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Proceedings Paper

Single dye molecules observed by total internal reflection fluorescence microscopy
Author(s): Jinyuan Wang; Guo Fu; Chen Wang; Li Liu; Guiying Wang
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Paper Abstract

Total internal reflection fluorescence microscopy (TIRFM) was used to image single molecules with evanescent waves. The molecules were excited by an evanescent wave with different intensities. Single molecules were imaged on low-noise high-quantum-yield charge-coupled device (CCD) cameras. Two-step photobleaching behavior was observed. Duration from fluorescent spots appearing to disappearing was counted. The duration was decided by the speed of photobleaching. Peak intensity was counted as exciting intensity change. The proportion relationship between the reciprocal of duration and exciting intensity was obtained. The emitted intensity and duration of fluorescein were compared with GFP. A single molecules emit the same number photons was proved.

Paper Details

Date Published: 1 May 2007
PDF: 6 pages
Proc. SPIE 6534, Fifth International Conference on Photonics and Imaging in Biology and Medicine, 65341B (1 May 2007); doi: 10.1117/12.741569
Show Author Affiliations
Jinyuan Wang, Shanghai Institute of Optics and Fine Mechanics (China)
Guo Fu, Shanghai Institute of Optics and Fine Mechanics (China)
Chen Wang, Shanghai Institute of Optics and Fine Mechanics (China)
Li Liu, Shanghai Institute of Optics and Fine Mechanics (China)
Guiying Wang, Shanghai Institute of Optics and Fine Mechanics (China)


Published in SPIE Proceedings Vol. 6534:
Fifth International Conference on Photonics and Imaging in Biology and Medicine

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