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Correction method of center of mass method used for calculating fluorescent lifetime

A technology of fluorescence lifetime and centroid algorithm, which is applied in computing, special data processing applications, instruments, etc., can solve problems such as low photon utilization rate, low precision, and difficult control of gate delay time, so as to improve accuracy, The effect of expanding the reduction range and lifting the limitation of laser frequency

Inactive Publication Date: 2016-12-21
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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  • Correction method of center of mass method used for calculating fluorescent lifetime
  • Correction method of center of mass method used for calculating fluorescent lifetime
  • Correction method of center of mass method used for calculating fluorescent lifetime

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Embodiment Construction

[0037] The technical scheme adopted in the present invention is that based on the two-step calibration method of RLD pre-estimation, the calibration method is processed and executed in a computer or FPGA, including the following steps:

[0038] The decay lifetime to be restored is τ, the laser frequency used by the time-domain TCSPC system is F (period is T), each laser period is regarded as a detection window, and at most one photon can be recorded in each laser period. The time period for 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 solving windows is m), and the photon count value of each part is N respectively 1 , N 2 , N m , the parameters needed to judge and compare in the two-step calibration process are the photon count ratio K obtained by the actual measurement N and the photon count ratio K obtained by theoretical calculation 0 , the total ...

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

Claims

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Application Information

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IPC IPC(8): G06F19/00
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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