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Sensors 2018, 18(2), 442; https://doi.org/10.3390/s18020442

Analysis of Flow Cytometric Fluorescence Lifetime with Time-Delay Estimation of Pulse Signals

1
Beijing Key Laboratory for Optoelectronics Measurement Technology, Beijing Information Science and Technology University, Beijing 100192, China
2
Key Lab of Microelectronic Devices & Integrated Technology, Institute of Microelectronics, Chinese Academy of Sciences, Beijing 100029, China
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Received: 24 December 2017 / Revised: 29 January 2018 / Accepted: 30 January 2018 / Published: 3 February 2018
(This article belongs to the Special Issue Optical Biochemical Sensor Systems and Applications)
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Abstract

The measurement of fluorescence lifetimes emerged in flow cytometry because it is not impacted by the non-linearity, which occurs in fluorescence intensity measurements. However, this significantly increases the cost and complexity of a traditional flow cytometer. This work reports a simple method of fluorescence lifetime measurement of a flow cytometer based on the cytometric fluorescence pulse time-delay estimation and hardware time-delay calibration. The modified chirp Z-transform (MCZT) algorithm, combined with the algorithm of fine interpolation of correlation peak (FICP), is applied to improve the temporal resolution of the cross-correlation function of the scattering and fluorescence signals, which in turn improves the time-delay estimation accuracy. The estimation accuracy is verified by Gauss fitting. Cells that were labeled simultaneously with three-color reagents are measured; the statistical results of 5000 cells are compared with reference values and are verified with the pulse width variation. The results show the potential of fluorescence lifetime measurements in the traditional flow cytometer. View Full-Text
Keywords: flow cytometry; time-delay estimation; fluorescence lifetime; pulse width variation flow cytometry; time-delay estimation; fluorescence lifetime; pulse width variation
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
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Zhu, L.; Zhang, W.; Dong, M.; Lou, X. Analysis of Flow Cytometric Fluorescence Lifetime with Time-Delay Estimation of Pulse Signals. Sensors 2018, 18, 442.

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