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Proceedings Paper

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
Author(s): Yuling Zhang; Steven A. Soper; Lyle Richard Middendorf; John A. Wrum; Rainer Erdmann; Michael Wahl
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Paper 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.

Paper Details

Date Published: 3 May 1999
PDF: 12 pages
Proc. SPIE 3602, Advances in Fluorescence Sensing Technology IV, (3 May 1999); doi: 10.1117/12.347539
Show Author Affiliations
Yuling Zhang, Louisiana State Univ. (United States)
Steven A. Soper, Louisiana State Univ. (United States)
Lyle Richard Middendorf, LI-COR, Inc. (United States)
John A. Wrum, LI-COR, Inc. (United States)
Rainer Erdmann, PicoQuant GmbH (Germany)
Michael Wahl, PicoQuant GmbH (Germany)

Published in SPIE Proceedings Vol. 3602:
Advances in Fluorescence Sensing Technology IV
Joseph R. Lakowicz; Steven A. Soper; Richard B. Thompson, Editor(s)

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