The Fluorescence Lifetime (FLIM) upgrade kit from PicoQuant has been further improved and is now also available for the new confocal microscope from Olympus, the FluoView FV1000. The upgrade kit extends the capabilities of the microscope towards time-resolved measurements with a resolution down to picoseconds and allows users to perform various analysis on this time scale.
The kit is build from state-of-the-art picosecond diode laser sources in the wavelength range from 375 nm to 470 nm and from 635 nm upwards for fluorescence excitation which are coupled in a single mode fibre for perfect Gaussian beam shape and easy handling. The fluorescence emission is guided to photon counting detectors, such as Photomultiplier Tubes (PMTs) or Single Photon Avalanche Diodes (SPADs) and is finally recorded by the TimeHarp 200 TCSPC system. In combination with a special router solution up to four individual detector channels are possible. The synchronization signals from the FluoView FV1000 are fed into the data stream recorded by the TimeHarp 200 via the unique Time-Tagged Time-Resolved (TTTR) data acquisition mode. In this mode each photon is recorded individually with its specific parameters as detector channel, picosecond timing, global arrival time and, in this special application, up to three additional markers. These markers, in combination with the global arrival time, allow the system software to reconstruct the complete image and subsequently fit the full fluorescence lifetime image. Furthermore the analysis of parameter dependencies is also possible in a multitude of ways and allows e.g. to calculate the fluorescence fluctuation correlation function (FCS) on any single spot of interest, to reconstruct the fluorescence decay of each image pixel and detector channel for the purpose of Fluorescence Lifetime Imaging (FLIM) or to perform advanced Fluorescence Resonance Energy Transfer (FRET) analysis.
The kit can be configured to different laser scanning microscopes, detector schemes, special demands in excitation and emission wavelength as well as installation or special training on FLIM and time-resolved techniques.