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Proceedings Paper

Fluorescence lifetime imaging system with nm-resolution and single-molecule sensitivity
Author(s): Michael Wahl; Hans-Juergen Rahn; Uwe Ortmann; Rainer Erdmann; Martin Boehmer; Joerg Enderlein
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Paper Abstract

Fluorescence lifetime measurement of organic fluorophores is a powerful tool for distinguishing molecules of interest from background or other species. This is of interest in sensitive analysis and Single Molecule Detection (SMD). A demand in many applications is to provide 2-D imaging together with lifetime information. The method of choice is then Time-Correlated Single Photon Counting (TCSPC). We have devloped a compact system on a single PC board that can perform TCSPC at high throughput, while synchronously driving a piezo scanner holding the immobilized sample. The system allows count rates up to 3 MHz and a resolution down to 30 ps. An overall Instrument Response Function down to 300ps is achieved with inexpensive detectors and diode lasers. The board is designed for the PCI bus, permitting high throughput without loss of counts. It is reconfigurable to operate in different modes. The Time-Tagged Time-Resolved (TTTR) mode permits the recording of all photon events with a real-time tag allowing data analysis with unlimited flexibility. We use the Time-Tag clock for an external piezo scanner that moves the sample. As the clock source is common for scanning and tagging, the individual photons can be matched to pixels. Demonstrating the capablities of the system we studied single molecule solutions. Lifetime imaging can be performed at high resolution with as few as 100 photons per pixel.

Paper Details

Date Published: 28 March 2002
PDF: 8 pages
Proc. SPIE 4634, Methods for Ultrasensitive Detection II, (28 March 2002); doi: 10.1117/12.463829
Show Author Affiliations
Michael Wahl, PicoQuant GmbH (Germany)
Hans-Juergen Rahn, PicoQuant GmbH (Germany)
Uwe Ortmann, PicoQuant GmbH (Germany)
Rainer Erdmann, PicoQuant GmbH (Germany)
Martin Boehmer, Univ. Regensburg (Germany)
Joerg Enderlein, Univ. Regensburg (Germany)

Published in SPIE Proceedings Vol. 4634:
Methods for Ultrasensitive Detection II
Charles W. Wilkerson Jr., Editor(s)

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