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.