Cross-talk reduction in confocal images of dual fluorescence labelled cell spheroids.
Dual fluorescence labelling is an advanced method to separate two individual specimens in a biological system using confocal microscopy. An inherent problem of this method is fluorescence channel cross-talk, which causes problems for the exact spatial determination and separation of the specimens. Using a parallel fluorescence detection and an image processing technique, based on an image subtraction method, we have developed a very straight forward method for correcting the dual channel fluorescence images. We successfully applied this method to a 3-dimensional cancer spheroid invasion assay and controlled the cross-talk compensation efficiency by a quality parameter.
Ahammer
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H
Institute of Medical Physics and Biophysics, University of Graz, Austria. helmut.ahammer@kfunigraz.ac.at
DeVaney
T T
TT
Hartbauer
M
M
Tritthart
H A
HA
D000818:Animals; D002460:Cell Line; D002642:Chick Embryo; D007089:Image Enhancement; D007091:Image Processing, Computer-Assisted; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D013382:Subtraction Technique