%0 Journal Article %1 Weston2002 %A Weston, KD %A Dyck, M %A Tinnefeld, P %A Muller, C %A Herten, DP %A Sauer, M %C 1155 16TH ST, NW, WASHINGTON, DC 20036 USA %D 2002 %I AMER CHEMICAL SOC %J ANALYTICAL CHEMISTRY %K sauer %N 20 %P 5342-5349 %T Measuring the number of independent emitters in single-molecule fluorescence images and trajectories using coincident photons %U http://dx.doi.org/10.1021/ac025730z %V 74 %X A simple new approach is described and demonstrated for measuring the number of independent emitters along with the fluorescence intensity, lifetime, and emission wavelength for trajectories and images of single molecules and multichromophoric systems using a single PC plug-in card for time-correlated single-photon counting. The number of independent emitters present in the detection volume can be determined using the interphoton times in a manner similar to classical antibunching experiments. In contrast to traditional coincidence analysis based on pulsed laser excitation and direct measurement of coincident photon pairs using a time-to-amplitude converter, the interphoton distances are retrieved afterward by recording the absolute arrival time of each photon with nanosecond time resolution on two spectrally separated detectors. Intensity changes that result from fluctuations of a photophysical parameter can be distinguished from fluctuations due to changes in the number of emitters (e.g., photobleaching) in single chromophore and multichromophore intensity trajectories. This is the first report to demonstrate imaging with contrast based on the number of independently emitting species within the detection volume.

@article{Weston2002, abstract = {A simple new approach is described and demonstrated for measuring the number of independent emitters along with the fluorescence intensity, lifetime, and emission wavelength for trajectories and images of single molecules and multichromophoric systems using a single PC plug-in card for time-correlated single-photon counting. The number of independent emitters present in the detection volume can be determined using the interphoton times in a manner similar to classical antibunching experiments. In contrast to traditional coincidence analysis based on pulsed laser excitation and direct measurement of coincident photon pairs using a time-to-amplitude converter, the interphoton distances are retrieved afterward by recording the absolute arrival time of each photon with nanosecond time resolution on two spectrally separated detectors. Intensity changes that result from fluctuations of a photophysical parameter can be distinguished from fluctuations due to changes in the number of emitters (e.g., photobleaching) in single chromophore and multichromophore intensity trajectories. This is the first report to demonstrate imaging with contrast based on the number of independently emitting species within the detection volume.}, added-at = {2011-03-01T10:57:13.000+0100}, address = {1155 16TH ST, NW, WASHINGTON, DC 20036 USA}, affiliation = {Sauer, M (Reprint Author), Univ Heidelberg, Inst Phys Chem, Neuenheimer Feld 253, D-69120 Heidelberg, Germany. Univ Heidelberg, Inst Phys Chem, D-69120 Heidelberg, Germany.}, author = {Weston, KD and Dyck, M and Tinnefeld, P and Muller, C and Herten, DP and Sauer, M}, biburl = {https://www.bibsonomy.org/bibtex/23132f8e5ef7c121d6303bdbb9bb3d0af/reichert}, cited-references = {AMBROSE WP, 1997, CHEM PHYS LETT, V269, P365. BETZIG E, 1993, SCIENCE, V262, P1422. BOPP MA, 1997, P NATL ACAD SCI USA, V94, P10630. BOPP MA, 1999, P NATL ACAD SCI USA, V96, P11271. BRUNCICARDI FC, 1999, J SURG RES, V83, P1. COTLET M, 2001, P NATL ACAD SCI USA, V98, P14398. DESCHENES LA, 2001, SCIENCE, V292, P255. GARCIAPARAJO MF, 2001, P NATL ACAD SCI USA, V98, P14392. GENSCH T, 1999, ANGEW CHEM INT EDIT, V38, P3752. HA T, 1996, PHYS REV LETT, V77, P3982. HA T, 1997, CHEM PHYS LETT, V271, P1. HA T, 1998, PHYS REV LETT, V80, P2093. HANBURYBROWN R, 1956, NATURE, V177, P27. HOFKENS J, 2000, J AM CHEM SOC, V122, P9278. HU DH, 1999, J AM CHEM SOC, V121, P6936. LEONHARDT H, 1998, J CELL BIOCHEM, V243, P3. LOUNIS B, 2000, NATURE, V407, P491. LOUNIS B, 2001, J PHYS CHEM B, V105, P5048. LU HP, 1997, NATURE, V385, P143. MACKLIN JJ, 1996, SCIENCE, V272, P255. NIE SM, 1994, SCIENCE, V266, P1018. POMBO A, 1999, EMBO J, V18, P2241. RUITER AGT, 1997, J PHYS CHEM A, V101, P7318. TELLINGHUISEN J, 1994, ANAL CHEM, V66, P64. TINNEFELD P, 2000, SINGLE MOL, V1, P215. TINNEFELD P, 2001, CHEM PHYS LETT, V343, P252. TINNEFELD P, 2001, J PHYS CHEM A, V105, P7989. VANDENBOUT DA, 1997, SCIENCE, V277, P1074. VEERMAN JA, 1999, PHYS REV LETT, V83, P2155. VOSCH T, 2001, ANGEW CHEM INT EDIT, V40, P4643. WESTON KD, 1998, J CHEM PHYS, V109, P7474. WESTON KD, 1999, CHEM PHYS LETT, V308, P58. WESTON KD, 2001, J PHYS CHEM B, V105, P3453. WU M, 1996, J PHYS CHEM-US, V100, P17406. YING LM, 1998, J PHYS CHEM B, V102, P10399.}, doc-delivery-number = {604UP}, groups = {public}, interhash = {c50d7a8c424d324808b315d24ed21328}, intrahash = {3132f8e5ef7c121d6303bdbb9bb3d0af}, issn = {0003-2700}, journal = {ANALYTICAL CHEMISTRY}, journal-iso = {Anal. Chem.}, keywords = {sauer}, keywords-plus = {SCANNING OPTICAL MICROSCOPY; LIGHT-HARVESTING COMPLEXES; ROOM-TEMPERATURE; DYE MOLECULES; DYNAMICS; SPECTROSCOPY; SURFACE; PROTEIN; JUMPS; PHOTOCHEMISTRY}, language = {English}, month = {OCT 15}, number = 20, number-of-cited-references = {35}, pages = {5342-5349}, publisher = {AMER CHEMICAL SOC}, subject-category = {Chemistry, Analytical}, times-cited = {59}, timestamp = {2011-03-09T14:16:08.000+0100}, title = {Measuring the number of independent emitters in single-molecule fluorescence images and trajectories using coincident photons}, type = {Article}, unique-id = {ISI:000178639700026}, url = {http://dx.doi.org/10.1021/ac025730z}, username = {reichert}, volume = 74, year = 2002 }