Fluorescence decay data analysis correcting for detector pulse pile-up at very high count rates

Using Time-Correlated Single Photon Counting (TCSPC) for the purpose of fluorescence lifetime measurements is usually limited in speed due to pile-up. With modern instrumentation this limitation can be lifted significantly but some artefacts due to frequent merging of closely spaced detector pulses (detector pulse pile-up) remains an issue to be addressed. We propose here a data analysis method correcting for this type of artefact and the resulting systematic errors. It physically models the photon losses due to detector pulse pile-up and incorporates the loss in the decay fit model employed to obtain fluorescence lifetimes and relative amplitudes of the decay components. Comparison of results with and without this correction show a significant reduction of systematic errors at count rates approaching the excitation rate. This allows quantitatively accurate fluorescense lifetime imaging (FLIM) at very high frame rates.Comment: 12 pages, 3 figures. v2: spelled out unexplained abbreviation FRET as requested by reviewer. v3: updated acknowledgements, accepted for publication in Opt. En

GHz bandwidth electro-optics of a single self-assembled quantum dot in a charge-tunable device

The response of a single InGaAs quantum dot, embedded in a miniaturized charge-tunable device, to an applied GHz bandwidth electrical pulse is investigated via its optical response. Quantum dot response times of 1.0 \pm 0.1 ns are characterized via several different measurement techniques, demonstrating GHz bandwidth electrical control. Furthermore a novel optical detection technique based on resonant electron-hole pair generation in the hybridization region is used to map fully the voltage pulse experienced by the quantum dot, showing in this case a simple exponential rise.Comment: 7 pages, 4 figure

Ultra-portable explosives sensor based on a CMOS fluorescence lifetime analysis micro-system

The paper published in AIP Advances (a new journal from the American Institute of Physics) had 1100 article downloads in the first 5 months after publication, and was on the journal’s “most read” list for 4 months. The work was featured by AIP in a Physics News Highlight and press release which resulted in >50 international internet articles and an article in Laser Focus World.This work explores the use of a green-light-emitting copolymer as a chemosensor to detect nitroaromatic-based explosive vapors by recording photoluminescence (PL) and time-resolved PL decay. We show successful detection of 10 ppb 1,4-dinitrobenzene (DNB) vapor. Both a conventional time-correlated single photon counting (TCSPC) device and CMOS time-resolved fluorescence lifetime microsystem are used in the DNB detection. An ultra-portable on-site explosive sensor based on the micro-system has also been demonstrated. This gives rise to the potential for real-time, reliable, inexpensive organic/inorganic hybrid explosives detection.Peer reviewe

Probing subtle fluorescence dynamics in cellular proteins by streak camera based Fluorescence Lifetime Imaging Microscopy

We report the cell biological applications of a recently developed multiphoton fluorescence lifetime imaging microscopy system using a streak camera (StreakFLIM). The system was calibrated with standard fluorophore specimens and was shown to have high accuracy and reproducibility. We demonstrate the applicability of this instrument in living cells for measuring the effects of protein targeting and point mutations in the protein sequence which are not obtainable in conventional intensity based fluorescence microscopy methods. We discuss the relevance of such time resolved information in quantitative energy transfer microscopy and in measurement of the parameters characterizing intracellular physiology

Measurement of the lifetime of the 7s2S1/27s^2S_{1/2} state in atomic cesium using asynchronous gated detection

We report a measurement of the lifetime of the cesium $7s\,^2S_{1/2}$ state using time-correlated single-photon counting spectroscopy in a vapor cell. We excite the atoms using a Doppler-free two-photon transition from the $6s\,^2S_{1/2}$ ground state, and detect the 1.47$\mu$m photons from the spontaneous decay of the $7s\,^2S_{1/2}$ to the $6p\,^2P_{3/2}$ state. We use a gated single photon detector in an asynchronous mode, allowing us to capture the fluorescence profile for a window much larger than the detector gate length. Analysis of the exponential decay of the photon count yields a $7s\,^2S_{1/2}$ lifetime of 48.28$\pm$0.07ns, an uncertainty of 0.14%. These measurements provide sensitive tests of theoretical models of the Cs atom, which play a central role in parity violation measurements.Comment: 5 pages, 5 figures, 2 table