Kinetoplast and nuclear DNA staining in Leishmania mexicana

Kinetoplast and nuclear DNA in kinetoplastids bind different DNA stains with different affinities. Staining of Leishmania mexicana promastigotes was optimised for five DNA stains (DAPI, propidium iodide, SYTO16, DRAQ5 and hoechst 33342). Maximum brightness (maximum pixel intensity) and total signal (sum pixel intensity) of the kinetoplast and nucleus were quantified from widefield epifluorescence micrographs for each stain.

Two stains were analysed from methanol fixed cells mounted in glycerol DABCO: DAPI and propidium iodide. The others were analysed from live cells in PBS. Propidium iodide staining included an RNase A digestion.

This resource lets the relative contribution of kinetoplast and nuclear DNA be determined from signal intensity data from fluorescence micrographs and flow cytometry.

This is a useful resource if working with the methods in one of my previous papers; see links.