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Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
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Fluorescence-based Broad Dynamic Range Viscosity Probes.

Anatoliy Dragan1, August E Graham1, Chris D Geddes2

  • 1Institute of Fluorescence and Department of Chemistry and Biochemistry, The Columbus Center, University of Maryland Baltimore County, 701 East Pratt Street, Baltimore, MD, 21202, USA.

Journal of Fluorescence
|November 19, 2013
PubMed
Summary
This summary is machine-generated.

Two new fluorescent probes, SYBR Green (SG) and PicoGreen (PG), show dramatically increased quantum yields in viscous environments. Their performance, sensitive to temperature, offers broad dynamic range viscosity measurements for life sciences.

Keywords:
Collagen sensingFluorescence probesFluorescent viscosity probeIntramolecular quenchingMolecular rotorsPicoGreenSYBR Green I

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Area of Science:

  • Biochemistry
  • Physical Chemistry
  • Molecular Biology

Background:

  • Fluorescent probes are essential tools for molecular detection and quantification.
  • Viscosity measurements are critical in biological and chemical systems.
  • Existing fluorescent viscosity probes have limitations in dynamic range and sensitivity.

Purpose of the Study:

  • To introduce and characterize two novel fluorescent viscosity probes: SYBR Green (SG) and PicoGreen (PG).
  • To evaluate the performance of SG and PG across a wide range of viscosities and in collagen solutions.
  • To explore the potential applications of these probes in life sciences for fluorescence-based viscosity measurements.

Main Methods:

  • Spectroscopic analysis of SYBR Green and PicoGreen quantum yields and excited state lifetimes.
  • Testing probe performance in various solvents with controlled viscosity.
  • Investigating probe behavior in complexation with DNA and in collagen solutions.
  • Analyzing the relationship between quantum yield and viscosity on a log-log scale, including temperature sensitivity.

Main Results:

  • Both SG and PG exhibited low quantum yields and lifetimes in water.
  • A dramatic 300-1000-fold increase in quantum yields and lifetimes was observed in viscous solvents and when complexed with DNA.
  • A linear relationship was established between the logarithm of quantum yield and the logarithm of viscosity.
  • The slope of this relationship demonstrated temperature sensitivity.

Conclusions:

  • SYBR Green and PicoGreen function as effective fluorescent probes for broad dynamic range viscosity measurements.
  • The observed changes in fluorescence properties are highly sensitive to the microenvironment's viscosity.
  • These probes offer a promising tool for real-time, non-invasive viscosity monitoring in life science applications.