Resources

Spectral Image Analysis - Example Code and Image Data

MATLAB Code:

Example Image Data:

  • Spectral Image
    • Spectral Library
      • Note - this spectral library assumes that lumps 2 FRET labels together as one component in the library (e.g., it assumes that FRET in the image is at a constant energy transfer efficiency).  This is for simplicity / demonstration purposes only.

Automated Image Analysis of FRET Signals for Subcellular cAMP Quantification - Example Data

Supplemental resources for the Methods in Molecular Biology book chapter: Automated Image Analysis of FRET Signals for Subcellular cAMP Quantification.  Please cite the book chapter when referencing this work:

Silas J. Leavesley, Arie Nakhmani, Yi Gao, Thomas C. Rich. “Automated image analysis of FRET signals for subcellular cAMP quantification” in cAMP Signaling: Methods and Protocols, Manuela Zaccolo (editor). Methods in Molecular Biology, vol. 1294, Ch. 5. ISBN: 978-1-4939-2536-0. Springer Science+Business Media, LLC, (2015)

For more information on Cell Profiler, please see: http://www.cellprofiler.org/

Note - images are 16-bit greyscale.

Spectral Unmixing, Processing, and Visualization of 4D (x,y,z,λ) FRET/cAMP Signals

Supplemental resources for the JoVE article: Measurement of 3-Dimensional cAMP Distributions in Living Cells Using 4-Dimensional (x, y, z, and λ) Hyperspectral FRET Imaging and Analysis. Please cite the JoVE article when referencing this work.  note that all files should be saved into the same folder.

Naga S. Annamdevula, Hayden Gunn, Rachel Sweat, John R. Griswold, Andrea L. Britain, Thomas C. Rich, Silas J. Leavesley. Measurement of 3-Dimensional cAMP Distributions in Living Cells Using 4-Dimensional (x, y, z, and λ) Hyperspectral FRET Imaging and Analysis. JoVE (submitted 2020)