Color deconvolution can separate kinetoplast and nuclear DNA signal in Trypanosoma brucei even when the organelles are closely apposed or abnormal in structure. 4', 6-Diamidino-2-phenylindole (DAPI) and propidium iodide (PI) fluorescence and color-deconvolved images of procyclic T. brucei 48 h after induction of expression of sister chromatid cohesion protein 1 (SCC1)-mutAB. (A) 1K1N cell. (B) 2K2N cell. Signal from the closely apposed anterior kinetoplast and posterior nucleus (arrowhead) can be separated. (C) Zoid (1K0N) cell illustrating a lack of nuclear DNA. (D) Cell with zoid-like DAPI staining which possess a fragment of nucleus next to the kinetoplast (arrowhead). (E,F) Cells undergoing aberrant cytokinesis, having failed to undergo mitosis, with a severely distorted nuclear structure. Color deconvolution can unambiguously identify kinetoplasts (arrowheads) among nuclear DNA fragments. (G) An out of focus 2K1N cell; kinetoplast and nucleus signal can still be accurately separated by color deconvolution despite being out of focus. Scale bar represents 5 μm.
Wheeler et al. BMC Biology 2012 10:1 doi:10.1186/1741-7007-10-1