Fig. 1

Example of follicles treated with neutral red dye and live/dead fluorochrome assay. a neutral red positive follicle; b B neutral red negative follicle. Neutral red dye is a viability stain which shows a deep red colour in acid cell structures like lysosomes. c V1, fully viable isolated follicle (granulosa cells as well as oocyte show bright green fluorescence) positive to Calcein AM, negative to ethidium homodimer-1; d V2, minimally damaged follicle with two cells positive (bright red colour) to ethidium homodimer-1; e negative control: dead follicle showing bright red fluorescence (positive to ethidium homodimer-1, negative to Calcein AM); f group of follicles, mostly V1, arrow indicates a follicle that was mechanically disrupted during the manipulation process. Bar = 50 μm