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Fluorometers and spectrofluorometers are two types of instruments used for measuring molecular fluorescence. These instruments differ in how they select excitation and emission wavelengths and the type of light sources they utilize. Fluorometers use absorption interference filters to choose excitation and emission wavelengths. The excitation source in a fluorometer is typically a low-pressure mercury vapor lamp that emits intense lines distributed throughout the ultraviolet and visible regions.
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Fluorescence Lifetime Macro Imager for Biomedical Applications
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Published on: April 7, 2023

Fluorescence lifetime-based optical molecular imaging.

Anand T N Kumar1

  • 1Athinoula A Martinos Center for Biomedical Imaging, Harvard Medical School, Massachusetts General Hospital, Charlestown, MA 02129, USA. ankumar@nmr.mgh.harvard.edu

Methods in Molecular Biology (Clifton, N.J.)
|December 15, 2010
PubMed
Summary
This summary is machine-generated.

This study introduces a time domain fluorescence tomography system for enhanced in vivo molecular imaging. The system improves target localization and detection of fluorescent signals, even with background autofluorescence.

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Area of Science:

  • Biomedical Optics
  • Molecular Imaging
  • Medical Physics

Background:

  • Fluorescence lifetime imaging (FLI) is a valuable technique for in vivo molecular imaging.
  • Conventional optical imaging methods face limitations in resolution and sensitivity for deep tissue targets.
  • Autofluorescence in biological tissues can obscure specific fluorescent signals.

Purpose of the Study:

  • To describe instrumentation and methods for optimal exploitation of lifetime contrast in time domain fluorescence tomography.
  • To enhance the resolution and sensitivity of in vivo molecular imaging.
  • To demonstrate the system's capability in detecting fluorescent signals in the presence of background autofluorescence.

Main Methods:

  • Utilized a time domain fluorescence tomography system with point excitation using ultrashort laser pulses.
  • Employed non-contact detection with a gated, intensified CCD camera.
  • Integrated a photogrammetric camera for surface boundary acquisition and incorporated into light propagation models.
  • Analyzed time domain data using a lifetime-based tomography approach, extracting lifetimes and decay amplitudes.

Main Results:

  • Achieved improved localization of in vivo targets with resolution superior to conventional optical methods.
  • Demonstrated effective detection of fluorescent protein signals in a breast adenocarcinoma mouse model, overcoming background autofluorescence.
  • Validated the system using phantoms and a tumor-bearing mouse model.

Conclusions:

  • The developed time domain fluorescence tomography system offers a powerful approach for in vivo molecular imaging.
  • The lifetime-based tomography method enhances target localization and signal detection in complex biological environments.
  • This technology holds potential for pre-clinical evaluation of drug treatment response and studying tumor physiology.