Spectral Unmixing of Bioluminescence Signals

Author: Seth Gammon (gammons at mir.wustl.edu)
History: 2006/04/14: First version
Source:Contained in the JAR file.
Installation: Download Spectral_Unmixing_Plugin.jar to the plugins folder, or subfolder, restart ImageJ, and there will be a new "Spectral Unmixing" command in the Plugins menu, or submenu.
Description: The program allows spectral deconvolution of bioluminescence images. There is no limit to the number of luciferases present in the image; nor is there an internal limit to the maxiumum number of filters that may be used during image aquisisiton. 8, 12, and 32 bit images are all acceptable. The strength/limitation of this program is that one area in each image must contain a known pure sample of each luciferase to serve as an internal calibration.

This program was not intented for use in living animals as attenuation of light due to depth will confound the quatification of the images.

If you use this program in a publication please cite and read the following reference: Gammon ST, Leevy WM, Gross S, Gokel GW, Piwnica-Worms D. Spectral unmixing of multicolored bioluminescence emitted from heterogeneous biological sources. Anal Chem. 2006 Mar 1;78(5):1520-7. PMID: 16503603 [PubMed - in process]. Download the publication from Analytical Chemistry

The linear unmixing algorithm utilizes the pseudoinverse function of the public domain JAMA matrix algebra library from the NIST, which is included in the JAR file.

Instructions in PDF format

Sample stack of images for deconvolution


See Also: Spectral Unmixing Plugins by Joachim Walter

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