Combined confocal and spectroscopic TPE architecture for the identification of single fluorescent molecules

Giuseppe Chirico; Fabio Cannone; Francesca Olivini; Sabrina Beretta; Giancarlo Baldini; Alberto Diaspro; Mauro Robello

doi:10.1117/12.424583

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24 April 2001 Combined confocal and spectroscopic TPE architecture for the identification of single fluorescent molecules

Giuseppe Chirico, Fabio Cannone, Francesca Olivini, Sabrina Beretta, Giancarlo Baldini, Alberto Diaspro, Mauro Robello

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Proceedings Volume 4262, Multiphoton Microscopy in the Biomedical Sciences; (2001) https://doi.org/10.1117/12.424583
Event: BiOS 2001 The International Symposium on Biomedical Optics, 2001, San Jose, CA, United States

Abstract

We have combined a confocal laser scanning head modified for TPE with some spectroscopic modules to study single molecules and molecular aggregates of rhodamine 6G and labeled proteins on glass substrates. The fluorescence intensity of the spots occurs at definite values that are multiples of a reference signal and extrapolate to the background level measured on the images. These properties suggest that these spots arise from single rhodamines. The discrete character of the intensity distribution can therefore be used as a simple, quantitative tool to discriminate between single molecules and molecular aggregates on single snapshots. These studies have been performed by using a combined confocal and spectroscopic architecture realized for two-photon excitation.

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Giuseppe Chirico, Fabio Cannone, Francesca Olivini, Sabrina Beretta, Giancarlo Baldini, Alberto Diaspro, and Mauro Robello "Combined confocal and spectroscopic TPE architecture for the identification of single fluorescent molecules", Proc. SPIE 4262, Multiphoton Microscopy in the Biomedical Sciences, (24 April 2001); https://doi.org/10.1117/12.424583

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