📋 Workflow
Full workflow, products add-to-cart directly1
Sample preparation and pretreatment
2 hFix cells/tissue sections, digest with protease to enhance probe accessibility
💡 Tip Optimize protease digestion per sample
⚠️ Caution Over-digestion ruins morphology
🧪 Materials for this step (click to shop by spec/brand)
2
Probe hybridization
OvernightDenature then add fluorescent probe, hybridize overnight at appropriate temperature
💡 Tip Hybridization temperature depends on probe GC/length
⚠️ Caution Moderate probe amount avoids background
🧪 Materials for this step (click to shop by spec/brand)
3
Washing
1 hWash with stringency-gradient buffers to remove non-specifically bound probes
💡 Tip Stringent washing improves signal/noise
⚠️ Caution Over-washing loses signal
🧪 Materials for this step (click to shop by spec/brand)
4
Counterstain and imaging
30 minCounterstain nuclei with DAPI, image by fluorescence/confocal microscopy and analyze signals
💡 Tip DAPI reveals chromosomes and nuclear morphology
⚠️ Caution Antifade mountant aids imaging
🧪 Materials for this step (click to shop by spec/brand)
⚠️ 5 Common Beginner Mistakes
- ① Over-digestion ruins morphology
- ② Wrong temp/buffer weakens signal
- ③ Over-washing loses specific signal
- ④ Probe-target mismatch causes false negatives
- ⑤ Photobleaching affects quantification
❓ FAQ
+FISH vs PCR detection?
FISH gives in situ location/copy number; PCR more sensitive but needs extracted DNA.
+How to boost signal?
Optimize digestion, longer hybridization or signal amplification (TSA).
+Can multiple targets be detected?
Yes, with multicolor probes or multiple fluorophores on one slide.