Advanced time-correlated single photon counting techniques for spectroscopy and imaging in biomedical systems

Becker, Wolfgang; Bergmann, Axel; Biscotti, Giovanni; Rueck, Angelika C.

doi:10.1117/12.529143

Cited by 40 publications

(24 citation statements)

References 38 publications

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“…The femtosecond (fs) pulses from these lasers are almost ideal for this purpose as they combine high excitation power during the pulse, which is essential for a good excitation yield, 4 with low-energy deposition to the sample preventing it from being photo damaged. Furthermore, pulsed excitation furnishes the basis for excellent time resolution using time-gated 5 or time-correlated single-photon-counting (TCSPC) 6 detection systems. Hence, a variety of time-resolved fluorescence techniques such as fluorescence lifetime imaging (FLIM) 5,[7][8][9] or fluorescence anisotropy imaging (Tr-FAIM) 8 have been established.…”

Section: Introductionmentioning

confidence: 99%

Two-Color Two-Photon Excitation Using Femtosecond Laser Pulses

et al. 2008

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The use of two-color two-photon (2c2p) excitation easily extends the wavelength range of Ti:sapphire lasers to the UV, widening the scope of its applications especially in biological sciences. We report observation of 2c2p excitation fluorescence of p-terphenyl (PTP), 2-methyl-5-t-butyl-p-quaterphenyl (DMQ) and tryptophan upon excitation with 400 and 800 nm wavelengths using the second harmonic and fundamental wavelength of a mode-locked Ti:sapphire femtosecond laser. This excitation is energetically equivalent to a one-photon excitation wavelength at 266 nm. The fluorescence signal is observed only when both wavelengths are spatially and temporally overlapping. Adjustment of the relative delay of the two laser pulses renders a cross correlation curve which is in good agreement with the pulse width of our laser. The fluorescence signal is linearly dependent on the intensity of each of the two colors but quadratically on the total incident illumination power of both colors. In fluorescence microscopy, the use of a combination of intense IR and low-intensity blue light as a substitute for UV light for excitation can have numerous advantages. Additionally, the effect of differently polarized excitation photons relative to each other is demonstrated. This offers information about different transition symmetries and yields deeper insight into the two-photon excitation process.

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Section: Introductionmentioning

confidence: 99%

Two-Color Two-Photon Excitation Using Femtosecond Laser Pulses

et al. 2008

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“…Excellent single photon detection performance has been reported for InGa 0.53 As 0.47 /InP avalanche photodiodes (APDs) [4][5][6][7][8][9]. For example, Hu et al have reported single photon detection efficiency (SPDE) of 30% with dark count probability of 1 10 -5 at 230K at 1.31 μm [8], while Itzler et al have shown a similar result.…”

Section: Introductionmentioning

confidence: 68%

“…Applications for single photon avalanche diodes (SPADs) in the wavelength range of 1.0-1.6 μm include optical time domain reflectometry [1], quantum key distribution (QKD) [2], laser ranging [3], three-dimensional imaging [4,5], and time resolved spectroscopy [6]. Excellent single photon detection performance has been reported for InGa 0.53 As 0.47 /InP avalanche photodiodes (APDs) [4][5][6][7][8][9].…”

Section: Introductionmentioning

confidence: 99%

Ge on Si and InP/InGaAs single photon avalanche diodes

Sun

et al. 2011

SPIE Proceedings

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This paper describes single photon detection for Ge on Si separate-absorption-charge-multiplication (SACM) avalanche photodiodes and advances in quenching for InP/InGaAs single photon avalanche diodes. INTRODUCTIONApplications for single photon avalanche diodes (SPADs) in the wavelength range of 1.0-1.6 μm include optical time domain reflectometry [1], quantum key distribution (QKD) [2], laser ranging [3], three-dimensional imaging [4, 5], and time resolved spectroscopy [6]. Excellent single photon detection performance has been reported for InGa 0.53 As 0.47 /InP avalanche photodiodes (APDs) [4-9]. For example, Hu et al. have reported single photon detection efficiency (SPDE) of 30% with dark count probability of 1 10 -5 at 230K at 1.31 μm [8], while Itzler et al. have shown a similar result. They reported SPDE of ~ 20% with dark count rate of a few kHz with moderate cooling [9]. Recently, at an operation wavelength of 1.55 μm, Tosi et al. have reported dark count rate as low as 400 s -1 with SPDE of 28% at 175K [10]. Si SPADs have exhibited high detection efficiency and low dark count probability [11][12][13], however, the band gap of Si restricts operation to wavelengths < 1 μm. One approach to extend the operating wavelength of Si-based SPADs is to utilize a separate absorption, charge, and multiplication (SACM) APD structure in which the multiplication region is Si with adjacent InGaAs [14] or Ge absorption regions [15]. The use of Si for the multiplication region is advantageous owing to its low excess noise factor and favorable avalanche breakdown probability. The benefits of Ge as the absorber include its compatibility with CMOS process technology and its long-wavelength cutoff (λ ~ 1.55 μm). An advantage of Si-based SPADs is that they can be integrated with CMOS circuits [12,16], which facilitates performance improvements in applications such as photon timing and monolithic focal plane array imaging. The challenge presented by Ge on Si for single photon detection is the relatively high dark current that results from the lattice mismatch between Ge and Si. This leads to high dark count rate (DCR). Recently, improved epitaxial growth techniques have enabled significant progress in the performance of Ge on Si APDs [15,17,18]. In this paper, we report Geiger-mode operation, i.e. single photon detection, of Ge on Si SACM APDs.InGaAs/InP single photon avalanche diodes (SPADs) have achieved high SPDE and low DCR in the near-infrared wavelength range between 1.0 and 1.7 μm with moderate cooling [19,20]. However, afterpulsing is a significant performance limitation for infrared SPADs, especially in high-speed applications. During an avalanche event, the current that flows through the diode will fill some of the deep level traps in the high-field multiplication region. These traps release carriers after the avalanche current is quenched. If these carriers are released during subsequent detection windows, they will give rise to excess dark counts. The release rate of the carriers from the traps is proportional to t...

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“…The photon data can either be used to build up photon distributions or to store the data of each individual photon. Thus, modern TCSPC modules can be configured to record either multiwavelength FLIM data (Becker et al, 2002, fast dynamic changes in the fluorescence decay behavior of a sample (Becker and Bergmann, 2005;Becker et al, 1999Becker et al, , 2004c, FCS and FIDA data combined with fluorescence lifetime data (Böhmer et al, 2002), and BIFL data (Eggeling et al, 2001;Prummer et al, 2004).…”

Section: Multidimensional Tcspcmentioning

confidence: 99%

Fluorescence lifetime images and correlation spectra obtained by multidimensional time‐correlated single photon counting

Becker

Bergmann

Haustein

et al. 2006

Microscopy Res & Technique

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Multidimensional time-correlated single photon counting (TCSPC) is based on the excitation of the sample by a high-repetition rate laser and the detection of single photons of the fluorescence signal in several detection channels. Each photon is characterized by its arrival time in the laser period, its detection channel number, and several additional variables such as the coordinates of an image area, or the time from the start of the experiment. Combined with a confocal or two-photon laser scanning microscope and a pulsed laser, multidimensional TCSPC makes a fluorescence lifetime technique with multiwavelength capability, near-ideal counting efficiency, and the capability to resolve multiexponential decay functions. We show that the same technique and the same hardware can be used for precision fluorescence decay analysis and fluorescence correlation spectroscopy (FCS) in selected spots of a sample.

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Advanced time-correlated single photon counting techniques for spectroscopy and imaging in biomedical systems

Cited by 40 publications

References 38 publications

Two-Color Two-Photon Excitation Using Femtosecond Laser Pulses

Two-Color Two-Photon Excitation Using Femtosecond Laser Pulses

Ge on Si and InP/InGaAs single photon avalanche diodes

Fluorescence lifetime images and correlation spectra obtained by multidimensional time‐correlated single photon counting

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