SP FLIM – The Method
The Leica TCS SP8 SMD can be supplied with spectral (SP) FLIM detectors. No emission filters in front of the FLIM detectors are necessary any more. This grants a much higher flexibility to the experimental design.
The SMD FLIM wizard allows to easily to select the FLIM detection range and to set up automated FLIM lambda stacks.
- Complete spectral freedom in selection of the FLIM detection range allows optimal adaptation to fluorescence properties of the sample
- The additional information dimension allows the separation of multiple dyes with overlapping spectra or lifetimes
- Optimization of FLIM-FRET experiments
- Recognition of metabolic states by auto- fluorescence characterization
- Spectral lifetime fingerprinting: Identification of characteristic lifetime-wavelength patterns
Spectral freedom with SP FLIM PMTs
SP FLIM PMTs, which are integrated into the SP module of the scan head, ensure optimal adjustment to experimental conditions and removal of autofluorescence. The fluorescence light is spit spectrally by a prism and hits the detector. Software-controlled mirror sliders in front of the detector select the wavelength range of interest. This gives the freedom and flexibility to choose the spectral detection range for FLIM.
A new dimension of knowledge – FLIM lambda stacks
Within the SMD FLIM wizard , SP FLIM detectors are used for automated acquisition of FLIM lambda stacks. That means, that FLIM image series at defined bands of the emission spectrum are acquired. Lifetime emission spectra are especially useful for characterization and identification of autofluorescence or new chromophores, for better separation of dyes with similar properties, and for identification of conformational states and aggregation of chromophores.