Opera for FLIM

Opera is the first fully automated FLIM instrument for High Content Screening (HCS). It analyzes spatial distribution of fluorescence with lifetime resolution. Fluorescence lifetime can be used in High Content Screening as  read-out of intermolecular effects like protein-protein interactions.

The Opera is a time domain fluorescence lifetime instrument using a pulsed laser source and a fast ICCD camera for image acquisition. Lifetime information measured for every CCD pixel. A multi component fit results in the amplitudes corresponding to the various lifetimes.

Figure: Time gated fluorescence lifetime measurement in the Opera. Excitation: Laser pulses every 12 ns (red vertical lines). Red curves: Fluorescence emission of an ensemble of dye molecules with characteristic lifetime t. Grey boxes: First detection window of lifetime camera (one frame with 80Mio shutter open/close per second) with fluorescence lifetime t1 and intensity i1. Blue boxes: Second detection window of lifetime camera  with fluorescence lifetime t2 and intensity i2.