Single dye molecules observed by total internal reflection fluorescence microscopy

Jinyuan Wang; Guo Fu; Chen Wang; Li Liu; Guiying Wang

doi:10.1117/12.741569

1 May 2007 Single dye molecules observed by total internal reflection fluorescence microscopy

Jinyuan Wang, Guo Fu, Chen Wang, Li Liu, Guiying Wang

Proceedings Volume 6534, Fifth International Conference on Photonics and Imaging in Biology and Medicine; 65341B (2007) https://doi.org/10.1117/12.741569
Event: Fifth International Conference on Photonics and Imaging in Biology and Medicine, 2006, Wuhan, China

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.

Citation Download Citation

Jinyuan Wang, Guo Fu, Chen Wang, Li Liu, and Guiying Wang "Single dye molecules observed by total internal reflection fluorescence microscopy", Proc. SPIE 6534, Fifth International Conference on Photonics and Imaging in Biology and Medicine, 65341B (1 May 2007); https://doi.org/10.1117/12.741569

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