Fig. 2: Dronpa-2 for characterization of illumination in wide-field fluorescence imaging.
a–i, Epifluorescence microscopy. a, Initial image of the Dronpa-2 solution under homogeneous illumination. b, Time fluorescence response (τ = 0.63 s), maps (c,e) and histograms (d,f) of the characteristic time τ (c,d) and light intensity (e,f) in the field of view. g–i, Initial image (g) and maps of light intensity (h,i) of Dronpa-2 in solution (h) or in polyacrylamide gel under patterned illumination. j–l, Macroscopic fluorescence imaging: initial image of the Dronpa-2 solution (j) and experimental (k) and simulated (l) maps of light intensity. The blue line shows the angle of the linear light gradient; the angle between the simulated gradient and the measured one is 3°. a–f,h,j–l, 10 μM Dronpa-2 solution or 19% polyacrylamide gel in Tris buffer pH 7.4 (50 mM Tris, 150 mM NaCl). Scale bars, 100 μm (a,c,e,g,h); 3 mm (j–l). T = 293 K. λexc = 470 nm; λem = 550 nm (text and Supplementary Tables 1 and 2). Independent repeats, more than 30 (a,c,e); 3 (g–i); 15 (j,k).
