Teagan’s Tech Tip: Avoiding Hybridization Artifacts

Teagan’s Tech Tip: Avoiding Hybridization Artifacts

Straight from the bench, our Field Application Scientist Teagan shares a few simple—but impactful—ways to reduce hybridization artifacts and improve signal-to-noise in your FISH workflow:

Start with properly conditioned reagents
Let your hybridization buffer fully equilibrate to room temperature before use. It’s more viscous than the probe concentrate, and when used too cold (straight from -20°C), salts can remain visible on the slide and contribute to background.

Protect your chemistry
Keep your hybridization buffer protected from light—especially when it’s stored separately from the probe. Light exposure can degrade the solution and impact overall performance.

Prep your slides for success
Cleaning blank slides with ice-cold 100% ethanol or Carnoy’s fixative before applying heme samples can significantly reduce artifacts. As an added benefit, this helps the coverslip lay flat and minimizes air bubbles.

Ensure full coverage of the target area
Make sure your probe mix fully covers the area of interest. Trapped air bubbles under the coverslip can lead to uneven hybridization and unwanted artifacts.

Control your environment
Hybridization performs best in a humid environment—around 65%. Maintaining proper humidity helps prevent evaporation and drying, both of which can negatively affect signal quality.

Sometimes, the smallest adjustments make the biggest difference. Keeping these tips in mind can help ensure cleaner slides, stronger signals, and more consistent results.

Have a question for Teagan or want help troubleshooting your workflow?

Connect directly with our FISH specialists here: https://empiregenomics.com/fish-specialist/