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CytoCell heamatology FISH protocol - Probe application

Watch our video for the probe application step of the CytoCell® haematology FISH protocol. Always refer to the instructions for use supplied with the product. Further validation is required for use in a diagnostic setting.

CytoCell heamatology FISH protocol - Probe application (2 minutes) Image
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CytoCell heamatology FISH protocol - Probe application (2 minutes)

Heamatology FISH protocol tips and tricks - Probe application

Watch quick video tips and tricks from our OGT experts for the probe application stage of the CytoCell haematology FISH protocol.

1

Try not to push down too hard on the coverslip when applying the FISH probe, as it can squeeze out of the sides and create a patchy hybridisation.

2

To avoid patchy hybridisations, make sure to press out visible bubbles when applying the coverslip on the probe mix.

3

Filtered pipette tips can reduce intermittent background issues from debris being expelled onto FISH slides.

Haematology FISH FAQs - 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 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:

IGH Plus Breakapart (LPH 070) using 10ul of probe

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

 

IGH Plus Breakapart (LPH 070) using 1ul of probe

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

 

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

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.

 

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

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

 

P53 (LPH 017) sealed with Parafilm

Example 2: P53 (LPH 017) sealed with Parafilm

 

P53 (LPH 017) sealed with Cling Film

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

 

P53 (LPH 017) not sealed with glue

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.

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