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Fig. 3. Schematic drawing of prism-type TIRM. (A) Schematic drawing of the optical setup. The green laser illuminates the surface in a TIR mode while the red laser is blocked. Both Cy3 and Cy5 fluorescence spectra are recorded independently by an intensified charge-coupled device. (B) Single-molecule images are obtained by the system. The two images show colocalization of Cy3- and Cy5-labeled nucleotides in the same template (scale bar 10 mm). (C) Schematic of primed DNA templates attached to the surface of a microscope slide via streptavidin-biotin. Adapted from a figure originally published in Braslavsky et al. (2003). Copyright (2003), reprinted with permission from National Academy of Sciences (USA).

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