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. 2015 Jun;79:125–131. doi: 10.1016/j.fgb.2015.03.022

Fig. 2.

Fig. 2

Signal intensity and bleaching behavior of GFP proteins in epi-fluorescence microscopy. (A) Images showing cytoplasmic expression of A. coerulescens GFP (AcGFP), enhanced GFP from A. victoria (eGFP) and codon-optimized enhanced GFP from A. victoria (ZtGFP). Note that Z. tritici shows virtually no auto fluorescence (inset shows extreme image processing, showing very low cytoplasmic background). All images were acquired using 150 ms exposure time and binning 1 and identically processed. Bar represents 10 μm. (B) Bar chart showing average intensity of cytoplasmic fluorescence of various GFPs. Autofluor.: background fluorescence without expressing a GFP; AcGFP: cells expressing A. coerulescens GFP; eGFP; cells expressing enhanced GFP from A. victoria; ZtGFP; cells expressing Z. tritici codon-optimized enhanced GFP from A. victoria. Mean ± standard error of the mean is shown, sample size n is indicated. Triple asterisk indicates significant difference at P < 0.0001, Student t-test. (C) Graph showing decay of fluorescent signals due to photo-bleaching. AcGFP: cells expressing A. coerulescens GFP; eGFP; cells expressing enhanced GFP from A. victoria; ZtGFP; cells expressing Z. tritici codon-optimized enhanced GFP from A. victoria. Each data point is given as mean ± standard error of the mean, sample size n is indicated. Note that little variation is found between experiments and that the standard error of the mean is very small.