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Novel Fluorescent Probes for the Oxidative Milieu

Figure 4. Single step photobleaching of SGFP2 in the ER. Cells expressing ss-SGFP2 were extensively fixed for 4 days with paraformaldehyde
as described in Materials and Methods and single molecules were directly visualized by total reflection fluorescence microscopy. (A) When 400 frames
were recorded at a frame rate of 250 Hz, 4 spots disappeared from the first frame in the recorded 1.6 sec (indicated by arrows). The width and height
of an image is 13.6 mm. (B) Kymograph of each spot (1–4) from panel A. A single pixel image of the Y axis is lined along progression of time. (C)
Quantification of the fluorescent signal in panel B. All spots were photobleached in a single step.
doi:10.1371/journal.pone.0037551.g004

fluorescence was inconsistent and in some cells (asterisk) only a very exposed to Cu2+ at pH 5 to induce cytotoxic damage [37], large
faint autofluorescence was observed. Most of the signal appeared aggregates were formed on the surface of cells, leading to apoptosis
to be in the Golgi and ER-like reticular structures. When COS7 of some cells (Supplementary movie).
cells expressing the ss-cfSGFP2-prion fusion protein were briefly

PLoS ONE | www.plosone.org 6 May 2012 | Volume 7 | Issue 5 | e37551