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| Fluorescence Spectra and Fluorescence Saturation Intensity Analysis of Hepatic Cell，Hepatoma Carcinoma Cell and Hepatic Fibrosis Cell |
| HU Yue, FU Yun*, LI Xin-yang, LI Yong-liang |
| School of Electro-Optical Engineering, Changchun University of Science and Technology, Changchun 130022, China |
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Abstract Researched the fluorescence spectral characteristics of hepatic cell, hepatoma carcinoma cell and hepatic fibrosis cell to provide spectroscopy basis for early screening of liver cancer. The purpose of the experiment included the detection of cells by fluorescence spectrometer to acquire specific fluorescence spectra; eliminate background noise by de-Raman scattering to acquire fluorescence spectra of five cell concentrations；the cell diameter was detected by flow cytometry, analysis of the diameter characteristics of four cells based on two-parameter scatterplot; combined with Gauss multi-peak fitting parameter analysis results. The experimental process began as hepatic cell, hepatic fibrosis cell and two hepatoma carcinoma cell were detected by fluorescence spectroscopy and analyzed by flow cytometry as well. Then, Gauss multi-peak was used to fit the fluorescence spectrum differences, and the difference among the cells was analyzed. Finally, the fluorescence saturation intensity nonlinear fitting curve was compared to analyze the effect of cell size on it. The results showed that there were two specific fluorescence peaks in the hepatic cell between 550 and 750 nm. Combined with Gaussian multi-peak fitting, the peak height, peak center and peak width were analyzed. The results showed, the first peak was at 592 nm and the second peak was at 682 nm, and the former was significantly higher than the latter. In hepatoma carcinoma cell and hepatic fibrosis cell there was a third specific fluorescence peak at 726 nm except for two peaks at the same position as hepatic cell, and the maximum excitation intensity was obtained at 592 nm, and the fluorescence peak at 726 nm was higher than the second peak at 682 nm. The width of hepatoma carcinoma cell and hepatic fibrosis cell were basically the same. The maximum excitation peak width of hepatic cell was slightly smaller than that of the other two cells, but the small peak width at 682 nm was slightly larger than that of the diseased cells. The results of flow cytometry showed that the diameter of hepatoma carcinoma cell was the largest diameter, and hepatic fibrosis cell was larger than hepatic cell. Through fitting the curve of the fluorescence saturation intensity trend of the cell with the concentration curve by the nonlinear curve and analyzing the slope of the curve, the results showed that the fluorescence saturation intensity trend of the four cells increased with the increase of the cell concentration, but gradually showed the fluorescence saturation state. As the cell diameter increased, the trend of maximum fluorescence saturation intensity was more obvious, and auto fluorescence spectrum efficiency of single cells decreased. The results showed that the rational combination of cell morphology and spectroscopy, combined with two methods of analysis, improved the accuracy and effectiveness of cell judgment. By studying the fluorescence spectrum characteristics of hepatic cell, hepatoma carcinoma cell, hepatic fibrosis cell, and analyzing the fluorescence saturation intensity in combination with cell diameter, it can provide a certain spectral basis for the study of liver disease cells.
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Received: 2019-01-04
Accepted: 2019-04-27
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Corresponding Authors:
FU Yun
E-mail: linda_fy@cust.edu.cn
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