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Whole-field optically sectioned fluorescence lifetime imaging.

M J Cole, J Siegel, S E Webb

    Optics Letters
    |December 11, 2007
    PubMed
    Summary
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    We developed a new 3D fluorescence microscope using structured illumination. This microscope achieves high-resolution, whole-field, sectioned fluorescence lifetime images.

    Area of Science:

    • Biophysics
    • Microscopy
    • Optical Imaging

    Background:

    • Fluorescence lifetime imaging (FLIM) provides molecular contrast.
    • Achieving high spatial resolution and optical sectioning in FLIM remains challenging.

    Purpose of the Study:

    • To introduce a novel three-dimensional fluorescence lifetime imaging microscope.
    • To demonstrate its capability for whole-field sectioned FLIM with micrometer resolution.

    Main Methods:

    • Utilizing structured illumination microscopy (SIM) principles.
    • Integrating SIM with time-correlated single-photon counting (TCSPC) for lifetime measurements.
    • Developing a system for whole-field, 3D FLIM acquisition.

    Main Results:

    Related Experiment Videos

  • Demonstrated whole-field sectioned fluorescence lifetime images.
  • Achieved a spatial resolution of a few micrometers.
  • Successfully implemented 3D FLIM using structured illumination.

    • Conclusions:

    • The novel microscope enables efficient 3D FLIM acquisition.
    • Structured illumination significantly enhances spatial resolution in FLIM.
    • This technique offers a powerful tool for biological imaging.