4 In Situ Nick End-Labeling: Electron Microscopical
43
Fig. 9. A necrotic cell labeled with the ISEL/TEM technique. The many immunogold particles are observed on the agglutinated chromatin showing the fine
granular pattern. X 12,000. The inset (X20,000) corresponds to the boxed area.
- Check the optimal dilution of the anti-digoxigenin antibody conjugated to colloidal gold.
• Immunogold particles are present throughout the ultra-thin
section, as background:
- The grids may have sunk at the bottom of the droplets on
the way, throughout the staining process; carefully blot the
excessive medium both at the tip of the forceps and on the
face of the grid, using small pieces of filter paper.
- Do not dry the sections during the staining process.
• Immunogold particles present on the chromatin or chromatin condensation are not counted via NIH Image program:
- Heavy metals such as gold and silver show the most intense
electron-density among substances on the electron micrographs. The NIH Image program recognizes and counts the
immunogolds based on the difference of the contrast be-
43
Fig. 9. A necrotic cell labeled with the ISEL/TEM technique. The many immunogold particles are observed on the agglutinated chromatin showing the fine
granular pattern. X 12,000. The inset (X20,000) corresponds to the boxed area.
- Check the optimal dilution of the anti-digoxigenin antibody conjugated to colloidal gold.
• Immunogold particles are present throughout the ultra-thin
section, as background:
- The grids may have sunk at the bottom of the droplets on
the way, throughout the staining process; carefully blot the
excessive medium both at the tip of the forceps and on the
face of the grid, using small pieces of filter paper.
- Do not dry the sections during the staining process.
• Immunogold particles present on the chromatin or chromatin condensation are not counted via NIH Image program:
- Heavy metals such as gold and silver show the most intense
electron-density among substances on the electron micrographs. The NIH Image program recognizes and counts the
immunogolds based on the difference of the contrast be-