Living Image 3.1

F. Fluorescent Imaging
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Figure F.17 Example of the autofluorescent subtraction technique using a background
excitation filter.
a) primary excitation filter (DsRed), b) blue-shifted background excitation filter (DsRed Bkg),
and c) corrected data. The corrected image was obtained by subtracting the scaled
background filter image (multiplied by 0.47) from the primary filter image. The 6-week old
female Nu/nu mouse was injected subcutaneously with 1× 10 6 HeLa-luc/PKH26 cells in the
left flank.
Figure F.18 Spectral data describing the autofluorescent subtraction technique using a
background filter.
The graph shows the excitation and emission spectrum of PKH26 and the autofluorescent
excitation spectrum of mouse tissue. Also included are the spectral passbands for the blueshifted
background filter (DsRed Bkg), the primary excitation filter (DsRed), and the emission
filter used with this dye.