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Animated 3D reconstruction depicting desmosomes in methanol/acetone-fixed MDCK cells of clone MDc-2 expressing fluorescent Dsc2a chimera Dsc2a-YFP. The reconstruction was prepared from epifluorescence micrographs that were recorded as a z-stack of eight consecutive focal planes.

Time-lapse fluorescence recording depicting the co-ordinated mobility of a selected array of desmosomes in PDc-13 cells that express Dsc2a chimera Dsc2a-GFP.

Time-lapse recording of double epifluorescence microscopy of living MDc-2 cells co-expressing Dsc2a chimera Dsc2a-YFP and cytokeratin 18 chimera HK18-CFP. Fluorescence is depicted in false colours for better visualisation, showing Dsc2a-YFP-containing desmosomes in red and HK18-CFP-positive cytokeratinfilaments in green. All frames of the movie consist of five superimposed focal recordings that were taken at each time point after excitation at 498 nm (YFP)and 436 nm (CFP).

Epifluorescence microscopy of live MDc-2 monolayer monitoring the distribution of desmosomes containing fluorescent chimera Dsc2a-YFP for 23.3 hours. Each frame represents a projection of seven focal planes.

Detail taken from Movie 6 to demonstrate the characteristic stages of desmosome distribution during mitosis at higher magnification. Each frame consists of seven projected focal planes. Note the transient increase of diffuse fluorescence, the increased fusion of desmosomes, the enrichment of fluorescent puncta around the cleavage furrow, and the continued presence of desmosomes at all time points.

Double epifluorescence microscopy of live MDc-2 cells showing the distribution of Dsc2a-YFP and HK18-CFP during mitosis. Images at each time point represent the projection of six pictures (z-distance of 0.5 mm) recording the fluorescence emitted after excitation at 498 nm (YFP chimera) and 436 nm (CFP chimera). Note the continued presence of Dsc2a-YFP-positive desmosomes throughout cell division and the considerable alterations in the HK18-CFP-containing cytokeratin filament system with some residual desmosome-associated material.

Time-lapse fluorescence microscopy of MDc-2 cells expressing fluorescent Dsc2a chimeras depicting alterations in desmosomal cadherin distribution upon reduction of calcium. Cells were transferred from standard calcium medium (+ Ca²⁺) to low calcium medium (- Ca²⁺) as indicated. A large pinhole was used for confocal laser scan microscopy. The movie shows the internalisation of Dsc2a-YFP-containing desmosomal particles and the continued presence of diffuse non-desmosomal Dsc2a-YFP fluorescence at the cell surface.

Time-lapse fluorescence microscopy of cells expressing fluorescent Dsc2a chimeras depicting alterations in desmosomal cadherin distribution upon reduction of calcium. Cells were transferred from standard calcium medium (+ Ca²⁺) to low calcium medium (- Ca²⁺) as indicated. 3D reconstructions of z-stacks each consisting of five epifluorescence micrographs show the disintegration of large Dsc2a-GFP-labeled desmosomal structures into smaller particles in PDc-13 cells after the reduction of the calcium concentration (arrows).

Time-lapse fluorescence microscopy of cells expressing fluorescent Dsc2a chimeras depicting alterations in desmosomal cadherin distribution upon reduction of calcium. Cells were transferred from standard calcium medium (+ Ca²⁺) to low calcium medium (- Ca²⁺) as indicated. Projection images of z-stacks are shown, each consisting of five focal planes recording the epifluorescence of Dsc2a-GFP in PDc-13 cells. Note the fusion of small fluorescent desmosomal particles after the reduction of calcium.

Time-lapse fluorescence microscopy of cells expressing fluorescent Dsc2a chimeras depicting alterations in desmosomal cadherin distribution upon reduction of calcium. Cells were transferred from standard calcium medium (+ Ca²⁺) to low calcium medium (- Ca²⁺) as indicated. Overlay of projected fluorescence pictures (five focal planes) and corresponding phase contrast micrographs were obtained from PDc-13 cells. Note the overall reduction of desmosomes together with the fusion of desmosomes (black arrows) and fission of desmosomes (white arrows) after removal of calcium.

Time-lapse epifluorescence microscopy of MDc-2 cells monitoring the distribution of Dsc2a-YFP in response to a transient 5 minute reduction of calcium. Note the uptake of fluorescent puncta into the cytoplasm shortly after the pulse of low calcium medium and their continued presence in this location, while small dots re-appear at the cell surface

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