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. 2007 May 1;64(4):886–903. doi: 10.1111/j.1365-2958.2007.05706.x

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© 2007 The Authors Journal compilation © 2007 Blackwell Publishing Ltd

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Fig. 2 — Single molecule studies on bacteriophage phi29 DNA packaging motor (adapted from (Shu et al., 2007) with permission from the European Molecular Biology Organization). A–C. Single molecule dual-view imaging for pRNA counting on the motor. (A) Experimental design. (B) Single molecule counting of Cy3 (green) or Cy5 (red)-labelled pRNA via step photobleaching. Each step represents the presence of single Cy3 or Cy5 on pRNA. (C) Single molecule dual imaging of procapsid containing Cy3-pRNA (green spots), Cy5-pRNA (red spots) or both Cy3 and Cy5-pRNA (yellow spots). The histograms of procapsid with Cy3 photobleaching steps were compared with the theoretical histogram predicted to have three Cy3 pRNA with a labelling efficiency of 70%. The stoichiometry of ferritin-conjugated pRNA on procapsid also revealed by EM. D. Direct observation of phi29 DNA translocation with gradual reduction of swinging range (distance) of the fluorescence microsphere at real time. The experimental design and the frames of the image are also shown.