Dual Color Enumeration
Two or more probes labeled with different color fluorophores that arehybridized to the same cells may be enumerated singly using singlebandpass filter sets or simultaneously using multi-bandpass filter setsdepending upon the signal intensity of the probes and the size of thenucleus. Cells with small or constricted nuclei may not be easilyenumerated using multi-bandpass filter sets due to the spacialarrangement of the probe signal in the nucleus or due to perceived dimprobe signals.
1. Do not evaluate cells that are touching or overlapping.
2. Count diffuse signals if they are separate from other signals.
3.Count split signals - two smaller signals in very close proximity - asone signal. The two signals represent a single chromosome complement.
4. Count two signals connected by a strand of fluorescence as one signal.
5. Do not count cells with zero signals unless they are surrounded by nuclei that contain signals.
6. Do not evaluate nuclei that are not intact.
7. Do not evaluate nuclei with overlapping signals of different color.
8.Do not count non-specific hybridization signals. These signals can berecognized by their lower intensity and different shape.
9. Do not evaluate nuclei with signals located on the periphery of the nucleus.
10. Count only nuclei in which a definite enumeration can be made; skip all others.
11. Record accurately. Recount in order to validate questionable counts.
1. Do not evaluate cells that are touching or overlapping.
2. Count diffuse signals if they are separate from other signals.
3.Count split signals - two smaller signals in very close proximity - asone signal. The two signals represent a single chromosome complement.
4. Count two signals connected by a strand of fluorescence as one signal.
5. Do not count cells with zero signals unless they are surrounded by nuclei that contain signals.
6. Do not evaluate nuclei that are not intact.
7. Do not evaluate nuclei with overlapping signals of different color.
8.Do not count non-specific hybridization signals. These signals can berecognized by their lower intensity and different shape.
9. Do not evaluate nuclei with signals located on the periphery of the nucleus.
10. Count only nuclei in which a definite enumeration can be made; skip all others.
11. Record accurately. Recount in order to validate questionable counts.
Two or more probes labeled with different color fluorophores that arehybridized to the same cells may be enumerated singly using singlebandpass filter sets or simultaneously using multi-bandpass filter setsdepending upon the signal intensity of the probes and the size of thenucleus. Cells with small or constricted nuclei may not be easilyenumerated using multi-bandpass filter sets due to the spacialarrangement of the probe signal in the nucleus or due to perceived dimprobe signals.
1. Do not evaluate cells that are touching or overlapping.
2. Count diffuse signals if they are separate from other signals.
3.Count split signals - two smaller signals in very close proximity - asone signal. The two signals represent a single chromosome complement.
4. Count two signals connected by a strand of fluorescence as one signal.
5. Do not count cells with zero signals unless they are surrounded by nuclei that contain signals.
6. Do not evaluate nuclei that are not intact.
7. Do not evaluate nuclei with overlapping signals of different color.
8.Do not count non-specific hybridization signals. These signals can berecognized by their lower intensity and different shape.
9. Do not evaluate nuclei with signals located on the periphery of the nucleus.
10. Count only nuclei in which a definite enumeration can be made; skip all others.
11. Record accurately. Recount in order to validate questionable counts.
1. Do not evaluate cells that are touching or overlapping.
2. Count diffuse signals if they are separate from other signals.
3.Count split signals - two smaller signals in very close proximity - asone signal. The two signals represent a single chromosome complement.
4. Count two signals connected by a strand of fluorescence as one signal.
5. Do not count cells with zero signals unless they are surrounded by nuclei that contain signals.
6. Do not evaluate nuclei that are not intact.
7. Do not evaluate nuclei with overlapping signals of different color.
8.Do not count non-specific hybridization signals. These signals can berecognized by their lower intensity and different shape.
9. Do not evaluate nuclei with signals located on the periphery of the nucleus.
10. Count only nuclei in which a definite enumeration can be made; skip all others.
11. Record accurately. Recount in order to validate questionable counts.
