Monitoring Actin Disassembly with Time-lapse Microscopy

Protocol

1. Construct a flow chamber as shown in the video.  Make sure that the parafilm seal is tight and use washed coverslips.
2. Incubate actin-binding agent in the chamber for 5-10'.
3. Block non-specific binding sites on the glass coverslip with a blocking protein.
4. Polymerize actin inside the chamber by flowing in G-actin in polymerizing buffer.
5. Washout unpolymerized actin by flowing in excess buffer.