3 May 1999 Simple scanning near-infrared time-correlated single-photon counting instrument with a pulsed diode laser and avalanche photodiode (APD) for time-resolved measurements in scanning applications with a DNA sequencer
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Proceedings Volume 3602, Advances in Fluorescence Sensing Technology IV; (1999) https://doi.org/10.1117/12.347539
Event: BiOS '99 International Biomedical Optics Symposium, 1999, San Jose, CA, United States
Abstract
A simple apparatus for time-correlated single photon counting (TCSPC) measurements in the near-infrared (near-IR) region for scanning-type applications has been constructed and examined. The apparatus consisted of five major components including a pulsed diode laser source (lasing wavelength equals 780 nm; repetition rate equals 80 MHz; power equals 5 mW; pulse width equals 150 ps), an integrated microscope, a large photoactive area avalanche photodiode (APD), a TCSPC PC-board including the electronics and a windows-based software package for accumulating the fluorescence decay profiles. The instrument response function of this assembly was found to be 460 ps, which is adequate for measuring lifetimes with (tau) f greater than or equal to 500 ps. Due to the small size of the device, it also allowed implementation into scanning experiments where lifetimes were measured. To demonstrate this capability, a three-well microscope slide containing a near-IR dye was scanned. The decay profile of the near-IR dye, aluminum 2,3-naphthalocyanine, was collected and analyzed to obtain its lifetime, which was found to be 2.73 ns, in close agreement to literature values for this particular dye. In addition, a three dimensional image of aluminum 2,3- naphthalocyanine fluorescence decays was acquired by scanning the microscope head over this three-well glass slide. In the scanning mode, the IRFs as well as the decays of the dyes were found to be very stable. The device demonstrated a concentration detection sensitivity of 2.33 nM, however, the dynamic range was limited due to the APD and its slow time constant (passive quenching). In addition, this microscope head was installed in a LI-COR DNA Sequencer 4000 for collection of scanning images.
© (1999) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Yuling Zhang, Yuling Zhang, Steven A. Soper, Steven A. Soper, Lyle Richard Middendorf, Lyle Richard Middendorf, John A. Wrum, John A. Wrum, Rainer Erdmann, Rainer Erdmann, Michael Wahl, Michael Wahl, "Simple scanning near-infrared time-correlated single-photon counting instrument with a pulsed diode laser and avalanche photodiode (APD) for time-resolved measurements in scanning applications with a DNA sequencer", Proc. SPIE 3602, Advances in Fluorescence Sensing Technology IV, (3 May 1999); doi: 10.1117/12.347539; https://doi.org/10.1117/12.347539
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