FISH protocol: Probe application
Cytocell probes (aside from Sub-telomeric and enumeration probes) are supplied ready to use with probe and hybridisation solution pre-mixed. This mix has been developed and optimised for the intended purpose using 10μl of probe and a 24mmx24mm coverslip. For this reason, we do not recommend dilution or reduction in volume. Both can produce weaker, more variable probe signals.
- Use a glass coverslip. Avoid using alternatives such as parafilm, clingfilm etc. These will not create a sufficient seal to allow for probe spreading.
- Ensure coverslips are sealed to prevent evaporation which can lead to weak, diffuse green signals.
- Seal correctly with rubber glue solution e.g. Weldite/Fixogum
Ensure pipette tips are long enough so that when the tip reaches the bottom of the vial, the pipette itself doesn’t come into contact. Pipettes entering a probe vial may carry over probe reagent from one vial to the next resulting in cross contamination.