Photon efficiency optimization in time-correlated single photon counting technique for fluorescence lifetime imaging systems. 2013

Lior Turgeman, and Dror Fixler
Faculty of Engineering and the Institute of Nanotechnology and Advanced Materials, Bar-Ilan University, Ramat Gan 52900, Israel. tur.lior@gmail.com

In time-correlated single photon counting (TCSPC) systems, the maximum signal throughput is limited by the occurrence of pile-up and other effects. In many biological applications that exhibit high levels of fluorescence intensity (FI), pile-up-related distortions yield serious distortions in the fluorescence lifetime (FLT) calculation as well as significant decrease in the signal-to-noise ratio (SNR). Recent developments that allow the use of high-repetition-rate light sources (in the range of 50-100 MHz) in fluorescence lifetime imaging (FLIM) experiments enable minimization of pile-up-related distortions. However, modern TCSPC configurations that use high-repetition-rate excitation sources for FLIM suffer from dead-time-related distortions that cause unpredictable distortions of the FI signal. In this study, the loss of SNR is described by F- value as it is typically done in FLIM systems. This F-value describes the relation of the relative standard deviation in the estimated FLT to the relative standard deviation in FI measurements. Optimization of the F-value allows minimization of signal distortion, as well as shortening of the acquisition time for certain samples. We applied this method for Fluorescein, Rhodamine B, and Erythrosine fluorescent solutions that have different FLT values (4 ns, 1.67 ns, and 140 ps, respectively).

UI MeSH Term Description Entries
D008856 Microscopy, Fluorescence Microscopy of specimens stained with fluorescent dye (usually fluorescein isothiocyanate) or of naturally fluorescent materials, which emit light when exposed to ultraviolet or blue light. Immunofluorescence microscopy utilizes antibodies that are labeled with fluorescent dye. Fluorescence Microscopy,Immunofluorescence Microscopy,Microscopy, Immunofluorescence,Fluorescence Microscopies,Immunofluorescence Microscopies,Microscopies, Fluorescence,Microscopies, Immunofluorescence
D004923 Erythrosine A tetraiodofluorescein used as a red coloring in some foods (cherries, fish), as a disclosure of DENTAL PLAQUE, and as a stain of some cell types. It has structural similarity to THYROXINE. FD & C Red No. 3,2',4',5',7'-Tetraiodofluorescein,Erythrosin,Erythrosin B,Erythrosine B,F D & C #3,FDC Red No. 3
D005456 Fluorescent Dyes Chemicals that emit light after excitation by light. The wave length of the emitted light is usually longer than that of the incident light. Fluorochromes are substances that cause fluorescence in other substances, i.e., dyes used to mark or label other compounds with fluorescent tags. Flourescent Agent,Fluorescent Dye,Fluorescent Probe,Fluorescent Probes,Fluorochrome,Fluorochromes,Fluorogenic Substrates,Fluorescence Agents,Fluorescent Agents,Fluorogenic Substrate,Agents, Fluorescence,Agents, Fluorescent,Dyes, Fluorescent,Probes, Fluorescent,Substrates, Fluorogenic
D012235 Rhodamines A family of 3,6-di(substituted-amino)-9-benzoate derivatives of xanthene that are used as dyes and as indicators for various metals; also used as fluorescent tracers in histochemistry. Rhodamine
D012815 Signal Processing, Computer-Assisted Computer-assisted processing of electric, ultrasonic, or electronic signals to interpret function and activity. Digital Signal Processing,Signal Interpretation, Computer-Assisted,Signal Processing, Digital,Computer-Assisted Signal Interpretation,Computer-Assisted Signal Interpretations,Computer-Assisted Signal Processing,Interpretation, Computer-Assisted Signal,Interpretations, Computer-Assisted Signal,Signal Interpretation, Computer Assisted,Signal Interpretations, Computer-Assisted,Signal Processing, Computer Assisted
D017785 Photons Discrete concentrations of energy, apparently massless elementary particles, that move at the speed of light. They are the unit or quantum of electromagnetic radiation. Photons are emitted when electrons move from one energy state to another. (From Hawley's Condensed Chemical Dictionary, 11th ed)
D059629 Signal-To-Noise Ratio The comparison of the quantity of meaningful data to the irrelevant or incorrect data. Ratio, Signal-To-Noise,Ratios, Signal-To-Noise,Signal To Noise Ratio,Signal-To-Noise Ratios
D019793 Fluorescein A phthalic indicator dye that appears yellow-green in normal tear film and bright green in a more alkaline medium such as the aqueous humor. Fluorescein Sodium,Sodium Fluorescein,C.I. 45350,Colircusi Fluoresceina,D & C Yellow No. 7,D & C Yellow No. 8,D and C Yellow No. 7,D and C Yellow No. 8,D&C Yellow No. 7,D&C Yellow No. 8,Diofluor,Disodium Fluorescein,Fluor-I-Strip A.T.,Fluorescein Dipotassium Salt,Fluorescein Disodium Salt,Fluorescein Monosodium Salt,Fluorescite,Fluorescéine sodique Faure,Fluorets,Ful-Glo,Funduscein,Minims Fluorescein Sodium,Minims Fluoresceine,Minims Stains,Optifluor Diba,Uranine,Dipotassium Salt, Fluorescein,Disodium Salt, Fluorescein,Fluor I Strip A.T.,Fluorescein Sodium, Minims,Fluorescein, Disodium,Fluorescein, Sodium,Fluoresceina, Colircusi,Fluoresceine, Minims,Ful Glo,Monosodium Salt, Fluorescein,Sodium, Fluorescein

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