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Related Concept Videos

Protein Dynamics in Living Cells01:19

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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
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Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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Related Experiment Video

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Mapping Molecular Diffusion in the Plasma Membrane by Multiple-Target Tracing MTT
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Locating and classifying fluorescent tags behind turbid layers using time-resolved inversion.

Guy Satat1, Barmak Heshmat1, Christopher Barsi1

  • 1Media Lab, Massachusetts Institute of Technology, 75 Amherst Street, Cambridge, Massachusetts 02139, USA.

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|April 14, 2015
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This study introduces a new method to locate and measure the lifetimes of fluorescent markers behind scattering material. This technique enables non-invasive medical imaging and analysis, even without direct line of sight.

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Area of Science:

  • Biomedical optics
  • Fluorescence imaging
  • Photonics

Background:

  • Fluorescence lifetime imaging microscopy (FLIM) offers valuable insights in biological and medical applications.
  • Imaging through turbid media, such as biological tissues, remains a significant challenge in optical imaging.

Purpose of the Study:

  • To develop and demonstrate a method for reconstructing the spatial locations and fluorescence lifetimes of probes obscured by a turbid layer.
  • To enable non-invasive diagnostic and analytical applications through scattering media.

Main Methods:

  • Utilizing time-resolved fluorescence measurements to capture both spatial and lifetime information.
  • Employing a sparse optimization framework to invert the time-resolved data for scene reconstruction.
  • Implementing a wide-angle imaging technique that does not require coherence or direct line of sight.

Main Results:

  • Successful experimental demonstration of reconstructing hidden fluorescent marker locations and lifetimes.
  • Validation of the sparse optimization approach for inverting complex optical measurements.
  • Proof of concept for non-invasive imaging through scattering materials.

Conclusions:

  • The proposed method advances fluorescence imaging capabilities by enabling visualization and analysis behind turbid layers.
  • This technique holds potential for diverse applications including non-invasive medical diagnosis, flowmetry, and inspection.
  • The wide-angle, coherence-independent approach broadens the scope for long-range and challenging imaging scenarios.