Steady-state fluorescence anisotropy and lifetime measurements of fluorophores and fluorescent-dye-loaded microspheres

POSTER

Abstract

We perform steady-state fluorescence anisotropy and lifetime measurements via illumination of fluorophores with a continuous intensity beam of light. In steady-state fluorescence anisotropy, fluorophore molecules are excited when the polarization of the incoming excitation light is parallel to the excitation axis of the fluorophore. Following a delay known as the fluorophore lifetime $\tau $ a molecule will return to its rest state by emitting photons polarized along the instantaneous orientation of the molecule. The steady-state anisotropy r is defined as the average change in orientation of the sample weighted by the average intensity of each polarization axis. Experimental results are used to determine the anisotropy r and the lifetime $\tau $ of the samples: freely diffusing rhodamine as well as yellow-green fluorescent-dye-loaded microspheres with sizes ranging from 0.51 $\mu$m to 6.2 $\mu$m. Experimental outcomes confirm that the custom-made steady-state fluorescence anisotropy optical system and analysis software are properly engineered and optimized. These outcomes include the fluorescence lifetime $\tau $ for each fluorophore type, the fluorescence lifetime $\tau $ for each fluorescent bead size, and the anisotropy r at various temperatures.

Authors

  • Jacob A. Cole

    Appalachian State University

  • Eva Andrei

    National Center for Biotechnology Information, NIH, Pennsylvania State Univ, Department of Physics, Temple University, CAS Key Laboratory of Materials for Energy Conversion, Shanghai Institute of Ceramics, Chinese Academy of Science, Department of Physics, Boise State University, National Synchrotron Light Source, Brookhaven National Laboratory, York Univ, Erciyes Univ, Chinese Academy of Sciences Hefei, Pennsylvania State Univ and USTC, Heidelberg Institute for Theoretical Studies, MIT, Harvard, Penn State University, Canadian Institute for Theoretical Astrophysics, The Pennsylvania State University, University Park, Pennsylvania State University, Penn State, Department of Physics, Penn State University, University Park, USA, National Institute for Material Science, 1-1 Namiki, Tsukuba, Japan, Carnegie Mellon University, IPICYT, Mexico, Shinshu University, Japan, Universidad Carlos III de Madrid, Spain, Universidade de Sao Paulo, Brazil, Tsinghua University, China, American Physical Society, Appalachian State University, West Chester Univ, Lehigh University, Drew University, University of Balearics Islands, Roanoke College, NIST, Tulane Univ, Shanghai Jiao Tong Univ, Zhejiang Univ, National Institute of Standards and Technology, Department of Physics, The Pennsylvania State University, Department of Electrical Engineering, The Pennsylvania State University, Department of Materials Science and Engineering, the Pennsylvania State University, Hefei National Laboratory for Physical Sciences at Microscale, Department of Physics, University of Science and Technology of China, International Center for Quantum Materials, Peking University, Villanova University, West Virginia University, Institut Lumiere Matiere, PSU, Oak Ridge National Laboratory, Towson Univ, CINVESTAV-Queretaro, Prince George Community College, Howard University, Towson University, The Pennsylvania State University, Hokkaido Univ, Cinvestav-Unidad Queretaro, Queretaro-76230, Mexico, Lawrence Livermore National Laboratory, Osaka University, Penn State Univ, Department of Mechanical and Nuclear Engineering, The Pennsylvania State University, Department of Anthropology, The Pennsylvania State University, ILM Universit\'{e} Lyon 1, Moravian College, Bloomsburg Univ, Universit\"at Bielefeld, Syracuse University, Alabama A\&M University, Indiana University of PA, University of Denver, IUP, University of Nebraska, University of Science and Technology of China, Seoul National University, Soochow University China, US Army Research Laboratory, Drexel University, University of Stuttgart, Youngstown State University, University of Surrey, Department of Chemistry and Biochemistry, University of Maryland, College Park, Department of Mathematics, West Virginia University, University of Delaware, Physics Department, The Pennsylvania State University, 104 Davey Lab, University Park, PA, 16802, USA, Department of Physics and Astronomy, West Virginia University, McMaster University, University of Toronto, The University of Texas at Austin, Army Research Lab, Adelphi MD, University of Connecticut, Notre Dame, University of Maryland, Department of Physics and Center for 2-Dimensional and Layered Materials, The Pennsylvania State University, Department of Physics, Applied Physics, and Astronomy, Rensselaer Polytechnic Institute, Rutgers University

  • Eva Andrei

    National Center for Biotechnology Information, NIH, Pennsylvania State Univ, Department of Physics, Temple University, CAS Key Laboratory of Materials for Energy Conversion, Shanghai Institute of Ceramics, Chinese Academy of Science, Department of Physics, Boise State University, National Synchrotron Light Source, Brookhaven National Laboratory, York Univ, Erciyes Univ, Chinese Academy of Sciences Hefei, Pennsylvania State Univ and USTC, Heidelberg Institute for Theoretical Studies, MIT, Harvard, Penn State University, Canadian Institute for Theoretical Astrophysics, The Pennsylvania State University, University Park, Pennsylvania State University, Penn State, Department of Physics, Penn State University, University Park, USA, National Institute for Material Science, 1-1 Namiki, Tsukuba, Japan, Carnegie Mellon University, IPICYT, Mexico, Shinshu University, Japan, Universidad Carlos III de Madrid, Spain, Universidade de Sao Paulo, Brazil, Tsinghua University, China, American Physical Society, Appalachian State University, West Chester Univ, Lehigh University, Drew University, University of Balearics Islands, Roanoke College, NIST, Tulane Univ, Shanghai Jiao Tong Univ, Zhejiang Univ, National Institute of Standards and Technology, Department of Physics, The Pennsylvania State University, Department of Electrical Engineering, The Pennsylvania State University, Department of Materials Science and Engineering, the Pennsylvania State University, Hefei National Laboratory for Physical Sciences at Microscale, Department of Physics, University of Science and Technology of China, International Center for Quantum Materials, Peking University, Villanova University, West Virginia University, Institut Lumiere Matiere, PSU, Oak Ridge National Laboratory, Towson Univ, CINVESTAV-Queretaro, Prince George Community College, Howard University, Towson University, The Pennsylvania State University, Hokkaido Univ, Cinvestav-Unidad Queretaro, Queretaro-76230, Mexico, Lawrence Livermore National Laboratory, Osaka University, Penn State Univ, Department of Mechanical and Nuclear Engineering, The Pennsylvania State University, Department of Anthropology, The Pennsylvania State University, ILM Universit\'{e} Lyon 1, Moravian College, Bloomsburg Univ, Universit\"at Bielefeld, Syracuse University, Alabama A\&M University, Indiana University of PA, University of Denver, IUP, University of Nebraska, University of Science and Technology of China, Seoul National University, Soochow University China, US Army Research Laboratory, Drexel University, University of Stuttgart, Youngstown State University, University of Surrey, Department of Chemistry and Biochemistry, University of Maryland, College Park, Department of Mathematics, West Virginia University, University of Delaware, Physics Department, The Pennsylvania State University, 104 Davey Lab, University Park, PA, 16802, USA, Department of Physics and Astronomy, West Virginia University, McMaster University, University of Toronto, The University of Texas at Austin, Army Research Lab, Adelphi MD, University of Connecticut, Notre Dame, University of Maryland, Department of Physics and Center for 2-Dimensional and Layered Materials, The Pennsylvania State University, Department of Physics, Applied Physics, and Astronomy, Rensselaer Polytechnic Institute, Rutgers University

  • Eva Andrei

    National Center for Biotechnology Information, NIH, Pennsylvania State Univ, Department of Physics, Temple University, CAS Key Laboratory of Materials for Energy Conversion, Shanghai Institute of Ceramics, Chinese Academy of Science, Department of Physics, Boise State University, National Synchrotron Light Source, Brookhaven National Laboratory, York Univ, Erciyes Univ, Chinese Academy of Sciences Hefei, Pennsylvania State Univ and USTC, Heidelberg Institute for Theoretical Studies, MIT, Harvard, Penn State University, Canadian Institute for Theoretical Astrophysics, The Pennsylvania State University, University Park, Pennsylvania State University, Penn State, Department of Physics, Penn State University, University Park, USA, National Institute for Material Science, 1-1 Namiki, Tsukuba, Japan, Carnegie Mellon University, IPICYT, Mexico, Shinshu University, Japan, Universidad Carlos III de Madrid, Spain, Universidade de Sao Paulo, Brazil, Tsinghua University, China, American Physical Society, Appalachian State University, West Chester Univ, Lehigh University, Drew University, University of Balearics Islands, Roanoke College, NIST, Tulane Univ, Shanghai Jiao Tong Univ, Zhejiang Univ, National Institute of Standards and Technology, Department of Physics, The Pennsylvania State University, Department of Electrical Engineering, The Pennsylvania State University, Department of Materials Science and Engineering, the Pennsylvania State University, Hefei National Laboratory for Physical Sciences at Microscale, Department of Physics, University of Science and Technology of China, International Center for Quantum Materials, Peking University, Villanova University, West Virginia University, Institut Lumiere Matiere, PSU, Oak Ridge National Laboratory, Towson Univ, CINVESTAV-Queretaro, Prince George Community College, Howard University, Towson University, The Pennsylvania State University, Hokkaido Univ, Cinvestav-Unidad Queretaro, Queretaro-76230, Mexico, Lawrence Livermore National Laboratory, Osaka University, Penn State Univ, Department of Mechanical and Nuclear Engineering, The Pennsylvania State University, Department of Anthropology, The Pennsylvania State University, ILM Universit\'{e} Lyon 1, Moravian College, Bloomsburg Univ, Universit\"at Bielefeld, Syracuse University, Alabama A\&M University, Indiana University of PA, University of Denver, IUP, University of Nebraska, University of Science and Technology of China, Seoul National University, Soochow University China, US Army Research Laboratory, Drexel University, University of Stuttgart, Youngstown State University, University of Surrey, Department of Chemistry and Biochemistry, University of Maryland, College Park, Department of Mathematics, West Virginia University, University of Delaware, Physics Department, The Pennsylvania State University, 104 Davey Lab, University Park, PA, 16802, USA, Department of Physics and Astronomy, West Virginia University, McMaster University, University of Toronto, The University of Texas at Austin, Army Research Lab, Adelphi MD, University of Connecticut, Notre Dame, University of Maryland, Department of Physics and Center for 2-Dimensional and Layered Materials, The Pennsylvania State University, Department of Physics, Applied Physics, and Astronomy, Rensselaer Polytechnic Institute, Rutgers University