The pancreas was inflated with 3ml cold collagenase (Sigma; St Louis, MO, USA) solution (0.3mg/ml) through the bile duct with a 20G needle, then dissected and transferred into a 2 ml collagenase solution (1 mg/ml) in a siliconised glass tube followed by at 37°C in a water bath for 12–15 min. Following 3 washes, islets were hand-picked under a light microscope. Islets were treated with Cell Dissociation Solution (Sigma) and the single-cell suspension was harvested for flow cytometry.
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