Download Advanced Time-Correlated Single Photon Counting Applications by Wolfgang Becker PDF

By Wolfgang Becker

This publication is an try to bridge the space among the instrumental ideas of multi-dimensional time-correlated unmarried photon counting (TCSPC) and commonplace purposes of the procedure. Written by way of an originator of the procedure and by way of sucessful clients, it covers the fundamental rules of the approach, its interplay with optical imaging equipment and its software to quite a lot of experimental initiatives in lifestyles sciences and scientific research.

The e-book is usually recommended for all clients of time-resolved detection options in biology, bio-chemistry, spectroscopy of stay structures, reside phone microscopy, medical imaging, spectroscopy of unmarried molecules, and different functions that require the detection of low-level gentle signs at single-photon sensitivity and picosecond time resolution.

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Even in simple applications as the ones shown above the modern devices have advantages over NIM-based instruments: They work at higher pulse repetition rates and higher count rates, achieve shorter acquisition times, and are fully computer controlled. The real advantage of the modern implementation is, however, that the recording process is multi-dimensional. As a result, experiments can be performed which are entirely out of the reach of the classic design. 1 Principle of Multi-dimensional Recording Development of multi-dimensional TCSPC goes back to the realisation that TCSPC records a photon distribution.

The technique is derived from spatial ‘Mosaic’ or ‘Tile’ imaging. A FLIM mosaic is a data array (either one- or two-dimensional) which has space for a large number of FLIM data sets. The recording process starts to record FLIM data in the first mosaic element. After a defined number of frames of the scan it switches to the next element. Thus, all 32 W. Becker Fig. 33 Time series recorded at a speed of 2 images per second. Chloroplasts in a moss leaf. Dual-memory recording, images 128 × 128 pixels elements of the mosaic are filled with data one after another.

In applications were only a single optical waveform has to be recorded, or the experiment allows a series of waveforms to be recorded sequentially the classic principle is still used to a large extend. The typical application is recording of fluorescence decay curves. Examples are shown in Fig. 6. 6, left, shows fluorescence decay curves of quinine sulphate quenched by Cl− ions. The fluorescence lifetime changes with the quencher concentration [92]. 6, right shows decay curves of DODCI (3,3′diethyloxadicarbocyanine iodide) for different wavelength.

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