References and Notes

The room temperature dynamics of single sulforhodamine 101 molecules dispersed on a glass surface are investigated on two different time scales with near-field optics. On the 10-2- to 102-second time scale, intensity fluctuations in the emission from single molecules are examined with polarization measurements, providing insight into their spectroscopic properties. On the nanosecond time scale, the fluorescence lifetimes of single molecules are measured, and their excited-state energy transfer to the aluminum coating of the near-field probe is characterized. A movie of the time-resolved emission demonstrates the feasibility of fluorescence lifetime imaging with single molecule sensitivity, picosecond temporal resolution, and a spatial resolving power beyond the diffraction limit.

[1]  E. Betzig,et al.  Combined shear force and near‐field scanning optical microscopy , 1992 .

[2]  Floyd E. Bloom,et al.  Advances in Optical and Electron Microscopy , 1967, The Yale Journal of Biology and Medicine.

[3]  W. Moerner,et al.  Optical detection and spectroscopy of single molecules in a solid. , 1989, Physical review letters.

[4]  W. Lukosz,et al.  Optical-environment-dependent effects on the fluorescence of submonomolecular dye layers on interfaces , 1987 .

[5]  H. Bethe.  Theory of Diffraction by Small Holes , 1944 .

[6]  B. Persson,et al.  Excited states at metal surfaces and their non-radiative relaxation , 1984 .

[7]  Robert J. Chichester,et al.  Single Molecules Observed by Near-Field Scanning Optical Microscopy , 1993, Science.

[8]  T. D. Harris,et al.  Breaking the Diffraction Barrier: Optical Microscopy on a Nanometric Scale , 1991, Science.

[9]  J. Lakowicz,et al.  Fluorescence lifetime imaging of free and protein-bound NADH. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[10]  Martin Grayson,et al.  Kirk-Othmer Concise encyclopedia of chemical technology , 1985 .

[11]  M. Orrit,et al.  Single pentacene molecules detected by fluorescence excitation in a p-terphenyl crystal. , 1990, Physical review letters.

[12]  U. Dürig,et al.  Near‐field optical‐scanning microscopy , 1986 .

[13]  E. Betzig,et al.  Near-Field Optics: Microscopy, Spectroscopy, and Surface Modification Beyond the Diffraction Limit , 1992, Science.

[14]  R. Toledo-Crow,et al.  Near‐field differential scanning optical microscope with atomic force regulation , 1992 .

[15]  Lloyd M. Davis,et al.  Single photon avalanche diode for single molecule detection , 1993, Optical Society of America Annual Meeting.

[16]  K. Stetson,et al.  Progress in optics , 1980, IEEE Journal of Quantum Electronics.

[17]  A. Renn,et al.  Single molecule spectroscopy. Fluorescence-lifetime measurements of pentacene in p-terphenyl , 1993 .

[18]  Thomas Basché,et al.  Optical Spectroscopy Of Single Impurity Molecules In Solids , 1993, Laser Applications to Chemical Analysis.

[19]  Robert C. Dunn,et al.  Near-field fluorescence imaging and fluorescence lifetime measurement of light harvesting complexes in intact photosynthetic membranes , 1994 .

[20]  E. Betzig,et al.  Near-field spectroscopy of single molecules at room temperature , 1994, Nature.

[21]  W. P. Ambrose,et al.  Alterations of Single Molecule Fluorescence Lifetimes in Near-Field Optical Microscopy , 1994, Science.

[22]  Steven A. Soper,et al.  Detection of single fluorescent molecules , 1990 .

[23]  Douwe A. Wiersma,et al.  Fluorescence properties of submonolayers of rhodamine 6G in front of a mirror , 1993 .

[24]  B. Persson.  Theory of the damping of excited molecules located above a metal surface , 1978 .

[25]  W. P. Ambrose,et al.  Fluorescence spectroscopy and spectral diffusion of single impurity molecules in a crystal , 1991, Nature.

[26]  J. Michael Ramsey,et al.  Detection of single Rhodamine 6G molecules in levitated microdroplets , 1993 .