1) Whole animals incubated with the dye for 30/60 minutes at 37, washed with the wash buffer.
2) Ventrally-dissected animals (light organ exposed) incubated with the dye for 30 and 60 minutes at 37, then washed with wash buffer.
3) Completely removed light organs incubated with the dye for 30 and 60 minutes at 37, then washed with wash buffer.
4) All of the above repeated, but permeabilized by 1 and 2% Triton X 100 for 30 minutes before staining.
Apoptosis in the light organ tissues was always verified before caspase staining by staining with acridine orange. For some reason, however, the FITC-conjugated inhibitor peptide was never observed to penetrate into the light organ. In the picture attached, the arms of the light organ extending outwards showed signs of apoptotic cell death when examined by arcridine orange (image not shown). However, nothing was visible with the caspase dye (image shown).
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Submitted Aug 08 2018