Import of Leica FLIM data does not display correct lifetimes

Hi there,

When importing a .lif file containing Leica FLIM data, the /FLIM/FlimDecayTime channel displays pixel values that do not correspond to the calculated lifetimes. There does not seem to be a linear relationship between the pixel values and the lifetime. Is there any conversion necessary or possible?

e.g. For a 2-component FLIM dye, in the Leica LASX software the calculated and separately saved lifetimes range between 0.05-0.836ns for lifetime 1 and between 3.195-6.373ns for lifetime 2. Saving the data as .tif in LASX gives me pixel values of 50-836 and 3195-6373, respectively.
However, when opening the /FLIM/FlimDecayTime 1 and 2 channel from a .lif file in ImageJ, it gives me pixel values of 13529-15073 and 17278-17917, respectively.

The example .tif files and the .lif project can be found here:


(Relevant files in the .lif project are "20200601_HEK_02.lif - 00_0nN_0mbar/FLIM/FlimDecayTime 1/2 ch1 - C=0)

Thanks a lot for your help,
Ines

Hi Ines, the pixel values being read from the LIF with Bio-Formats do appear to match the values stored in the file. Do you know if when you are exporting the files is there any different processing happening for the TIFF files compared to the LIF?