A calibration approach for the rapid estimation of fluorophore excited-state lifetimes with applications in biomedicine

POSTER

Abstract

Nanosecond-pulse-width lasers, such as nitrogen lasers, are relatively inexpensive and are commonly used for biological and biomedical applications. Because the laser pulse width is comparable to fluorophore excited-state lifetimes ($\sim $1 -- 10 ns), deconvolution of the temporal system response is needed for accurate lifetime determination. Such deconvolution analysis can be time consuming, and so we report a method for rapid lifetime estimation based on geometric features of the time-resolved emission signal. We find that the time-integrated signal, normalized to the peak intensity, is linear with the lifetime, and can be used for calibration of the measured emission signal to the excited-state lifetime. The approach is accurate to $\sim $15{\%} despite using emission signals with low signal-to-noise ratios ($\sim $10). The approach has applications ranging from flow cytometry to fluorescence lifetime imaging microscopy. Results from fluorescence-based pH sensing are presented.

Authors

  • William H. Rauckhorst

    Gothenburg University, Denison University, Bowling Green State University, NASA Glenn Research Center, Ohio Wesleyan University, Nitronex Corporation, 2305 Presidential Drive, Durham, NC, AFRL/MLPS, Wright-Patterson AFB, OH, Miami University, Ursinus College, Purdue University Calumet, GSI, Florida State University, Carnegie Mellon University, Dept. of Health Sciences, Cleveland State University, Physics Dept., Cleveland State University, University of Rochester, OMI College of Applied Science, University of Cincinnati, Davis and Elkins College, WV, Schlumberger-Doll, Korea Institute of Science and Technology, University of Cincinnati, Lock Haven University of Pennsylvania, Oak Ridge National Laboratory, Oak Ridge, TN 37831-6372, Albion College, Albion, MI 49224, University of Toledo, Toledo, OH 43606, Youngstown State University, Joint Institute for Nuclear Research (Dubna), Naval Research Laboratory, ManTech SRS Technologies, Inc., Ecopulse, Inc., Army Research Laboratory, University of Surrey, The Ohio State University, Univ. Akron, Owens Community College, Toledo, OH

  • William H. Rauckhorst

    Gothenburg University, Denison University, Bowling Green State University, NASA Glenn Research Center, Ohio Wesleyan University, Nitronex Corporation, 2305 Presidential Drive, Durham, NC, AFRL/MLPS, Wright-Patterson AFB, OH, Miami University, Ursinus College, Purdue University Calumet, GSI, Florida State University, Carnegie Mellon University, Dept. of Health Sciences, Cleveland State University, Physics Dept., Cleveland State University, University of Rochester, OMI College of Applied Science, University of Cincinnati, Davis and Elkins College, WV, Schlumberger-Doll, Korea Institute of Science and Technology, University of Cincinnati, Lock Haven University of Pennsylvania, Oak Ridge National Laboratory, Oak Ridge, TN 37831-6372, Albion College, Albion, MI 49224, University of Toledo, Toledo, OH 43606, Youngstown State University, Joint Institute for Nuclear Research (Dubna), Naval Research Laboratory, ManTech SRS Technologies, Inc., Ecopulse, Inc., Army Research Laboratory, University of Surrey, The Ohio State University, Univ. Akron, Owens Community College, Toledo, OH

  • William H. Rauckhorst

    Gothenburg University, Denison University, Bowling Green State University, NASA Glenn Research Center, Ohio Wesleyan University, Nitronex Corporation, 2305 Presidential Drive, Durham, NC, AFRL/MLPS, Wright-Patterson AFB, OH, Miami University, Ursinus College, Purdue University Calumet, GSI, Florida State University, Carnegie Mellon University, Dept. of Health Sciences, Cleveland State University, Physics Dept., Cleveland State University, University of Rochester, OMI College of Applied Science, University of Cincinnati, Davis and Elkins College, WV, Schlumberger-Doll, Korea Institute of Science and Technology, University of Cincinnati, Lock Haven University of Pennsylvania, Oak Ridge National Laboratory, Oak Ridge, TN 37831-6372, Albion College, Albion, MI 49224, University of Toledo, Toledo, OH 43606, Youngstown State University, Joint Institute for Nuclear Research (Dubna), Naval Research Laboratory, ManTech SRS Technologies, Inc., Ecopulse, Inc., Army Research Laboratory, University of Surrey, The Ohio State University, Univ. Akron, Owens Community College, Toledo, OH

  • William H. Rauckhorst

    Gothenburg University, Denison University, Bowling Green State University, NASA Glenn Research Center, Ohio Wesleyan University, Nitronex Corporation, 2305 Presidential Drive, Durham, NC, AFRL/MLPS, Wright-Patterson AFB, OH, Miami University, Ursinus College, Purdue University Calumet, GSI, Florida State University, Carnegie Mellon University, Dept. of Health Sciences, Cleveland State University, Physics Dept., Cleveland State University, University of Rochester, OMI College of Applied Science, University of Cincinnati, Davis and Elkins College, WV, Schlumberger-Doll, Korea Institute of Science and Technology, University of Cincinnati, Lock Haven University of Pennsylvania, Oak Ridge National Laboratory, Oak Ridge, TN 37831-6372, Albion College, Albion, MI 49224, University of Toledo, Toledo, OH 43606, Youngstown State University, Joint Institute for Nuclear Research (Dubna), Naval Research Laboratory, ManTech SRS Technologies, Inc., Ecopulse, Inc., Army Research Laboratory, University of Surrey, The Ohio State University, Univ. Akron, Owens Community College, Toledo, OH

  • Paul Urayama

    Miami University