Acute osmotic challenge
Cells expressing tagged proteins were grown to log growth phase, then pelleted by centrifugation in a clinical centrifuge for 1 min and gently resuspended in 22°C 10 mM Tris-HCl buffer, pH 7.4 (hypoosmotic) or 10 mM sorbitol (hyperosmotic). The cells were then immediately immobilized for live imaging.