Looking at single molecules of proteins inside living cells using live-cell smFRET laser microscopy setup

Most complex molecules in living cells mediate their involvement in biological processes by performing series of conformational changes. Understanding these conformational changes requires their measurement within living cells. Here, we combine Förster resonance energy transfer (FRET) and a live-cell smFRET HILO microscopy setup to observe conformational changes of the heat shock protein Hsp90 within the cell. This is a crucial step towards an understanding of the complex dynamic processes in living cells and will ultimately guide us to manipulate cellular processes in a defined and specific way.