Quantitative Fluorescence Resonance Energy Transfer Measurement Based on Spectral Unmixing

Yin Ao; Chen Tongsheng

doi:10.3788/CJL202047.0207009

Chinese Journal of Lasers, Volume. 47, Issue 2, 207009(2020)

Quantitative Fluorescence Resonance Energy Transfer Measurement Based on Spectral Unmixing

Yin Ao^1,2 and Chen Tongsheng^1,2、*

¹Key Laboratory of Laser Life Science, Ministry of Education, College of Biophotonics, South China Normal University,Guangzhou, Guangdong 510631, China

²Guangdong Key Laboratory of Laser Life Science, College of Biophotonics, South China Normal University,Guangzhou, Guangdong 510631, China

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Figures & Tables(3)

Fig. 1. Emission spectra and excitation emission spectra of living HepG2 cells CTV, C5V, CVC, VCV, C+V and C32V+C^[62]. (a) Fluorescence images of cells with different FRET constructs. Scale bar is 10 μm; (b) unit-area normalized emission spectra measured from the corresponding cell 1 and cell 2 at Ex405 or Ex436 excitation; (c) unit-volume normalized excitation emission spectra measured from the corresponding cell 1 and cell 2 at Ex436 or Ex470 excitatio

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Fig. 2. Statistical E and R_C of HepG2 cells CTV, C5V, CVC, VCV, C+V and C32V+C with different SNR^[62]. (a) Statistical E with R_SN>9; (b) statistical R_C with R_SN>9; (c) statistical E with 3<R_SN<9; (d) statistical R_C with 3<R_SN<9; (e) statistical E with R_SN<3; (f) statistical R

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Table 1. Statistical E and R_C values in cell 1 and cell 2 measured by ExEm-spFRET and IIem-spFRET method^[62]

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Table 1. Statistical E and R_C values in cell 1 and cell 2 measured by ExEm-spFRET and IIem-spFRET method^[62]

Construct	Condition	E		R_C
Construct	Condition	ExEm-spFRET	IIem-spFRET	ExEm-spFRET	IIem-spFRET
	R_SN>9	0.04±0.01	0.04±0.01	0.94±0.03	0.95±0.03
CTV	9>R_SN>3	0.04±0.01	0.04±0.01	0.92±0.03	0.94±0.05
	R_SN<3	0.02±0.01	0.02±0.01	0.91±0.04	0.98±0.12
	R_SN>9	0.47±0.02	0.47±0.02	1.00±0.03	1.05±0.05
C5V	9>R_SN>3	0.45±0.01	0.45±0.01	0.98±0.03	1.04±0.14
	R_SN<3	0.42±0.02	0.41±0.02	0.95±0.07	0.98±0.20
	R_SN>9	0.41±0.02	0.42±0.02	0.52±0.01	0.44±0.05
CVC	9>R_SN>3	0.40±0.01	0.40±0.01	0.53±0.01	0.46±0.06
	R_SN<3	0.41±0.02	0.40±0.02	0.49±0.01	0.46±0.29
	R_SN>9	0.69±0.02	0.69±0.03	2.27±0.16	2.25±0.24
VCV	9>R_SN>3	0.67±0.02	0.66±0.02	2.28±0.08	3.12±0.50
	R_SN<3	0.70±0.02	0.64±0.03	2.31±0.02	4.28±1.37
	R_SN>9	0.04±0.01	0.03±0.01	0.73±0.22	0.76±0.23
C+V	9>R_SN>3	0.04±0.02	0.03±0.02	0.81±0.22	0.85±0.23
	R_SN<3	0.04±0.01	0.01±0.01	0.83±0.48	0.83±0.52
	R_SN>9	0.23±0.05	0.23±0.05	0.67±0.13	0.76±0.18
C32V+C	9>R_SN>3	0.23±0.04	0.23±0.05	0.70±0.13	0.68±0.21
	R_SN<3	0.22±0.06	0.22±0.06	0.66±0.19	0.63±0.23

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Yin Ao, Chen Tongsheng. Quantitative Fluorescence Resonance Energy Transfer Measurement Based on Spectral Unmixing[J]. Chinese Journal of Lasers, 2020, 47(2): 207009

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Paper Information

Category: biomedical photonics and laser medicine

Received: Oct. 8, 2019

Accepted: --

Published Online: Feb. 21, 2020

The Author Email: Tongsheng Chen (chentsh@ scnu.edu.cn)

DOI:10.3788/CJL202047.0207009

Topics

laser devices and laser physics

Lasers and Laser Optics

Laser physics

laser manufacturing

Instrumentation, Measurement and Metrology

Table 1. Statistical E and RC values in cell 1 and cell 2 measured by ExEm-spFRET and IIem-spFRET method[62]

Table 1. Statistical E and RC values in cell 1 and cell 2 measured by ExEm-spFRET and IIem-spFRET method[62]

Table 1. Statistical E and R_C values in cell 1 and cell 2 measured by ExEm-spFRET and IIem-spFRET method^[62]

Table 1. Statistical E and R_C values in cell 1 and cell 2 measured by ExEm-spFRET and IIem-spFRET method^[62]