Supplementary Figure 6: Spectral properties, pH sensitivity and photostabilities of mEosEM.
From: mEosEM withstands osmium staining and Epon embedding for super-resolution CLEM
a, Absorption spectra of mEosEM. The absorption peak of mEosEM is at 504 nm. b, Excitation and emission spectra of mEosEM in the green state. The excitation peak and emission peak of mEosEM are at 503 nm and 513 nm, respectively. c, Excitation and emission spectra of mEosEM in the red state. The excitation peak and the emission peak of mEosEM are at 569 nm and 579 nm, respectively. d, The pH sensitivity of mEosEM in the green state. The fluorescence intensity of mEosEM was plotted against the pH and fitted using Equation 1 (described in Methods). The calculated pKa is 6.0. e, The pH sensitivity of mEosEM in the red state. The fluorescence intensity of mEosEM was plotted against the pH and fitted using Equation 1 (described in Methods). The calculated pKa is 6.5. The experiments of a-g were performed three times with similar results. f, Photobleaching curves of mEosEM and mEos4b in the green state. Fluorescence intensity was plotted against time and presented as the mean ± s.d., n = 6 cells. Illumination intensity (488-nm light), 3 W/cm2. g, Photobleaching curves of mEosEM and mEos4b in the red state. Fluorescence intensity was plotted against time and presented as the mean ± s.d., n = 5 cells. Illumination intensity (561-nm light), 300 W/cm2. h, Statistics of the time when the green fluorescence decreased to half of its original signal. Bars represent mean ± s.d., n = 6 cells. Pair-wised two-tailed t-test was performed. i, Statistics of the time when red fluorescence decreased to half of its original signal. Bars represent mean ± s.d., n = 5 cells. Pair-wised two-tailed t-test was performed.
