General Guidelines For Slide Preparation for FISH Analysis

FISH slide preparation of CTC samples recovered from the IsoFlux system

Print Friendly and PDF

Preparation of slides

Slides can be purchased from Polysciences Inc. These are poly-L-Lysine coated microscope slides CAT # 22247.

  • Draw a small circle on the slide ~1cm in diameter with a hydrophobic pen (PAP pen). Allow to air dry
  • Immediately after removing the CellSpot from the IsoFlux plate, transfer the sample from the CellSpot in 20uL PBS onto the slide slowly and completely using a pipette. Use additional volume to clean the CellSpot at your discretion. (Optional: place the slide with the sample centered on a round magnet 1cm in diameter)
  • Allow the sample to dry in the hood

Use hood for fixation- both fixatives are significantly toxic/corrosive, use nitrile gloves.

Fixation version 1

Once the sample is dry (approximately 1 hour), fix the sample with freshly prepared 95% methanol for 10 minutes.

Fixation version 2

Once the sample is dry (approximately 1 hour), fix the sample with freshly prepared, ice cold, 3:1 methanol: acetic acid for 15 minutes.

After Fixation

Allow the sample to air dry in hood.
Pack slides in slide boxes and keep in a desiccated environment.