Islet cell analysis and purification by light scatter and autofluorescence

Rat pancreatic A- and B-cells differ in light scatter and flavin-adenine-dinucleotide (FAD)-related fluorescence and are thus represented by two easily distinguishable populations in a fluorescence-activated cell sorter (FACS). Sorting of dissociated islet cells yields highly purified single A- and B-cell preparations. FACS-analysis of islet cells also indicated that FAD-fluorescence in 3-cells is reduced within a 5 minute exposure to 20 mM glucose, whereas no variations were observed in A-cell fluorescence nor with 3-0-methylglucose or fructose. FACS-analysis of blood cells and of dissociated liver, parotid, pituitary and pancreatic exocrine cells demonstrated a wide variation in the respective FAD-fluorescence intensities, which could be used for their purification as viable single cells as well as in studying their metabolic redox state.