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《Acta Optica Sinica》 2014-01
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Multicolor Fluorescence Detection in the Multiplex Quantitative PCR System and Spectra Crosstalk Correction Method

Zang Liuqin;Zhang Zhenxi;Miao Baogang;Peng Niancai;Li Zheng;Key Laboratory of Biomedical Information Engineering,Ministry of Education,School of Life Science and Technology,Xi′an Jiaotong University;State Key Laboratory for Manufacturing System Engineering,School of Mechanical Engineering,Xi′an Jiaotong University;  
Optical design for multicolor fluorescence detection is the core part of multiplex quantitative polymerase chain reaction(PCR)system.According to the system requirements for excitation uniform,detection time and sensitivity,an optical system is presented,which is based on magneto-optical switch and photomultiplier tube(PMT) detector.Four LEDs excitation unit and PMT detection unit are adopted to improve system sensitivity.Electronic optical switch combined with the one-dimensional scanning mechanism and filter to implement four-color fluorescence scanning on a 96-well standard PCR plate.The design avoids the crosstalk among the four color optical paths. Whereas the crosstalk generated by the spectral characteristics of the fluorescent dye itself can be estimated by standard iterative four-dimensional clustering analysis algorithm.We finally evaluate the systematical fluorescent spectra crosstalk by the experimental study of four commonly used dyes.
【Fund】: 国家重大科学仪器设备开发专项(2012YQ030261);; 国家自然科学基金(61120106013);; 国家科技支撑计划(2012BAI19B057)
【CateGory Index】: O433
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