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Probe application

Can I reduce or dilute the probe volume prior to application?

Cytocell probes (aside from Sub-telomeric and enumeration probes) are supplied ready to use with probe and hybridzsation 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:

LPH070 IGH Breakapart using 10uL of probe
IGH Plus Breakapart (LPH 070) using 10ul of probe
LPH070 IGH Breakapart using 1uL of probe
IGH Plus Breakapart (LPH 070) using 1ul of probe
LPH070 IGH Breakapart using 5uL probe 5uL Cytocell Hybridisation A (HA500L)
IGH Plus Breakapart (LPH 070) using 5ul probe + 5ul Cytocell Hybridization A (HA500L)

Can you use an alternative to a glass coverslip?

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

LPH017 P53 sealed with a glass coverslip and Weldite glue
P53 (LPH 017) sealed with a glass coverslip and Weldite glue
LPH017 P53 sealed with Parafilm
P53 (LPH 017) sealed with Parafilm
LPH017 P53 sealed with Cling Film
P53 (LPH 017) sealed with Cling Film
LPH017 P53 not sealed with glue
P53 (LPH 017) not sealed with glue

How can I reduce the risk of probe contamination when using different probes during my preparation?

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.