Automated image analysis of live/dead staining of the fungus Aureobasidiumpullulans on microscope slides and leaf surfaces

An image analysis program and protocol for the identification and enumerati on of live versus dead cells of the yeast-like fungus Aureobasidium pullula ns was developed for both populations on microscope slides and leaf surface s. Live cells took up CellTracker Blue(TM), while nonviable cells stained w ith DEAD Red(TM). Image analysis macro programs running under Optimas(TM) s oftware were used to acquire images and to differentiate and enumerate viab le from nonviable cells. The software was capable of discriminating green a s a third parameter for identification and quantification of green fluoresc ent protein-expressing cells in a wild-type population.