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Fluorescence imaging of single molecules and photosynthetic membranes with two-photon excitation

Conference ·
OSTI ID:560345
; ;  [1]
  1. Pacific Northwest National Lab., Richland, WA (United States)
We report the imaging of single-molecule fluorescence induced by two-photon excitation in ambient conditions. Using an inverted fluorescence microscope, we obtained the two-photon images of different single fluorophores (Rhodamine B, Sulforhodamine 101, Coumarin 535 on poly-methyl methacrylate films) and biological membrane fragments by Faster scanning the sample with respect to a diffraction limited focus of a mode-locked Ti: sapphire laser beam. The signal to background ratio was as high as 50:1 and the full width at half maximum (250nm) of a single-molecule peak was significantly shorter than that for one photon excitation. With its high sensitivity and simplicity, the two-photon experiment offers a valuable approach for spectroscopic studies on individual immobilized molecules.
OSTI ID:
560345
Report Number(s):
CONF-970443--
Country of Publication:
United States
Language:
English

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