TABLE 2.

Time-resolved fluorescence anisotropy of EGFP and DsRed in both solution (pH 7.2) and in live RBL cells as a function of excitation and detection wavelengths

Protein	Environment	λx/nm	λfl/nm	φ1/ns	A1	φ2/ns	A2	r0	n*
EGFP	Solution	485	585 ± 75	—	—	17(1)	—	0.383(8)	6
		740	515 ± 15	—	—	17.9(2)	0.45(1)	0.45(1)†	2
			450 ± 40	0.58(4)	0.03	11.6(9)	0.40(1)	0.43(1)†	2
EGFP	Cytoplasm	485	580 ± 75	1.6(5)	0.018(6)	42(4)	0.36(2)	0.38(1)	7
LynB-EGFP	Plasma membrane	485	585 ± 75	1.2(8)	0.017(5)	80(14)	0.35(2)	0.37(1)	8
DsRed	Solution	485	580 ± 75	0.29(3)	0.06(1)	76(5)	0.26(2)	0.32(1)	5
		740	585 ± 20	0.9(1)	0.05(1)	70(5)	0.34(1)	0.39(1)†	3
			480 ± 45	0.21(3)	0.06(1)	11.6(5)	0.34(1)	0.40(1)†	2
DsRed	Cytoplasm	485	585 ± 20	0.29(7)	0.05(1)	134(31)	0.26(1)	0.31(1)	4
LynB-DsRed	Plasma membrane‡	485	585 ± 75	0.21(2)	0.04(1)	3.5(9)	0.04(1)	0.29(2)	3
DsRed2	Cytoplasm	485	580 ± 75	0.39(7)	0.04(1)	111(11)	0.21(1)	0.25(1)	3

The fitting parameters are defined in Eq. 6.

^*The number of measurements (n), in either solution or live RBL cells, includes different cells and transfections.

^†The maximal theoretical value for the initial 2P-anisotropy is 0.57 as compared with 0.4 for 1P-excitation.

^‡The anisotropy decays as a biexponential plus a residual anisotropy r_∞ = 0.21 ± 0.02.