Lambert Instruments

The spectrally resolved LIFA is an imaging system for fluorescence microscopy that preserves the information required to determine the position, spectrum, and lifetime of the observed fluorescence. This is done by combining several modular components. These consist of the typical LIFA (modulated ICCD camera, modulated LED excitation) and a prism-based imaging spectrograph.

Lifetime (pseudo colours)	Intensity (grey scale)
		spectrally  resolved
		2D image

The spectrally resolved images show one line (y-axis) out of the 2D intensity image of typical dicot root. The emission wavelengths at the x-axis start at 515nm. The fluorescence lifetime is shown in pseudo colors.
The higher wavelength components have a shorter lifetime (blue) than the short wavelength components (red).

Lifetime (pseudo colours)	Intensity (grey scale)
		spectrally resolved  GFP transfected  cells (donor only)
		spectrally resolved  GFP-RFP transfected  cell (FRET)

The spectrally resolved images show one line (y-axis) out of a sample of GFP transfected cells or GFP-RFP transfected cells. The emission wavelengths at the x-axis start at 515nm; the first peak is the GFP emission peak and the second the RFP emission peak. The fluorescence lifetime is shown in pseudo colors.
In the FRET sample (GFP-RFP) the fluorescence lifetime of the GFP has decreased from 2,3ns (red) to 2,0ns (yellow).