Single-photon fluorescence

Figure C1.5.12.(A) Fluorescence decay of a single molecule of cresyl violet on an indium tin oxide (ITO) surface measured by time-correlated single photon counting. The solid line is tire fitted decay, a single exponential of 480 5 ps convolved witli tire instmment response function of 160 ps fwiim. The decay, which is considerably faster tlian tire natural fluorescence lifetime of cresyl violet, is due to electron transfer from tire excited cresyl violet (D ) to tire conduction band or energetically accessible surface electronic states of ITO. (B) Distribution of lifetimes for 40 different single molecules showing a broad distribution of electron transfer rates. Reprinted witli pennission from Lu andXie [1381. Copyright 1997 American Chemical Society.

Brunei C, Lounis B, Tamarat P and Orrit M 1999 Triggered source of single photons based on controlled single molecule fluorescence Phys. Rev. Lett. 83 2722-5... 

Cline-Love L J and Shaver L A 1976 Time correlated single photon technique fluorescence lifetime measurements Anal. Chem. 48 370A-371A... 

Birch D J S and Imhof R E 1977 A single-photon counting fluorescence decay-time spectrometer J. Phys. E Sol. Instrum. 10 1044-9... 

Figure 4.6 shows an apparatus for the fluorescence depolarization measurement. The linearly polarized excitation pulse from a mode-locked Ti-Sapphire laser illuminated a polymer brush sample through a microscope objective. The fluorescence from a specimen was collected by the same objective and input to a polarizing beam splitter to detect 7 and I by photomultipliers (PMTs). The photon signal from the PMT was fed to a time-correlated single photon counting electronics to obtain the time profiles of 7 and I simultaneously. The experimental data of the fluorescence anisotropy was fitted to a double exponential function. 

The principal advantages of this technique are its very good time resolution, allowing the determination of lifetimes ranging from 10-e to 10-10 sec, and the fact that single photons are counted. Thus good results can be obtained even with very weakly fluorescent materials. 

Harris, J. M. and Lytle, F. E. (1977). Measurement of subnanosecond fluorescence decays by sampled single-photon detection. Rev. Sci. Instrum. 48, 1469-76. 

Becker, W., Bergmann, A., Hink, M. A., Konig, K., Benndorf, K. and Biskup, C. (2004b). Fluorescence lifetime imaging by time-correlated single-photon counting. Microsc. Res. Tech. 63, 58-66. 

Mosconi, D., Stoppa, D., Pancheri, L., Gonzo, L. and Simoni, A. (2006). CMOS single-photon avalanche diode array for time-resolved fluorescence detection. IEEE ESSCIRC. 564-67. 

Volkmer, A., Hatrick, D. A. and Birch, D. J. S. (1997). Time-resolved nonlinear fluorescence spectroscopy using femtosecond multiphoton excitation and single-photon timing detection. Meas. Sci. Technol. 8, 1339 19. 

The introduction and diversification of genetically encoded fluorescent proteins (FPs) [1] and the expansion of available biological fluorophores have propelled biomedical fluorescent imaging forward into new era of development [2], Particular excitement surrounds the advances in microscopy, for example, inexpensive time-correlated single photon counting (TCSPC) cards for desktop computers that do away with the need for expensive and complex racks of equipment and compact infrared femtosecond pulse length semiconductor lasers, like the Mai Tai, mode locked titanium sapphire laser from Spectra physics, or the similar Chameleon manufactured by Coherent, Inc., that enable multiphoton excitation. 

Fluorescent chemical sensors occupy nowadays a prominent place among the optical devices due to its superb sensitivity (just a single photon sometimes suffices for quantifying luminescence compared to detecting the intensity difference between two beams of light in absorption techniques), combined with the required selectivity that photo- or chemi-luminescence impart to the electronic excitation. This is due to the fact that the excitation and emission wavelengths can be selected from those of the absorption and luminescence bands of the luminophore molecule in addition, the emission kinetics and anisotropy features of the latter add specificity to luminescent measurements8 10. 

Merkel OM, Librizzi D, Pfestroff A, Schurrat T, Buyens K, Sanders NN, De Smedt SC, Behe M, Kissel T (2009) Stability of siRNA polyplexes from poly(ethylenimine) and poly (ethylenimine)-g-poly(ethylene glycol) under in vivo conditions effects on pharmacokinetics and biodistribution measured by Fluorescence Fluctuation Spectroscopy and Single Photon Emission Computed Tomography (SPECT) imaging. J Control Release 138 148-159... 

Summary. This Chapter focuses on the investigation of fast electron transport studies in solids irradiated at relativistic laser intensities. Experimental techniques based upon space-resolved spectroscopy are presented in view of their application to both ultrashort Ka X-ray sources and fast ignition studies. Spectroscopy based upon single-photon detection is unveiled as a complementary diagnostic technique, alternative to well established techniques based upon bent crystals. Application of this technique to the study of X-ray fluorescence emission from fast electron propagation in multilayer targets is reported and explored as an example case. 

For fluorescence decay curves of the J-aggregate LB films of [CI-MC] mixed with various matrix agents, measured with a picosecond time-resolved single photon counting system, three components of the the lifetimes fitting to exponential terms in the following equation ... 

The dynamics of the SHL intensity after subpicosecond UV laser excitation of RuC18B LB films is shown in Figure 32[115,116]. The SHL intensity decreased to 70 % of its initial value upon excitation and returned to almost the initial value within several hundred picoseconds as shown by a bold line. The fluorescence decay of RuC18B LB films measured by the single photon-counting... 

For EPy-doped PMMA film, a 308 nm excimer laser (Lumonics TE 430T-2, 6ns) was used as as exposure source. We used a tine-correlated single photon counting systen (18) for measuring fluorescence spectra and rise as well as decay curves of a snail ablated area. The excitation was a frequency-doubled laser pulse (295 nm, lOps) generated from a synchronously punped cavity-dumped dye laser (Spectra Physics 375B) and a CW mode-locked YAG laser (Spectra Physics 3000). Decay curves under a fluorescence microscope were measured by the same systen as used before (19). 

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