FISH FAQs for probe application

CytoCell probes (aside from Sub-telomeric and enumeration probes) are supplied ready to use with probe and hybridization solution pre-mixed. This mix has been developed and optimized for the intended purpose using 10μl of probe and a 24mm x 24mm coverslip. For this reason, we do not recommend dilution or reduction in volume. Both can produce weaker, more variable probe signals.

Please see the example images below for guidance:

Example 1: IGH Plus Breakapart (LPH 070) using 10ul of probe.

Example 2: IGH Plus Breakapart (LPH 070) using 1ul of probe.

Example 3: IGH Plus Breakapart (LPH 070) using 5ul probe + 5ul Cytocell Hybridisation A (HA500L)

Recommendations:

• 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.

Example 1: P53 (LPH 017) sealed with a glass coverslip and Weldite glue

Example 2: P53 (LPH 017) sealed with Parafilm

Example 3: P53 (LPH 017) sealed with Cling Film

Example 4: P53 (LPH 017) not sealed with glue.

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.