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Calibration method for the centroid algorithm used to calculate fluorescence lifetime

A technology of fluorescence lifetime and correction method, which is applied in complex mathematical operations, instruments, informatics, etc., can solve the problems of difficult control of gate delay time, low photon utilization rate, and low precision, and achieve the removal of laser frequency restrictions , improve accuracy, and expand the effect of restoring range

Inactive Publication Date: 2019-12-10
TIANJIN UNIV
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Problems solved by technology

The advantage of the RLD algorithm is that the algorithm is simple and fast, but the disadvantages are obvious, the accuracy is not high, the photon utilization rate is very low, and the gate delay time is difficult to control

Method used

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  • Calibration method for the centroid algorithm used to calculate fluorescence lifetime
  • Calibration method for the centroid algorithm used to calculate fluorescence lifetime
  • Calibration method for the centroid algorithm used to calculate fluorescence lifetime

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Embodiment Construction

[0036] The technical scheme adopted in the present invention is a two-step correction method based on RLD pre-estimation, and the correction method is processed and executed in a computer or FPGA, including the following steps:

[0037] The attenuation lifetime to be restored is τ, the laser frequency used by the time-domain TCSPC system is F (period T), each laser cycle is regarded as a detection window, and at most one photon can be recorded in each laser cycle, the CMM algorithm The time period of data extraction is T T (T T The time length is p laser cycles T), the software processing system divides each detection time window into m parts (that is, the number of solution windows is m), and the photon count value of each part is N 1 , N 2 , N m , the parameter of judgment and comparison required in the two-step calibration process is the measured photon count ratio K N and the photon count ratio K obtained by theoretical calculation 0 , the total number of photons reco...

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Abstract

The invention relates to the field of fluorescent lifetime imaging, and the lifetime estimation accuracy is improved for expanding the fluorescent lifetime estimation range under the same laser frequency condition. An RLD (Rapid Lifetime Determination) pre-estimation method is improved, so that the iterative operation is avoided. According to the adopted technical scheme, a correction method of a center of mass method used for calculating fluorescent lifetime comprises the following steps that to-be-recovered decay lifetime is tau, the laser frequency adopted by a time-correlated single-photon counting (TCSPC) system is F, the recorded total photon number is NC, and the photon arrival time summation obtained by quantification is NT; a threshold is defined and determination is carried out; when the lifetime is smaller than the threshold, a lifetime deviation is compensated by adopting nonlinear one-iteration; or otherwise, the fluorescent lifetime is estimated through the RLD method; and correction is carried out through nonlinear compensation to obtain final fluorescent lifetime. The method is mainly applied to the occasion of designing and manufacturing fluorescent lifetime imaging products.

Description

technical field [0001] The invention relates to the field of fluorescence lifetime imaging, in particular to an estimation algorithm of real-time fluorescence decay lifetime. Suitable for high-speed fluorescence lifetime analysis. Specifically, it relates to a hardware-executable center of mass algorithm (Center of Mass Method, CMM), in particular to a correction method for the center of mass algorithm used to calculate fluorescence lifetime. Background technique [0002] The fluorescence lifetime is not affected by factors such as changes in excitation light intensity, fluorophore concentration, and environmental absorption, and has very high sensitivity and molecular specificity. Fluorescence lifetime imaging quantitatively measures the microenvironment through the difference in the decay time of fluorescent tracer molecules excited in different environments. Fluorescence imaging analysis technology is widely used in life sciences, medicine and other fields, such as clin...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G06F17/10
CPCG16Z99/00
Inventor 徐江涛乔俊高静史再峰高志远聂凯明
Owner TIANJIN UNIV
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