A system for fluorescence lifetime imaging with submicron spatial resolution
Fluorescence lifetime imaging of intratissue chloroplasts excited at 810 nm (A) and 405 nm (B)
TauMap combines high resolution three-dimensional tomography with fluorescence lifetime imaging (FLIM). This results in 4D-x,y,z,τ-mapping of fluorophores with submicron spatial and sub nanosecond temporal resolution. Using this novel imaging system, in situ drug monitoring and fluorescence resonance energy transfer (FRET) can be performed in living cells and histological sections. The device is based on a conventional fluorescence microscope. It is equipped with a high-speed galvo-scanning module as well as an ultrafast detection unit. High numerical aperture (NA1.3) objectives enable fluorescence imaging with sub-cellular spatial resolution. TauMap uses a time-correlated single photon counting (TCSPC) module for data acquisition. The detection unit consists of multichannel plates (MCP) or fast photomultiplier tubes (PMT) for temporal resolutions of 50 ps and 250 ps, respectively. The compact system TauMap is fully computer-controlled and includes JenLab Scan Software and SPCImage Data Analysis Software for data acquisition and image processing.
Fluorescence intensity image (left) and fluorescence lifetime image (right) of human stratum corneum (z=10 μm)
In vivo multiphoton FLIM in human stratum basale (z=60 μm) red: SHG in collagen