Hardware solution for continuous time-resolved burst detection of single molecules in flow

Michael Wahl; Rainer Erdmann; Kristian Lauritsen; Hans-Juergen Rahn

doi:10.1117/12.307325

10 April 1998 Hardware solution for continuous time-resolved burst detection of single molecules in flow

Michael Wahl, Rainer Erdmann, Kristian Lauritsen, Hans-Juergen Rahn

Proceedings Volume 3259, Systems and Technologies for Clinical Diagnostics and Drug Discovery; (1998) https://doi.org/10.1117/12.307325
Event: BiOS '98 International Biomedical Optics Symposium, 1998, San Jose, CA, United States

Abstract

Time Correlated Single Photon Counting (TCSPC) is a valuable tool for Single Molecule Detection (SMD). However, existing TCSPC systems did not support continuous data collection and processing as is desirable for applications such as SMD for e.g. DNA-sequencing in a liquid flow. First attempts at using existing instrumentation in this kind of operation mode required additional routing hardware to switch between several memory banks and were not truly continuous. We have designed a hard- and software system to perform continuous real-time TCSPC based upon a modern solid state Time to Digital Converter (TDC). Short dead times of the fully digital TDC design combined with fast Field Programmable Gay Array logic permit a continuous data throughput as high as 3 Mcounts/sec. The histogramming time may be set as short as 100 microsecond(s) . Every histogram or every single fluorescence photon can be real-time tagged at 200 ns resolution in addition to recording its arrival time relative to the excitation pulse. Continuous switching between memory banks permits concurrent histogramming and data read-out. The instrument provides a time resolution of 60 ps and up to 4096 histogram channels. The overall instrument response function in combination with a low cost picosecond diode laser and an inexpensive photomultiplier tube was found to be 180 ps and well sufficient to measure sub-nanosecond fluorescence lifetimes.

Citation Download Citation

Michael Wahl, Rainer Erdmann, Kristian Lauritsen, and Hans-Juergen Rahn "Hardware solution for continuous time-resolved burst detection of single molecules in flow", Proc. SPIE 3259, Systems and Technologies for Clinical Diagnostics and Drug Discovery, (10 April 1998); https://doi.org/10.1117/12.307325

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