Cross-talk reduction in confocal images of dual fluorescence labelled cell spheroids. |
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| Authors: | H Ahammer T T DeVaney M Hartbauer H A Tritthart |
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| Institution: | Institute of Medical Physics and Biophysics, University of Graz, Austria. helmut.ahammer@kfunigraz.ac.at |
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| Abstract: | 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. |
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