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Energy transfer rates in FRET and BRET modes. ( a – c ) Schemes of the estimation of energy transfer rates and radiative decay rates. The fluorescent ( a ) and bioluminescent ( b ) spectra of hyBRET sensors were fitted with the spectra of CFP, YFP, and RLuc8 ( c ) by the <t>nlinfit</t> function of <t>MATLAB</t> to estimate the parameters. ( d ) Overlay of the measured (blue) and fitted (red) spectra of hyBRET-ERK before and after TPA stimulation. Envelopes with light red show the 95% confidence interval in the fitted curves. The y-axis indicates the emission intensity normalized by the area of spectrum. ( e ) Summary of the energy flow absorbed by RLuc8. Rates of energy transfer and radiative decay are shown. E RC , E RY , and E CY are the energy transfer rates from RLuc8 to CFP, RLuc8 to YFP, and YFP to CFP, respectively. ϕ C , ϕ Y , and ϕ R are the quantum efficiencies of CFP, YFP, and RLuc8, respectively.
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Energy transfer rates in FRET and BRET modes. ( a – c ) Schemes of the estimation of energy transfer rates and radiative decay rates. The fluorescent ( a ) and bioluminescent ( b ) spectra of hyBRET sensors were fitted with the spectra of CFP, YFP, and RLuc8 ( c ) by the <t>nlinfit</t> function of <t>MATLAB</t> to estimate the parameters. ( d ) Overlay of the measured (blue) and fitted (red) spectra of hyBRET-ERK before and after TPA stimulation. Envelopes with light red show the 95% confidence interval in the fitted curves. The y-axis indicates the emission intensity normalized by the area of spectrum. ( e ) Summary of the energy flow absorbed by RLuc8. Rates of energy transfer and radiative decay are shown. E RC , E RY , and E CY are the energy transfer rates from RLuc8 to CFP, RLuc8 to YFP, and YFP to CFP, respectively. ϕ C , ϕ Y , and ϕ R are the quantum efficiencies of CFP, YFP, and RLuc8, respectively.
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Energy transfer rates in FRET and BRET modes. ( a – c ) Schemes of the estimation of energy transfer rates and radiative decay rates. The fluorescent ( a ) and bioluminescent ( b ) spectra of hyBRET sensors were fitted with the spectra of CFP, YFP, and RLuc8 ( c ) by the <t>nlinfit</t> function of <t>MATLAB</t> to estimate the parameters. ( d ) Overlay of the measured (blue) and fitted (red) spectra of hyBRET-ERK before and after TPA stimulation. Envelopes with light red show the 95% confidence interval in the fitted curves. The y-axis indicates the emission intensity normalized by the area of spectrum. ( e ) Summary of the energy flow absorbed by RLuc8. Rates of energy transfer and radiative decay are shown. E RC , E RY , and E CY are the energy transfer rates from RLuc8 to CFP, RLuc8 to YFP, and YFP to CFP, respectively. ϕ C , ϕ Y , and ϕ R are the quantum efficiencies of CFP, YFP, and RLuc8, respectively.
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Energy transfer rates in FRET and BRET modes. ( a – c ) Schemes of the estimation of energy transfer rates and radiative decay rates. The fluorescent ( a ) and bioluminescent ( b ) spectra of hyBRET sensors were fitted with the spectra of CFP, YFP, and RLuc8 ( c ) by the <t>nlinfit</t> function of <t>MATLAB</t> to estimate the parameters. ( d ) Overlay of the measured (blue) and fitted (red) spectra of hyBRET-ERK before and after TPA stimulation. Envelopes with light red show the 95% confidence interval in the fitted curves. The y-axis indicates the emission intensity normalized by the area of spectrum. ( e ) Summary of the energy flow absorbed by RLuc8. Rates of energy transfer and radiative decay are shown. E RC , E RY , and E CY are the energy transfer rates from RLuc8 to CFP, RLuc8 to YFP, and YFP to CFP, respectively. ϕ C , ϕ Y , and ϕ R are the quantum efficiencies of CFP, YFP, and RLuc8, respectively.
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MathWorks Inc nonlinear least-squares regression procedure
Energy transfer rates in FRET and BRET modes. ( a – c ) Schemes of the estimation of energy transfer rates and radiative decay rates. The fluorescent ( a ) and bioluminescent ( b ) spectra of hyBRET sensors were fitted with the spectra of CFP, YFP, and RLuc8 ( c ) by the <t>nlinfit</t> function of <t>MATLAB</t> to estimate the parameters. ( d ) Overlay of the measured (blue) and fitted (red) spectra of hyBRET-ERK before and after TPA stimulation. Envelopes with light red show the 95% confidence interval in the fitted curves. The y-axis indicates the emission intensity normalized by the area of spectrum. ( e ) Summary of the energy flow absorbed by RLuc8. Rates of energy transfer and radiative decay are shown. E RC , E RY , and E CY are the energy transfer rates from RLuc8 to CFP, RLuc8 to YFP, and YFP to CFP, respectively. ϕ C , ϕ Y , and ϕ R are the quantum efficiencies of CFP, YFP, and RLuc8, respectively.
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Energy transfer rates in FRET and BRET modes. ( a – c ) Schemes of the estimation of energy transfer rates and radiative decay rates. The fluorescent ( a ) and bioluminescent ( b ) spectra of hyBRET sensors were fitted with the spectra of CFP, YFP, and RLuc8 ( c ) by the nlinfit function of MATLAB to estimate the parameters. ( d ) Overlay of the measured (blue) and fitted (red) spectra of hyBRET-ERK before and after TPA stimulation. Envelopes with light red show the 95% confidence interval in the fitted curves. The y-axis indicates the emission intensity normalized by the area of spectrum. ( e ) Summary of the energy flow absorbed by RLuc8. Rates of energy transfer and radiative decay are shown. E RC , E RY , and E CY are the energy transfer rates from RLuc8 to CFP, RLuc8 to YFP, and YFP to CFP, respectively. ϕ C , ϕ Y , and ϕ R are the quantum efficiencies of CFP, YFP, and RLuc8, respectively.

Journal: Scientific Reports

Article Title: A platform of BRET-FRET hybrid biosensors for optogenetics, chemical screening, and in vivo imaging

doi: 10.1038/s41598-018-27174-x

Figure Lengend Snippet: Energy transfer rates in FRET and BRET modes. ( a – c ) Schemes of the estimation of energy transfer rates and radiative decay rates. The fluorescent ( a ) and bioluminescent ( b ) spectra of hyBRET sensors were fitted with the spectra of CFP, YFP, and RLuc8 ( c ) by the nlinfit function of MATLAB to estimate the parameters. ( d ) Overlay of the measured (blue) and fitted (red) spectra of hyBRET-ERK before and after TPA stimulation. Envelopes with light red show the 95% confidence interval in the fitted curves. The y-axis indicates the emission intensity normalized by the area of spectrum. ( e ) Summary of the energy flow absorbed by RLuc8. Rates of energy transfer and radiative decay are shown. E RC , E RY , and E CY are the energy transfer rates from RLuc8 to CFP, RLuc8 to YFP, and YFP to CFP, respectively. ϕ C , ϕ Y , and ϕ R are the quantum efficiencies of CFP, YFP, and RLuc8, respectively.

Article Snippet: The Lerbenberg-Marquardt method implemented in the nlinfit function in MATLAB (MathWorks, Natick, MA) and solver function in Microsoft Excel (Microsoft Corporation, Redmond, WA) were used for the non-linear regression.

Techniques: