By Wolfgang Becker
This e-book is an try and bridge the distance among the instrumental ideas of multi-dimensional time-correlated unmarried photon counting (TCSPC) and normal functions of the method. Written through an originator of the method and through sucessful clients, it covers the fundamental rules of the procedure, its interplay with optical imaging equipment and its software to quite a lot of experimental initiatives in existence sciences and medical research.
The publication is usually recommended for all clients of time-resolved detection suggestions in biology, bio-chemistry, spectroscopy of reside platforms, stay cellphone microscopy, medical imaging, spectroscopy of unmarried molecules, and different purposes that require the detection of low-level mild signs at single-photon sensitivity and picosecond time resolution.
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Time per curve 100 µs, 10,000 on/off cycles were accumulated. Time-series starts from the front 1 Introduction to Multi-dimensional TCSPC 21 Memory Bank Swapping Memory Memory Bank 1 Bank 1 Time Measurement Bus Interface Sequencer Readout Time Measurement Readout Bus Inter- Sequencer Memory Bank 2 face Memory Bank 2 Fig. 21 Unlimited sequential recording by memory bank swapping. When one memory bank is full, the sequencer swaps the banks. While the sequencer writes into one bank, the other bank is read by the computer memory bank are ﬁlled with data.
Bird et al. demonstrated the technique for lifetime imaging of stained kidney tissue samples . Rück et al. used of multi-wavelength FLIM for monitoring the conversion of photosensitisers for PDT and the generation of photoproducts [126, 127]. FRET measurements by multi-wavelength FLIM were described in [30, 31, 76]. The majority of multi-wavelength FLIM applications are in autofluorescence imaging, especially imaging of the coenzymes NADH and FAD [42, 98, 128, 150]. Chorvatova and Chorvat worked out spectral unmixing techniques based on multiwavelength FLIM data and used them to determine metabolic parameters in cardiomyocytes [40–46] and investigate their response to drugs and stress conditions.
17. A leaf was excited by two multiplexed diode lasers of 405 and 650 nm wavelength. Fluorescence was detected by two detectors at 510 and 700 nm and recorded in one TCSPC module via a router. The result are three fluorescence decay curves for different combination of excitation and emission wavelength: 405/510, 405/700, and 650/700 nm. The forth combination (650/ 510 nm, not shown) does not contain photons because the detection wavelength is shorter than the excitation wavelength. It should be noted here that multiplexing of lasers can also be obtained by ‘pulseinterleaved excitation’, or PIE, see Sect.