F-techniques: FRET, FLIM, FRAP, FCS

F-techniques take advantage of particular aspects of the fluorescence process by which fluorochromes are excited and emit fluorescent light, are damaged during repetitive excitation, or undergo non-radiative decay prior to light emission.

The CCI offers FRET (fluorescence resonance energy transfer), FLIM (fluorescence lifetime imaging microscopy), FRAP (fluorescence recovery after photobleaching), and FCS (fluorescence correlation spectroscopy).

Labelled cells were imaged as a sequence of images showing movement of the cell over time. Cells attach to surfaces using protein complexes called focal adhesions. The focal adhesions of this cell were labelled with Talin-TS-GFP/RFP, a tension sensor enabling the analysis of focal adhesion force transmission and mechanosensitivity. Data courtesy of Tom Waring (Zech lab).

Microscopes:

• 3i Marianas spinning disk (FRAP, FRET)
• LSM 900 (Nuffield, FRAP)
• LSM 900 (William Henry Duncan)
• LSM 800 (FRAP)
• LSM 780 (FRAP, FRET, FLIM, FCS)