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Fluorescence-sensing methods.

Zygmunt Gryczynski1, Ignacy Gryczynski, Joseph R Lakowicz

  • 1Department of Biochemistry and Molecular Biology, Center for Fluorescence Spectroscopy, University of Maryland School of Medicine, Baltimore 21201, USA.

Methods in Enzymology
|March 8, 2003
PubMed
Summary
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Novel fluorescence sensing methods utilize internal standards for accurate measurements in various formats. These techniques show promise for transdermal diagnostics using red-emitting dyes like indocyanine green (IcG).

Area of Science:

  • Biomedical Engineering
  • Analytical Chemistry
  • Optical Sensing

Background:

  • Fluorescence sensing offers versatile detection capabilities.
  • Internal standards enhance accuracy and reliability in fluorescence measurements.
  • Existing methods face challenges in complex biological media and transdermal applications.

Purpose of the Study:

  • To develop novel fluorescence sensing strategies using internal standards.
  • To explore the application of these methods in cuvettes, tissues, and high-throughput formats.
  • To assess the potential for transdermal diagnostics using red-emitting fluorophores.

Main Methods:

  • Utilized internal standards with specific spectral properties for combined emission detection.
  • Employed modulation-sensing and polarization-sensing techniques.

Related Experiment Videos

  • Investigated the use of long-lifetime references and oriented films.
  • Evaluated performance in scattering media like Intralipid and tissue.

    • Main Results:

    • Successfully detected combined emission from reference and analyte-sensitive fluorophores.
    • Demonstrated effective sensing of pH, O2, pCO2, glucose, calcium, and lactate.
    • Showcased the efficacy of modulation and polarization sensing in scattering media.
    • Identified indocyanine green (IcG) as a potential red fluorophore for in vivo transdermal sensing.

    Conclusions:

    • Novel internal standard-based fluorescence sensing offers new possibilities for simple device design.
    • Modulation and polarization sensing are effective even in highly scattering biological samples.
    • Indocyanine green (IcG) shows potential for transdermal diagnostics, paving the way for practical biomedical devices.