition system is transmitted to computer, can be in existing TCSPC
The SPAD of higher pixel is used in system.
Specific embodiment
Polymerization processing is carried out to the data of TCSPC output, obtains new histogram data
Obtain smoothed out autocorrelation matrix data
Eigenvalues Decomposition is carried out to smoothed out autocorrelation matrix, obtains matrix Un=[uP+1 uP+2 … uM], according to can
The fluorescence lifetime of energy, construction vector A (τ)=| 1 eh/τ … e(M-1)h/τ|, it obtains
Search QsearchThe P peak value corresponding time of (τ), the fluorescence lifetime of each component as estimated；
According to each component fluorescence lifetime estimated, according to
S=(BHB)-1BHX
Obtain the content estimation of each component.
Claims (1)
1. a kind of multicomponent fluorescence lifetime and component ratio estimation method for Single Photon Counting, requires no knowledge about
Multicomponent fluorescence lifetime and constituent content information once just calculate the fluorescence lifetime information of all components, in fluorescence lifetime
On the basis of, once calculate each component content information, which is characterized in that specific step is as follows：
Step 1：It is combined collected histogram data to form new histogram first, to eliminate the influence of noise and subtract
Small algorithm operation quantity carries out auto-correlation computation to new histogram interval censored data, the characteristics of according to TCSPC, to autocorrelation matrix into
Row smoothing processing carries out Eigenvalues Decomposition to its autocorrelation matrix, obtains signal subspace,
Step 2：According to possible fluorescence lifetime range, locating vector is formed, fluorescence lifetime is obtained according to the spectrum peak position of search
Estimation,
Step 3：Least square method is recycled, each component content is estimated；
The step 1 is specially to three：
Assuming that shared P kind component, the data modeling of TCSPC at a time t output is as follows
Wherein fDjFor the content of j-th of component, τjFor the fluorescence lifetime of j-th of component, n (t) is additivity shot noise, then exists
The output data in the mh section is in the histogram of TCSPC output
Wherein h is the resolution ratio of TCSPC, and m is the serial number of discretization sequence；
The data in all sections of histogram are listed in together
Wherein M is the section number of histogram,
For AS+N will be denoted as on the right of above formula equal sign convenient for simplified formula
Its autocorrelation matrix is
RI=E [YYH]=E [(AS+N) (AS+N)H]=ARSAH+σ2Σ0
Wherein ()HIndicate conjugate transposition, Σ0For the autocorrelation matrix of noise, RS=E [SSH], E [NNH]=σ2∑0；σ2To make an uproar
Acoustic energy；
Enable λiAnd uiFor equation RxU=λ Σ0The solution of u, wherein i=1,2 ..., M, λiIt is arranged according to descending, then λiAnd uiFor pencil of matrix
(Rx,Σ0) characteristic value and feature vector, if RSFor non-singular matrix, then each column of matrix A are all orthogonal to matrix Un=
[uP+1 uP+2 … uM]
Therefore, matrix U is being obtainedn, enable
Wherein A (τ)=| 1 eh/τ … e(M-1)h/τ|, then QsearchThe maximum P peak value corresponding time of (τ) is exactly the fluorescence longevity
The estimation time of life,
Firstly, carrying out data aggregate to the histogram data of TCSPC output, b adjacent data are added up, it is assumed that polymerization
Afterwards, K section is shared, data are as follows
Wherein：
In order to eliminate RSThe case where by scarce order, autocorrelation matrix is smoothed, using following smoothing processing：
Wherein, XiFor a series of continuous datas taken out from X, each XiIsometric, l is fluorescent component number；
Eigenvalues Decomposition is carried out to R, solves the estimation of each fluorescence lifetime,
After the estimated result for obtaining fluorescence lifetime, the content information of each component is obtained with following formula：
S=(BHB)-1BHB
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610052282.9A CN105675568B (en) | 2016-01-27 | 2016-01-27 | A kind of multicomponent fluorescence lifetime and component ratio estimation method for Single Photon Counting |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610052282.9A CN105675568B (en) | 2016-01-27 | 2016-01-27 | A kind of multicomponent fluorescence lifetime and component ratio estimation method for Single Photon Counting |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105675568A CN105675568A (en) | 2016-06-15 |
| CN105675568B true CN105675568B (en) | 2018-11-23 |
Family
ID=56303742
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610052282.9A Expired - Fee Related CN105675568B (en) | 2016-01-27 | 2016-01-27 | A kind of multicomponent fluorescence lifetime and component ratio estimation method for Single Photon Counting |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105675568B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106250683B (en) * | 2016-07-21 | 2019-12-10 | 天津大学 | Correction method of centroid algorithm for calculating fluorescence lifetime |
| CN106841143B (en) * | 2017-02-24 | 2019-02-19 | 国家纳米科学中心 | The acquisition methods of each species brightness and concentration |
| KR101886764B1 (en) * | 2017-03-31 | 2018-08-08 | 연세대학교 산학협력단 | Measuring apparatus to obtain high-speed data analysis method for multiple exponential decaying functions and measuring method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102597746A (en) * | 2009-09-29 | 2012-07-18 | 三井造船株式会社 | Method and device for FRET measurement |
| DE102012019471A1 (en) * | 2012-09-28 | 2014-04-03 | Carl Zeiss Microscopy Gmbh | Method for determining e.g. fluorescence lifetime of paper to identify banknote, involves recording phase difference values of sample light by runtime camera, and determining material characteristic based on difference values |
| CN104035122A (en) * | 2014-05-22 | 2014-09-10 | 沈阳东软医疗系统有限公司 | Method and device for correcting energy value |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20150042954A1 (en) * | 2013-08-08 | 2015-02-12 | University Of Rochester | System and Method for Fluorescence Lifetime Imaging Aided by Adaptive Optics |
-
2016
- 2016-01-27 CN CN201610052282.9A patent/CN105675568B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102597746A (en) * | 2009-09-29 | 2012-07-18 | 三井造船株式会社 | Method and device for FRET measurement |
| DE102012019471A1 (en) * | 2012-09-28 | 2014-04-03 | Carl Zeiss Microscopy Gmbh | Method for determining e.g. fluorescence lifetime of paper to identify banknote, involves recording phase difference values of sample light by runtime camera, and determining material characteristic based on difference values |
| CN104035122A (en) * | 2014-05-22 | 2014-09-10 | 沈阳东软医疗系统有限公司 | Method and device for correcting energy value |
Non-Patent Citations (5)
| Title |
|---|
| Fluorescence Lifetime Correlation Spectroscopy (FLCS): Concepts, Applications and Outlook;Peter Kapusta et al;《International Journal of Molecular Sciences》;20121009;第13卷;第12890-12910页 * |
| Fluorescence Lifetime Correlation Spectroscopy;Peter Kapusta et al;《J Fluoresc》;20071231;第17卷;第43-48页 * |
| 时间分辨荧光技术与荧光寿命测量;李东旭 等;《大学化学》;20080831;第23卷(第4期);第1-11页 * |
| 时间相关单光子计数法测量荧光寿命;张兴康 等;《化学通报》;19821231(第1期);第30-33页 * |
| 荧光寿命测定的现代方法与应用;房喻 等;《化学通报》;20011231(第10期);第631-636页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105675568A (en) | 2016-06-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chambers et al. | Star formation activity of cores within infrared dark clouds | |
| Savage et al. | Bayesian methods of astronomical source extraction | |
| Budavari et al. | Angular clustering with photometric redshifts in the Sloan Digital Sky Survey: Bimodality in the clustering properties of galaxies | |
| Bartels et al. | Bayesian model comparison and analysis of the Galactic disc population of gamma-ray millisecond pulsars | |
| CN105675568B (en) | A kind of multicomponent fluorescence lifetime and component ratio estimation method for Single Photon Counting | |
| Kistler et al. | Tomography of massive stars from core collapse to supernova shock breakout | |
| Prochaska et al. | The SDSS-DR5 Survey for Proximate Damped Lyα Systems | |
| Vacca et al. | Using rotation measure grids to detect cosmological magnetic fields: A Bayesian approach | |
| Witzemann et al. | Model-independent curvature determination with 21 cm intensity mapping experiments | |
| Cappelluti et al. | Clustering of x-ray-selected AGN | |
| US20150220488A1 (en) | System and method for interferometrically tracking objects using a low-antenna-count antenna array | |
| Allison et al. | The First Large Absorption Survey in H i (FLASH): I. Science goals and survey design | |
| US20210156991A1 (en) | System, device and method for imaging of objects using signal clustering | |
| Hu et al. | Anisotropic Turbulence in Position–Position–Velocity Space: Probing Three-dimensional Magnetic Fields | |
| Rogerson | Optimal geographic scales for local spatial statistics | |
| D’Amico et al. | Signal estimation in on/off measurements including event-by-event variables | |
| Duffy | Probing the nature of dark energy through galaxy redshift surveys with radio telescopes | |
| CN105548120B (en) | A kind of fluorescence resonance energy transfer multicomponent fluorescence lifetime method of estimation | |
| Schüssler et al. | The HESS multi-messenger program | |
| Sclavo et al. | Italian seas wave extremes: a preliminary assessment | |
| Gong et al. | Pairwise kinematic Sunyaev-Zel’dovich signal extraction efficacy and optical depth estimation | |
| Mayers et al. | Correlations between X-ray properties and Black Hole Mass in AGN: towards a new method to estimate black hole mass from short exposure X-ray observations | |
| Fang et al. | A Two-Stage Strategy for Retrieving Two-Dimensional Ocean Wave Spectra from Chinese Gaofen-3 SAR Wave Mode Products | |
| Gong et al. | Pairwise kSZ signal extraction efficacy and optical depth estimation | |
| Ghosh et al. | Bayesian framework to infer the Hubble constant from cross-correlation of individual gravitational wave events with galaxies |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20220919 Address after: National University of Defense Technology, Deya Road, Kaifu District, Changsha City, Hunan Province 410073 Patentee after: National University of Defense Technology Address before: Room 606, Building A3, Lugu Yuyuan, No. 27 Wenxuan Road, Changsha High-tech Development Zone, Changsha, Hunan Province, 410205 Patentee before: HUNAN JUNHAN INFORMATION TECHNOLOGY Co.,Ltd. |
|
| TR01 | Transfer of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20181123 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |