Cross-talk reduction in confocal images of dual fluorescence labelled cell spheroids

Micron. 1999 Aug;30(4):309-17. doi: 10.1016/s0968-4328(99)00029-3.


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.

MeSH terms

  • Animals
  • Cell Line
  • Chick Embryo
  • Image Enhancement
  • Image Processing, Computer-Assisted
  • Microscopy, Confocal / methods*
  • Microscopy, Fluorescence / methods*
  • Subtraction Technique