If probes are selected so that both location and color contribute to identification, it is theoretically possible to combine all our fluorescent dUTPs in multicolor in situ hybridizations (MFISH). An ideal pairing for 4-color-plexing would be comprised of Aqua 431 dUTP, Green 496 dUTP, Red 580 dUTP, and Red 650 dUTP – allowing for maximal separation of the channels and the use of frequently available filter sets. Combining these dUTPs in multicolored probes can be used to generate many more colors. Chromosome painting with these types of labels has been used to generate more than 20 colors – with in silico mathematical analysis, an MFLISH multiplicity of 48-96 has been achieved.

Leave a comment