The fixed 3D thin-layer cultures of ReN cells were washed 3 times with saline and incubated with 0.001% (v/v) Amylo-Glo solution for 5 minutes in a dark environment. The cells were then washed with saline followed by three washings with DDW. To avoid fluorescence quenching, a drop of anti-fade gold (Life Technologies) was added on top of the stained cells. The Amylo-Glo fluorescence was measured by Olympus DSU confocal microscope with Metamorph image analysis software (Olympus).
