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Fig. 4. 3D-iFRAP. (A) Overview of part of a wing disc with cross-hair on the
fluorescent locus selected for iFRAP. Fluorescence depletion was calculated
from a recorded series of z-stack images over time after bleaching
and image registration. (B) Magnified XY confocal image of the nucleus before
bleaching. Cross-hair set in the bleach region next to the unbleached locus.
The images have been registered to correct for the movement of the chromatin,
and the nucleus itself and the fluorescent locus is centered in the final
analysis image time stack. (C,D) X-time and time-Y views, respectively, of the
registered images at the slices in the XY image corresponding to the
cross-hair in B. In C, dissociation of the fluorescent molecules from the
spared locus can be seen over time. In D, a bleached region is shown over
time. The transition from before bleaching to after bleaching is indicated by
the arrows in the time planes.
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