Results

The first step was to have good metaphase spreads that the probes can attach to.

Figure 8: Metaphase Spread #1

Figure 9: Metaphase Spread #2

In the above two figures, a comparison in the quality of the metaphase spreads can be seen. In Figure 8, the chromosomes are bunched and hard to tell apart, whereas in Figure 9, the chromosomes are longer and well spread.

The second step was to make sure the probes were exactly similar, and the colouring worked well.

Figure 10: Results from Gel #1

Figure 11: Results from Gel #2

In these two gel images, the probes can be seen quite clearly. In figure 10, all the probes are the same length. The ladders on the side clarify that information. However in Figure 11, the template in some of the wells were not amplified since no signal is visible.

Figure 12: Wavelengths of the fluors used (Molecular Probes, 1996-2004)

The five fluors, and the counterstain, DAPI, are the paints that are used in the creation of the chromosomal 8 barcode. Other fluors such as Alexa 488 and Cy- 5 did not work.

The final step was the attaching of the fluor containing probes to the harvested cells that were on slides.

Figure 13: FISHed slide – Alexa 594 probe

Figure 14: FISHed – FITC & Alexa 594 fluors

These images that were derived from the imaging microscopes are the results of the experiment. In Figure 14, it is evident that the red colour is more dominant that the green.

FIGURE 15: THE PROPOSED COLOUR SCHEME FOR CHROMOSOME 8.

Figure 16: The Final Coloured Barcode for Chromosome 8