On the right: Fluorescence Lifetime Image of stained mouse kidney section measured with a conventional laser scanning microsocpe equipped with PicoQuants' FLIM Upgrade Kit (Molecular Probes, Fluo Cells Slide 3, DAPI, Alexa Fluor 488 WGA, Alexa Fluor 568 phalloidin). Different colours represent different fluorescence lifetimes (blue: 0.8 ns, green: 2.1 ns, red: 4.5 ns). The image is a raster scan recorded with a pulsed laser at 435 nm excitation wavelength and a laser repetition rate of 20 MHz. Fluorescence was detected through a 510 nm longpass filter with a PMA 182 PMT photomultiplier. Resolution of this image is 512 x 512 pixels which corresponds to a field of view of approximately 100 µm x 100 µm in the sample plane.

On the left: Raster-scanned image of immobilized single molecules, measured with the MicroTime 200. Polarization-resolved fluorescence of isolated, single Cy5 molecules on top of a standard glass coverslip. The collected fluorescence light was divided with a polarizing beamsplitter cube and simultaneously detected with two SPAD detectors. The image contains all molecules which exhibit either a predominant parallel (yellow) or perpendicular (blue) polarized emission. The data acquistion time was 5 ms for each of the 200 × 200 pixels. The color coding was realized after combining the informations from both channels using the image calculator tool.

On the right: Fluorescence Lifetime Image of a part of the membrane of a living hepatocyte cell, stained with the dye NBD (7-nitrobenz-2-oxa-1,3-diazole), whose lifetime is depending on the hydrophobicity of the environment (courtesy of Dr. Tannert, Humboldt-University Berlin, Molecular Biophysics). Every pixel shows the result of an exponential fit to the fluorescence decay built from all photons contained in this pixel. The sample was measured with the MicroTime 200, excited with a picosecond diode laser at 467 nm using a 100×, 1.3 N.A. oil immersion microscope objective. The image consists of 300 × 300 pixels with an acquisition time of 2 ms/pixel.

On the left: Fluorescence Lifetime Image of a slice of apple seed from young fruit (pirus malus). The sample was measured with the MicroTime 200 excited with a picosecond diode laser at 635 nm at a repetition rate of 10 MHz. The fluorescence emission was collected through a 695 longpassfilter by a PMA 182 photomultiplier. The recording time was approx. 4 minutes.