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YOSHINORI OTSUKI AND YUKO ITO
• The conventional ISEL technique is positive not only for
apoptotic cells but also for necrotic cells (Ansari et al.
1993; Wijsman et al. 1993; Inoki et al. 1997).
• The ISEL using TEM (ISELITEM) technique offers more considerable advantages than the conventional ISEL technique
and agarose gel electrophoresis.
• When the sample contains only a few apoptotic cells, the
ISELITEM technique enables to not only discriminate the
apoptotic cells among many cells but also to demonstrate
the ultrastructural characteristics of apoptosis.
• Necrotic cells labeled with the ISELITEM technique are easily
differenciated from the labeled apoptoic cells due to the
ultrastructural characteristics between them.
• The ISELITEM technique combined with image analysis can
semi-quantify the sites of free 3' -OH DNA ends in apoptotic
cells, when immunogold particles are used in the ISELITEM
technique, as a substitution for the immunoperoxidase ill the
conventional ISEL technique.
Outline
The entire protocol of the ISELITEM technique is outlined in Fig. 1 .
Procedure for the in situ nick end labeling method (ISELlTEM)
etching with sodium metll periodllte
P5'---------------5 ' P
HO-3' _ _ _ _ _ _ _
Fig.!. The entire protocol of the ISELITEM technique as a simplified schema.